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Sample records for myogenic factor myod

  1. In situ detection of transcripts of the myogenic factor MyoD in whole chicken embryos

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    Jane E. Gabriel

    2000-03-01

    Full Text Available In situ hybridization has provided insights into the molecular basis of skeletal myogenesis during embryonic development. In situ detection of different muscle-specific regulatory factors in whole embryos has been described. Spatial and temporal expression patterns of these factors differed among species. The expression pattern of MyoD in whole chicken embryos was studied via in situ hybridization using a probe obtained by the reverse transcription - polymerase chain reaction (RT-PCR technique. In newly formed somites (embryos of stage 12, MyoD mRNA transcripts were detected along the anterior to posterior axis of somites immediately adjacent to the neural tube, whereas in mature somites (embryos of stage 24, MyoD transcripts were detected throughout the entire somite. These results indicate that MyoD expression is important for initiating and maintaining the avian myogenic system.A detecção in situ de diferentes fatores regulatórios músculo-específicos vem sendo uma prática muito empregada para o entendimento das bases moleculares envolvidas no processo de formação do tecido muscular esquelético durante o desenvolvimento embrionário em diferentes organismos. O objetivo deste estudo foi investigar o padrão de expressão dos transcritos do fator miogênico MyoD em embriões inteiros de galinha por análises de hibridização in situ, usando uma sonda obtida a partir de ensaios de transcrição reversa e reação em cadeia da polimerase (RT-PCR. Nos somitos recém-formados (embriões no estádio 12, os transcritos de MyoD foram detectados na porção mediana dos somitos posicionados imediatamente adjacentes ao tubo neural, enquanto nos somitos maduros (embriões no estádio 24, a expressão do gene do MyoD foi observada em toda a região dos somitos. Tais resultados indicam que a expressão do MyoD é importante no processo de determinação e manutenção da miogênese nas aves.

  2. Regulation of eukaryotic initiation factor 4AII by MyoD during murine myogenic cell differentiation.

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    Gabriela Galicia-Vázquez

    Full Text Available Gene expression during muscle cell differentiation is tightly regulated at multiple levels, including translation initiation. The PI3K/mTOR signalling pathway exerts control over protein synthesis by regulating assembly of eukaryotic initiation factor (eIF 4F, a heterotrimeric complex that stimulates recruitment of ribosomes to mRNA templates. One of the subunits of eIF4F, eIF4A, supplies essential helicase function during this phase of translation. The presence of two cellular eIF4A isoforms, eIF4AI and eIF4AII, has long thought to impart equivalent functions to eIF4F. However, recent experiments have alluded to distinct activities between them. Herein, we characterize distinct regulatory mechanisms between the eIF4A isoforms during muscle cell differentiation. We find that eIF4AI levels decrease during differentiation whereas eIF4AII levels increase during myofiber formation in a MyoD-dependent manner. This study characterizes a previously undefined mechanism for eIF4AII regulation in differentiation and highlights functional differences between eIF4AI and eIF4AII. Finally, RNAi-mediated alterations in eIF4AI and eIF4AII levels indicate that the myogenic process can tolerate short term reductions in eIF4AI or eIF4AII levels, but not both.

  3. Small activating RNA induces myogenic differentiation of rat adipose-derived stem cells by upregulating MyoD

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    Chenghe Wang

    2015-08-01

    Full Text Available ABSTRACTPurpose:RNA activation (RNAa is a mechanism of gene activation triggered by promoter-targeted small double stranded RNAs (dsRNAs, also known as small activating RNAs (saRNAs. Myogenic regulatory factor MyoD is regarded as the master activator of myogenic differentiation cascade by binding to enhancer of muscle specific genes. Stress urinary incontinence (SUI is a condition primarily resulted from urethral sphincter deficiency. It is thus expected that by promoting differentiation of adipose-derived stem cells (ADSCs into myoblasts by activating MyoD gene through RNAa may offer benefits to SUI.Materials and Methods:Rats ADSCs were isolated, proliferated in vitro, and identified by flow cytometry. Purified ADSCs were then transfected with a MyoD saRNA or control transfected. Real-time polymerase chain reaction (RT-PCR and western blotting were used to detect MyoD mRNA and protein expression, respectively. Immunocytochemical staining was applied to determine the expression of desmin protein in transfected cells. Cell viability was measured by using CellTiter 96® AQueous One Solution Cell Proliferation Assay kit.Results:Transfection of a MyoD saRNA (dsMyoD into ADSCs significantly induced the expression of MyoD at both the mRNA and protein levels, and inhibited cell proliferation. Desmin protein expression was detected in dsMyoD treated ADSCs 2 weeks later.Conclusion:Our findings show that RNAa mediated overexpression of MyoD can promote transdifferentiation of ADSCs into myoblasts and may help treat stress urinary incontinence (SUI–a condition primarily resulted from urethral sphincter deficiency.

  4. Efficient myogenic differentiation of human adipose-derived stem cells by the transduction of engineered MyoD protein.

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    Sung, Min Sun; Mun, Ji-Young; Kwon, Ohsuk; Kwon, Ki-Sun; Oh, Doo-Byoung

    2013-07-19

    Human adipose-derived stem cells (hASCs) have great potential as cell sources for the treatment of muscle disorders. To provide a safe method for the myogenic differentiation of hASCs, we engineered the MyoD protein, a key transcription factor for myogenesis. The engineered MyoD (MyoD-IT) was designed to contain the TAT protein transduction domain for cell penetration and the membrane-disrupting INF7 peptide, which is an improved version of the HA2 peptide derived from influenza. MyoD-IT showed greatly improved nuclear targeting ability through an efficient endosomal escape induced by the pH-sensitive membrane disruption of the INF7 peptide. By applying MyoD-IT to a culture, hASCs were efficiently differentiated into long spindle-shaped myogenic cells expressing myosin heavy chains. Moreover, these cells differentiated by an application of MyoD-IT fused to myotubes with high efficiency through co-culturing with mouse C2C12 myoblasts. Because internalized proteins can be degraded in cells without altering the genome, the myogenic differentiation of hASCs using MyoD-IT would be a safe and clinically applicable method. Copyright © 2013 Elsevier Inc. All rights reserved.

  5. Characterization of the Methylation Status of and Myogenic Regulator Factors in Cell Myogenic Differentiation

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    Zhe Chao

    2016-07-01

    Full Text Available Epigenetic processes in the development of skeletal muscle have been appreciated for over a decade. DNA methylation is a major epigenetic modification important for regulating gene expression and suppressing spurious transcription. Up to now, the importance of epigenetic marks in the regulation of Pax7 and myogenic regulatory factors (MRFs expression is far less explored. In the present study, semi-quantitative the real-time polymerase chain reaction (RT-PCR analyses showed MyoD and Myf5 were expressed in activated and quiescent C2C12 cells. MyoG was expressed in a later stage of myogenesis. Pax7 was weakly expressed in differentiated C2C12 cells. To further understand the regulation of expression of these genes, the DNA methylation status of Pax7, MyoD, and Myf5 was determined by bisulfite sequencing PCR. During the C2C12 myoblasts fusion process, the changes of promoter and exon 1 methylation of Pax7, MyoD, and Myf5 genes were observed. In addition, an inverse relationship of low methylation and high expression was found. These results suggest that DNA methylation may be an important mechanism regulating Pax7 and MRFs transcription in cell myogenic differentiation.

  6. Long non-coding RNA Linc-RAM enhances myogenic differentiation by interacting with MyoD

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    Yu, Xiaohua; Zhang, Yong; Li, Tingting; Ma, Zhao; Jia, Haixue; Chen, Qian; Zhao, Yixia; Zhai, Lili; Zhong, Ran; Li, Changyin; Zou, Xiaoting; Meng, Jiao; Chen, Antony K.; Puri, Pier Lorenzo; Chen, Meihong; Zhu, Dahai

    2017-01-01

    Long non-coding RNAs (lncRNAs) are important regulators of diverse biological processes. Here we report on functional identification and characterization of a novel long intergenic non-coding RNA with MyoD-regulated and skeletal muscle-restricted expression that promotes the activation of the myogenic program, and is therefore termed Linc-RAM (Linc-RNA Activator of Myogenesis). Linc-RAM is transcribed from an intergenic region of myogenic cells and its expression is upregulated during myogenesis. Notably, in vivo functional studies show that Linc-RAM knockout mice display impaired muscle regeneration due to the differentiation defect of satellite cells. Mechanistically, Linc-RAM regulates expression of myogenic genes by directly binding MyoD, which in turn promotes the assembly of the MyoD–Baf60c–Brg1 complex on the regulatory elements of target genes. Collectively, our findings reveal the functional role and molecular mechanism of a lineage-specific Linc-RAM as a regulatory lncRNA required for tissues-specific chromatin remodelling and gene expression. PMID:28091529

  7. Patterns of Positive Selection of the Myogenic Regulatory Factor Gene Family in Vertebrates

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    Zhao, Xiao; Yu, Qi; Huang, Ling; Liu, Qing-Xin

    2014-01-01

    The functional divergence of transcriptional factors is critical in the evolution of transcriptional regulation. However, the mechanism of functional divergence among these factors remains unclear. Here, we performed an evolutionary analysis for positive selection in members of the myogenic regulatory factor (MRF) gene family of vertebrates. We selected 153 complete vertebrate MRF nucleotide sequences from our analyses, which revealed substantial evidence of positive selection. Here, we show that sites under positive selection were more frequently detected and identified from the genes encoding the myogenic differentiation factors (MyoG and Myf6) than the genes encoding myogenic determination factors (Myf5 and MyoD). Additionally, the functional divergence within the myogenic determination factors or differentiation factors was also under positive selection pressure. The positive selection sites were more frequently detected from MyoG and MyoD than Myf6 and Myf5, respectively. Amino acid residues under positive selection were identified mainly in their transcription activation domains and on the surface of protein three-dimensional structures. These data suggest that the functional gain and divergence of myogenic regulatory factors were driven by distinct positive selection of their transcription activation domains, whereas the function of the DNA binding domains was conserved in evolution. Our study evaluated the mechanism of functional divergence of the transcriptional regulation factors within a family, whereby the functions of their transcription activation domains diverged under positive selection during evolution. PMID:24651579

  8. Quantitative comparison of the expression of myogenic regulatory factors in flounder(Paralichthys olivaceus) embryos and adult tissues

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    张玉青; 谭训刚; 徐芃; 孙威; 徐永立; 张培军

    2010-01-01

    MyoD,Myf5,and myogenin are myogenic regulatory factors that play important roles during myogenesis.It is thought that MyoD and Myf5 are required for myogenic determination,while myogenin is important for terminal differentiation and lineage maintenance.To better understand the function of myogenic regulatory factors in muscle development of flounder,an important economic fish in Asia,real-time quantitative RT-PCR was used to characterize the expression patterns of MyoD,Myf5, and myogenin at early stages of ...

  9. Coordinated vascular endothelial growth factor expression and signaling during skeletal myogenic differentiation.

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    Bryan, Brad A; Walshe, Tony E; Mitchell, Dianne C; Havumaki, Josh S; Saint-Geniez, Magali; Maharaj, Arindel S; Maldonado, Angel E; D'Amore, Patricia A

    2008-03-01

    Angiogenesis is largely controlled by hypoxia-driven transcriptional up-regulation and secretion of vascular endothelial growth factor (VEGF) and its binding to the endothelial cell tyrosine receptor kinases, VEGFR1 and VEGFR2. Recent expression analysis suggests that VEGF is expressed in a cell-specific manner in normoxic adult tissue; however, the transcriptional regulation and role of VEGF in these tissues remains fundamentally unknown. In this report we demonstrate that VEGF is coordinately up-regulated during terminal skeletal muscle differentiation. We reveal that this regulation is mediated in part by MyoD homo- and hetero-dimeric transcriptional mechanisms. Serial deletions of the VEGF promoter elucidated a region containing three tandem CANNTG consensus MyoD sites serving as essential sites of direct interaction for MyoD-mediated up-regulation of VEGF transcription. VEGF-null embryonic stem (ES) cells exhibited reduced myogenic differentiation compared with wild-type ES cells, suggesting that VEGF may serve a role in skeletal muscle differentiation. We demonstrate that VEGFR1 and VEGFR2 are expressed at low levels in myogenic precursor cells and are robustly activated upon VEGF stimulation and that their expression is coordinately regulated during skeletal muscle differentiation. VEGF stimulation of differentiating C2C12 cells promoted myotube hypertrophy and increased myogenic differentiation, whereas addition of sFlt1, a VEGF inhibitor, resulted in myotube hypotrophy and inhibited myogenic differentiation. We further provide evidence indicating VEGF-mediated myogenic marker expression, mitogenic activity, migration, and prosurvival functions may contribute to increased myogenesis. These data suggest a novel mechanism whereby VEGF is coordinately regulated as part of the myogenic differentiation program and serves an autocrine function regulating skeletal myogenesis.

  10. Expression Pattern of Myogenic Regulatory Transcription Factor mRNAs in the Embryo and Adult Labeo rohita (Hamilton, 1822

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    Archya Sengupta

    2014-01-01

    Full Text Available Understanding the regulation of skeletal muscle development is important to meet the increasing demand of Indian major carp Labeo rohita. Myogenic regulatory factors (MRFs along with myocyte specific enhancer factor 2 (MEF2 play the pivotal role in the determination and differentiation of skeletal muscle. The majority of skeletal muscle genes require both MRFs and MEF2 family members to activate their transcription. In this study, the expression pattern of MyoD, myf-5, myogenin, and MEF2A was observed from 6 h after fertilization to 12 months of age using semiquantitative RT-PCR as well as real-time PCR method. MyoD and myf-5 mRNAs were expressed at high level at the early embryonic stages. Myogenin and MEF2A were expressed after MyoD and myf-5 and remained active up to adult stage. Expression of MyoD was lower than that of Myf-5 after the 5th month. Partial sequencing of MyoD, myf-5, and MEF2A was done to draw phylogeny. In phylogenetic study, Labeo MyoD, MEF2A and myf-5 were found to be closely related to those of common carp. The present investigation suggests that the four transcription factors play pivotal role in the regulation of muscle growth of Labeo rohita in an overlapping and interconnected way.

  11. Restricted maternal nutrition alters myogenic regulatory factor expression in satellite cells of ovine offspring.

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    Raja, J S; Hoffman, M L; Govoni, K E; Zinn, S A; Reed, S A

    2016-07-01

    Poor maternal nutrition inhibits muscle development and postnatal muscle growth. Satellite cells are myogenic precursor cells that contribute to postnatal muscle growth, and their activity can be evaluated by the expression of several transcription factors. Paired-box (Pax)7 is expressed in quiescent and active satellite cells. MyoD is expressed in activated and proliferating satellite cells and myogenin is expressed in terminally differentiating cells. Disruption in the expression pattern or timing of expression of myogenic regulatory factors negatively affects muscle development and growth. We hypothesized that poor maternal nutrition during gestation would alter the in vitro temporal expression of MyoD and myogenin in satellite cells from offspring at birth and 3 months of age. Ewes were fed 100% or 60% of NRC requirements from day 31±1.3 of gestation. Lambs from control-fed (CON) or restricted-fed (RES) ewes were euthanized within 24 h of birth (birth; n=5) or were fed a control diet until 3 months of age (n=5). Satellite cells isolated from the semitendinosus muscle were used for gene expression analysis or cultured for 24, 48 or 72 h and immunostained for Pax7, MyoD or myogenin. Fusion index was calculated from a subset of cells allowed to differentiate. Compared with CON, temporal expression of MyoD and myogenin was altered in cultured satellite cells isolated from RES lambs at birth. The percent of cells expressing MyoD was greater in RES than CON (P=0.03) after 24 h in culture. After 48 h of culture, there was a greater percent of cells expressing myogenin in RES compared with CON (P0.05). In satellite cells from RES lambs at 3 months of age, the percent of cells expressing MyoD and myogenin were greater than CON after 72 h in culture (Psatellite cells of the offspring, which may reduce the pool of myoblasts, decrease myoblast fusion and contribute to the poor postnatal muscle growth previously observed in these animals.

  12. Growth and differentiation of C2 myogenic cells are dependent on serum response factor.

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    Soulez, M; Rouviere, C G; Chafey, P; Hentzen, D; Vandromme, M; Lautredou, N; Lamb, N; Kahn, A; Tuil, D

    1996-01-01

    In order to study to what extent and at which stage serum response factor (SRF) is indispensable for myogenesis, we stably transfected C2 myogenic cells with, successively, a glucocorticoid receptor expression vector and a construct allowing for the expression of an SRF antisense RNA under the direction of the mouse mammary tumor virus long terminal repeat. In the clones obtained, SRF synthesis is reversibly down-regulated by induction of SRF antisense RNA expression by dexamethasone, whose effect is antagonized by the anti-hormone RU486. Two kinds of proliferation and differentiation patterns have been obtained in the resulting clones. Some clones with a high level of constitutive SRF antisense RNA expression are unable to differentiate into myotubes; their growth can be blocked by further induction of SRF antisense RNA expression by dexamethasone. Other clones are able to differentiate and are able to synthesize SRF, MyoD, myogenin, and myosin heavy chain at confluency. When SRF antisense RNA expression is induced in proliferating myoblasts by dexamethasone treatment, cell growth is blocked and cyclin A concentration drops. When SRF antisense RNA synthesis is induced in arrested confluent myoblasts cultured in a differentiation medium, cell fusion is blocked and synthesis of not only SRF but also MyoD, myogenin, and myosin heavy chain is inhibited. Our results show, therefore, that SRF synthesis is indispensable for both myoblast proliferation and myogenic differentiation. PMID:8887636

  13. Conversion of MyoD to a Neurogenic Factor: Binding Site Specificity Determines Lineage

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    Abraham P. Fong

    2015-03-01

    Full Text Available MyoD and NeuroD2, master regulators of myogenesis and neurogenesis, bind to a “shared” E-box sequence (CAGCTG and a “private” sequence (CAGGTG or CAGATG, respectively. To determine whether private-site recognition is sufficient to confer lineage specification, we generated a MyoD mutant with the DNA-binding specificity of NeuroD2. This chimeric mutant gained binding to NeuroD2 private sites but maintained binding to a subset of MyoD-specific sites, activating part of both the muscle and neuronal programs. Sequence analysis revealed an enrichment for PBX/MEIS motifs at the subset of MyoD-specific sites bound by the chimera, and point mutations that prevent MyoD interaction with PBX/MEIS converted the chimera to a pure neurogenic factor. Therefore, redirecting MyoD binding from MyoD private sites to NeuroD2 private sites, despite preserved binding to the MyoD/NeuroD2 shared sites, is sufficient to change MyoD from a master regulator of myogenesis to a master regulator of neurogenesis.

  14. Ectopic Myf5 or MyoD prevents the neuronal differentiation program in addition to inducing skeletal muscle differentiation, in the chick neural tube.

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    Delfini, Marie-Claire; Duprez, Delphine

    2004-02-01

    Forced expression of the bHLH myogenic factors, Myf5 and MyoD, in various mammalian cell lines induces the full program of myogenic differentiation. However, this property has not been extensively explored in vivo. We have taken advantage of the chick model to investigate the effect of electroporation of the mouse Myf5 and MyoD genes in the embryonic neural tube. We found that misexpression of either mouse Myf5 or MyoD in the chick neural tube leads to ectopic skeletal muscle differentiation, assayed by the expression of the myosin heavy chains in the neural tube and neural crest derivatives. We also showed that the endogenous neuronal differentiation program is inhibited under the influence of either ectopic mouse Myf5 or MyoD. We used this new system to analyse, in vivo, the transcriptional regulation between the myogenic factors. We found that MyoD and Myogenin expression can be activated by ectopic mouse Myf5 or MyoD, while Myf5 expression cannot be activated either by mouse MyoD or by itself. We also analysed the transcriptional regulation between the myogenic factors and the different genes involved in myogenesis, such as Mef2c, Pax3, Paraxis, Six1, Mox1, Mox2 and FgfR4. We established the existence of an unexpected regulatory loop between MyoD and FgfR4. The consequences for myogenesis are discussed.

  15. Skeletal muscle hypertrophy and regeneration: interplay between the myogenic regulatory factors (MRFs) and insulin-like growth factors (IGFs) pathways.

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    Zanou, Nadège; Gailly, Philippe

    2013-11-01

    Adult skeletal muscle can regenerate in response to muscle damage. This ability is conferred by the presence of myogenic stem cells called satellite cells. In response to stimuli such as injury or exercise, these cells become activated and express myogenic regulatory factors (MRFs), i.e., transcription factors of the myogenic lineage including Myf5, MyoD, myogenin, and Mrf4 to proliferate and differentiate into myofibers. The MRF family of proteins controls the transcription of important muscle-specific proteins such as myosin heavy chain and muscle creatine kinase. Different growth factors are secreted during muscle repair among which insulin-like growth factors (IGFs) are the only ones that promote both muscle cell proliferation and differentiation and that play a key role in muscle regeneration and hypertrophy. Different isoforms of IGFs are expressed during muscle repair: IGF-IEa, IGF-IEb, or IGF-IEc (also known as mechano growth factor, MGF) and IGF-II. MGF is expressed first and is observed in satellite cells and in proliferating myoblasts whereas IGF-Ia and IGF-II expression occurs at the state of muscle fiber formation. Interestingly, several studies report the induction of MRFs in response to IGFs stimulation. Inversely, IGFs expression may also be regulated by MRFs. Various mechanisms are proposed to support these interactions. In this review, we describe the general process of muscle hypertrophy and regeneration and decipher the interactions between the two groups of factors involved in the process.

  16. Nitric oxide drives embryonic myogenesis in chicken through the upregulation of myogenic differentiation factors.

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    Cazzato, Denise; Assi, Emma; Moscheni, Claudia; Brunelli, Silvia; De Palma, Clara; Cervia, Davide; Perrotta, Cristiana; Clementi, Emilio

    2014-01-15

    The muscle-specific variant of neuronal nitric oxide (NO) synthase (NOS-I), is developmentally regulated in mouse suggesting a role of NO during myogenesis. In chick embryo, a good model of development, we found that the expression of NOS-I is up-regulated, but only in the early phase of development. Through a pharmacological intervention in ovo we found that NO signalling plays a relevant role during embryonic development. The inhibition of NOS-I decreased the growth of embryo, in particular of muscle tissue, while the restoring of physiological NO levels, via administration of a NO donor, reversed this effect. We found a selective action of NO, produced by NOS-I, on regulatory factors involved in myogenic differentiation in the early phase of chick embryo development: inhibition of NO generation leads to a decreased expression of the Myocyte enhancer factor 2a (Mef2a), Mef2c, Myogenin and Myosin, which was reversed by the administration of a NO donor. NO had no effects on Myf5 and MyoD, the myogenic regulatory factors necessary for myogenic determination. The action of NO on the myogenic regulatory factors was mediated via generation of cyclic GMP (cGMP) and activation of the cGMP-dependent protein kinase G (PKG). Finally we found in myoblasts in vitro that the activation of Mef2c was the key event mediating the NO-induced modulation of myogenesis. Our results identify NO produced by NOS-I as a key messenger in the early phase of embryonic development of chicken, acting as a critical determinant of myogenesis through its physiological cGMP/PKG pathway.

  17. The Krüppel-like Factor 15 as a Molecular Link between Myogenic Factors and a Chromosome 4q Transcriptional Enhancer Implicated in Facioscapulohumeral Dystrophy*

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    Dmitriev, Petr; Petrov, Andrei; Ansseau, Eugenie; Stankevicins, Luiza; Charron, Sébastien; Kim, Elena; Bos, Tomas Jan; Robert, Thomas; Turki, Ahmed; Coppée, Frédérique; Belayew, Alexandra; Lazar, Vladimir; Carnac, Gilles; Laoudj, Dalila; Lipinski, Marc; Vassetzky, Yegor S.

    2011-01-01

    Facioscapulohumeral muscular dystrophy (FSHD), a dominant hereditary disease with a prevalence of 7 per 100,000 individuals, is associated with a partial deletion in the subtelomeric D4Z4 repeat array on chromosome 4q. The D4Z4 repeat contains a strong transcriptional enhancer that activates promoters of several FSHD-related genes. We report here that the enhancer within the D4Z4 repeat binds the Krüppel-like factor KLF15. KLF15 was found to be up-regulated during myogenic differentiation induced by serum starvation or by overexpression of the myogenic differentiation factor MYOD. When overexpressed, KLF15 activated the D4Z4 enhancer and led to overexpression of DUX4c (Double homeobox 4, centromeric) and FRG2 (FSHD region gene 2) genes, whereas its silencing caused inactivation of the D4Z4 enhancer. In immortalized human myoblasts, the D4Z4 enhancer was activated by the myogenic factor MYOD, an effect that was abolished upon KLF15 silencing or when the KLF15-binding sites within the D4Z4 enhancer were mutated, indicating that the myogenesis-related activation of the D4Z4 enhancer was mediated by KLF15. KLF15 and several myogenesis-related factors were found to be expressed at higher levels in myoblasts, myotubes, and muscle biopsies from FSHD patients than in healthy controls. We propose that KLF15 serves as a molecular link between myogenic factors and the activity of the D4Z4 enhancer, and it thus contributes to the overexpression of the DUX4c and FRG2 genes during normal myogenic differentiation and in FSHD. PMID:21937448

  18. Beta-catenin regulates myogenesis by relieving I-mfa-mediated suppression of myogenic regulatory factors in P19 cells.

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    Pan, Weijun; Jia, Yingying; Wang, Jiyong; Tao, Donglei; Gan, Xiaoqing; Tsiokas, Leonidas; Jing, Naihe; Wu, Dianqing; Li, Lin

    2005-11-29

    Wnt/beta-catenin signaling plays a critical role in embryonic myogenesis. Here we show that, in P19 embryonic carcinoma stem cells, Wnt/beta-catenin signaling initiates the myogenic process depends on beta-catenin-mediated relief of I-mfa (inhibitor of MyoD Family a) suppression of myogenic regulatory factors (MRFs). We found that beta-catenin interacted with I-mfa and that the interaction was enhanced by Wnt3a. In addition, we found that the interaction between beta-catenin and I-mfa was able to attenuate the interaction of I-mfa with MRFs, relieve I-mfa-mediated suppression of the transcriptional activity and cytosolic sequestration of MRFs, and initiate myogenesis in a P19 myogenic model system that expresses exogenous myogenin. This work reveals a mechanism for the regulation of MRFs during myogenesis by elucidating a beta-catenin-mediated, but lymphoid enhancing factor-1/T cell factor independent, mechanism in regulation of myogenic fate specification and differentiation of P19 mouse stem cells.

  19. Characterization of flounder ( Paralichthys olivaceus) FoxD5 and its function in regulating myogenic regulatory factor

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    Tan, Xungang; Zhang, Yuqing; Sun, Wei; Zhang, Peijun; Xu, Yongli

    2012-03-01

    As one member of winged helix domain transcription factors, FoxD5 was reported to be a trunk organizer. Recent study showed that zebrafish foxd5 is expressed in the somites. To further understand the function of FoxD5 in fish muscle development, the FoxD5 gene was isolated from flounder. Its expression pattern was analyzed by in situ hybridization, while its function in regulating myogenic regulatory factor, MyoD, was analyzed by ectopic expression. It showed that flounder FoxD5 was firstly expressed in the tailbud, adaxial cells, and neural plate of the head. In flounder embryo, FoxD5 is expressed not only in forebrain but also in somite cells that will form muscle in the future. When flounder FoxD5 was over-expressed in zebrafish by microinjection, the expression of zebrafish MyoD in the somites was reduced, suggesting that FoxD5 is involved in myogenesis by regulating the expression of MyoD.

  20. Insulin-like growth factor-1 suppresses the Myostatin signaling pathway during myogenic differentiation

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    Retamales, A.; Zuloaga, R.; Valenzuela, C.A. [Laboratorio de Biotecnología Molecular, Facultad de Ciencias Biológicas, Universidad Andrés Bello, Santiago (Chile); Gallardo-Escarate, C. [Laboratory of Biotechnology and Aquatic Genomics, Universidad de Concepción, Concepción (Chile); Interdisciplinary Center for Aquaculture Research (INCAR), P.O. Box 160-C, Concepción (Chile); Molina, A. [Laboratorio de Biotecnología Molecular, Facultad de Ciencias Biológicas, Universidad Andrés Bello, Santiago (Chile); Interdisciplinary Center for Aquaculture Research (INCAR), P.O. Box 160-C, Concepción (Chile); Valdés, J.A., E-mail: jvaldes@unab.cl [Laboratorio de Biotecnología Molecular, Facultad de Ciencias Biológicas, Universidad Andrés Bello, Santiago (Chile); Interdisciplinary Center for Aquaculture Research (INCAR), P.O. Box 160-C, Concepción (Chile)

    2015-08-21

    Myogenic differentiation is a complex and well-coordinated process for generating mature skeletal muscle fibers. This event is autocrine/paracrine regulated by growth factors, principally Myostatin (MSTN) and Insulin-like Growth Factor-1 (IGF-1). Myostatin, a member of the transforming growth factor-β superfamily, is a negative regulator of skeletal muscle growth in vertebrates that exerts its inhibitory function by activating Smad transcription factors. In contrast, IGF-1 promotes the differentiation of skeletal myoblasts by activating the PI3K/Akt signaling pathway. This study reports on a novel functional crosstalk between the IGF-1 and MSTN signaling pathways, as mediated through interaction between PI3K/Akt and Smad3. Stimulation of skeletal myoblasts with MSTN resulted in a transient increase in the pSmad3:Smad3 ratio and Smad-dependent transcription. Moreover, MSTN inhibited myod gene expression and myoblast fusion in an Activin receptor-like kinase/Smad3-dependent manner. Preincubation of skeletal myoblasts with IGF-1 blocked MSTN-induced Smad3 activation, promoting myod expression and myoblast differentiation. This inhibitory effect of IGF-1 on the MSTN signaling pathway was dependent on IGF-1 receptor, PI3K, and Akt activities. Finally, immunoprecipitation assay analysis determined that IGF-1 pretreatment increased Akt and Smad3 interaction. These results demonstrate that the IGF-1/PI3K/Akt pathway may inhibit MSTN signaling during myoblast differentiation, providing new insight to existing knowledge on the complex crosstalk between both growth factors. - Highlights: • IGF-1 inhibits Myostatin canonical signaling pathway through IGF-1R/PI3K/Akt pathway. • IGF-1 promotes myoblast differentiation through a direct blocking of Myostatin signaling pathway. • IGF-1 induces the interaction of Akt with Smad3 in skeletal myoblast.

  1. Polymorphism of MyoD1 and Myf6 genes and associations with carcass and meat quality traits in beef cattle.

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    Du, X H; Gan, Q F; Yuan, Z R; Gao, X; Zhang, L P; Gao, H J; Li, J Y; Xu, S Z

    2013-12-13

    Myogenic determination factor 1 (MyoD1) and myogenic factor 6 (Myf6) genes belong to the myogenic differentiation (MyoD) gene family, which play key roles in growth and muscle development. The study aimed to investigate the effects of variants in cattle MyoD1 and Myf6 on carcass and meat traits. We screened single nucleotide polymorphisms (SNPs) of both genes in 8 cattle populations, including Simmental, Angus, Hereford, Charolais, Limousin, Qinchuan, Luxi, and Jinnan by sequencing. The G782A locus was identified in exon 1 of MyoD1 (MyoD1-BglI) as well as the T186C locus in exon 1 of Myf6 (Myf6-ApaLI). For the two SNPs, the A allele was significantly more frequent than the B allele in the populations tested. The χ(2) test showed that the MyoD1-BglI locus conformed to Hardy-Weinberg equilibrium in the 8 populations, as did the Myf6-ApaLI locus, with the exception of the Simmental population (P > 0.05). Association analysis revealed that the MyoD1-BglI locus was significantly associated with loin muscle area (LMA) (P genetic markers, opening up new possibilities for cattle breeding and improvements in gene-assisted selection.

  2. Effects of oral creatine and resistance training on myogenic regulatory factor expression.

    Science.gov (United States)

    Willoughby, Darryn S; Rosene, John M

    2003-06-01

    This study examined 12 wk of creatine (Cr) supplementation and heavy resistance training on skeletal muscle creatine kinase (M-CK) mRNA expression and the mRNA and protein expression of the myogenic regulatory factors Myo-D, myogenin, MFR-4, and Myf5. Twenty-two untrained males were randomly assigned to either a control (CON), placebo (PLC), or Cr (CRT) group in a double-blind fashion. Muscle biopsies were obtained before and after training. PLC and CRT trained thrice weekly using 3 sets of 6-8 repetitions at 85-90% 1-RM on the leg press, knee extension, and knee curl exercises. CRT ingested 6 g.d-1 of Cr for 12 wk while PLC consumed the equal amount of placebo. After training, M-CK mRNA expression, as well as myogenin and MRF-4 mRNA and protein expression, were found to be significantly greater for CRT compared with PLC and CON, whereas PLC was also significantly different from CON (P 0.05). M-CK mRNA was correlated with myogenin (r = 0.916) and MRF-4 (r = 0.883) protein (P resistance training, Cr supplementation increases M-CK mRNA expression, likely due to concomitant increases in the expression of myogenin and MRF-4. Therefore, increases in myogenin and MRF-4 mRNA and protein may play a role in increasing myosin heavy chain expression, already shown to occur with Cr supplementation.

  3. Myogenic factors that regulate expression of muscle-specific microRNAs.

    Science.gov (United States)

    Rao, Prakash K; Kumar, Roshan M; Farkhondeh, Mina; Baskerville, Scott; Lodish, Harvey F

    2006-06-01

    Since their discovery as key regulators of early animal development, microRNAs now are recognized as widespread regulators of gene expression. Despite their abundance, little is known regarding the regulation of microRNA biogenesis. We show that three highly conserved muscle-specific microRNAs, miR-1, miR-133 and miR-206, are robustly induced during the myoblast-myotube transition, both in primary human myoblasts and in the mouse mesenchymal C2C12 stem cell line. These microRNAs were not induced during osteogenic conversion of C2C12 cells. Moreover, both loci encoding miR-1, miR-1-1, and miR-1-2, and two of the three encoding miR-133, miR-133a-1 and miR-133a-2, are strongly induced during myogenesis. Some of the induced microRNAs are in intergenic regions, whereas two are transcribed in the opposite direction to the nonmuscle-specific gene in which they are embedded. By using CHIP analysis, we demonstrate that the myogenic factors Myogenin and MyoD bind to regions upstream of these microRNAs and, therefore, are likely to regulate their expression. Because miR-1 and miR-206 are predicted to repress similar mRNA targets, our work suggests that induction of these microRNAs is important in regulating the expression of muscle-specific proteins.

  4. Expression of Myod and myogenin in muscles of mice experimentally infected with Trichinella spiralis or Trichinella pseudospiralis

    Directory of Open Access Journals (Sweden)

    Wu Z.

    2001-06-01

    Full Text Available We developed a detection system for myogenic regulatory factors such as MyoD and myogenin. Adapting the method we performed a longitudinal analysis of such regulatory factors after infection with T. spiralis and T. pseudospiralis. MyoD and myogenin were expressed from the early phase of cystogenesis in T. spiralis infection. The expression returned to the normal level after 18 days from the infection when the cyst was complete. In T. pseudospiralis infection, they were also expressed from the early phase of cystogenesis, but continuously expressed at least up to 43 days post infection.

  5. [Regional differences in the level of ERK1/2 phosphorylation and expression of the myogenic regulatory factors following electrostimulation with different mechanic and metabolic action on the gastrocnemius muscle].

    Science.gov (United States)

    Borzykh, A A; Kuz'min, I V; Lysenko, E A; Vinogradova, O L

    2014-01-01

    Effect of high-frequency electrical stimulation of the sciatic nerve on ERK1/2 kinase phosphorylation and mRNA expression in MyoD (myogenic regulation factor) and myogenin in the red (RGM) and white (WGM) parts of the medial head in rat's m. gastrocnemius was studied. Two stimulation regimes were equalized both lengthwise and in total effort but differed in duration and number of contractions and, therefore, in mechanic and metabolic effects on the muscle. It was shown that growth of the number of phosphorylated ERK1/2 was particularly high in WCM due to application of the protocol for multiple short-time contractions. Whatever the stimulation regime, MyoD mRNA expression in RGM and WGM increases to the same extent, whereas myogenin mRNA expression does not change. Consequently, the regime with the predominantly mechanic effect is favorable to activation of the ERK signaling pathway in glycolytic myofibers.

  6. Activation of muscle enhancers by MyoD and epigenetic modifiers.

    Science.gov (United States)

    Blum, Roy

    2014-11-01

    The early 1980s revelation of cis-acting genomic elements, known as transcriptional enhancers, is still regarded as one of the fundamental discoveries in the genomic field. However, only with the emergence of genome-wide techniques has the genuine biological scope of enhancers begun to be fully uncovered. Massive scientific efforts of multiple laboratories rapidly advanced the overall perception that enhancers are typified by common epigenetic characteristics that distinguish their activating potential. Broadly, chromatin modifiers and transcriptional regulators lay down the essential foundations necessary for constituting enhancers in their activated form. Basing on genome-wide ChIP-sequencing of enhancer-related marks we identified myogenic enhancers before and after muscle differentiation and discovered that MyoD was bound to nearly a third of condition-specific enhancers. Experimental studies that tested the deposition patterns of enhancer-related epigenetic marks in MyoD-null myoblasts revealed the high dependency that a specific set of muscle enhancers have towards this transcriptional regulator. Re-expression of MyoD restored the deposition of enhancer-related marks at myotube-specific enhancers and partially at myoblasts-specific enhancers. Our proposed mechanistic model suggests that MyoD is involved in recruitment of methyltransferase Set7, acetyltransferase p300 and deposition of H3K4me1 and H3K27ac at myogenic enhancers. In addition, MyoD binding at enhancers is associated with PolII occupancy and with local noncoding transcription. Modulation of muscle enhancers is suggested to be coordinated via transcription factors docking, including c-Jun and Jdp2 that bind to muscle enhancers in a MyoD-dependent manner. We hypothesize that distinct transcription factors may act as placeholders and mediate the assembly of newly formed myogenic enhancers.

  7. Differential Expression of Myogenic Regulatory Factor Genes in the Skeletal Muscles of Tambaqui Colossoma macropomum (Cuvier 1818 from Amazonian Black and Clear Water

    Directory of Open Access Journals (Sweden)

    F. A. Alves-Costa

    2013-01-01

    Full Text Available Hypothesizing that the Amazonian water system differences would affect the expression of muscle growth-related genes in juvenile tambaqui Colossoma macropomum (Cuvier 1818, this study aimed to analyze the morphometric data and expression of myogenic regulatory factors (MRFs in the white and red muscle from tambaqui obtained from clear and black Amazonian water systems. All of the MRF transcript levels (myod, myf5, myogenin, and mrf4 were significantly lower in the red muscle from black water fish in comparison to clear water fish. However, in white muscle, only the myod transcript level was significantly decreased in the black water tambaqui. The changes in MRFs gene expression in muscle fibers of tambaqui from black water system provide relevant information about the environmental influence as that of water systems on gene expression of muscle growth related genes in the C. macropomum. Our results showed that the physical and chemical water characteristics change the expression of genes that promote muscle growth, and these results may be also widely applicable to future projects that aim to enhance muscle growth in fish that are of substantial interest to the aquaculture.

  8. Passive Repetitive Stretching for a Short Duration within a Week Increases Myogenic Regulatory Factors and Myosin Heavy Chain mRNA in Rats' Skeletal Muscles

    Directory of Open Access Journals (Sweden)

    Yurie Kamikawa

    2013-01-01

    Full Text Available Stretching is a stimulation of muscle growth. Stretching for hours or days has an effect on muscle hypertrophy. However, differences of continuous stretching and repetitive stretching to affect muscle growth are not well known. To clarify the difference of continuous and repetitive stretching within a short duration, we investigated the gene expression of muscle-related genes on stretched skeletal muscles. We used 8-week-old male Wistar rats ( for this study. Animals medial gastrocnemius muscle was stretched continuously or repetitively for 15 min daily and 4 times/week under anesthesia. After stretching, muscles were removed and total RNA was extracted. Then, reverse transcriptional quantitative real-time PCR was done to evaluate the mRNA expression of MyoD, myogenin, and embryonic myosin heavy chain (MyHC. Muscles, either stretched continuously or repetitively, increased mRNA expression of MyoD, myogenin, and embryonic MyHC more than unstretched muscles. Notably, repetitive stretching resulted in more substantial effects on embryonic MyHC gene expression than continuous stretching. In conclusion, passive stretching for a short duration within a week is effective in increasing myogenic factor expression, and repetitive stretching had more effects than continuous stretching for skeletal muscle on muscle growth. These findings are applicable in clinical muscle-strengthening therapy.

  9. Klf5 regulates muscle differentiation by directly targeting muscle-specific genes in cooperation with MyoD in mice

    Science.gov (United States)

    Hayashi, Shinichiro; Manabe, Ichiro; Suzuki, Yumi; Relaix, Frédéric; Oishi, Yumiko

    2016-01-01

    Krüppel-like factor 5 (Klf5) is a zinc-finger transcription factor that controls various biological processes, including cell proliferation and differentiation. We show that Klf5 is also an essential mediator of skeletal muscle regeneration and myogenic differentiation. During muscle regeneration after injury (cardiotoxin injection), Klf5 was induced in the nuclei of differentiating myoblasts and newly formed myofibers expressing myogenin in vivo. Satellite cell-specific Klf5 deletion severely impaired muscle regeneration, and myotube formation was suppressed in Klf5-deleted cultured C2C12 myoblasts and satellite cells. Klf5 knockdown suppressed induction of muscle differentiation-related genes, including myogenin. Klf5 ChIP-seq revealed that Klf5 binding overlaps that of MyoD and Mef2, and Klf5 physically associates with both MyoD and Mef2. In addition, MyoD recruitment was greatly reduced in the absence of Klf5. These results indicate that Klf5 is an essential regulator of skeletal muscle differentiation, acting in concert with myogenic transcription factors such as MyoD and Mef2. DOI: http://dx.doi.org/10.7554/eLife.17462.001 PMID:27743478

  10. Effect of cyclooxygenase-2 inhibition by meloxicam, on atrogin-1 and myogenic regulatory factors in skeletal muscle of rats injected with endotoxin.

    Science.gov (United States)

    Martin, A I; Nieto-Bona, M P; Castillero, E; Fernandez-Galaz, C; Lopez-Menduina, M; Gomez-Sanmiguel, A B; Gomez-Moreira, C; Villanua, M Angeles; Lopez-Calderon, A

    2012-12-01

    Cyclooxygenase-2-induction by inflammatory stimuli has been proposed as a mediator of inflammatory cachexia. We analyse whether cyclooxygenase-2 inhibition by meloxicam administration is able to modify the response of skeletal muscle to inflammation induced by lipopolysaccharide endotoxin (LPS). Male rats were injected with 1 mg kg(-1) LPS at 17:00 h and at 10:00 h the following day, and euthanized 4, 24 or 72 hours later. Atrogin-1, MuRF1, myogenic regulatory factors and cyclooxygenase-2 in the gastrocnemius were determined by real time-PCR (mRNA) and Western blot (protein). In a second experiment the effect of meloxicam administration (1 mg kg⁻¹) was analyzed. Meloxicam was administered either in a preventive manner, 1 hour before each endotoxin injection, or in a therapeutic manner, starting 2 hours after the second LPS injection and at 24 and 48 hours afterwards. There was a marked increase in MuRF1 mRNA (Pmeloxicam treatment blocked LPS-induced increase in atrogin-1. Preventive, but not therapeutic, meloxicam decreased myostatin (Pmeloxicam treatment decreased gastrocnemius myogenin. These data suggest that cyclooxygenase-2 inhibition by meloxicam administration can prevent the increase in atrogin-1 and the decrease in MyoD induced by LPS administration. However, prolonged therapeutic meloxicam treatment seems to be less effective, since it can inhibit myogenic regulatory factors.

  11. PPARγ and MyoD are differentially regulated by myostatin in adipose-derived stem cells and muscle satellite cells

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Feng [Key Laboratory of Swine Genetics and Breeding of the Agricultural Ministry and Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of the Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan, 430070 (China); Deng, Bing [Wuhan Institute of Animal Science and Veterinary Medicine, Wuhan Academy of Agricultural Science and Technology, Wuhan, Hubei, 430208 (China); Wen, Jianghui [Wu Han University of Technology, Wuhan 430074 (China); Chen, Kun [Key Laboratory of Swine Genetics and Breeding of the Agricultural Ministry and Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of the Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan, 430070 (China); Liu, Wu; Ye, Shengqiang; Huang, Haijun [Wuhan Institute of Animal Science and Veterinary Medicine, Wuhan Academy of Agricultural Science and Technology, Wuhan, Hubei, 430208 (China); Jiang, Siwen, E-mail: jiangsiwen@mail.hzau.edu.cn [Key Laboratory of Swine Genetics and Breeding of the Agricultural Ministry and Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of the Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan, 430070 (China); Xiong, Yuanzhu, E-mail: xiongyzhu@163.com [Key Laboratory of Swine Genetics and Breeding of the Agricultural Ministry and Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of the Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan, 430070 (China)

    2015-03-06

    Myostatin (MSTN) is a secreted protein belonging to the transforming growth factor-β (TGF-β) family that is primarily expressed in skeletal muscle and also functions in adipocyte maturation. Studies have shown that MSTN can inhibit adipogenesis in muscle satellite cells (MSCs) but not in adipose-derived stem cells (ADSCs). However, the mechanism by which MSTN differently regulates adipogenesis in these two cell types remains unknown. Peroxisome proliferator-activated receptor-γ (PPARγ) and myogenic differentiation factor (MyoD) are two key transcription factors in fat and muscle cell development that influence adipogenesis. To investigate whether MSTN differentially regulates PPARγ and MyoD, we analyzed PPARγ and MyoD expression by assessing mRNA, protein and methylation levels in ADSCs and MSCs after treatment with 100 ng/mL MSTN for 0, 24, and 48 h. PPARγ mRNA levels were downregulated after 24 h and upregulated after 48 h of treatment in ADSCs, whereas in MSCs, PPARγ levels were downregulated at both time points. MyoD expression was significantly increased in ADSCs and decreased in MSCs. PPARγ and MyoD protein levels were upregulated in ADSCs and downregulated in MSCs. The CpG methylation levels of the PPARγ and MyoD promoters were decreased in ADSCs and increased in MSCs. Therefore, this study demonstrated that the different regulatory adipogenic roles of MSTN in ADSCs and MSCs act by differentially regulating PPARγ and MyoD expression. - Highlights: • PPARγ and MyoD mRNA and protein levels are upregulated by myostatin in ADSCs. • PPARγ and MyoD mRNA and protein levels are downregulated by myostatin in MSCs. • PPARγ exhibited different methylation levels in myostatin-treated ADSCs and MSCs. • MyoD exhibited different methylation levels in myostatin-treated ADSCs and MSCs. • PPARγ and MyoD are differentially regulated by myostatin in ADSCs and MSCs.

  12. 17β-Estradiol Increases Non-CpG Methylation in Exon 1 of the Rainbow Trout (Oncorhynchus mykiss) MyoD Gene.

    Science.gov (United States)

    Koganti, Prasanthi P; Wang, Jian; Cleveland, Beth; Yao, Jianbo

    2017-08-01

    MyoD is an important myogenic transcription factor necessary for the differentiation of myogenic precursor cells (MPC) to form mature myotubes, a process essential for muscle growth. Epigenetic markers such as CpH methylation are known gene regulators that are important for the differentiation process. In this study, we investigated whether DNA methylation is a potential mechanism associated with the ability of 17β-estradiol (E2) to reduce MyoD gene expression and muscle growth in rainbow trout. Rainbow trout received a single intraperitoneal injection of E2 or the injection vehicle (control). Skeletal muscle was collected 24 h post injection and analyzed for DNA methylation within the MyoD gene and the expression of DNA methyltransferases. CpG islands of the MyoD gene were predicted using MethPrimer software, and these regions were PCR amplified and analyzed using bisulfite sequencing. The percent methylation of the targeted CpG did not differ between control and E2-treated fish. However, percent CpH methylation in the MyoD exon 1 region was elevated with E2 treatment. Two of the methylated CpH sites were located in conserved transcription factor binding motifs, estrogen response element (ERE), and Myc binding site. Quantitative real-time PCR analysis revealed a significant increase in expression of DNA methyltransferases, Dnmt3a and Dnmt3b, in E2-treated muscle, suggesting an increased genome methylation. Differential CpH methylation in MyoD gene of control and E2-treated fish suggests an epigenetic mechanism through which E2 decreases MyoD gene expression and contributes to reduced muscle growth.

  13. Response of growth and myogenic factors in human skeletal muscle to strength training

    NARCIS (Netherlands)

    Liu, Y.; Heinichen, M.; Wirth, K.; Schmidtbleicher, D.; Steinacker, J. M.

    2008-01-01

    Objective: To investigate the response to different strength training techniques of growth and myogenic factors in human skeletal muscle, with particular emphasis on satellite cell (SC) activation. Methods: 24 volunteers were divided into two groups and performed a 6-week strength training (group A

  14. Differential expression of B-crystallin and Hsp27 in skeletal muscle during continuous contractile activity. Relationship to myogenic regulatory factors.

    Science.gov (United States)

    Neufer, P D; Benjamin, I J

    1996-09-27

    AlphaB-crystallin (alphaBC) is a major structural protein (22 kDa) of the ocular lens as well as a bona fide heat shock protein in non-lens tissue. The alphaBC gene is abundantly expressed in tissues with high oxidative capacity, including the heart and type I skeletal muscle fibers, and is regulated by the MyoD family of basic helix-loop-helix transcription factors during myogenesis. To test the hypothesis that alphaBC expression may be directly regulated by the demand for oxidative metabolism, we examined the expression of alphaBC and the ancestral-related Hsp27 in rabbit tibialis anterior muscle subjected to continuous low frequency motor nerve stimulation (3 V, 10 Hz). alphaBC mRNA and protein increased within the 1st day of continuous contractile activity (5- and 2.5-fold, respectively) and achieved maximum levels (>20-and 4-fold, respectively) after 21 d of stimulation. Hsp27 mRNA and protein levels also increased with stimulation, but with a less specific and dramatic induction pattern. In agreement with the Northern analysis, in situ hybridization performed on cross sections from tibialis anterior muscle revealed progressively increasing alphaBC transcript signal, localized in a ringlet pattern, from 1 through 21 days of stimulation. Serial sections subjected to myosin immunohistochemistry revealed that alphaBC expression was confined to slow-twitch type I and a subpopulation of fast twitch type II fibers after 1 day but present in nearly all fibers after 21 days of stimulation. Transcript levels of all four myogenic regulatory factors (MyoD, myogenin, myf-5, and MRF4) also increased with stimulation in a pattern temporally similar with alphaBC, suggesting that expression of alphaBC in response to stimulation may, in part, be regulated through myogenic regulatory factor(s) interaction with the canonical E-box element located within the alphaBC promotor. These data demonstrate that expression of the small heat shock protein, alphaBC, is rapidly induced

  15. Dietary cholecalciferol regulates the recruitment and growth of skeletal muscle fibers and the expressions of myogenic regulatory factors and the myosin heavy chain in European sea bass larvae.

    Science.gov (United States)

    Alami-Durante, Hélène; Cluzeaud, Marianne; Bazin, Didier; Mazurais, David; Zambonino-Infante, José L

    2011-12-01

    The aim of this study was to determine whether dietary cholecalciferol affects the recruitment and growth of axial skeletal muscle fibers in first-feeding European sea bass. Larvae were fed diets containing 0.28 (VD-L, low dose), 0.69 (VD-C, control dose), or 3.00 (VD-H, high dose) mg cholecalciferol/kg from 9 to 44 d posthatching (dph). Larvae were sampled at 44 dph for quantification of somatic growth, muscle growth, and muscle growth dynamics and at 22 and 44 dph for the relative quantification of transcripts encoded by genes involved in myogenesis, cell proliferation, and muscle structure. The weight increase of the VD-L-fed larvae was less than that of the VD-H-fed group, whereas that of VD-C-fed larvae was intermediate. The level of expression of genes involved in cell proliferation (PCNA) and early myogenesis (Myf5) decreased between 22 and 44 dph, whereas that of the myogenic determination factor MyoD1 and that of genes involved in muscle structure and function (myosin heavy chain, myosin light chains 2 and 3) increased. Dietary cholecalciferol regulated Myf5, MyoD1, myogenin, and myosin heavy chain gene expression, with a gene-specific shape of response. The maximum hypertrophy of white muscle fibers was higher in larvae fed the VD-C and VD-H diets than in larvae fed the VD-L diet. White muscle hyperplasia was highly stimulated in VD-H-fed larvae compared to VD-L- and VD-C-fed ones. These findings demonstrate a dietary cholecalciferol effect on skeletal muscle growth mechanisms of a Teleost species.

  16. Variation in myogenic differentiation 1 mRNA abundance is associated with beef tenderness in Nelore cattle.

    Science.gov (United States)

    Tizioto, P C; Coutinho, L L; Mourão, G B; Gasparin, G; Malagó-Jr, W; Bressani, F A; Tullio, R R; Nassu, R T; Taylor, J F; Regitano, L C A

    2016-08-01

    The myogenic differentiation 1 gene (MYOD1) has a key role in skeletal muscle differentiation and composition through its regulation of the expression of several muscle-specific genes. We first used a general linear mixed model approach to evaluate the association of MYOD1 expression levels on individual beef tenderness phenotypes. MYOD1mRNA levels measured by quantitative polymerase chain reactions in 136 Nelore steers were significantly associated (P ≤ 0.01) with Warner-Bratzler shear force, measured on the longissimus dorsi muscle after 7 and 14 days of beef aging. Transcript abundance for the muscle regulatory gene MYOD1 was lower in animals with more tender beef. We also performed a co-expression network analysis using whole transcriptome sequence data generated from 30 samples of longissimus muscle tissue to identify genes that are potentially regulated by MYOD1. The effect of MYOD1 gene expression on beef tenderness may emerge from its function as an activator of muscle-specific gene transcription such as for the serum response factor (C-fos serum response element-binding transcription factor) gene (SRF), which determines muscle tissue development, composition, growth and maturation. © 2016 Stichting International Foundation for Animal Genetics.

  17. Sp3 controls fibroblast growth factor receptor 4 gene activity during myogenic differentiation.

    Science.gov (United States)

    Cavanaugh, Eric; DiMario, Joseph X

    2017-03-27

    Fibroblast growth factor/fibroblast growth factor receptor (FGF/FGFR) signaling is a critical component in the regulation of myoblast proliferation and differentiation. The transient FGFR4 gene expression during the transition from proliferating myoblasts to differentiated myotubes indicates that FGFR4 regulates this critical phase of myogenesis. The Specificity Protein (SP) family of transcription factors controls FGFR family member gene activity. We sought to determine if members of the Sp family regulate mouse FGFR4 gene activity during myogenic differentiation. RT-PCR and western blot analysis of FGFR4 mRNA and protein revealed transient expression over 72h, with peak expression between 24 and 36h after addition of differentiation medium to C2C12 myogenic cultures. Sp3 also displayed a transient expression pattern with peak expression occurring after 6h of differentiation. We cloned a 1527bp fragment of the mouse FGFR4 promoter into a luciferase reporter. This FGFR4 promoter contains eight putative Sp binding sites and directed luciferase gene activity comparable to native FGFR4 expression. Overexpression of Sp1 and Sp3 showed that Sp1 repressed FGFR4 gene activity, and Sp3 activated FGFR4 gene activity during myogenic differentiation. Mutational analyses of multiple Sp binding sites within the FGFR4 promoter revealed that three of these sites were transcriptionally active. Electromobility shift assays and chromatin immunoprecipitation of the area containing the activator sites showed that Sp3 bound to this promoter location.

  18. Myogenic regulatory factor (MRF) expression is affected by exercise in postnatal chicken skeletal muscles.

    Science.gov (United States)

    Yin, Huadong; Li, Diyan; Wang, Yan; Zhao, Xiaoling; Liu, Yiping; Yang, Zhiqin; Zhu, Qing

    2015-05-01

    The MyoD1, MyoG, Myf5, and Mrf4 proteins belong to the family of muscle regulatory factors (MRFs) and play important roles in skeletal muscle hyperplasia and hypertrophy. We hypothesized that exercise would affect MRF mRNA and protein abundance in postnatal chicken skeletal muscle driving molecular changes that could ultimately lead to increased muscle fiber diameter. At day (d) 43, twelve hundred chickens with similar body weight were randomly assigned to cage, pen, and free-range groups. The MRF mRNA abundance was measured in the pectoralis major and thigh muscle at d56, d70, and d84, and the protein levels of MRFs were determined from the thigh muscle at d84. The results showed no significant difference in mRNA of the MRFs among the three groups at d56 (P>0.05). At d84, chicken in the pen and free-range group showed higher MyoD1, MyoG, Myf5, and Mrf4 mRNA abundance compared to the caged chickens (Pchickens had higher Mrf4 and MyoG expression than those in penned ones (Pchickens (Pchickens.

  19. Integrated functions of Pax3 and Pax7 in the regulation of proliferation, cell size and myogenic differentiation.

    Directory of Open Access Journals (Sweden)

    Charlotte A Collins

    Full Text Available Pax3 and Pax7 are paired-box transcription factors with roles in developmental and adult regenerative myogenesis. Pax3 and Pax7 are expressed by postnatal satellite cells or their progeny but are down regulated during myogenic differentiation. We now show that constitutive expression of Pax3 or Pax7 in either satellite cells or C2C12 myoblasts results in an increased proliferative rate and decreased cell size. Conversely, expression of dominant-negative constructs leads to slowing of cell division, a dramatic increase in cell size and altered morphology. Similarly to the effects of Pax7, retroviral expression of Pax3 increases levels of Myf5 mRNA and MyoD protein, but does not result in sustained inhibition of myogenic differentiation. However, expression of Pax3 or Pax7 dominant-negative constructs inhibits expression of Myf5, MyoD and myogenin, and prevents differentiation from proceeding. In fibroblasts, expression of Pax3 or Pax7, or dominant-negative inhibition of these factors, reproduce the effects on cell size, morphology and proliferation seen in myoblasts. Our results show that in muscle progenitor cells, Pax3 and Pax7 function to maintain expression of myogenic regulatory factors, and promote population expansion, but are also required for myogenic differentiation to proceed.

  20. Selective androgen receptor modulator, YK11, regulates myogenic differentiation of C2C12 myoblasts by follistatin expression.

    Science.gov (United States)

    Kanno, Yuichiro; Ota, Rumi; Someya, Kousuke; Kusakabe, Taichi; Kato, Keisuke; Inouye, Yoshio

    2013-01-01

    The myogenic differentiation of C2C12 myoblast cells is induced by the novel androgen receptor (AR) partial agonist, (17α,20E)-17,20-[(1-methoxyethylidene)bis-(oxy)]-3-oxo-19-norpregna-4,20-diene-21-carboxylic acid methyl ester (YK11), as well as by dihydrotestosterone (DHT). YK11 is a selective androgen receptor modulator (SARM), which activates AR without the N/C interaction. In this study, we further investigated the mechanism by which YK11 induces myogenic differentiation of C2C12 cells. The induction of key myogenic regulatory factors (MRFs), such as myogenic differentiation factor (MyoD), myogenic factor 5 (Myf5) and myogenin, was more significant in the presence of YK11 than in the presence of DHT. YK11 treatment of C2C12 cells, but not DHT, induced the expression of follistatin (Fst), and the YK11-mediated myogenic differentiation was reversed by anti-Fst antibody. These results suggest that the induction of Fst is important for the anabolic effect of YK11.

  1. Muscle hypertrophy in heavy weight Japanese quail line: delayed muscle maturation and continued muscle growth with prolonged upregulation of myogenic regulatory factors.

    Science.gov (United States)

    Choi, Y M; Suh, Y; Ahn, J; Lee, K

    2014-09-01

    The objective of this study was to compare the temporal expression of myosin heavy chain (MyHC) isoforms, Pax7, and myogenic regulatory factors (MRF) between heavy weight (HW) and random bred control (RBC) Japanese quail lines during muscle development to better understand the mechanisms leading to increased skeletal muscle mass in the HW quail line selected for a greater BW at 4 wk of age separated from RBC quail. Expression of neonatal MyHC isoform began at 3 and 7 d posthatch in RBC and HW quail lines, respectively. In the RBC quail line, adult MyHC isoform, as a marker for muscle maturation, was expressed at 28 d posthatch with sustained expression through 75 d posthatch, whereas this protein was detected only at 75 d posthatch in the HW quail line. Moreover, Pax7 expression continued from embryonic ages to 14 d posthatch in the HW quail line and to 7 d posthatch in the RBC quail line. These expression patterns of MyHC isoforms and Pax7 in the HW quail line were accompanied by delayed muscle maturation and prolonged growth compared with the RBC quail line. Temporal expressions of the primary MRF showed that higher expression levels of MyoD and Myf-5 were observed at 9 and 11 d embryo in the HW quail line compared with the RBC quail line (P muscle development is associated with delayed maturation and continued muscle growth, which consequently would permit muscle hypertrophic potentials in the HW quail line compared with the RBC quail line. © 2014 Poultry Science Association Inc.

  2. The effect of nutritional status and myogenic satellite cell age on turkey satellite cell proliferation, differentiation, and expression of myogenic transcriptional regulatory factors and heparan sulfate proteoglycans syndecan-4 and glypican-1.

    Science.gov (United States)

    Harthan, Laura B; McFarland, Douglas C; Velleman, Sandra G

    2014-01-01

    Posthatch satellite cell mitotic activity is a critical component of muscle development and growth. Satellite cells are myogenic stem cells that can be induced by nutrition to follow other cellular developmental pathways, and whose mitotic activity declines with age. The objective of the current study was to determine the effect of restricting protein synthesis on the proliferation and differentiation, expression of myogenic transcriptional regulatory factors myogenic determination factor 1, myogenin, and myogenic regulatory factor 4, and expression of the heparan sulfate proteoglycans syndecan-4 and glypican-1 in satellite cells isolated from 1-d-, 7-wk-, and 16-wk-old turkey pectoralis major muscle (1 d, 7 wk, and 16 wk cells, respectively) by using variable concentrations of Met and Cys. Four Met concentrations-30 (control), 7.5, 3, or 0 mg/L with 3.2 mg/L of Cys per 1 mg/L of Met-were used for culture of satellite cells to determine the effect of nutrition and age on satellite cell behavior during proliferation and differentiation. Proliferation was reduced by lower Met and Cys concentrations in all ages at 96 h of proliferation. Differentiation was increased in the 1 d Met-restricted cells, whereas the 7 wk cells treated with 3 mg/L of Met had decreased differentiation. Reduced Met and Cys levels from the control did not significantly affect the 16 wk cells at 72 h of differentiation. However, medium with no Met or Cys suppressed differentiation at all ages. The expression of myogenic determination factor 1, myogenin, myogenic regulatory factor 4, syndecan-4, and glypican-1 was differentially affected by age and Met or Cys treatment. These data demonstrate the age-specific manner in which turkey pectoralis major muscle satellite cells respond to nutritional availability and the importance of defining optimal nutrition to maximize satellite cell proliferation and differentiation for subsequent muscle mass accretion.

  3. Wnt3a signal pathways activate MyoD expression by targeting cis-elements inside and outside its distal enhancer.

    Science.gov (United States)

    Pan, Yu Chih; Wang, Xiao Wen; Teng, Han Feng; Wu, Yi Ju; Chang, Hsuan Chia; Chen, Shen Liang

    2015-03-18

    Wnt proteins are secreted cytokines and several Wnts are expressed in the developing somites and surrounding tissues. Without proper Wnt stimulation, the organization of the dermomyotome and myotome can become defective. These Wnt signals received by somitic cells can lead to activation of Pax3/Pax7 and myogenic regulatory factors (MRFs), especially Myf5 and MyoD. However, it is currently unknown whether Wnts activate Myf5 and MyoD through direct targeting of their cis-regulatory elements or via indirect pathways. To clarify this issue, in the present study, we tested the regulation of MyoD cis-regulatory elements by Wnt3a secreted from human embryonic kidney (HEK)-293T cells. We found that Wnt3a activated the MyoD proximal 6.0k promoter (P6P) only marginally, but highly enhanced the activity of the composite P6P plus distal enhancer (DE) reporter through canonical and non-canonical pathways. Further screening of the intervening fragments between the DE and the P6P identified a strong Wnt-response element (WRE) in the upstream -8 to -9k region (L fragment) that acted independently of the DE, but was dependent on the P6P. Deletion of a Pax3/Pax7-targeted site in the L fragment significantly reduced its response to Wnt3a, implying that Wnt3a activates the L fragment partially through Pax3/Pax7 action. Binding of β-catenin and Pax7 to their target sites in the DE and the L fragment respectively was also demonstrated by ChIP. These observations demonstrated the first time that Wnt3a can directly activate MyoD expression through targeting cis-elements in the DE and the L fragment.

  4. The Effect of Myogenic Factor 5 Polymorphism on the Meat Quality in Chinese Bos Taurus

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    J. A. Ujan

    2011-12-01

    Full Text Available In the present study, we evaluated polymorphism of myogenic factor 5, involved in growth and meat quality traits. Based on PCR-SSCP technology, a novel missense substitution SNP (single-nucleotide polymorphism g.1142 A > G was identified in the intron1 region of the MyF-5 gene, it causes an amino acid substitution (1142Glutamine/ Glycine1142. Allele frequencies, gene heterozygosity, effective allele number and polymorphism information content of the bovine MyF-5 SNP in three population breeds were determined and evaluated by the χ2 test. Results showed that the polymorphism distribution was not similar in all of the three Bos taurus breeds, the genotype distributions of two cattle breeds Jia xian red and Nanyang did not agree with Hardy–Weinberg equilibrium (P 0.05. The A/G allelic frequencies in these breeds were 0.797/0.202, 0.770/0.229, 0.863/0.136 respectively. The genotype frequencies in Jia xian red and Nanyang cattle breeds showed moderate diversity (0.25< polymorphism information content <0.5. Furthermore, least squares analysis revealed significant effects of genotype on intramuscular fat, rib area and water holding capacity in 510 individuals (P < 0.05. Our result suggests that A1142G SNP can be used as an efficacious genetic marker for meat quality traits in native Chinese cattle breeds (Bos taurus but a much large number of animals are required for Marker assisted selection.

  5. Heterogeneity of myotubes generated by the MyoD and E12 basic helix-loop-helix transcription factors in otherwise non-differentiation growth conditions.

    Science.gov (United States)

    Grubišić, Vladimir; Gottipati, Manoj K; Stout, Randy F; Grammer, J Robert; Parpura, Vladimir

    2014-02-01

    We used a synthetic biology approach to produce myotubes from mammalian C2C12 myoblasts in non-differentiation growth conditions using the expression of basic helix-loop-helix transcription factors, MyoD and E12, in various combinations and configurations. Our approach not only recapitulated the basics of muscle development and physiology, as the obtained myotubes showed qualities similar to those seen in striated muscle fibers in vivo, but also allowed for the synthesis of populations of myotubes which assumed distinct morphology, myofibrillar development and Ca(2+) dynamics. This fashioned class of biomaterials is suitable for the building blocks of soft actuators in micro-scale biomimetic robotics. This production line strategy can be embraced in reparative medicine as synthetic human myotubes with predetermined morphological/functional properties could be obtained using this very approach. This methodology can be adopted beyond striated muscle for the engineering of other tissue components/cells whose differentiation is governed by the principles of basic helix-loop-helix transcription factors, as in the case, for example, of neural or immune cell types.

  6. Differential mesodermal expression of two amphioxus MyoD family members (AmphiMRF1 and AmphiMRF2)

    Science.gov (United States)

    Schubert, Michael; Meulemans, Daniel; Bronner-Fraser, Marianne; Holland, Linda Z.; Holland, Nicholas D.

    2003-01-01

    To explore the evolution of myogenic regulatory factors in chordates, we isolated two MyoD family genes (AmphiMRF1 and AmphiMRF2) from amphioxus. AmphiMRF1 is first expressed at the late gastrula in the paraxial mesoderm. As the first somites form, expression is restricted to their myotomal region. In the early larva, expression is strongest in the most anterior and most posterior somites. AmphiMRF2 transcription begins at mid/late gastrula in the paraxial mesoderm, but never spreads into its most anterior region. Through much of the neurula stage, AmphiMRF2 expression is strong in the myotomal region of all somites except the most anterior pair; by late neurula expression is downregulated except in the most posterior somites forming just rostral to the tail bud. These two MRF genes of amphioxus have partly overlapping patterns of mesodermal expression and evidently duplicated independent of the diversification of the vertebrate MRF family.

  7. Magic-factor 1, a partial agonist of Met, induces muscle hypertrophy by protecting myogenic progenitors from apoptosis.

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    Marco Cassano

    Full Text Available BACKGROUND: Hepatocyte Growth Factor (HGF is a pleiotropic cytokine of mesenchymal origin that mediates a characteristic array of biological activities including cell proliferation, survival, motility and morphogenesis. Its high affinity receptor, the tyrosine kinase Met, is expressed by a wide range of tissues and can be activated by either paracrine or autocrine stimulation. Adult myogenic precursor cells, the so called satellite cells, express both HGF and Met. Following muscle injury, autocrine HGF-Met stimulation plays a key role in promoting activation and early division of satellite cells, but is shut off in a second phase to allow myogenic differentiation. In culture, HGF stimulation promotes proliferation of muscle precursors thereby inhibiting their differentiation. METHODOLOGY/PRINCIPAL FINDINGS: Magic-Factor 1 (Met-Activating Genetically Improved Chimeric Factor-1 or Magic-F1 is an HGF-derived, engineered protein that contains two Met-binding domains repeated in tandem. It has a reduced affinity for Met and, in contrast to HGF it elicits activation of the AKT but not the ERK signaling pathway. As a result, Magic-F1 is not mitogenic but conserves the ability to promote cell survival. Here we show that Magic-F1 protects myogenic precursors against apoptosis, thus increasing their fusion ability and enhancing muscular differentiation. Electrotransfer of Magic-F1 gene into adult mice promoted muscular hypertrophy and decreased myocyte apoptosis. Magic-F1 transgenic mice displayed constitutive muscular hypertrophy, improved running performance and accelerated muscle regeneration following injury. Crossing of Magic-F1 transgenic mice with alpha-sarcoglycan knock-out mice -a mouse model of muscular dystrophy- or adenovirus-mediated Magic-F1 gene delivery resulted in amelioration of the dystrophic phenotype as measured by both anatomical/histological analysis and functional tests. CONCLUSIONS/SIGNIFICANCE: Because of these features Magic-F1

  8. Integrative analysis of the zinc finger transcription factor Lame duck in the Drosophila myogenic gene regulatory network.

    Science.gov (United States)

    Busser, Brian W; Huang, Di; Rogacki, Kevin R; Lane, Elizabeth A; Shokri, Leila; Ni, Ting; Gamble, Caitlin E; Gisselbrecht, Stephen S; Zhu, Jun; Bulyk, Martha L; Ovcharenko, Ivan; Michelson, Alan M

    2012-12-11

    Contemporary high-throughput technologies permit the rapid identification of transcription factor (TF) target genes on a genome-wide scale, yet the functional significance of TFs requires knowledge of target gene expression patterns, cooperating TFs, and cis-regulatory element (CRE) structures. Here we investigated the myogenic regulatory network downstream of the Drosophila zinc finger TF Lame duck (Lmd) by combining both previously published and newly performed genomic data sets, including ChIP sequencing (ChIP-seq), genome-wide mRNA profiling, cell-specific expression patterns of putative transcriptional targets, analysis of histone mark signatures, studies of TF cooccupancy by additional mesodermal regulators, TF binding site determination using protein binding microarrays (PBMs), and machine learning of candidate CRE motif compositions. Our findings suggest that Lmd orchestrates an extensive myogenic regulatory network, a conclusion supported by the identification of Lmd-dependent genes, histone signatures of Lmd-bound genomic regions, and the relationship of these features to cell-specific gene expression patterns. The heterogeneous cooccupancy of Lmd-bound regions with additional mesodermal regulators revealed that different transcriptional inputs are used to mediate similar myogenic gene expression patterns. Machine learning further demonstrated diverse combinatorial motif patterns within tissue-specific Lmd-bound regions. PBM analysis established the complete spectrum of Lmd DNA binding specificities, and site-directed mutagenesis of Lmd and additional newly discovered motifs in known enhancers demonstrated the critical role of these TF binding sites in supporting full enhancer activity. Collectively, these findings provide insights into the transcriptional codes regulating muscle gene expression and offer a generalizable approach for similar studies in other systems.

  9. Insulin-like growth factor 1 regulation of proliferation and differentiation of Xenopus laevis myogenic cells in vitro.

    Science.gov (United States)

    Miyata, Sairi; Yada, Tomotaka; Ishikawa, Natsuko; Taheruzzaman, Kazi; Hara, Ryohei; Matsuzaki, Takashi; Nishikawa, Akio

    2017-03-01

    To understand the mechanism of muscle remodeling during Xenopus laevis metamorphosis, we examined the in vitro effect of insulin-like growth factor 1 (IGF-1) on growth and differentiation of three different-fate myogenic cell populations: tadpole tail, tadpole dorsal, and young adult leg muscle. IGF-1 promoted growth and differentiation of both tail and leg myogenic cells only under conditions where these cells could proliferate. Inhibition of cell proliferation by DNA synthesis inhibitor cytosine arabinoside completely canceled the IGF-1's cell differentiation promotion, suggesting the possibility that IGF-1's differentiation-promotion effect is an indirect effect via IGF-1's cell proliferation promotion. IGF-1 promoted differentiation dose dependently with maximum effect at 100-500 ng/ml. RT-PCR analysis revealed the upregulation (11-fold) of ifg1 mRNA expression in developing limbs, suggesting that IGF-1 plays a role in promoting muscle differentiation during limb development. The combined effect of triiodo-L-thyronine (T3) and IGF-1 was also examined. In adult leg cells, IGF-1 promoted growth and differentiation irrespective of the presence of T3. In larval tail cells, cell count was 76% lower in the presence of T3, and IGF-1 did not promote proliferation and differentiation in T3-containing medium. In larval dorsal cells, cell count was also lower in the presence of T3, but IGF-1 enhanced proliferation and differentiation in T3-containing medium. This result is likely due to the presence among dorsal cells of both adult and larval types (1:1). Thus, IGF-1 affects only adult-type myogenic cells in the presence of T3 and helps accelerate dorsal muscle remodeling during metamorphosis.

  10. Positive Control Mutations in the MyoD Basic Region Fail to Show Cooperative DNA Binding and Transcriptional Activation in vitro

    Science.gov (United States)

    Bengal, Eyal; Flores, Osvaldo; Rangarajan, Pundi N.; Chen, Amy; Weintraub, Harold; Verma, Inder M.

    1994-06-01

    An in vitro transcription system from HeLa cells has been established in which MyoD and E47 proteins activate transcription both as homodimers and heterodimers. However, heterodimers activate transcription more efficiently than homodimers, and function synergistically from multiple binding sites. Positive control mutants in the basic region of MyoD that have previously been shown to be defective in initiating the myogenic program, can bind DNA but have lost their ability to function as transcriptional activators in vitro. Additionally, positive control mutants, unlike wild-type MyoD, fail to bind cooperatively to DNA. We propose that binding of MyoD complexes to high affinity MyoD binding sites induces conformational changes that facilitate cooperative binding to multiple sites and promote transcriptional activation.

  11. Myospheres are composed of two cell types: one that is myogenic and a second that is mesenchymal.

    Science.gov (United States)

    Westerman, Karen A

    2015-01-01

    Previously, in an attempt to isolate stem cells that would be capable of regenerating injured skeletal muscle, we cultured cells derived from muscle, non-adherently, in serum-free media. As a result of the culture conditions used, these cells formed spheres, and thus were referred to as myospheres. It was found that myosphere-derived cells expressed Sca-1, a marker that is not typically associated with myogenic cells, and as a result has generated some questions as to the origin of these cells. The goal of this study was to clearly determine the origin of myosphere-derived cells, and in particular to answer the question of whether myospheres contain myogenic cells. To determine if myospheres were composed of myogenic cells without altering the structure of myospheres or the culture conditions used to maintain myospheres, I isolated these cells from yellow fluorescent protein (YFP)-Myf5, YFP-MyoD, and ZsGreen-Pax7 lineage-tracing mice and monitored their growth over time. I found that myospheres do contain myogenic cells, but that these cells are gradually lost over time (within 2 months). Additionally, the use of the lineage-tracing mice gave an interesting perspective into the composition of myospheres. I found that myospheres were composed of two distinct cell types, one that is myogenic (α7 integrin+) and contains cells expressing Myf5, MyoD, and Pax7, and a second that is non-myogenic (α7 integrin-) expressing platelet-derived growth factor receptor alpha (PDGFRα) and Sca-1, both of which have been associated with fibro/adipocyte mesenchymal cells.

  12. Investigation of the Expression of Myogenic Transcription Factors, microRNAs and Muscle-Specific E3 Ubiquitin Ligases in the Medial Gastrocnemius and Soleus Muscles following Peripheral Nerve Injury.

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    Rebecca Wiberg

    Full Text Available Despite surgical innovation, the sensory and motor outcome after a peripheral nerve injury remains incomplete. One contributing factor to the poor outcome is prolonged denervation of the target organ, leading to apoptosis of both mature myofibres and satellite cells with subsequent replacement of the muscle tissue with fibrotic scar and adipose tissue. In this study, we investigated the expression of myogenic transcription factors, muscle specific microRNAs and muscle-specific E3 ubiquitin ligases at several time points following denervation in two different muscles, the gastrocnemius (containing predominantly fast type fibres and soleus (slow type muscles, since these molecules may influence the degree of atrophy following denervation. Both muscles exhibited significant atrophy (compared with the contra-lateral sides at 7 days following either a nerve transection or crush injury. In the crush model, the soleus muscle showed significantly increased muscle weights at days 14 and 28 which was not the case for the gastrocnemius muscle which continued to atrophy. There was a significantly more pronounced up-regulation of MyoD expression in the denervated soleus muscle compared with the gastrocnemius muscle. Conversely, myogenin was more markedly elevated in the gastrocnemius versus soleus muscles. The muscles also showed significantly contrasting transcriptional regulation of the microRNAs miR-1 and miR-206. MuRF1 and Atrogin-1 showed the highest levels of expression in the denervated gastrocnemius muscle. This study provides further insights regarding the intracellular regulatory molecules that generate and maintain distinct patterns of gene expression in different fibre types following peripheral nerve injury.

  13. The role of MyoD1 and histone modifications in the activation of muscle enhancers.

    Science.gov (United States)

    Blum, Roy; Dynlacht, Brian D

    2013-08-01

    MyoD1 is a key regulator that orchestrates skeletal muscle differentiation through the regulation of gene expression. Although many studies have focused on its role in transcriptional control at gene promoters, less is known regarding the role of MyoD1 in the assembly of active enhancers. Here, we discuss novel data that point to the ability of MyoD1 to mediate the assembly of active enhancers that augment the transcription of genes essential for muscle development and lineage specification. Based on genome-wide studies of epigenetic marks that typify active enhancers, we recently identified the compendium of distal regulatory elements that dictate transcriptional programs during myogenesis. Superimposition of MyoD1 binding sites upon the locations of muscle enhancers revealed its unequivocal binding to a core region of nearly a third of condition-specific muscle enhancers. Further studies exploring deposition of enhancer-related epigenetic marks in myoblasts lacking MyoD1 demonstrate the dependence of muscle enhancer assembly on the presence of MyoD1. We propose a model wherein MyoD1 mediates recruitment of Set7, H3K4me1, H3K27ac, p300, and RNAP II to MyoD1-bound enhancers to establish condition-specific activation of muscle genes. Moreover, muscle enhancers are modulated through coordinated binding of transcription factors, including c-Jun, Jdp2, Meis, and Runx1, which are recruited to muscle enhancers in a MyoD1-dependent manner. Thus, MyoD1 and enhancer-associated transcription factors function coordinately to assemble and regulate enhancers, thereby augmenting expression of muscle-related genes.

  14. Testosterone inhibits transforming growth factor-β signaling during myogenic differentiation and proliferation of mouse satellite cells: potential role of follistatin in mediating testosterone action.

    Science.gov (United States)

    Braga, Melissa; Bhasin, Shalender; Jasuja, Ravi; Pervin, Shehla; Singh, Rajan

    2012-03-05

    Testosterone (T) administration is associated with increased satellite cell number and skeletal muscle hypertrophy, although there is considerable heterogeneity in the response of different skeletal muscle groups to T in vivo. We investigated the effects of T on the growth and differentiation of satellite cells isolated from levator ani (LA) and gastrocnemius (gastroc) muscles. T up regulated follistatin (Fst) expression, but down regulated the mRNA and protein expression of a number of genes in the transforming growth factor-beta (TGF-β)-signaling pathway. Inhibition of Fst expression by small interfering RNA (siRNA) inhibited myogenic differentiation and blocked the pro-myogenic effects of T. Treatment of satellite cells with T or Fst up regulated the expression of Pax7 and PCNA, and increased their proliferation. T and Fst blocked TGF-β induced inhibition of growth and myogenic differentiation and down regulated TGF-β-dependent transcriptome in both LA and gastroc cells. We conclude that T stimulation of satellite cell proliferation and myogenic differentiation are associated with up regulation of Fst and inhibition of TGF-β-signaling.

  15. Myogenic potential of human alveolar mucosa derived cells.

    Science.gov (United States)

    Zorin, Vadim L; Pulin, Andrey A; Eremin, Ilya I; Korsakov, Ivan N; Zorina, Alla I; Khromova, Natalia V; Sokova, Olga I; Kotenko, Konstantin V; Kopnin, Pavel B

    2017-03-19

    Difficulties related to the obtainment of stem/progenitor cells from skeletal muscle tissue make the search for new sources of myogenic cells highly relevant. Alveolar mucosa might be considered as a perspective candidate due to availability and high proliferative capacity of its cells. Human alveolar mucosa cells (AMC) were obtained from gingival biopsy samples collected from 10 healthy donors and cultured up to 10 passages. AMC matched the generally accepted multipotent mesenchymal stromal cells criteria and possess population doubling time, caryotype and immunophenotype stability during long-term cultivation. The single myogenic induction of primary cell cultures resulted in differentiation of AMC into multinucleated myotubes. The myogenic differentiation was associated with expression of skeletal muscle markers: skeletal myosin, skeletal actin, myogenin and MyoD1. Efficiency of myogenic differentiation in AMC cultures was similar to that in skeletal muscle cells. Furthermore, some of differentiated myotubes exhibited contractions in vitro. Our data confirms the sufficiently high myogenic potential and proliferative capacity of AMC and their ability to maintain in vitro proliferation-competent myogenic precursor cells regardless of the passage number.

  16. Codependent activators direct myoblast-specific MyoD transcription.

    Science.gov (United States)

    Hu, Ping; Geles, Kenneth G; Paik, Ji-Hye; DePinho, Ronald A; Tjian, Robert

    2008-10-01

    Although FoxO and Pax proteins represent two important families of transcription factors in determining cell fate, they had not been functionally or physically linked together in mediating regulation of a common target gene during normal cellular transcription programs. Here, we identify MyoD, a key regulator of myogenesis, as a direct target of FoxO3 and Pax3/7 in myoblasts. Our cell-based assays and in vitro studies reveal a tight codependent partnership between FoxO3 and Pax3/7 to coordinately recruit RNA polymerase II and form a preinitiation complex (PIC) to activate MyoD transcription in myoblasts. The role of FoxO3 in regulating muscle differentiation is confirmed in vivo by observed defects in muscle regeneration caused by MyoD downregulation in FoxO3 null mice. These data establish a mutual interdependence and functional link between two families of transcription activators serving as potential signaling sensors and regulators of cell fate commitment in directing tissue specific MyoD transcription.

  17. Downhill exercise-induced changes in gene expression related with macrophage polarization and myogenic cells in the triceps long head of rats.

    Science.gov (United States)

    Minari, André Luis Araujo; Oyama, Lila Missae; Dos Santos, Ronaldo Vagner Thomatieli

    2015-02-01

    Macrophages are one of the most heterogenic immune cells involved in skeletal muscle regeneration. After skeletal muscle damage, M1 phenotypes exhibit pro-inflammatory reaction. In a later stage, they are converted to M2 phenotypes with anti-inflammatory properties. To study when gene expressions of macrophage polarization are changed after damage induced by downhill exercise to exhaustion is the objective of this paper. Before (CTRL) and 0 h (G0), 24 h (G24), 48 h (G48) and 72 h (G72) after 18 bouts of downhill exercise, the animals were euthanised, and the triceps were dissected. We measured gene expression of macrophages (CD68 and CD163), myogenic cells (MyoD and myogenin) and quantified cytokine secretion (interleukin (IL)-6, IL-10 and tumour necrosis factor alpha (TNF-α)). The CD68 expression was lower in G72 compared with G24 (P = 0.005) while CD163 was higher in G48 compared with G24 (P = 0.04). The MyoD expression was higher in G72 compared with G0 (P = 0.04). The myogenin expression was lower in G24 compared with CTRL (P = 0.01) and restored in G72 compared with G24 (P = 0.007). The TNF-α was significantly higher at all times after 24 h (all compared with CTRL, with P = 0.03). The CD68 and CD163 expressions behaved distinctly after exercise, which indicates macrophage polarization between 24 and 48 h. The distinct expression of myogenin, concomitantly with MyoD elevation in G72, indicates that myogenic cell differentiation and the significant change of TNF-α level show an important role of this cytokine in these processes.

  18. Spontaneous and specific myogenic differentiation of human mesenchymal stem cells on polyethylene glycol-linked multi-walled carbon nanotube films for skeletal muscle engineering

    Science.gov (United States)

    Zhao, Chunyan; Andersen, Henrik; Ozyilmaz, Barbaros; Ramaprabhu, Sundara; Pastorin, Giorgia; Ho, Han Kiat

    2015-10-01

    This study explored the influence of polyethylene glycol-linked multi-walled carbon nanotube (PEG-CNT) films on skeletal myogenic differentiation of human mesenchymal stem cells (hMSCs). PEG-CNT films were prepared with nanoscale surface roughness, orderly arrangement of PEG-CNTs, high hydrophilicity and high mechanical strength. Notably, PEG-CNT films alone could direct the skeletal myogenic differentiation of hMSCs in the absence of myogenic induction factors. The quantitative real-time polymerase chain reaction (RT-PCR) showed that the non-induced hMSCs plated on the PEG-CNT films, compared to the negative control, presented significant up-regulation of general myogenic markers including early commitment markers of myoblast differentiation protein-1 (MyoD) and desmin, as well as a late phase marker of myosin heavy chain-2 (MHC). Corresponding protein analysis by immunoblot assays corroborated these results. Skeletal muscle-specific markers, fast skeletal troponin-C (TnC) and ryanodine receptor-1 (Ryr) were also significantly increased in the non-induced hMSCs on PEG-CNT films by RT-PCR. For these cells, the commitment to specific skeletal myoblasts was further proved by the absence of enhanced adipogenic, chondrogenic and osteogenic markers. This study elucidated that PEG-CNT films supported a dedicated differentiation of hMSCs into a skeletal myogenic lineage and can work as a promising material towards skeletal muscle injury repair.This study explored the influence of polyethylene glycol-linked multi-walled carbon nanotube (PEG-CNT) films on skeletal myogenic differentiation of human mesenchymal stem cells (hMSCs). PEG-CNT films were prepared with nanoscale surface roughness, orderly arrangement of PEG-CNTs, high hydrophilicity and high mechanical strength. Notably, PEG-CNT films alone could direct the skeletal myogenic differentiation of hMSCs in the absence of myogenic induction factors. The quantitative real-time polymerase chain reaction (RT-PCR) showed

  19. Efeito da natação na expressão de fatores regulatórios miogênicos durante o reparo do musculoesquelético de rato Swimming influence on the expression of myogenic regulatory factors during skeletal muscle repair of rats

    Directory of Open Access Journals (Sweden)

    Paulo Roberto Dantas Pestana

    2012-12-01

    addition, it has demonstrated positive results in the therapeutic process and preventing several diseases.. OBJECTIVE: The aim of the present study was to analyze the effect of swimming on the expression of the myogenic regulatory factors MyoD and myogenin during the skeletal muscle repair process in rats following cryoinjury. METHODS: Forty Wistar rats were randomly divided into four groups: 1 Control; 2 Sham - non-muscle damaged, submitted to procedure for exposure of the tibialis anterior (TA muscle; 3 Cryoinjured; and 4 Cryoinjured and submitted to swimming. Analyses were carried out at 7, 14 and 21 days. Cryoinjury was performed with two applications of the flat end of a metal rod previously cooled in liquid nitrogen directly to the belly of the TA muscle. The protocol consisted of 90-minute swimming sessions six times a week. At the end of the protocol, the animals were euthanized and the TA muscles were removed. Total RNA was extracted using the TRIzol reagent. Next, cDNA was obtained to perform real-time PCR using specific primers for MyoD and myogenin. RESULTS: The results showed a reduction in the expression of myogenin in the groups cryoinjury with p 0.05 and without (p>0.05 swimming. Regarding the expression of MyoD there was no difference between the groups. CONCLUSION: Swimming did not affect the expression of myogenic regulatory factors during the skeletal muscle repair process in rats following cryoinjury.

  20. The Tocotrienol-Rich Fraction Is Superior to Tocopherol in Promoting Myogenic Differentiation in the Prevention of Replicative Senescence of Myoblasts.

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    Shy Cian Khor

    Full Text Available Aging results in a loss of muscle mass and strength. Myoblasts play an important role in maintaining muscle mass through regenerative processes, which are impaired during aging. Vitamin E potentially ameliorates age-related phenotypes. Hence, this study aimed to determine the effects of the tocotrienol-rich fraction (TRF and α-tocopherol (ATF in protecting myoblasts from replicative senescence and promoting myogenic differentiation. Primary human myoblasts were cultured into young and senescent stages and were then treated with TRF or ATF for 24 h, followed by an analysis of cell proliferation, senescence biomarkers, cellular morphology and differentiation. Our data showed that replicative senescence impaired the normal regenerative processes of myoblasts, resulting in changes in cellular morphology, cell proliferation, senescence-associated β-galactosidase (SA-β-gal expression, myogenic differentiation and myogenic regulatory factors (MRFs expression. Treatment with both TRF and ATF was beneficial to senescent myoblasts in reclaiming the morphology of young cells, improved cell viability and decreased SA-β-gal expression. However, only TRF treatment increased BrdU incorporation in senescent myoblasts, as well as promoted myogenic differentiation through the modulation of MRFs at the mRNA and protein levels. MYOD1 and MYOG gene expression and myogenin protein expression were modulated in the early phases of myogenic differentiation. In conclusion, the tocotrienol-rich fraction is superior to α-tocopherol in ameliorating replicative senescence-related aberration and promoting differentiation via modulation of MRFs expression, indicating vitamin E potential in modulating replicative senescence of myoblasts.

  1. Effects of Sunphenon and Polyphenon 60 on proteolytic pathways, inflammatory cytokines and myogenic markers in H22-treated C2C12 cells

    Indian Academy of Sciences (India)

    Allur Subramaniyan Sivakumar; Inho Hwang

    2015-03-01

    The effect of Sunphenon and Polyphenon 60 in oxidative stress response, myogenic regulatory factors, inflammatory cytokines, apoptotic and proteolytic pathways on H2O2-induced myotube atrophy was addressed. Cellular responses of H2O2-induced C2C12cells were examined, including mRNA expression of myogenic regulatory factors, such as MyoD and myogenin, inflammatory pathways, such as TNF- and NF-kB, as well as proteolytic enzymes, such as -calpain and m-calpain. The pre-treatment of Sunphenon (50 g/mL)/Polyphenon 60 (50 g/mL) on H2O2-treated C2C12 cells significantly down-regulated the mRNA expression of myogenin and MyoD when compared to those treated with H2O2-induced alone. Additionally, the mRNA expression of -calpain and m-calpain were significantly ( < 0.05) increased in H2O2-treated C2C12 cells, whereas pre-treatment with Sunphenon/Polyphenon significantly down-regulated the above genes, namely -calpain and m-calpain. Furthermore, the mRNA expression of TNF- and NF-kB were significantly increased in H2O2-treated C2C12 cells, while pre-treatment with Sunphenon (50 g/mL)/Polyphenon 60 (50 g/mL) significantly ( < 0.05) down-regulated it when compared to the untreated control group. Subsequent analysis of DNA degeneration and caspase activation revealed that Sunphenon (50 g/mL)/Polyphenon 60 (50 g/mL) inhibited activation of caspase-3 and showed an inhibitory effect on DNA degradation. From this result, we know that, in stress conditions, -calpain may be involved in the muscle atrophy through the suppression of myogenin and MyoD. Moreover, Sunphenon may regulate the skeletal muscle genes/promote skeletal muscle recovery by the up-regulation of myogenin and MyoD and suppression of -calpain and inflammatory pathways and may regulate the apoptosis pathways. Our findings suggest that dietary supplementation of Sunphenon might reduce inflammatory events in muscle-associated diseases, such as myotube atrophy.

  2. Comparative effects of low-level laser therapy pre- and post-injury on mRNA expression of MyoD, myogenin, and IL-6 during the skeletal muscle repair.

    Science.gov (United States)

    Alves, Agnelo Neves; Ribeiro, Beatriz Guimarães; Fernandes, Kristianne Porta Santos; Souza, Nadhia Helena Costa; Rocha, Lília Alves; Nunes, Fabio Daumas; Bussadori, Sandra Kalil; Mesquita-Ferrari, Raquel Agnelli

    2016-05-01

    This study analyzed the effect of pre-injury and post-injury irradiation with low-level laser therapy (LLLT) on the mRNA expression of myogenic regulatory factors and interleukin 6 (IL-6) during the skeletal muscle repair. Male rats were divided into six groups: control group, sham group, LLLT group, injury group; pre-injury LLLT group, and post-injury LLLT group. LLLT was performed with a diode laser (wavelength 780 nm; output power 40 mW' and total energy 3.2 J). Cryoinjury was induced by two applications of a metal probe cooled in liquid nitrogen directly onto the belly of the tibialis anterior (TA) muscle. After euthanasia, the TA muscle was removed for the isolation of total RNA and analysis of MyoD, myogenin, and IL-6 using real-time quantitative PCR. Significant increases were found in the expression of MyoD mRNA at 3 and 7 days as well as the expression of myogenin mRNA at 14 days in the post-injury LLLT group in comparison to injury group. A significant reduction was found in the expression of IL-6 mRNA at 3 and 7 days in the pre-injury LLLT and post-injury LLLT groups. A significant increase in IL-6 mRNA was found at 14 days in the post-injury LLLT group in comparison to the injury group. LLLT administered following muscle injury modulates the mRNA expression of MyoD and myogenin. Moreover, the both forms of LLLT administration were able to modulate the mRNA expression of IL-6 during the muscle repair process.

  3. Protein O-fucosyltransferase 1 expression impacts myogenic C2C12 cell commitment via the Notch signaling pathway.

    Science.gov (United States)

    Der Vartanian, Audrey; Audfray, Aymeric; Al Jaam, Bilal; Janot, Mathilde; Legardinier, Sébastien; Maftah, Abderrahman; Germot, Agnès

    2015-01-01

    The Notch signaling pathway plays a crucial role in skeletal muscle regeneration in mammals by controlling the transition of satellite cells from quiescence to an activated state, their proliferation, and their commitment toward myotubes or self-renewal. O-fucosylation on Notch receptor epidermal growth factor (EGF)-like repeats is catalyzed by the protein O-fucosyltransferase 1 (Pofut1) and primarily controls Notch interaction with its ligands. To approach the role of O-fucosylation in myogenesis, we analyzed a murine myoblastic C2C12 cell line downregulated for Pofut1 expression by short hairpin RNA (shRNA) inhibition during the time course of differentiation. Knockdown of Pofut1 affected the signaling pathway activation by a reduction of the amount of cleaved Notch intracellular domain and a decrease in downstream Notch target gene expression. Depletion in Pax7(+)/MyoD(-) cells and earlier myogenic program entrance were observed, leading to an increase in myotube quantity with a small number of nuclei, reflecting fusion defects. The rescue of Pofut1 expression in knockdown cells restored Notch signaling activation and a normal course in C2C12 differentiation. Our results establish the critical role of Pofut1 on Notch pathway activation during myogenic differentiation.

  4. Analysis list: Myod1 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Myod1 Adipocyte,Embryonic fibroblast,Muscle,Pluripotent stem cell + mm9 http://dbar...chive.biosciencedbc.jp/kyushu-u/mm9/target/Myod1.1.tsv http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/target/My...od1.5.tsv http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/target/Myod1.10.tsv http://dbarchive.bioscienced...bc.jp/kyushu-u/mm9/colo/Myod1.Adipocyte.tsv,http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/colo/My...od1.Embryonic_fibroblast.tsv,http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/colo/Myod1.Musc

  5. An update on MyoD evolution in teleosts and a proposed consensus nomenclature to accommodate the tetraploidization of different vertebrate genomes.

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    Daniel J Macqueen

    Full Text Available BACKGROUND: MyoD is a muscle specific transcription factor that is essential for vertebrate myogenesis. In several teleost species, including representatives of the Salmonidae and Acanthopterygii, but not zebrafish, two or more MyoD paralogues are conserved that are thought to have arisen from distinct, possibly lineage-specific duplication events. Additionally, two MyoD paralogues have been characterised in the allotetraploid frog, Xenopus laevis. This has lead to a confusing nomenclature since MyoD paralogues have been named outside of an appropriate phylogenetic framework. METHODS AND PRINCIPAL FINDINGS: Here we initially show that directly depicting the evolutionary relationships of teleost MyoD orthologues and paralogues is hindered by the asymmetric evolutionary rate of Acanthopterygian MyoD2 relative to other MyoD proteins. Thus our aim was to confidently position the event from which teleost paralogues arose in different lineages by a comparative investigation of genes neighbouring myod across the vertebrates. To this end, we show that genes on the single myod-containing chromosome of mammals and birds are retained in both zebrafish and Acanthopterygian teleosts in a striking pattern of double conserved synteny. Further, phylogenetic reconstruction of these neighbouring genes using Bayesian and maximum likelihood methods supported a common origin for teleost paralogues following the split of the Actinopterygii and Sarcopterygii. CONCLUSION: Our results strongly suggest that myod was duplicated during the basal teleost whole genome duplication event, but was subsequently lost in the Ostariophysi (zebrafish and Protacanthopterygii lineages. We propose a sensible consensus nomenclature for vertebrate myod genes that accommodates polyploidization events in teleost and tetrapod lineages and is justified from a phylogenetic perspective.

  6. Myogenic repressor I-mfa interferes with the function of Zic family proteins.

    Science.gov (United States)

    Mizugishi, Kiyomi; Hatayama, Minoru; Tohmonda, Takahide; Ogawa, Miyuki; Inoue, Takashi; Mikoshiba, Katsuhiko; Aruga, Jun

    2004-07-16

    Zinc finger proteins belonging to the Zic family control several developmental processes such as patterning of the axial skeleton. Here we mapped the transcriptional regulatory domains in Zic2 protein and identified a protein which specifically binds to one of them. In the mapping experiments, an amino-terminal region was identified as transcriptional regulatory domains. A search for proteins binding to the amino terminal domain of Zic2 revealed that inhibitor of MyoD family (I-mfa) protein, which has been identified as a repressor of myogenic helix-loop-helix class transcription factors, can physically interact with the amino terminal domain. When Zic1-3 and I-mfa proteins were co-expressed in cultured cells, nuclear import of the Zic proteins was inhibited. Consequently, I-mfa inhibited transcriptional activation by the Zic proteins in cultured cells. These results suggest that the physical and functional interaction between Zic and I-mfa proteins can play a role in the vertebrate development.

  7. Early inflammatory and myogenic responses to resistance exercise in the elderly.

    Science.gov (United States)

    Mathers, Jessica L; Farnfield, Michelle M; Garnham, Andrew P; Caldow, Marissa K; Cameron-Smith, David; Peake, Jonathan M

    2012-09-01

    This study compared changes in myokine and myogenic genes following resistance exercise (3 sets of 12 repetitions of maximal unilateral knee extension) in 20 elderly men (67.8 ± 1.0 years) and 15 elderly women (67.2 ± 1.5 years). Monocyte chemotactic protein (MCP)-1, macrophage inhibitory protein (MIP)-1β, interleukin (IL)-6 and MyoD mRNA increased significantly (P exercise in both groups. Macrophage-1 (Mac-1) and MCP-3 mRNA did not change significantly after exercise in either group. MIP-1β, Mac-1 and myostatin mRNA were significantly higher before and after exercise in men compared with women. In contrast, MCP-3 and myogenin mRNA were significantly higher before and after exercise in the women compared with the men. In elderly individuals, gender influences the mRNA expression of certain myokines and growth factors, both at rest and after resistance exercise. These differences may influence muscle regeneration following muscle injury. Copyright © 2012 Wiley Periodicals, Inc.

  8. TEAD transcription factors are required for normal primary myoblast differentiation in vitro and muscle regeneration in vivo.

    Science.gov (United States)

    Joshi, Shilpy; Davidson, Guillaume; Le Gras, Stéphanie; Watanabe, Shuichi; Braun, Thomas; Mengus, Gabrielle; Davidson, Irwin

    2017-02-01

    The TEAD family of transcription factors (TEAD1-4) bind the MCAT element in the regulatory elements of both growth promoting and myogenic differentiation genes. Defining TEAD transcription factor function in myogenesis has proved elusive due to overlapping expression of family members and their functional redundancy. We show that silencing of either Tead1, Tead2 or Tead4 did not effect primary myoblast (PM) differentiation, but that their simultaneous knockdown strongly impaired differentiation. In contrast, Tead1 or Tead4 silencing impaired C2C12 differentiation showing their different contributions in PMs and C2C12 cells. Chromatin immunoprecipitation identified enhancers associated with myogenic genes bound by combinations of Tead4, Myod1 or Myog. Tead4 regulated distinct gene sets in C2C12 cells and PMs involving both activation of the myogenic program and repression of growth and signaling pathways. ChIP-seq from mature mouse muscle fibres in vivo identified a set of highly transcribed muscle cell-identity genes and sites bound by Tead1 and Tead4. Although inactivation of Tead4 in mature muscle fibres caused no obvious phenotype under normal conditions, notexin-induced muscle regeneration was delayed in Tead4 mutants suggesting an important role in myogenic differentiation in vivo. By combining knockdown in cell models in vitro with Tead4 inactivation in muscle in vivo, we provide the first comprehensive description of the specific and redundant roles of Tead factors in myogenic differentiation.

  9. TEAD transcription factors are required for normal primary myoblast differentiation in vitro and muscle regeneration in vivo.

    Directory of Open Access Journals (Sweden)

    Shilpy Joshi

    2017-02-01

    Full Text Available The TEAD family of transcription factors (TEAD1-4 bind the MCAT element in the regulatory elements of both growth promoting and myogenic differentiation genes. Defining TEAD transcription factor function in myogenesis has proved elusive due to overlapping expression of family members and their functional redundancy. We show that silencing of either Tead1, Tead2 or Tead4 did not effect primary myoblast (PM differentiation, but that their simultaneous knockdown strongly impaired differentiation. In contrast, Tead1 or Tead4 silencing impaired C2C12 differentiation showing their different contributions in PMs and C2C12 cells. Chromatin immunoprecipitation identified enhancers associated with myogenic genes bound by combinations of Tead4, Myod1 or Myog. Tead4 regulated distinct gene sets in C2C12 cells and PMs involving both activation of the myogenic program and repression of growth and signaling pathways. ChIP-seq from mature mouse muscle fibres in vivo identified a set of highly transcribed muscle cell-identity genes and sites bound by Tead1 and Tead4. Although inactivation of Tead4 in mature muscle fibres caused no obvious phenotype under normal conditions, notexin-induced muscle regeneration was delayed in Tead4 mutants suggesting an important role in myogenic differentiation in vivo. By combining knockdown in cell models in vitro with Tead4 inactivation in muscle in vivo, we provide the first comprehensive description of the specific and redundant roles of Tead factors in myogenic differentiation.

  10. Inhibition of connective tissue growth factor (CTGF/CCN2) expression decreases the survival and myogenic differentiation of human rhabdomyosarcoma cells.

    Science.gov (United States)

    Croci, Stefania; Landuzzi, Lorena; Astolfi, Annalisa; Nicoletti, Giordano; Rosolen, Angelo; Sartori, Francesca; Follo, Matilde Y; Oliver, Noelynn; De Giovanni, Carla; Nanni, Patrizia; Lollini, Pier-Luigi

    2004-03-01

    Connective tissue growth factor (CTGF/CCN2), a cysteine-rich protein of the CCN (Cyr61, CTGF, Nov) family of genes, emerged from a microarray screen of genes expressed by human rhabdomyosarcoma cells. Rhabdomyosarcoma is a soft tissue sarcoma of childhood deriving from skeletal muscle cells. In this study, we investigated the role of CTGF in rhabdomyosarcoma. Human rhabdomyosarcoma cells of the embryonal (RD/12, RD/18, CCA) and the alveolar histotype (RMZ-RC2, SJ-RH4, SJ-RH30), rhabdomyosarcoma tumor specimens, and normal skeletal muscle cells expressed CTGF. To determine the function of CTGF, we treated rhabdomyosarcoma cells with a CTGF antisense oligonucleotide or with a CTGF small interfering RNA (siRNA). Both treatments inhibited rhabdomyosarcoma cell growth, suggesting the existence of a new autocrine loop based on CTGF. CTGF antisense oligonucleotide-mediated growth inhibition was specifically due to a significant increase in apoptosis, whereas cell proliferation was unchanged. CTGF antisense oligonucleotide induced a strong decrease in the level of myogenic differentiation of rhabdomyosarcoma cells, whereas the addition of recombinant CTGF significantly increased the proportion of myosin-positive cells. CTGF emerges as a survival and differentiation factor and could be a new therapeutic target in human rhabdomyosarcoma.

  11. Co-dependent Activators Direct Myoblast Specific MyoD Transcription

    Science.gov (United States)

    Hu, Ping; Geles, Kenneth G.; Paik, Ji-Hye; DePinho, Ronald A.; Tjian, Robert

    2008-01-01

    Summary Although FoxO and Pax proteins represent two important families of transcription factors in determining cell fate, they had not been functionally or physically linked together in mediating regulation of a common target gene during normal cellular transcription programs. Here we identify MyoD, a key regulator of myogenesis, as a direct target of FoxO3 and Pax3/7 in myoblasts. Our cell based assays and in vitro studies reveal a tight co-dependent partnership between FoxO3 and Pax3/7 to coordinately recruit RNA polymerase II and form a pre-initiation complex (PIC) to activate MyoD transcription in myoblasts. The role of FoxO3 in regulating muscle differentiation is confirmed in vivo by observed defects in muscle regeneration caused by MyoD down-regulation in FoxO3 null mice. These data establish a mutual interdependence and functional link between two families of transcription activators serving as potential signaling sensors and regulators of cell fate commitment in directing tissue specific MyoD transcription. PMID:18854138

  12. Effects of matrix metalloproteinase-1 on the myogenic differentiation of bone marrow-derived mesenchymal stem cells in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Zheng, Zhenyang [Department of Neurology, The First Affiliated Hospital, Sun Yat-sen University, No. 58, Zhongshan 2nd Road, Guangzhou 510080, Guangdong Province (China); Leng, Yan [Department of Rehabilitation, The First Affiliated Hospital, Sun Yat-sen University, No. 58, Zhongshan 2nd Road, Guangzhou 510080, Guangdong Province (China); Zhou, Chen; Ma, Zhenyu; Zhong, Zhigang [Department of Neurology, The First Affiliated Hospital, Sun Yat-sen University, No. 58, Zhongshan 2nd Road, Guangzhou 510080, Guangdong Province (China); Shi, Xing-Ming [Institute of Molecular Medicine and Genetics, Georgia Health Sciences University, Augusta, GA 30912 (United States); Zhang, Weixi, E-mail: weixizhang@qq.com [Department of Neurology, The First Affiliated Hospital, Sun Yat-sen University, No. 58, Zhongshan 2nd Road, Guangzhou 510080, Guangdong Province (China)

    2012-11-16

    Highlights: Black-Right-Pointing-Pointer MMP-1 is a member of the zinc-dependent endopeptidase family. Black-Right-Pointing-Pointer MMP-1 has no cytotoxic effects on BMSCs. Black-Right-Pointing-Pointer MMP-1 can promote the myogenic differentiation of BMSCs. Black-Right-Pointing-Pointer MyoD and desmin were chosen as myogenic markers in this study. -- Abstract: Matrix metalloproteinase-1 (MMP-1) is a member of the family of zinc-dependent endopeptidases that are capable of degrading extracellular matrix (ECM) and certain non-matrix proteins. It has been shown that MMP-1 can enhance muscle regeneration by improving the differentiation and migration of myoblasts. However, it is still not known whether MMP-1 can promote the myogenesis of bone marrow-derived mesenchymal stem cells (BMSCs). To address this question, we isolated BMSCs from C57BL/6J mice and investigated the effects of MMP-1 on their proliferation and myogenic differentiation. Our results showed that MMP-1 treatment, which had no cytotoxic effects on BMSCs, increased the mRNA and protein levels of MyoD and desmin in a dose-dependent manner, indicating that MMP-1 promoted myogenic differentiation of BMSCs in vitro. These results suggest that BMSCs may have a therapeutic potential for treating muscular disorders.

  13. Myogenic skeletal muscle satellite cells communicate by tunnelling nanotubes.

    Science.gov (United States)

    Tavi, Pasi; Korhonen, Topi; Hänninen, Sandra L; Bruton, Joseph D; Lööf, Sara; Simon, Andras; Westerblad, Håkan

    2010-05-01

    Quiescent satellite cells sit on the surface of the muscle fibres under the basal lamina and are activated by a variety of stimuli to disengage, divide and differentiate into myoblasts that can regenerate or repair muscle fibres. Satellite cells adopt their parent's fibre type and must have some means of communication with the parent fibre. The mechanisms behind this communication are not known. We show here that satellite cells form dynamic connections with muscle fibres and other satellite cells by F-actin based tunnelling nanotubes (TNTs). Our results show that TNTs readily develop between satellite cells and muscle fibres. Once developed, TNTs permit transport of intracellular material, and even cellular organelles such as mitochondria between the muscle fibre and satellite cells. The onset of satellite cell differentiation markers Pax-7 and MyoD expression was slower in satellite cells cultured in the absence than in the presence of muscle cells. Furthermore physical contact between myofibre and satellite cell progeny is required to maintain subtype identity. Our data establish that TNTs constitute an integral part of myogenic cell communication and that physical cellular interaction control myogenic cell fate determination.

  14. Influence of single nucleotide polymorphisms in the myostatin and myogenic factor 5 muscle growth-related genes on the performance traits of Marchigiana beef cattle.

    Science.gov (United States)

    Sarti, F M; Lasagna, E; Ceccobelli, S; Di Lorenzo, P; Filippini, F; Sbarra, F; Giontella, A; Pieramati, C; Panella, F

    2014-09-01

    The Marchigiana is famous for its large body size and favorable dressing percentage. A myostatin (MSTN) gene mutation (a G to T transversion) was identified in the breed. The homozygote "GG" yields a "normal" phenotype, the homozygote "TT" yields a double muscled body shape but sometimes causes survival problems, and the heterozygote genotype produces an extremely muscled body without defects. In practice, Marchigiana "TT" homozygotes are culled from reproduction, but the heterozygotes are chosen as sires. The objective of this study was to assess genes involved in Marchigiana muscle development to improve selection procedures. The effects of the MSTN and myogenic factor 5 (MYF5) genes on the growth and muscle traits in the Marchigiana breed were assessed. The effects of MSTN together with the genotype of the causative mutation (g.874G > T) and the effects of the two SNP in the promoter were studied (g.-371T > A and g.-805G > C). The SNP effects were evaluated in a comparison between the means of the several genotypes or for the average gene substitution and dominance effect. Two hundred forty-nine bullocks were evaluated using a performance test. At the beginning and end of the trial, the animals were weighed and their bodies were measured every 21 d up to 12 mo of age. In addition to these observations, morphological scores and the BLUP indices were estimated at the end of the performance test. The obtained results suggested that the MSTN g.874G > T and MYF5 SNP could be considered in the selection program of the Marchigiana breed. A MSTN g.874G > T genotyping service for the breeders could help to avoid the "TT" genotype and to select for the "GT" genotype. The "AA" MYF5 SNP genotype could also be selected for even if good muscle development yields a certain size reduction.

  15. [Recording cervical and ocular vestibular evoked myogenic potentials. Part 2: influencing factors, evaluation of findings and clinical significance].

    Science.gov (United States)

    Walther, L E; Hörmann, K; Pfaar, O

    2010-11-01

    VEMP measurements are subject to various influencing factors: patient age, threshold, sound intensity and frequency. Using air (AC) and bone conduction (BC) the vestibular receptors and afferents of the otolith organs can be activated to varying degrees. Recordings of cervical (cVEMP) and ocular VEMP (oVEMP) are clinically possible. AC-cVEMP are primarily an indicator of the sacculocollic reflex pathway. Together with findings on the vestibuloocular reflex (VOR) and complimentary otolith tests, VEMP enable otolith function analysis of each side separately. In addition, the distinction between combined or isolated canal and otolith dysfunction in terms of subtyping and patterns of damage in mono- and bilateral disorders, such as vestibular neuritis or bilateral vestibulopathy, is possible. Moreover, VEMP is relevant in terms of prognostic and therapeutic considerations as well as expert assessments.

  16. Myogenous temporomandibular disorders: diagnostic and management considerations.

    Science.gov (United States)

    Fricton, James

    2007-01-01

    Myogenous temporomandibular disorders (or masticatory myalgia) are characterized by pain and dysfunction that arise from pathologic and functional processes in the masticatory muscles. There are several distinct muscle disorder subtypes in the masticatory system, including myofascial pain, myositis, muscle spasm, and muscle contracture. The major characteristics of masticatory myalgia include pain, muscle tenderness, limited range of motion, and other symptoms (eg, fatigability, stiffness, subjective weakness). Comorbid conditions and complicating factors also are common and are discussed. Management follows with stretching, posture, and relaxation exercises, physical therapy, reduction of contributing factors, and as necessary, muscle injections.

  17. Promoting effect of 5-azacytidine on the myogenic differentiation of bone marrow stromal cells.

    Science.gov (United States)

    Burlacu, Alexandrina; Rosca, Ana-Maria; Maniu, Horia; Titorencu, Irina; Dragan, Emanuel; Jinga, Victor; Simionescu, Maya

    2008-03-01

    Bone marrow stromal cells (BMSC) can differentiate into various cell types including myocytes, which may be valuable in cellular therapy of myocardial infarction. In an attempt to increase the myogenic commitment of BMSC, we investigated the extent of conversion induced by the demethylation agent 5-azacytidine. BMSC isolated from the adult rat tibia were exposed in culture to 5microM 5-azacytidine for 24h, 1 day after seeding. The treatment was repeated at weekly intervals and the expression of muscle-specific proteins and genes was assessed. The results revealed that cultured cells lost the native expression of osteocalcin and alkaline phosphatase as a function of time and began to express connexin 43. Exposure to 5-azacytidine of BMSC induced, at 14 days, a myocyte-resembling phenotype that included the expression of muscle-specific proteins (sarcomeric alpha-actin, troponin T, desmin, alpha-actinin, and GATA-4) and genes (GATA-4, myoD, desmin, and alpha-actinin), numerous mitochondria and myofilaments; however, the latter did not form sarcomeres. Although some of these myogenic markers also appeared in untreated cells, exposure to 5-azacytidine induced an enhanced response of calcium channels, as well as a threefold increase in desmin and myoD gene expression and a twofold increase in alpha-actinin gene and protein expression above the control values. In conclusion, the results demonstrate a promoting effect of 5-azacytidine on the expression of muscle-specific proteins and genes in BMSC in culture. Notably, the myogenic differentiation takes place over a short period of time. Priming of mesenchymal cells to cardiomyogenic differentiation may have significant applications in cellular approaches to ameliorate muscle loss after myocardial ischemia.

  18. Characteristics of myogenic response and ankle torque recovery after lengthening contraction-induced rat gastrocnemius injury

    Directory of Open Access Journals (Sweden)

    Song Hongsun

    2012-10-01

    Full Text Available Abstract Background Although muscle dysfunction caused by unfamiliar lengthening contraction is one of most important issues in sports medicine, there is little known about the molecular events on regeneration process. The purpose of this study was to investigate the temporal and spatial expression patterns of myogenin, myoD, pax7, and myostatin after acute lengthening contraction (LC-induced injury in the rat hindlimb. Methods We employed our originally developed device with LC in rat gastrocnemius muscle (n = 24. Male Wistar rats were anesthetized with isoflurane (aspiration rate, 450 ml/min, concentration, 2.0%. The triceps surae muscle of the right hindlimb was then electrically stimulated with forced isokinetic dorsi-flexion (180°/sec and from 0 to 45°. Tissue contents of myoD, myogenin, pax7, myostatin were measured by western blotting and localizations of myoD and pax7 was measured by immunohistochemistry. After measuring isometric tetanic torque, a single bout of LC was performed in vivo. Results The torque was significantly decreased on days 2 and 5 as compared to the pre-treatment value, and recovered by day 7. The content of myoD and pax7 showed significant increases on day 2. Myogenin showed an increase from day 2 to 5. Myostatin on days 5 and 7 were significantly increased. Immunohistochemical analysis showed that myoD-positive/pax7-positive cells increased on day 2, suggesting that activated satellite cells play a role in the destruction and the early recovery phases. Conclusion We, thus, conclude that myogenic events associate with torque recovery after LC-induced injury.

  19. NFATc1 Controls Skeletal Muscle Fiber Type and Is a Negative Regulator of MyoD Activity

    Directory of Open Access Journals (Sweden)

    Melissa L. Ehlers

    2014-09-01

    Full Text Available Skeletal muscle comprises a heterogeneous population of fibers with important physiological differences. Fast fibers are glycolytic and fatigue rapidly. Slow fibers utilize oxidative metabolism and are fatigue resistant. Muscle diseases such as sarcopenia and atrophy selectively affect fast fibers, but the molecular mechanisms regulating fiber type-specific gene expression remain incompletely understood. Here, we show that the transcription factor NFATc1 controls fiber type composition and is required for fast-to-slow fiber type switching in response to exercise in vivo. Moreover, MyoD is a crucial transcriptional effector of the fast fiber phenotype, and we show that NFATc1 inhibits MyoD-dependent fast fiber gene promoters by physically interacting with the N-terminal activation domain of MyoD and blocking recruitment of the essential transcriptional coactivator p300. These studies establish a molecular mechanism for fiber type switching through direct inhibition of MyoD to control the opposing roles of MyoD and NFATc1 in fast versus slow fiber phenotypes.

  20. IPMK and β-catenin mediate PLC-β1-dependent signaling in myogenic differentiation.

    Science.gov (United States)

    Ramazzotti, Giulia; Billi, Anna Maria; Manzoli, Lucia; Mazzetti, Cristina; Ruggeri, Alessandra; Erneux, Christophe; Kim, Seyun; Suh, Pann-Ghill; Cocco, Lucio; Faenza, Irene

    2016-12-20

    In previous studies, we have reported that phospholipase C (PLC)-β1 plays a crucial role in myogenic differentiation and we determined the importance of its catalytic activity for the initiation of this process. Here we define the effectors that take part to its signaling pathway. We show that the Inositol Polyphosphate Multikinase (IPMK) is able to promote myogenic differentiation since its overexpression determines the up-regulation of several myogenic markers. Moreover, we demonstrate that IPMK activates the same cyclin D3 promoter region targeted by PLC-β1 and that IPMK-induced promoter activation relies upon c-jun binding to the promoter, as we have shown previously for PLC-β1. Furthermore, our data shows that IPMK overexpression causes an increase in β-catenin translocation and accumulation to the nuclei of differentiating myoblasts resulting in higher MyoD activation. Finally, we describe that PLC-β1 overexpression determines too an increase in β-catenin translocation and that PLC-β1, IPMK and β-catenin are mediators of the same signaling pathway since their overexpression results in cyclin D3 and myosin heavy chain (MYH) induction.

  1. Efficient and Reproducible Myogenic Differentiation from Human iPS Cells: Prospects for Modeling Miyoshi Myopathy In Vitro

    Science.gov (United States)

    Tanaka, Akihito; Woltjen, Knut; Miyake, Katsuya; Hotta, Akitsu; Ikeya, Makoto; Yamamoto, Takuya; Nishino, Tokiko; Shoji, Emi; Sehara-Fujisawa, Atsuko; Manabe, Yasuko; Fujii, Nobuharu; Hanaoka, Kazunori; Era, Takumi; Yamashita, Satoshi; Isobe, Ken-ichi; Kimura, En; Sakurai, Hidetoshi

    2013-01-01

    The establishment of human induced pluripotent stem cells (hiPSCs) has enabled the production of in vitro, patient-specific cell models of human disease. In vitro recreation of disease pathology from patient-derived hiPSCs depends on efficient differentiation protocols producing relevant adult cell types. However, myogenic differentiation of hiPSCs has faced obstacles, namely, low efficiency and/or poor reproducibility. Here, we report the rapid, efficient, and reproducible differentiation of hiPSCs into mature myocytes. We demonstrated that inducible expression of myogenic differentiation1 (MYOD1) in immature hiPSCs for at least 5 days drives cells along the myogenic lineage, with efficiencies reaching 70–90%. Myogenic differentiation driven by MYOD1 occurred even in immature, almost completely undifferentiated hiPSCs, without mesodermal transition. Myocytes induced in this manner reach maturity within 2 weeks of differentiation as assessed by marker gene expression and functional properties, including in vitro and in vivo cell fusion and twitching in response to electrical stimulation. Miyoshi Myopathy (MM) is a congenital distal myopathy caused by defective muscle membrane repair due to mutations in DYSFERLIN. Using our induced differentiation technique, we successfully recreated the pathological condition of MM in vitro, demonstrating defective membrane repair in hiPSC-derived myotubes from an MM patient and phenotypic rescue by expression of full-length DYSFERLIN (DYSF). These findings not only facilitate the pathological investigation of MM, but could potentially be applied in modeling of other human muscular diseases by using patient-derived hiPSCs. PMID:23626698

  2. Dose-dependent Effect of Boric Acid on Myogenic Differentiation of Human Adipose-derived Stem Cells (hADSCs).

    Science.gov (United States)

    Apdik, Hüseyin; Doğan, Ayşegül; Demirci, Selami; Aydın, Safa; Şahin, Fikrettin

    2015-06-01

    Boron, a vital micronutrient for plant metabolism, is not fully elucidated for embryonic and adult body development, and tissue regeneration. Although optimized amount of boron supplement has been shown to be essential for normal gestational development in zebrafish and frog and beneficial for bone regeneration in higher animals, effects of boron on myogenesis and myo-regeneration remains to be solved. In the current study, we investigated dose-dependent activity of boric acid on myogenic differentiation of human adipose-derived stem cells (hADSCs) using immunocytochemical, gene, and protein expression analysis. The results revealed that while low- (81.9 μM) and high-dose (819.6 μM) boron treatment increased myogenic gene expression levels such as myosin heavy chain (MYH), MyoD, myogenin, and desmin at day 4 of differentiation, high-dose treatment decreased myogenic-related gene and protein levels at day 21 of differentiation, confirmed by immunocytochemical analysis. The findings of the study present not only an understanding of boron's effect on myogenic differentiation but also an opportunity for the development of scaffolds to be used in skeletal tissue engineering and supplements for embryonic muscle growth. However, fine dose tuning and treatment period arranging are highly warranted as boron treatment over required concentrations and time might result in detrimental outcomes to myogenesis and myo-regeneration.

  3. Grazing-induced changes in muscle microRNA-206 and -208b expression in association with myogenic gene expression in cattle.

    Science.gov (United States)

    Horikawa, Akihiko; Ogasawara, Hideki; Okada, Kaito; Kobayashi, Misato; Muroya, Susumu; Hojito, Masayuki

    2015-11-01

    To investigate the roles of microRNAs (miRNAs) in muscle type conversion, the effects of 4 months of grazing on the expression levels of miRNAs and mRNAs associated with skeletal muscle development were analyzed by quantitative RT-PCR using the Biceps femoris muscle of Japanese Shorthorn cattle. After 4 months of grazing, the expression of muscle fiber type-associated miR-208b was higher in the grazed cattle than in the housed. In concordance with the pattern in miR-208b expression, the expression of MyoD, a myogenic regulatory factor associated with the shifting of muscle property to the fast type, was lower in the grazed cattle after 4 months of grazing than in the housed cattle. In addition, the expression of MyHC-2x (a fast type) was higher in the housed cattle than in the grazed, after 4 months of grazing. During the grazing period, miR-206 expression decreased in the housed cattle, whereas expression in the grazed cattle did not change, but rather remained higher than that of the housed cattle even at 3 months after the grazing ended. These miRNAs including miR-206 persisting with muscles of grazed cattle may be associated with regulation of muscle gene expression during skeletal muscle adaptation to grazing.

  4. File list: Oth.Myo.10.Myod1.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Myo.10.Myod1.AllCell mm9 TFs and others Myod1 Muscle SRX142536,SRX142537,SRX142...534,SRX328690,SRX497483,SRX328691,SRX029146,SRX246928,SRX497482,SRX142516,SRX029145,SRX029144 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Myo.10.Myod1.AllCell.bed ...

  5. File list: Oth.Myo.05.Myod1.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Myo.05.Myod1.AllCell mm9 TFs and others Myod1 Muscle SRX142537,SRX328690,SRX142...536,SRX246928,SRX328691,SRX142534,SRX497483,SRX497482,SRX029146,SRX142516,SRX029145,SRX029144 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Myo.05.Myod1.AllCell.bed ...

  6. Potential Role of Omega-3 Fatty Acids on the Myogenic Program of Satellite Cells.

    Science.gov (United States)

    Bhullar, Amritpal S; Putman, Charles T; Mazurak, Vera C

    2016-01-01

    Skeletal muscle loss is associated with aging as well as pathological conditions. Satellite cells (SCs) play an important role in muscle regeneration. Omega-3 fatty acids are widely studied in a variety of muscle wasting diseases; however, little is known about their impact on skeletal muscle regeneration. The aim of this review is to evaluate studies examining the effect of omega-3 fatty acids, α-linolenic acid, eicosapentaenoic acid, and docosahexaenoic acid on the regulation of SC proliferation and differentiation. This review highlights mechanisms by which omega-3 fatty acids may modulate the myogenic program of the stem cell population within skeletal muscles and identifies considerations for future studies. It is proposed that minimally three myogenic transcriptional regulatory factors, paired box 7 (Pax7), myogenic differentiation 1 protein, and myogenin, should be measured to confirm the stage of SCs within the myogenic program affected by omega-3 fatty acids.

  7. Ocular Vestibular Evoked Myogenic Potentials

    Directory of Open Access Journals (Sweden)

    Felipe, Lilian

    2014-01-01

    Full Text Available Introduction Diagnostic testing of the vestibular system is an essential component of treating patients with balance dysfunction. Until recently, testing methods primarily evaluated the integrity of the horizontal semicircular canal, which is only a portion of the vestibular system. Recent advances in technology have afforded clinicians the ability to assess otolith function through vestibular evoked myogenic potential (VEMP testing. VEMP testing from the inferior extraocular muscles of the eye has been the subject of interest of recent research. Objective To summarize recent developments in ocular VEMP testing. Results Recent studies suggest that the ocular VEMP is produced by otolith afferents in the superior division of the vestibular nerve. The ocular VEMP is a short latency potential, composed of extraocular myogenic responses activated by sound stimulation and registered by surface electromyography via ipsilateral otolithic and contralateral extraocular muscle activation. The inferior oblique muscle is the most superficial of the six extraocular muscles responsible for eye movement. Therefore, measurement of ocular VEMPs can be performed easily by using surface electrodes on the skin below the eyes contralateral to the stimulated side. Conclusion This new variation of the VEMP procedure may supplement conventional testing in difficult to test populations. It may also be possible to use this technique to evaluate previously inaccessible information on the vestibular system.

  8. Trenbolone enhances myogenic differentiation by enhancing β-catenin signaling in muscle-derived stem cells of cattle.

    Science.gov (United States)

    Zhao, J-X; Hu, J; Zhu, M-J; Du, M

    2011-05-01

    Testosterone is a key hormone regulating animal growth and development, which promotes skeletal muscle growth and inhibits fat deposition; however, the underlying mechanisms remain poorly defined. Because canonical Wingless and Int/β-catenin signaling promotes myogenesis, we hypothesized that testosterone regulates myogenesis through enhancing the β-catenin signaling pathway and the expression of its targeted genes. Muscle-derived stem cells were prepared from the skeletal muscle of fetal calf at day 180 of gestation and treated with or without trenbolone (10 nM), a synthetic analog of testosterone, in a myogenic medium. Trenbolone treatment increased the protein levels of MyoD and myosin heavy chain, as well as the androgen receptor content. The myogenic effect of trenbolone was blocked by cyproterone acetate, a specific inhibitor of androgen receptor, showing that the myogenic effect of trenbolone was mediated by the androgen receptor. Immunoprecipitation showed that androgen receptor and β-catenin formed a complex, which was increased by trenbolone treatment. Trenbolone activated adenosine monophosphate-activated protein kinase, which might phosphorylate β-catenin at Ser552, stabilizing β-catenin. Indeed, both cytoplasmic and nuclear β-catenin levels were increased after trenbolone treatment. As a result, β-catenin-mediated transcriptional activity was enhanced by trenbolone treatment. In conclusion, these data provide evidence that testosterone increases cellular β-catenin content which promotes the expression of β-catenin-targeted genes and myogenesis in the muscle-derived stem cells of cattle.

  9. Mammalian Skeletal Muscle Fibres Promote Non-Muscle Stem Cells and Non-Stem Cells to Adopt Myogenic Characteristics

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    Taryn Morash

    2017-01-01

    Full Text Available Skeletal muscle fibres are unique cells in large animals, often composed of thousands of post-mitotic nuclei. Following skeletal muscle damage, resident stem cells, called satellite cells, commit to myogenic differentiation and migrate to carry out repair. Satellite stem cells migrate on muscle fibres through amoeboid movement, which relies on dynamic cell membrane extension and retraction (blebbing. It is not known whether blebbing is due to the intrinsic properties of satellite cells, or induced by features of the myofibre surface. Here, we determined the influence of the muscle fibre matrix on two important features of muscle regeneration: the ability to migrate and to differentiate down a myogenic lineage. We show that the muscle fibre is able to induce amoeboid movement in non-muscle stem cells and non-stem cells. Secondly, we show that prolonged co-culture on myofibres caused amniotic fluid stem cells and breast cancer cells to express MyoD, a key myogenic determinant. Finally, we show that amniotic fluid stem cells co-cultured on myofibres are able to fuse and make myotubes that express Myosin Heavy Chain.

  10. Vestibular-evoked myogenic potentials.

    Science.gov (United States)

    Colebatch, J G; Rosengren, S M; Welgampola, M S

    2016-01-01

    The vestibular-evoked myogenic potential (VEMP) is a short-latency potential evoked through activation of vestibular receptors using sound or vibration. It is generated by modulated electromyographic signals either from the sternocleidomastoid muscle for the cervical VEMP (cVEMP) or the inferior oblique muscle for the ocular VEMP (oVEMP). These reflexes appear to originate from the otolith organs and thus complement existing methods of vestibular assessment, which are mainly based upon canal function. This review considers the basis, methodology, and current applications of the cVEMP and oVEMP in the assessment and diagnosis of vestibular disorders, both peripheral and central. © 2016 Elsevier B.V. All rights reserved.

  11. Loss of emerin alters myogenic signaling and miRNA expression in mouse myogenic progenitors.

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    Adam J Koch

    Full Text Available Emerin is an integral membrane protein of the inner nuclear membrane. Mutations in emerin cause X-linked Emery-Dreifuss muscular dystrophy (EDMD, a disease characterized by skeletal muscle wasting and dilated cardiomyopathy. Current evidence suggests the muscle wasting phenotype of EDMD is caused by defective myogenic progenitor cell differentiation and impaired muscle regeneration. We obtained genome-wide expression data for both mRNA and micro-RNA (miRNA in wildtype and emerin-null mouse myogenic progenitor cells. We report here that emerin-null myogenic progenitors exhibit differential expression of multiple signaling pathway components required for normal muscle development and regeneration. Components of the Wnt, IGF-1, TGF-β, and Notch signaling pathways are misexpressed in emerin-null myogenic progenitors at both the mRNA and protein levels. We also report significant perturbations in the expression and activation of p38/Mapk14 in emerin-null myogenic progenitors, showing that perturbed expression of Wnt, IGF-1, TGF-β, and Notch signaling components disrupts normal downstream myogenic signaling in these cells. Collectively, these data support the hypothesis that emerin is essential for proper myogenic signaling in myogenic progenitors, which is necessary for myogenic differentiation and muscle regeneration.

  12. Muscle Tissue Engineering Using Gingival Mesenchymal Stem Cells Encapsulated in Alginate Hydrogels Containing Multiple Growth Factors.

    Science.gov (United States)

    Ansari, Sahar; Chen, Chider; Xu, Xingtian; Annabi, Nasim; Zadeh, Homayoun H; Wu, Benjamin M; Khademhosseini, Ali; Shi, Songtao; Moshaverinia, Alireza

    2016-06-01

    Repair and regeneration of muscle tissue following traumatic injuries or muscle diseases often presents a challenging clinical situation. If a significant amount of tissue is lost the native regenerative potential of skeletal muscle will not be able to grow to fill the defect site completely. Dental-derived mesenchymal stem cells (MSCs) in combination with appropriate scaffold material, present an advantageous alternative therapeutic option for muscle tissue engineering in comparison to current treatment modalities available. To date, there has been no report on application of gingival mesenchymal stem cells (GMSCs) in three-dimensional scaffolds for muscle tissue engineering. The objectives of the current study were to develop an injectable 3D RGD-coupled alginate scaffold with multiple growth factor delivery capacity for encapsulating GMSCs, and to evaluate the capacity of encapsulated GMSCs to differentiate into myogenic tissue in vitro and in vivo where encapsulated GMSCs were transplanted subcutaneously into immunocompromised mice. The results demonstrate that after 4 weeks of differentiation in vitro, GMSCs as well as the positive control human bone marrow mesenchymal stem cells (hBMMSCs) exhibited muscle cell-like morphology with high levels of mRNA expression for gene markers related to muscle regeneration (MyoD, Myf5, and MyoG) via qPCR measurement. Our quantitative PCR analyzes revealed that the stiffness of the RGD-coupled alginate regulates the myogenic differentiation of encapsulated GMSCs. Histological and immunohistochemical/fluorescence staining for protein markers specific for myogenic tissue confirmed muscle regeneration in subcutaneous transplantation in our in vivo animal model. GMSCs showed significantly greater capacity for myogenic regeneration in comparison to hBMMSCs (p engineering.

  13. Pannexin channels mediate the acquisition of myogenic commitment in C2C12 reserve cells promoted by P2 receptor activation

    Science.gov (United States)

    Riquelme, Manuel A.; Cea, Luis A.; Vega, José L.; Puebla, Carlos; Vargas, Aníbal A.; Shoji, Kenji F.; Subiabre, Mario; Sáez, Juan C.

    2015-01-01

    The acquisition of myoblast commitment to the myogenic linage requires rises in intracellular free Ca2+ concentration ([Ca2+]i). Putative cell membrane pathways involved in these [Ca2+]i increments are P2 receptors (P2Rs) as well as connexin (Cx) and/or pannexin (Panx) hemichannels and channels (Cx HChs and Panx Chs), respectively, which are known to permeate Ca2+. Reserve cells (RCs) are uncommitted myoblasts obtained from differentiated C2C12 cell cultures, which acquire commitment upon replating. Regarding these cells, we found that extracellular ATP increases the [Ca2+]i via P2Rs. Moreover, ATP increases the plasma membrane permeability to small molecules and a non-selective membrane current, both of which were inhibited by Cx HCh/Panx1Ch blockers. However, RCs exposed to divalent cation-free saline solution, which is known to activate Cx HChs (but not Panx Chs), did not enhance membrane permeability, thus ruling out the possible involvement of Cx HChs. Moreover, ATP-induced membrane permeability was inhibited with blockers of P2Rs that activate Panx Chs. In addition, exogenous ATP induced the expression of myogenic commitment and increased MyoD levels, which was prevented by the inhibition of P2Rs or knockdown of Panx1 Chs. Similarly, increases in MyoD levels induced by ATP released by RCs were inhibited by Panx Ch/Cx HCh blockers. Myogenic commitment acquisition thus requires a feed-forward mechanism mediated by extracellular ATP, P2Rs, and Panx Chs. PMID:26000275

  14. Pannexin Channels Mediate the Acquisition of Myogenic Commitment in C2C12 Reserve Cells Promoted by P2 Receptor Activation

    Directory of Open Access Journals (Sweden)

    Manuel Antonio Riquelme

    2015-05-01

    Full Text Available The acquisition of myoblast commitment to the myogenic linage requires rises in intracellular free Ca2+ concentration ([Ca2+]i. Putative cell membrane pathways involved in these [Ca2+]i increments are P2 receptors (P2Rs as well as connexin (Cx and/or pannexin (Panx hemichannels and channels (Cx HChs and Panx Chs, respectively, which are known to permeate Ca2+. Reserve cells (RCs are uncommitted myoblasts obtained from differentiated C2C12 cell cultures, which acquire commitment upon replating. Regarding these cells, we found that extracellular ATP increases the [Ca2+]i via P2Rs. Moreover, ATP increases the plasma membrane permeability to small molecules and a non-selective membrane current, both of which were inhibited by Cx HCh/Panx1Ch blockers. However, RCs exposed to divalent cation-free saline solution, which is known to activate Cx HChs (but not Panx Chs, did not enhance membrane permeability, thus ruling out the possible involvement of Cx HChs. Moreover, ATP-induced membrane permeability was inhibited with blockers of P2Rs that activate Panx Chs. In addition, exogenous ATP induced the expression of myogenic commitment and increased MyoD levels, which was prevented by the inhibition of P2Rs or knockdown of Panx1 Chs. Similarly, increases in MyoD levels induced by ATP released by RCs were inhibited by Panx Ch/Cx HCh blockers. Myogenic commitment acquisition thus requires a feed-forward mechanism mediated by extracellular ATP, P2Rs and Panx Chs.

  15. The production of fluorescent transgenic trout to study in vitro myogenic cell differentiation

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    Rescan Pierre-Yves

    2010-05-01

    Full Text Available Abstract Background Fish skeletal muscle growth involves the activation of a resident myogenic stem cell population, referred to as satellite cells, that can fuse with pre-existing muscle fibers or among themselves to generate a new fiber. In order to monitor the regulation of myogenic cell differentiation and fusion by various extrinsic factors, we generated transgenic trout (Oncorhynchus mykiss carrying a construct containing the green fluorescent protein reporter gene driven by a fast myosin light chain 2 (MlC2f promoter, and cultivated genetically modified myogenic cells derived from these fish. Results In transgenic trout, green fluorescence appeared in fast muscle fibers as early as the somitogenesis stage and persisted throughout life. Using an in vitro myogenesis system we observed that satellite cells isolated from the myotomal muscle of transgenic trout expressed GFP about 5 days post-plating as they started to fuse. GFP fluorescence persisted subsequently in myosatellite cell-derived myotubes. Using this in vitro myogenesis system, we showed that the rate of muscle cell differentiation was strongly dependent on temperature, one of the most important environmental factors in the muscle growth of poikilotherms. Conclusions We produced MLC2f-gfp transgenic trout that exhibited fluorescence in their fast muscle fibers. The culture of muscle cells extracted from these trout enabled the real-time monitoring of myogenic differentiation. This in vitro myogenesis system could have numerous applications in fish physiology to evaluate the myogenic activity of circulating growth factors, to test interfering RNA and to assess the myogenic potential of fish mesenchymal stem cells. In ecotoxicology, this system could be useful to assess the impact of environmental factors and marine pollutants on fish muscle growth.

  16. Introgression of mitochondrial DNA among Myodes voles: consequences for energetics?

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    Boratyński Zbyszek

    2011-12-01

    Full Text Available Abstract Background Introgression of mitochondrial DNA (mtDNA is among the most frequently described cases of reticulate evolution. The tendency of mtDNA to cross interspecific barriers is somewhat counter-intuitive considering the key function of enzymes that it encodes in the oxidative-phosphorylation process, which could give rise to hybrid dysfunction. How mtDNA reticulation affects the evolution of metabolic functions is, however, uncertain. Here we investigated how morpho-physiological traits vary in natural populations of a common rodent (the bank vole, Myodes glareolus and whether this variation could be associated with mtDNA introgression. First, we confirmed that M. glareolus harbour mtDNA introgressed from M. rutilus by analyzing mtDNA (cytochrome b, 954 bp and nuclear DNA (four markers; 2333 bp in total sequence variation and reconstructing loci phylogenies among six natural populations in Finland. We then studied geographic variation in body size and basal metabolic rate (BMR among the populations of M. glareolus and tested its relationship with mtDNA type. Results Myodes glareolus and its arctic neighbour, M. rutilus, are reciprocally monophyletic at the analyzed nuclear DNA loci. In contrast, the two northernmost populations of M. glareolus have a fixed mitotype that is shared with M. rutilus, likely due to introgressive hybridization. The analyses of phenotypic traits revealed that the body mass and whole-body, but not mass corrected, BMR are significantly reduced in M. glareolus females from northern Finland that also have the introgressed mitotype. Restricting the analysis to the single population where the mitotypes coexist, the association of mtDNA type with whole-body BMR remained but those with mass corrected BMR and body mass did not. Mitochondrial sequence variation in the introgressed haplotypes is compatible with demographic growth of the populations, but may also be a result of positive selection. Conclusion Our

  17. Lrrc75b is a novel negative regulator of C2C12 myogenic differentiation

    Science.gov (United States)

    Zhong, Yuechun; Zou, Liyi; Wang, Zonggui; Pan, Yaqiong; Dai, Zhong; Liu, Xinguang; Cui, Liao; Zuo, Changqing

    2016-01-01

    Many transcription factors and signaling molecules involved in the guidance of myogenic differentiation have been investigated in previous studies. However, the precise molecular mechanisms of myogenic differentiation remain largely unknown. In the present study, by performing a meta-analysis of C2C12 myogenic differentiation microarray data, we found that leucine-rich repeat-containing 75B (Lrrc75b), also known as AI646023, a molecule of unknown biological function, was downregulated during C2C12 myogenic differentiation. The knockdown of Lrrc75b using specific siRNA in C2C12 myoblasts markedly enhanced the expression of muscle-specific myogenin and increased myoblast fusion and the myotube diameter. By contrast, the adenovirus-mediated overexpression of Lrrc75b in C2C12 cells markedly inhibited myoblast differentiation accompanied by a decrease in myogenin expression. In addition, the phosphorylation of extracellular signal-regulated kinase 1/2 (Erk1/2) was suppressed in the cells in which Lrrc75b was silenced. Taken together, our results demonstrate that Lrrc75b is a novel suppressor of C2C12 myogenic differentiation by modulating myogenin and Erk1/2 signaling. PMID:27633041

  18. Lichen compounds restrain lichen feeding by bank voles (Myodes glareolus).

    Science.gov (United States)

    Nybakken, Line; Helmersen, Anne-Marit; Gauslaa, Yngvar; Selås, Vidar

    2010-03-01

    Some lichen compounds are known to deter feeding by invertebrate herbivores. We attempted to quantify the deterring efficiency of lichen compounds against a generalist vertebrate, the bank vole (Myodes glareolus). In two separate experiments, caged bank voles had the choice to feed on lichens with natural or reduced concentrations of secondary compounds. We rinsed air-dry intact lichens in 100% acetone to remove extracellular compounds non-destructively. In the first experiment, pairs of control and rinsed lichen thalli were hydrated and offered to the bank voles. Because the lichens desiccated fast, we ran a second experiment with pairs of ground control and compound-deficient thalli, each mixed with water to porridge. Eight and six lichen species were tested in the first and second experiment, respectively. In the first, bank voles preferred compound-deficient thalli of Cladonia stellaris and Lobaria pulmonaria, but did not discriminate between the other thallus pairs. This was likely a result of deterring levels of usnic and stictic acid in the control thalli. When lichens were served as porridge, significant preference was found for acetone-rinsed pieces of Cladonia arbuscula, C. rangiferina, Platismatia glauca, and Evernia prunastri. The increased preference was caused mainly by lower consumption of control thalli. Grinding and mixing of thallus structures prevented bank voles from selecting thallus parts with lower concentration of secondary compounds and/or strengthened their deterring capacity. We conclude that some lichen secondary compounds deter feeding by bank voles.

  19. Differentially activated macrophages orchestrate myogenic precursor cell fate during human skeletal muscle regeneration

    DEFF Research Database (Denmark)

    Saclier, Marielle; Yacoub-Youssef, Houda; Mackey, Abigail;

    2013-01-01

    Macrophages (MPs) exert either beneficial or deleterious effects on tissue repair, depending on their activation/polarization state. They are crucial for adult skeletal muscle repair, notably by acting on myogenic precursor cells. However, these interactions have not been fully characterized. Here......, we explored both in vitro and in vivo, in human, the interactions of differentially activated MPs with myogenic precursor cells (MPCs) during adult myogenesis and skeletal muscle regeneration. We showed in vitro that through the differential secretion of cytokines and growth factors, proinflammatory...... MPs inhibited MPC fusion while anti-inflammatory MPs strongly promoted MPC differentiation by increasing their commitment into differentiated myocytes and the formation of mature myotubes. Furthermore, the in vivo time course of expression of myogenic and MP markers was studied in regenerating human...

  20. Calcium and mechanotransduction of the myogenic response.

    Science.gov (United States)

    D'Angelo, G; Davis, M J; Meininger, G A

    1997-07-01

    The purpose of this study was to measure vascular smooth muscle (VSM) cytosolic calcium ([Ca2+]i) during the myogenic response. We examined the temporal and steady-state relationships between lumen diameter and VSM [Ca2+]i in isolated arterioles exposed to step changes in intravascular pressure. We also studied the relationship between step sizes in intravascular pressure and changes in [Ca2+]i. First-order arterioles from the hamster cheek pouch were isolated, cannulated, and pressurized. [Ca2+]i was quantified using the ratio of emitted fluorescence intensity (R340/380) during alternate excitation of fura 2-loaded vessels at 340 and 380 nm. Stepwise increases in transmural pressure elicited corresponding increases in steady-state [Ca2+]i and myogenic constriction. From a common baseline pressure, the initial rise in [Ca2+]i after a step change in pressure was directly related to the magnitude of the step size and of the distension caused by that pressure step. This supports the theory that there is a relationship between the initial distension of the vessel and the initial [Ca2+]i change. Also, increasing the size of the step change in pressure resulted in a greater myogenic response, yet no difference in the steady-state [Ca2+]i was detected, which suggests that Ca2+ is not the principal or only determinant of steady-state constriction. Finally, larger increases in [Ca2+]i do not necessarily augment the myogenic response, which suggests that some minimal rise in [Ca2+]i is required to elicit myogenic vasoconstriction. Collectively, these data suggest the presence of a separate, Ca(2+)-independent regulatory system.

  1. Multiple phosphorylation events control mitotic degradation of the muscle transcription factor Myf5

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    Lorca Thierry

    2005-12-01

    Full Text Available Abstract Background The two myogenic regulatory factors Myf5 and MyoD are basic helix-loop-helix muscle transcription factors undergoing differential cell cycle dependent proteolysis in proliferating myoblasts. This regulated degradation results in the striking expression of these two factors at distinct phases of the cell cycle, and suggests that their precise and alternated disappearance is an important feature of myoblasts, maybe connected to the maintenance of the proliferative status and/or commitment to the myogenic lineage of these cells. One way to understand the biological function(s of the cyclic expression of these proteins is to specifically alter their degradation, and to analyze the effects of their stabilization on cells. To this aim, we undertook the biochemical analysis of the mechanisms governing Myf5 mitotic degradation, using heterologous systems. Results We show here that mitotic degradation of Myf5 is conserved in non-myogenic cells, and is thus strictly under the control of the cell cycle apparatus. Using Xenopus egg extracts as an in vitro system to dissect the main steps of Myf5 mitotic proteolysis, we show that (1 Myf5 stability is regulated by a complex interplay of phosphorylation/dephosphorylation, probably involving various kinases and phosphatases, (2 Myf5 is ubiquitylated in mitotic extracts, and this is a prerequisite to its degradation by the proteasome and (3 at least in the Xenopus system, the E3 responsible for its mitotic degradation is not the APC/C (the major E3 during mitosis. Conclusion Altogether, our data strongly suggest that the mitotic degradation of Myf5 by the ubiquitin-proteasome system is precisely controlled by multiple phosphorylation of the protein, and that the APC/C is not involved in this process.

  2. Myogenin, MyoD, and myosin expression after pharmacologically and surgically induced hypertrophy

    Science.gov (United States)

    Mozdziak, P. E.; Greaser, M. L.; Schultz, E.

    1998-01-01

    The relationship between myogenin or MyoD expression and hypertrophy of the rat soleus produced either by clenbuterol and 3,3', 5-triiodo-L-thyronine (CT) treatment or by surgical overload was examined. Mature female rats were subjected to surgical overload of the right soleus with the left soleus serving as a control. Another group received the same surgical treatment but were administered CT. Soleus muscles were harvested 4 wk after surgical overload and weighed. Myosin heavy chain isoforms were separated by using polyacrylamide gel electrophoresis while myogenin and MyoD expression were evaluated by Northern analysis. CT and functional overload increased soleus muscle weight. CT treatment induced the appearance of the fast type IIX myosin heavy chain isoform, depressed myogenin expression, and induced MyoD expression. However, functional overload did not alter myogenin or MyoD expression in CT-treated or non-CT-treated rats. Thus pharmacologically and surgically induced hypertrophy have differing effects on myogenin and MyoD expression, because their levels were associated with changes in myosin heavy chain composition (especially type IIX) rather than changes in muscle mass.

  3. Myogenic Differentiation of Mouse Embryonic Stem Cells That Lack a Functional Pax7 Gene

    Science.gov (United States)

    Czerwinska, Areta M.; Grabowska, Iwona; Archacka, Karolina; Bem, Joanna; Swierczek, Barbara; Helinska, Anita; Streminska, Wladyslawa; Fogtman, Anna; Iwanicka-Nowicka, Roksana; Koblowska, Marta

    2016-01-01

    The transcription factor Pax7 plays a key role during embryonic myogenesis and sustains the proper function of satellite cells, which serve as adult skeletal muscle stem cells. Overexpression of Pax7 has been shown to promote the myogenic differentiation of pluripotent stem cells. However, the effects of the absence of functional Pax7 in differentiating embryonic stem cells (ESCs) have not yet been directly tested. Herein, we studied mouse stem cells that lacked a functional Pax7 gene and characterized the differentiation of these stem cells under conditions that promoted the derivation of myoblasts in vitro. We analyzed the expression of myogenic factors, such as myogenic regulatory factors and muscle-specific microRNAs, in wild-type and mutant cells. Finally, we compared the transcriptome of both types of cells and did not find substantial differences in the expression of genes related to the regulation of myogenesis. As a result, we showed that the absence of functional Pax7 does not prevent the in vitro myogenic differentiation of ESCs. PMID:26649785

  4. Histone H3 Methyltransferase Suv39h1 Prevents Myogenic Terminal Differentiation by Repressing MEF2 Activity in Muscle Cells

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    Wei Jin

    2016-11-01

    Full Text Available The myogenic regulatory factors (MRFs and myocyte enhancer factor 2 (MEF2 transcription factors have been extensively studied as key transcription factors that regulate myogenic gene expression. However, few reports on the molecular mechanism that modulates chromatin remodeling during skeletal muscle differentiation are available. We reported here that the expression of the H3-K9 methyltransferase Suv39h1 was decreased during myoblast differentiation. Ectopic expression of Suv39h1 could inhibit myoblast differentiation, increasing H3-K9 methylation levels, whereas knockdown of Suv39h1 stimulated myoblast differentiation. Furthermore, Suv39h1 interacted with MEF2C directly and inhibited MEF2 transcription activity in a dose-dependent manner. Together, our studies revealed a molecular mechanism wherein Suv39h1 modulated myogenic gene expression and activation during skeletal muscle differentiation.

  5. DNMT3B in vitro knocking-down is able to reverse embryonal rhabdomyosarcoma cell phenotype through inhibition of proliferation and induction of myogenic differentiation.

    Science.gov (United States)

    Megiorni, Francesca; Camero, Simona; Ceccarelli, Simona; McDowell, Heather P; Mannarino, Olga; Marampon, Francesco; Pizer, Barry; Shukla, Rajeev; Pizzuti, Antonio; Marchese, Cinzia; Clerico, Anna; Dominici, Carlo

    2016-11-29

    Aberrant DNA methylation has been frequently observed in many human cancers, including rhabdomyosarcoma (RMS), the most common soft tissue sarcoma in children. To date, the expression and function of the de novo DNA methyltransferase (DNMT) 3B in RMS have not yet been investigated. Our study show for the first time a significant up-regulation of DNMT3B levels in 14 RMS tumour samples and 4 RMS cell lines in comparison to normal skeletal muscle. Transfection of RD and TE671 cells, two in vitro models of embryonal RMS (ERMS), with a synthetic DNMT3B siRNA decreased cell proliferation by arresting cell cycle at G1 phase, as demonstrated by the reduced expression of Cyclin B1, Cyclin D1 and Cyclin E2, and by the concomitant up-regulation of the checkpoint regulators p21 and p27. DNMT3B depletion also impaired RB phosphorylation status and decreased migratory capacity and clonogenic potential. Interestingly, DNMT3B knock-down was able to commit ERMS cells towards myogenic terminal differentiation, as confirmed by the acquisition of a myogenic-like phenotype and by the increased expression of the myogenic markers MYOD1, Myogenin and MyHC. Finally, inhibition of MEK/ERK signalling by U0126 resulted in a reduction of DNMT3B protein, giving evidence that DNMT3B is a down-stream molecule of this oncogenic pathway.Taken together, our data indicate that altered expression of DNMT3B plays a key role in ERMS development since its silencing is able to reverse cell cancer phenotype by rescuing myogenic program. Epigenetic therapy, by targeting the DNA methylation machinery, may represent a novel therapeutic strategy against RMS.

  6. Neonatal epicardial-derived progenitors aquire myogenic traits in skeletal muscle, but not cardiac muscle

    DEFF Research Database (Denmark)

    Andersen, Ditte C; Jensen, Charlotte H; Skovrind, Ida;

    2016-01-01

    BACKGROUND/OBJECTIVES: Epicardium-derived progenitor cells (EPDCs) differentiate into all heart cell types in the embryonic heart, yet their differentiation into cardiomyocytes in the adult heart is limited and poorly described. This may be due to EPDCs lacking myogenic potential or the inert adu...... that EPDCs may be more myogenic than previously anticipated. But, the heart may lack factors for induction of myogenesis of EPDCs, a scenario that should be taken into consideration when aiming for repair of damaged myocardium by stem cell transplantation.......BACKGROUND/OBJECTIVES: Epicardium-derived progenitor cells (EPDCs) differentiate into all heart cell types in the embryonic heart, yet their differentiation into cardiomyocytes in the adult heart is limited and poorly described. This may be due to EPDCs lacking myogenic potential or the inert adult...... heart missing regenerative signals essential for directed differentiation of EPDCs. Herein, we aimed to evaluate the myogenic potential of neonatal EPDCs in adult and neonatal mouse myocardium, as well as in skeletal muscle. The two latter tissues have an intrinsic capability to develop and regenerate...

  7. Life-long shedding of Puumala hantavirus in wild bank voles (Myodes glareolus).

    Science.gov (United States)

    Voutilainen, Liina; Sironen, Tarja; Tonteri, Elina; Bäck, Anne Tuiskunen; Razzauti, Maria; Karlsson, Malin; Wahlström, Maria; Niemimaa, Jukka; Henttonen, Heikki; Lundkvist, Åke

    2015-06-01

    The knowledge of viral shedding patterns and viraemia in the reservoir host species is a key factor in assessing the human risk of zoonotic viruses. The shedding of hantaviruses (family Bunyaviridae) by their host rodents has widely been studied experimentally, but rarely in natural settings. Here we present the dynamics of Puumala hantavirus (PUUV) shedding and viraemia in naturally infected wild bank voles (Myodes glareolus). In a monthly capture-mark-recapture study, we analysed 18 bank voles for the presence and relative quantity of PUUV RNA in the excreta and blood from 2 months before up to 8 months after seroconversion. The proportion of animals shedding PUUV RNA in saliva, urine and faeces peaked during the first month after seroconversion, but continued throughout the study period with only a slight decline. The quantity of shed PUUV in reverse transcription quantitative PCR (RT-qPCR) positive excreta was constant over time. In blood, PUUV RNA was present for up to 7 months but both the probability of viraemia and the virus load declined with time. Our findings contradict the current view of a decline in virus shedding after the acute phase and a short viraemic period in hantavirus infection - an assumption widely adopted in current epidemiological models. We suggest the life-long shedding as a means of hantaviruses to survive over host population bottlenecks, and to disperse in fragmented habitats where local host and/or virus populations face temporary extinctions. Our results indicate that the kinetics of pathogens in wild hosts may differ considerably from those observed in laboratory settings.

  8. Interaural difference values of vestibular evoked myogenic.

    Directory of Open Access Journals (Sweden)

    Marziyeh Moallemi

    2015-01-01

    Full Text Available Migraine is a neurologic disease, which often is associated with a unilateral headache. Vestibular abnormalities are common in migraine. Vestibular evoked myogenic potentials (VEMPs assess otolith function in particular functional integrity of the saccule and the inferior vestibular nerve. We used VEMP to evaluate if the migraine headache can affect VEMP asymmetry parameters. A total of 25 patients with migraine (22 females and 3 males who were diagnosed according to the criteria of IHS-1988 were enrolled in this cross-sectional study. Control group consisted of 26 healthy participants (18 female and 8 male, without neurotological symptoms and history of migraine. The short tone burst (95 dB nHL, 500 Hz was presented to ears. VEMP was recorded with surface electromyography over the contracted ipsilateral sternocleidomastoid (SCM muscle. Although current results showed that the amplitude ratio is greater in migraine patients than normal group, there was no statistical difference between two groups in mean asymmetry parameters of VEMP. Asymmetry measurements in vestibular evoked myogenic potentials probably are not indicators of unilateral deficient in saccular pathways of migraine patients.

  9. Premature myogenic differentiation and depletion of progenitor cells cause severe muscle hypotrophy in Delta1 mutants

    OpenAIRE

    Schuster-Gossler, Karin; Cordes, Ralf; Gossler, Achim

    2006-01-01

    In vertebrates, skeletal myogenesis is initiated by the generation of myoblasts followed by their differentiation to myocytes and the formation of myofibers. The determination of myoblasts and their differentiation are controlled by muscle regulatory factors that are activated at specific stages during myogenesis. During late embryonic and fetal stages a distinct population of resident proliferating progenitor cells is the major source of myogenic cells. How the differentiation of myoblasts a...

  10. Study of myogenic spontaneous contractile activities in the detrusor instability rats

    Institute of Scientific and Technical Information of China (English)

    JIANG Hai-hong; WEN Qian-jun; SONG Bo

    2006-01-01

    Objective: To explore a myogenic basis of the spontaneous contractions on the rat bladder smooth muscle strip in a detrusor instability (DI) model in vitro, and to study a nerve blocker's cocktail affecting the spontaneous contractions as well as electrical stimulated contractile response. Methods: DI model rats were made by partial bladder outlet obstruction (BOO) and confirmed by the filling cystometry. Detrusor strip was dissected from fresh bladder, fixed for an isometric tension trial. The contractions were recorded during electrical stimulation or exposure to some agents. Results: The cocktail diminished the nerve-mediated contractile response effectively in DI preparation. DI's spontaneous contractions remained during the presence of the cocktail with a significant change in its contractile amplitude. Conclusion: With the local nerve-concerned factors abolishment by the cocktail, the DI bladder preparations still have the spontaneous contractions, indicating a myogenic basis from themselves.

  11. Tuina Manipulation on Infantile Myogenic Torticollis

    Institute of Scientific and Technical Information of China (English)

    LU Jian-zhong

    2004-01-01

    With the knowledge handed down by his father, the author practices both acupuncture and Tuina and obtained good effects for infantile myogenic torticollis by Tuina manipulations, which relieved suffering of many kids and prevented them from the operative injury. Among 48 cases, the total effective rate reached 95.8% through the treatment of once everyday and 10 days a course.%笔者师承父学从事针灸,推拿临床工作,运用推拿手法治疗小儿肌性斜颈,每日1次,10d为1个疗程,在48例患儿中,总有效率95.8%,给众多的患儿解除了病痛,亦免于手术带来的创伤.

  12. RECORDING OF VESTIBULAR EVOKED MYOGENIC POTENTIALS

    Directory of Open Access Journals (Sweden)

    A. A. Sazgar

    2006-05-01

    Full Text Available It has been shown recently that loud clicks evoke myogenic potentials in the tonically contracting sternocleidomastoid muscles. Studies have suggested that these potentials are of vestibular origin, especially of the saccule and inferior vestibular nerve. A pilot study was undertaken in our hospital to record vestibular evoked myogenic potentials (VEMP for the first time in Iran. Eighteen healthy volunteers (32 ears without history of otologic or vestibular disorders were subjected to the VEMP test. Twenty-one patients (26 ears with unilateral (6 patients and bilateral (5 patients high frequency sensorineural hearing loss with unknown etiology, acoustic neuroma (1 patient, Meniere’s disease (4 patients and unilateral low frequency sensorineural hearing loss without vestibular complaint (5 patients were also enrolled in this study. VEMP response to clicks was obtained from 84.4% of ears of healthy subjects. These subjects demonstrated short latency waves to click stimuli during tonic neck flexor activation. Mean latencies of first positive (p13 and first negative (n23 potentials in healthy subjects were 12.45 ± 1.9 ms and 20.8 ± 3.5 ms, respectively. Median latencies of these two potentials were 12.1 and 19.3 ms, respectively. We could record VEMP in 5 patients with unilateral and all patients with high and low frequency sensorineural hearing loss without vestibular complaint. In the patient with acoustic neuroma VEMP was absent on the affected side. This technique may offer a new method to evaluate otolith and sacculocollic pathways in human.

  13. Premature myogenic differentiation and depletion of progenitor cells cause severe muscle hypotrophy in Delta1 mutants.

    Science.gov (United States)

    Schuster-Gossler, Karin; Cordes, Ralf; Gossler, Achim

    2007-01-09

    In vertebrates, skeletal myogenesis is initiated by the generation of myoblasts followed by their differentiation to myocytes and the formation of myofibers. The determination of myoblasts and their differentiation are controlled by muscle regulatory factors that are activated at specific stages during myogenesis. During late embryonic and fetal stages a distinct population of resident proliferating progenitor cells is the major source of myogenic cells. How the differentiation of myoblasts and progenitor cells is regulated is not clear. We show that in mouse embryos the Notch ligand Delta1 (Dll1) controls both differentiation of early myoblasts and maintenance of myogenic progenitor cells. Early dermomyotome-derived myoblasts are determined normally in Dll1 mutant embryos, but their differentiation is accelerated, leading to a transient excess of myotomal muscle fibers. Similarly, migratory hypaxial myogenic cells colonize the limb buds and activate muscle regulatory factor expression normally, but muscle differentiation progresses more rapidly. Resident progenitor cells defined by Pax3/Pax7 expression are formed initially, but they are progressively lost and virtually absent at embryonic day 14.5. Muscle growth declines beginning around embryonic day 12, leading to subsequent severe muscle hypotrophy in hypomorphic Dll1 fetuses. We suggest that premature and excessive differentiation leads to depletion of progenitor cells and cessation of muscle growth, and we conclude that Dll1 provides essential signals that are required to prevent uncontrolled differentiation early and ensure sustained muscle differentiation during development.

  14. FGFR4 and its novel splice form in myogenic cells: Interplay of glycosylation and tyrosine phosphorylation.

    Science.gov (United States)

    Kwiatkowski, Boguslaw A; Kirillova, Irina; Richard, Robert E; Israeli, David; Yablonka-Reuveni, Zipora

    2008-06-01

    The family of fibroblast growth factor receptors (FGFRs) is encoded by four distinct genes. FGFR1 and FGFR4 are both expressed during myogenesis, but whereas the function of FGFR1 in myoblast proliferation has been documented, the role of FGFR4 remains unknown. Here, we report on a new splice form of FGFR4 cloned from primary cultures of mouse satellite cells. This form, named FGFR4(-16), lacks the entire exon 16, resulting in a deletion within the FGFR kinase domain. Expression of FGFR4(-16) coincided with that of wild-type FGFR4 in all FGFR4-expressing tissues examined. Moreover, expression of both FGFR4 forms correlated with the onset of myogenic differentiation, as determined in mouse C2C12 cells and in the inducible myogenic system of 10T(1/2)-MyoD-ER cell line. Both endogenous and overexpressed forms of FGFR4 exhibited N-glycosylation. In contrast to FGFR1, induced homodimerization of FGFR4 proteins did not result in receptor tyrosine phosphorylation. Surprisingly, coexpression of FGFR4 forms and a chimeric FGFR1 protein resulted in FGFR4 tyrosine phosphorylation, raising the possibility that FGFR4 phosphorylation might be enabled by a heterologous tyrosine kinase activity. Collectively, the present study reveals novel characteristics of mouse FGFR4 gene products and delineates their expression pattern during myogenesis. Our findings suggest that FGFR4 functions in a distinctly different manner than the prototype FGFR during myogenic differentiation.

  15. New perspectives on vestibular evoked myogenic potentials.

    Science.gov (United States)

    Rosengren, Sally M; Kingma, Herman

    2013-02-01

    Although the vestibular evoked myogenic potential (VEMP) measured from the cervical muscles (cVEMP, cervical VEMP) is well described and has documented clinical utility, its analogue recorded from the extraocular muscles (oVEMP, ocular VEMP) has been described only recently and is currently emerging as an additional test of otolith function. This review will, therefore, summarize recent developments in VEMP research with a focus on the oVEMP. Recent studies suggest that the oVEMP is produced by otolith afferents in the superior vestibular nerve division, whereas the cVEMP evoked by sound is thought to be an inferior vestibular nerve reflex. Correspondingly, the oVEMP correlates better with caloric and subjective visual vertical tests than sound-cVEMPs. cVEMPs are more complicated than often thought, as shown by the presence of crossed responses and conflicting results of recent vibration studies. Altered inner ear mechanics produced by the vestibular diseases superior semicircular canal dehiscence and Ménière's disease lead to changes in the preferred frequency of the oVEMP and cVEMP. The oVEMP provides complementary diagnostic information to the cVEMP and is likely to be a useful addition to the diagnostic test battery in neuro-otology.

  16. Functional and Activity Analysis of Cattle UCP3 Promoter with MRFs-Related Factors

    Directory of Open Access Journals (Sweden)

    Wei Chen

    2016-05-01

    Full Text Available Uncoupling protein 3 (UCP3 is mainly expressed in muscle. It plays an important role in muscle, but less research on the regulation of cattle UCP3 has been performed. In order to elucidate whether cattle UCP3 can be regulated by muscle-related factors, deletion of cattle UCP3 promoter was amplified and cloned into pGL3-basic, pGL3-promoter and PEGFP-N3 vector, respectively, then transfected into C2C12 myoblasts cells and UCP3 promoter activity was measured using the dual-Luciferase reporter assay system. The results showed that there is some negative-regulatory element from −620 to −433 bp, and there is some positive-regulatory element between −433 and −385 bp. The fragment (1.08 kb of UCP3 promoter was cotransfected with muscle-related transcription factor myogenic regulatory factors (MRFs and myocyte-specific enhancer factor 2A (MEF2A. We found that UCP3 promoter could be upregulated by Myf5, Myf6 and MyoD and downregulated by MyoG and MEF2A.

  17. Effects of variation in porcine MYOD1 gene on muscle fiber characteristics, lean meat production, and meat quality traits.

    Science.gov (United States)

    Lee, E A; Kim, J M; Lim, K S; Ryu, Y C; Jeon, W M; Hong, K C

    2012-09-01

    Three single nucleotide polymorphisms (SNPs) in the porcine MYOD1 gene were used for association analysis and haplotype construction to evaluate the effects of their substitution. Four hundred and three pigs of Yorkshire and Berkshire breeds were used. The mRNA expression levels of MYOD1 were examined. The g.489C>T and g.1264C>A SNPs were significantly associated with several muscle fiber characteristics, the loin eye area, and lightness. Particularly, animals having hetero-genotypes of both sites showed good performance both in lean meat production and meat quality traits. The results of haplotype substitution were similar to the associations of individual SNPs. Moreover, the 2 SNPs had significant effects on mRNA expression. Therefore, the g.489C>T and g.1264C>A SNPs in MYOD1 may be meaningful DNA markers that can be used for improving important porcine economic traits.

  18. Cervical and ocular vestibular evoked myogenic potentials in multiple sclerosis participants

    OpenAIRE

    Parsa, Maryam Sadat; Mohammadkhani, Ghassem; Hajabolhassani, Fahimeh; Jalaee, Shohreh; Zakeri, Hassanali

    2015-01-01

    Background: Multiple sclerosis (MS) is a chronic neurological disease that affects brain and spinal cord. The infratentorial region contains the cerebellum and brainstem. Vestibular evoked myogenic potentials (VEMPs) are short-latency myogenic responses. Cervical vestibular evoked myogenic potential (cVEMP) is a manifestation of vestibulocolic reflex and ocular vestibular evoked myogenic potential (oVEMP) contributes to the linear vestibular?ocular reflex. The aim of this study was to evaluat...

  19. Genetic analysis of hantaviruses carried by Myodes and Microtus rodents in Buryatia

    Directory of Open Access Journals (Sweden)

    Lundkvist Åke

    2008-01-01

    Full Text Available Abstract Hantavirus genome sequences were recovered from tissue samples of Myodes rufocanus, Microtus fortis and Microtus oeconomus captured in the Baikal area of Buryatia, Russian Federation. Genetic analysis of S- and M-segment sequences of Buryatian hantavirus strains showed that Myodes-associated strains belong to Hokkaido virus (HOKV type while Microtus-associated strains belong to Vladivostok virus (VLAV type. On phylogenetic trees Buryatian HOKV strains were clustered together with M. rufocanus- originated strains from Japan, China and Far-East Russia (Primorsky region. Buryatian Microtus- originated strains shared a common recent ancestor with M. fortis- originated VLAV strain from Far-East Russia (Vladivostok area. Our data (i confirm that M. rufocanus carries a hantavirus which is similar to but distinct from both Puumala virus carried by M. glareolus and Muju virus associated with M. regulus, (ii confirm that M. fortis is the natural host for VLAV, and (iii suggest M. oeconomus as an alternative host for VLAV.

  20. Asymmetric vestibular evoked myogenic potentials in unilateral Meniere patients

    NARCIS (Netherlands)

    Kingma, C. M.; Wit, H. P.

    2011-01-01

    Vestibular evoked myogenic potentials (VEMPs) were measured in 22 unilateral MeniSre patients with monaural and binaural stimulation with 250 and 500 Hz tone bursts. For all measurement situations significantly lower VEMP amplitudes were on average measured at the affected side compared to the unaff

  1. Bovine gallbladder muscularis: Source of a myogenic receptor for cholecystokinin

    Energy Technology Data Exchange (ETDEWEB)

    Schjoldager, B.; Shaw, M.J.; Powers, S.P.; Schmalz, P.E.; Szurszewski, J.; Miller, L.J. (Mayo Clinic and Foundation, Rochester, MN (USA))

    1988-03-01

    Despite being a classic target for the gastrointestinal peptide hormone, cholecystokinin (CCK), the gallbladder CCK receptor is not well characterized. Pharmacological studies of small species suggest that CCK action can be mediated by direct myogenic or by both myogenic and neurogenic receptors. To prepare for the biochemical characterization of a gallbladder CCK receptor and to define the subtype of the receptor being studied. The authors have performed autoradiographic localization and pharmacological characterization of CCK receptors on bovine gallbladder. Autoradiography demonstrated high-affinity specific CCK-binding sites only on the muscularis. CCK-8 stimulated tonic contraction of longitudinal strips of gallbladder muscularis in a concentration-dependent manner. Antagonism at the cholinergic receptor with 1{mu}M atropine or axonal transmission with 1{mu}M tetrodotoxin did not modify CCK-induced contraction, supporting a direct myogenic effect of this hormone. Optimal electrical field stimulation to elicit a neuronal response resulted in muscle strip relaxation, which was abolished with adrenergic blockade. Although acetylcholine administration stimulated contraction, electrical field stimulation did not, even in the presence of phentolamine, propranolol, and/or CCK. Thus, in bovine gallbladder muscularis, there is evidence for a functional CCK receptor only on smooth muscle cells. Demonstration of a single, high-affinity specific CCK-binding site on an enriched plasma membrane preparation of bovine gallbladder muscularis is consistent with this representing a myogenic CCK receptor.

  2. CD36 is required for myoblast fusion during myogenic differentiation

    Energy Technology Data Exchange (ETDEWEB)

    Park, Seung-Yoon [Department of Biochemistry, College of Medicine, Dongguk University and Medical Institute of Dongguk University, Gyeongju 780-714 (Korea, Republic of); Yun, Youngeun [Department of Biochemistry and Cell Biology, Cell and Matrix Research Institute, School of Medicine, Kyungpook National University, Daegu 700-422 (Korea, Republic of); Kim, In-San, E-mail: iskim@knu.ac.kr [Department of Biochemistry and Cell Biology, Cell and Matrix Research Institute, School of Medicine, Kyungpook National University, Daegu 700-422 (Korea, Republic of); Biomedical Research Institute, Korea Institute Science and Technology, Seoul (Korea, Republic of)

    2012-11-02

    Highlights: Black-Right-Pointing-Pointer CD36 expression was induced during myogenic differentiation. Black-Right-Pointing-Pointer CD36 expression was localized in multinucleated myotubes. Black-Right-Pointing-Pointer The expression of myogenic markers is attenuated in CD36 knockdown C2C12 cells. Black-Right-Pointing-Pointer Knockdown of CD36 significantly inhibited myotube formation during differentiation. -- Abstract: Recently, CD36 has been found to be involved in the cytokine-induced fusion of macrophage. Myoblast fusion to form multinucleated myotubes is required for myogenesis and muscle regeneration. Because a search of gene expression database revealed the attenuation of CD36 expression in the muscles of muscular dystrophy patients, the possibility that CD36 could be required for myoblast fusion was investigated. CD36 expression was markedly up-regulated during myoblast differentiation and localized in multinucleated myotubes. Knockdown of endogenous CD36 significantly decreased the expression of myogenic markers as well as myotube formation. These results support the notion that CD36 plays an important role in cell fusion during myogenic differentiation. Our finding will aid the elucidation of the common mechanism governing cell-to-cell fusion in various fusion models.

  3. Analysis of genetic distribution and population genetic structure of the MyoD gene in 10 pig breeds

    Institute of Scientific and Technical Information of China (English)

    Li ZHU; Xuewei LI; Surong SHUAI; Mingzhou LI; Fangqiong LI; Lei CHEN

    2008-01-01

    Restriction fragment length polymorphism (RFLP) data was applied to analyze the distribution of the MyoD gene in 10 pig breeds and pig breed crosses.The population genetic information about genetic distribution,variation,and heterozygosity of the MyoD gene in different breed populations were analyzed.Based on the allele frequency,genetic distance and evolution distance among each breed populations were calculated and Unweighted Pair Group Method with Arithmetic mean (UPGMA) phylogenetic tree was gained based on the evolution distances between populations.The results indicated that the distribution of the MyoD genotype kept in Hardy-Weinberg equilibrium in most tested groups but not in Duroc (D) and Duroc × (Landrance × Yorkshire)(DLY) population.Generally,the genetic diversity of the MyoD gene was abundant and these tested breed populations had high genetic variations.The evolution of the MyoD gene was under natural selection pressure.On the phylogenetic tree,10 pig breeds were divided into 4 clusters.The first cluster consisted of four breeds developed from Landrace.The second cluster was two indigenous Chinese pig breeds.The third cluster was three breeds developed from Duroc.The fourth cluster was a Tibetan pig breed.The constitution of the topology of the phylogenetic tree was consistent with the breeding history of each pig breed.From this experiment,we can conclude that some RFLP data obtained from functional gene can be used in the genetic deviation research between some closely related species or between different populations in certain species.

  4. Direct laser machining-induced topographic pattern promotes up-regulation of myogenic markers in human mesenchymal stem cells.

    Science.gov (United States)

    Li, Huaqiong; Wen, Feng; Wong, Yee Shan; Boey, Freddy Yin Chiang; Subbu, Venkatraman S; Leong, David Tai; Ng, Kee Woei; Ng, Gary Ka Lai; Tan, Lay Poh

    2012-02-01

    The engineering of tissue is preferably done with stem cells, which can be differentiated into the tissue of interest using biochemical or physical cues. While much effort has been focused on using biological factors to regulate stem cell differentiation, recently interest in the contribution of physical factors has increased. In this work, three-dimensional (3-D) microchannels with topographic micropatterns were fabricated by femtosecond laser machining on a biodegradable polymer (poly(L-lactide-co-ε-caprolactone)) substrate. Two substrates with narrow and wide channels respectively were created. Human mesenchymal stem cells (hMSCs) were cultured on the scaffolds for cell proliferation and cellular organization. Gene expression and the immunostaining of myogenic and neurogenic markers were studied. Both scaffolds improved the cell alignment along the channels as compared to the control group. Microfilaments within hMSCs were more significantly aligned and elongated on the narrower microchannels. The gene expression study revealed significant up-regulation of several hallmark markers associated with myogenesis for hMSCs cultured on the scaffold with narrow microchannels, while osteogenic and neurogenic markers were down-regulated or remained similar to the control at day 14. Immunostaining of myogen- and neurogen-specific differentiation markers were used to further confirm the specific differentiation towards a myogenic lineage. This study demonstrates that femtosecond laser machining is a versatile tool for generating controllable 3-D microchannels with topographic features that can be used to induce specific myogenic differentiation of hMSCs in vitro, even in the absence of biological factors.

  5. Insulin-Like Growth Factor Binding Protein-6 Alters Skeletal Muscle Differentiation of Human Mesenchymal Stem Cells

    Directory of Open Access Journals (Sweden)

    Doaa Aboalola

    2017-01-01

    Full Text Available Insulin-like growth factor binding protein-6 (IGFBP-6, the main regulator of insulin-like growth factor-2 (IGF-2, is a component of the stem cell niche in developing muscle cells. However, its role in muscle development has not been clearly defined. In this study, we investigated the role of IGFBP-6 in muscle commitment and differentiation of human mesenchymal stem cells derived from the placenta. We showed that placental mesenchymal stem cells (PMSCs have the ability to differentiate into muscle cells when exposed to a specific culture medium by expressing muscle markers Pax3/7, MyoD, myogenin, and myosin heavy chain in a stage-dependent manner with the ultimate formation of multinucleated fibers and losing pluripotency-associated markers, OCT4 and SOX2. The addition of IGFBP-6 significantly increased pluripotency-associated markers as well as muscle differentiation markers at earlier time points, but the latter decreased with time. On the other hand, silencing IGFBP-6 decreased both pluripotent and differentiation markers at early time points. The levels of these markers increased as IGFBP-6 levels were restored. These findings indicate that IGFBP-6 influences MSC pluripotency and myogenic differentiation, with more prominent effects observed at the beginning of the differentiation process before muscle commitment.

  6. Effect of transforming growth factor-beta1 on embryonic and posthatch muscle growth and development in normal and low score normal chicken.

    Science.gov (United States)

    Li, X; Velleman, S G

    2009-02-01

    During skeletal muscle development, transforming growth factor-beta1 (TGF-beta1) is a potent inhibitor of muscle cell proliferation and differentiation. The TGF-beta1 signal is carried by Smad proteins into the cell nucleus, inhibiting the expression of key myogenic regulatory factors including MyoD and myogenin. However, the molecular mechanism by which TGF-beta1 inhibits muscle cell proliferation and differentiation has not been well documented in vivo. The present study investigated the effect of TGF-beta1 on in vivo skeletal muscle growth and development. A chicken line, Low Score Normal (LSN) with reduced muscling and upregulated TGF-beta1 expression, was used and compared to a normal chicken line. The injection of TGF-beta1 at embryonic day (ED) 3 significantly reduced the pectoralis major (p. major) muscle weight in the normal birds at 1 wk posthatch, whereas no significant difference was observed in the LSN birds. The difference between normal and LSN birds in response to TGF-beta1 is likely due to different levels of endogenous TGF-beta1 where the LSN birds have increased TGF-beta1 expression in their p. major muscle at both 17 ED and 6 wk posthatch. Smad3 expression was reduced by TGF-beta1 from 10 ED to 1 wk posthatch in normal p. major muscle. Unlike Smad3, Smad7 expression was not significantly affected by TGF-beta1 until posthatch in both normal and LSN p. major muscle. Expression of MyoD was reduced 35% by TGF-beta1 during embryonic development in normal p. major muscle, whereas LSN p. major muscle showed a delayed decrease at 1 d posthatch in MyoD expression in response to the TGF-beta1 treatment. Myogenin expression was reduced 29% by TGF-beta1 after hatch in normal p. major muscle. In LSN p. major muscle, TGF-beta1 treatment significantly decreased myogenin expression by 43% at 1 d posthatch and 32% at 1 wk posthatch. These data suggested that TGF-beta1 reduced p. major muscle growth by inhibiting MyoD and myogenin expression during both embryonic

  7. Effects of pulsed electromagnetic fields on the expression of MyoD after acute skeletal muscle contusion%脉冲电磁场对大鼠骨骼肌急性挫伤后成肌分化因子D表达的促进作用

    Institute of Scientific and Technical Information of China (English)

    王晶; 张长杰; 顾旭东; 姚云海

    2009-01-01

    目的 观察脉冲电磁场(PEMFs)对大鼠骨骼肌急性挫伤(ASMC)后组织学及成肌调节因子D(MyoD)表达的影响,并探讨脉冲电磁场在大鼠骨骼肌急性挫伤早期治疗中的应用.方法 将42只Sprague-Dawley大鼠随机等分为治疗组(造模+PEMFs干预)、造模对照组(造模)、空白对照组(不予任何处理),每组大鼠14只.治疗组与造模对照组大鼠均选取大鼠左后肢小腿三头肌,采用金属砝码在PVC管引导下垂直下落建立ASMC模型.治疗组造模成功后即刻予以PEMFs干预1次.每组分别在造模成功后第12小时和第18小时随机处死7只大鼠,取小腿三头肌进行HE染色观察,测定MyoD荧光强度.结果 造模对照组在两个时间点均出现胞浆淡染、胞核多形态改变,在治疗组,其程度明显减轻,但较空白对照组严重;治疗组及造模对照组的MyoD荧光强度,第18小时均高于第12小时(P<0.01),且两组在各时间点的表达均高于空白对照组(P<0.01),第18小时,治疗组高于造模对照组(P<0.01).结论 大鼠骨骼肌急性挫伤后早期应用PEMFs,可以在干预后18 h出现MyoD的表达上调,这可能是PEMFs促进骨骼肌再生的机理之一.%Objective To observe the effects of pulsed electromagnetic fields (PEMFs) on histological changes and myogenic differentiation factor D (MyoD) expression in rats with acute skeletal muscle contusion ( ASMC), and to explore the effects of PEMF therapy on rats with ASMC in its very early stages. Methods Forty-two rats were randomly divided into three groups : a treatment group, a control group and a blank control group. ASMC models were established with all the animals in the treatment and control groups. PEMF treatment was admin-istered to the treatment group immediately after the establishment of the ASMC model. Seven rats in each group were sacrificed at the 12th and 18th h after the models were set up. Their triceps surae muscles were sampled and treated with haematoxylin

  8. Biocompatible Elastic Conductive Films Significantly Enhanced Myogenic Differentiation of Myoblast for Skeletal Muscle Regeneration.

    Science.gov (United States)

    Dong, Ruonan; Zhao, Xin; Guo, Baolin; Ma, Peter X

    2017-09-11

    The key factor in skeletal muscle tissue engineering is regeneration of the functional skeletal muscles. Materials that could promote the myoblast proliferation and myogenic differentiation are promising candidates in skeletal muscle tissue engineering. Herein, we developed an elastic conductive poly(ethylene glycol)-co-poly(glycerol sebacate) (PEGS) grafted aniline pentamer (AP) copolymer that could promote the formation of myotubes by differentiating the C2C12 myoblast cells. The results of hydration behavior and water contact angle suggested that by adjusting the poly(ethylene glycol) (PEG) and AP content, this film showed a proper surface hydrophilicity for cell attachment. Additionally, these films showed tunable conductivity and mechanical properties that can be altered by changing the AP content. The maximum conductivity of the films was 1.84 × 10(-4) S/cm and the Young's modulus of these films ranged from 14.58 ± 1.35 MPa to 24.62 ± 0.61 MPa. Our findings indicate that the PEGS-AP films promote the proliferation and myogenic differentiation of C2C12 cells, suggesting that they are promising biomaterials for skeletal muscle tissue engineering.

  9. Non-myogenic tumors display altered expression of dystrophin (DMD) and a high frequency of genetic alterations

    Science.gov (United States)

    Luce, Leonela N.; Abbate, Mercedes

    2017-01-01

    DMD gene mutations have been associated with the development of Dystrophinopathies. Interestingly, it has been recently reported that DMD is involved in the development and progression of myogenic tumors, assigning DMD a tumor suppressor activity in these types of cancer. However, there are only few reports that analyze DMD in non-myogenic tumors. Our study was designed to examine DMD expression and genetic alterations in non-myogenic tumors using public repositories. We also evaluated the overall survival of patients with and without DMD mutations. We studied 59 gene expression microarrays (GEO database) and RNAseq (cBioPortal) datasets that included 9817 human samples. We found reduced DMD expression in 15/27 (56%) pairwise comparisons performed (Fold-Change (FC) ≤ 0.70; p-value range = 0.04-1.5×10−20). The analysis of RNAseq studies revealed a median frequency of DMD genetic alterations of 3.4%, higher or similar to other well-known tumor suppressor genes. In addition, we observed significant poorer overall survival for patients with DMD mutations. The analyses of paired tumor/normal tissues showed that the majority of tumor specimens had lower DMD expression compared to their normal adjacent counterpart. Interestingly, statistical significant over-expression of DMD was found in 6/27 studies (FC ≥ 1.4; p-value range = 0.03-3.4×10−15). These results support that DMD expression and genetic alterations are frequent and relevant in non-myogenic tumors. The study and validation of DMD as a new player in tumor development and as a new prognostic factor for tumor progression and survival are warranted. PMID:27391342

  10. Lentivirus Live Cell Array for Quantitative Assessment of Gene and Pathway Activation during Myogenic Differentiation of Mesenchymal Stem Cells.

    Directory of Open Access Journals (Sweden)

    Janhavi Moharil

    Full Text Available Stem cell differentiation involves multiple cascades of transcriptional regulation that govern the cell fate. To study the real-time dynamics of this complex process, quantitative and high throughput live cell assays are required. Herein, we developed a lentiviral library of promoters and transcription factor binding sites to quantitatively capture the gene expression dynamics over a period of several days during myogenic differentiation of human mesenchymal stem cells (MSCs harvested from two different anatomic locations, bone marrow and hair follicle. Our results enabled us to monitor the sequential activation of signaling pathways and myogenic gene promoters at various stages of differentiation. In conjunction with chemical inhibitors, the lentiviral array (LVA results also revealed the relative contribution of key signaling pathways that regulate the myogenic differentiation. Our study demonstrates the potential of LVA to monitor the dynamics of gene and pathway activation during MSC differentiation as well as serve as a platform for discovery of novel molecules, genes and pathways that promote or inhibit complex biological processes.

  11. Myogenic potential of chick limb bud mesenchyme in micromass culture.

    Science.gov (United States)

    Archer, C W; Langille, R M; Teran, M A; Solursh, M

    1992-01-01

    The myogenic potential of chick limb mesenchyme from stages 18-25 was assessed by micromass culture under conditions conductive to myogenesis, and was measured as the proportion of differentiated (muscle myosin-positive) mononucleated cells detected. It was found that similar myogenic potentials existed in mesenchyme from whole limbs between stages 18 and 19, but this potential was halved by stage 20. At stage 21, proximal mesenchyme showed significantly more myogenesis than distal mesenchyme, but this difference was abolished by stage 22. Thereafter, myogenesis was increasingly restricted from the distal mesenchyme, whilst the potential in more proximal regions did not significantly increase after stage 23. When the ratio between total limb myoblasts which differentiated on days 1 and 4 of culture was analysed, it was found that two distinct peaks existed at stages 20 and 23. The significance of these ratio peaks is unclear, but may be related to different proliferative potentials of the pre-myoblasts at these stages.

  12. Vestibular-evoked myogenic potentials in miniature pigs

    Directory of Open Access Journals (Sweden)

    Xi Shi

    2016-06-01

    Conclusion: The latencies and thresholds of VEMPs recorded from the neck extensor muscle and the masseter muscle appear to be comparable in normal adult Bama miniature pigs, although the amplitude recorded from the neck extensor muscle seems to be higher than that from the masseter muscle. However, because of their usually relatively superficial and easily accessible location, as well as their large volume and strong contractions, masseter muscles may be better target muscles for recording myogenic potentials.

  13. Antisense oligonucleotide induction of progerin in human myogenic cells.

    Directory of Open Access Journals (Sweden)

    Yue-Bei Luo

    Full Text Available We sought to use splice-switching antisense oligonucleotides to produce a model of accelerated ageing by enhancing expression of progerin, translated from a mis-spliced lamin A gene (LMNA transcript in human myogenic cells. The progerin transcript (LMNA Δ150 lacks the last 150 bases of exon 11, and is translated into a truncated protein associated with the severe premature ageing disease, Hutchinson-Gilford progeria syndrome (HGPS. HGPS arises from de novo mutations that activate a cryptic splice site in exon 11 of LMNA and result in progerin accumulation in tissues of mesodermal origin. Progerin has also been proposed to play a role in the 'natural' ageing process in tissues. We sought to test this hypothesis by producing a model of accelerated muscle ageing in human myogenic cells. A panel of splice-switching antisense oligonucleotides were designed to anneal across exon 11 of the LMNA pre-mRNA, and these compounds were transfected into primary human myogenic cells. RT-PCR showed that the majority of oligonucleotides were able to modify LMNA transcript processing. Oligonucleotides that annealed within the 150 base region of exon 11 that is missing in the progerin transcript, as well as those that targeted the normal exon 11 donor site induced the LMNA Δ150 transcript, but most oligonucleotides also generated variable levels of LMNA transcript missing the entire exon 11. Upon evaluation of different oligomer chemistries, the morpholino phosphorodiamidate oligonucleotides were found to be more efficient than the equivalent sequences prepared as oligonucleotides with 2'-O-methyl modified bases on a phosphorothioate backbone. The morpholino oligonucleotides induced nuclear localised progerin, demonstrated by immunostaining, and morphological nuclear changes typical of HGPS cells. We show that it is possible to induce progerin expression in myogenic cells using splice-switching oligonucleotides to redirect splicing of LMNA. This may offer a model

  14. Clinical Testing of Otolith Function: Perceptual Thresholds and Myogenic Potentials

    OpenAIRE

    Agrawal, Yuri; BREMOVA, TATIANA; Kremmyda, Olympia; Strupp, Michael; MacNeilage, Paul R.

    2013-01-01

    Cervical and ocular vestibular-evoked myogenic potential (cVEMP/oVEMP) tests are widely used clinical tests of otolith function. However, VEMP testing may not be the ideal measure of otolith function given the significant inter-individual variability in responses and given that the stimuli used to elicit VEMPs are not physiological. We therefore evaluated linear motion perceptual threshold testing compared with cVEMP and oVEMP testing as measures of saccular and utricular function, respective...

  15. Cervical and ocular vestibular evoked myogenic potentials in Behcet's disease.

    Science.gov (United States)

    Bayram, Ali; Doğan, Murat; Koç, Ali; Kalkan, Mehmet; Akçadağ, Alper; Özcan, İbrahim

    2015-01-01

    To investigate vestibular evoked myogenic potentials combined with audiologic status in Behcet's disease (BD) and to compare the results with normal healthy subjects. Cervical vestibular evoked myogenic potential (cVEMP) test, ocular vestibular evoked myogenic potential (oVEMP) test, Dix-Hallpike test, conventional pure tone audiometry (cPTA) and high frequency audiometry (HFA), and 226 and 1000Hz tympanometry were performed to each subject of the study. Cranial magnetic resonance imaging (MRI) with contrast enhancement was also performed to evaluate the central nervous system (CNS) in patients with BD. VEMP parameters including the mean peak latencies of p13-n23 and n10-p15, AR values and thresholds were not statistically different both in cVEMP and oVEMP between the BD and control groups. Except for 250Hz, mean audiological thresholds were significantly higher in the BD group. Five of the 20 patients had pathological cranial MRI findings that may be compatible with central nervous system involvement. To our knowledge, the present study is the first report investigating oVEMP and cVEMP responses combined with MRI findings in patients with BD. The presence of high frequency hearing loss is a common finding in BD and HFA may help early detection of hearing loss in patients with BD when combined with cPTA. Copyright © 2015 Elsevier Inc. All rights reserved.

  16. It is raining mice and voles: which weather conditions influence the activity of Apodemus flavicollis and Myodes glareolus?

    OpenAIRE

    Wróbel, Aleksandra; Bogdziewicz, Michał

    2015-01-01

    Rodents constitute a crucial part of food chains in many ecosystems; thus, changes in their activity might influence many other species in the community. Moreover, daily variations in activity appear to be an important adaptation, helping rodents to cope with fluctuating intensity of predation pressure and food availability. We investigated how the nightly activity of the yellow-necked mouse (Apodemus flavicollis) and the bank vole (Myodes glareolus) changes with weather conditions. Increased...

  17. [Helminth fauna of the bank vole myodes glareolus (Schreber, 1780) in the Kizhi Archipelago].

    Science.gov (United States)

    Bugmyrin, S V; Korosov, A V; Bespyatova, L A; Ieshko, E P

    2015-01-01

    The present study was aimed to examine the specific features of the helminth fauna in insular populations of the bank vole (Myodes glareolus) in the north of the species range. The material was collected in and nearby the Kizhi Archipelago (Lake Onega, 62°1' N 35°12' E) during August 1997, 2005-2007, 2012 and 2013. Small mammals were trapped on 23 islands (varying from 2 to 15,000 ha) and on the mainland. Helminthological met- hods were applied to examine 301 specimens of M glareolus. Fourteen helminth species were found: trematodes--Skrjabinoplagiorchis vigisi; cestodes--Paranoplocephala omphalodes, P. gracilis, Catenotaenia henttoneni, Taenia mustelae, Cladotaenia globife- ra, Spirometra erinacei; nematodes--Trichocephalus muris, Aonchotheca murissylvatici, Hepaticola hepatica, Heligmosomum mixtum, Heligmosomoides glareoli, Longistriata minuta, Syphacia petrusewiczi. The parasites S. vigisi, S. erinaci, H. hepatica and T. muris were identified in the bank vole in Karelia for the first time. Significant differences were detected between the helminth faunas of local insular populations of the bank vole. A distinctive feature of all small islands was that samples from them lacked the widespread pa- rasitic nematode Heligmosomum mixtum. The studies have confirmed the general trends observed in the parasite fauna of most isolated populations of small mammals: a poorer species diversity and high infestation rates with certain species of parasites. The Kizhi Archipelago is characterized by the specific high abundance of regionally rare parasite species (H hepatica, A. murissylvatici), and by the absence of common parasites (H. mixtum, H. glareoli).

  18. Adaptive evolution during an ongoing range expansion: the invasive bank vole (Myodes glareolus) in Ireland.

    Science.gov (United States)

    White, Thomas A; Perkins, Sarah E; Heckel, Gerald; Searle, Jeremy B

    2013-06-01

    Range expansions are extremely common, but have only recently begun to attract attention in terms of their genetic consequences. As populations expand, demes at the wave front experience strong genetic drift, which is expected to reduce genetic diversity and potentially cause 'allele surfing', where alleles may become fixed over a wide geographical area even if their effects are deleterious. Previous simulation models show that range expansions can generate very strong selective gradients on dispersal, reproduction, competition and immunity. To investigate the effects of range expansion on genetic diversity and adaptation, we studied the population genomics of the bank vole (Myodes glareolus) in Ireland. The bank vole was likely introduced in the late 1920s and is expanding its range at a rate of ~2.5 km/year. Using genotyping-by-sequencing, we genotyped 281 bank voles at 5979 SNP loci. Fourteen sample sites were arranged in three transects running from the introduction site to the wave front of the expansion. We found significant declines in genetic diversity along all three transects. However, there was no evidence that sites at the wave front had accumulated more deleterious mutations. We looked for outlier loci with strong correlations between allele frequency and distance from the introduction site, where the direction of correlation was the same in all three transects. Amongst these outliers, we found significant enrichment for genic SNPs, suggesting the action of selection. Candidates for selection included several genes with immunological functions and several genes that could influence behaviour.

  19. Effect of copper exposure on reproductive ability in the bank vole (Myodes glareolus).

    Science.gov (United States)

    Miska-Schramm, Agata; Kruczek, Małgorzata; Kapusta, Joanna

    2014-10-01

    The amount of copper in natural ecosystems is steadily increasing, due to human activities. It accumulates in plants, posing a threat to herbivores. In polluted areas the population density of small rodents is observed to be lower. The decline in rodent numbers may be caused by increased mortality or diminished fertility. This study examined the effect of copper on the reproductive activity of the bank vole (Myodes glareolus), a small rodent which during foraging often wanders into fields where it might be exposed to pollution. The animals were treated with solutions of 0, 150 or 600 ppm Cu. After 12 weeks of exposure the quality and quantity of the male's sperm was tested. To assess morphological development we compared the experimental groups for body weight, the weight of the male's testes and accessory sex glands, the female's uterus, and the number of matured ovary follicles in tested females. At both doses, copper administration led to lower sperm count and caused sperm head anomalies. The higher dose compromised sperm tail membrane integrity, viability and motility. No effect of copper on morphological development was observed in males, and only the lower dose increased testes weight. In females the higher dose had a negative effect on morphological development, and the lower dose increased uterus weight. No effect of copper on ovarian follicle number was found. For the first time, the morphology of the most typical ovarian follicles of the bank vole is presented.

  20. Analyses of the differentiation potential of satellite cells from myoD-/-, mdx, and PMP22 C22 mice

    Directory of Open Access Journals (Sweden)

    Huxley Clare

    2005-03-01

    Full Text Available Abstract Background Sporadic and sometimes contradictory studies have indicated changes in satellite cell behaviour associated with the progressive nature of human Duchenne muscular dystrophy (DMD. Satellite cell proliferation and number are reportedly altered in DMD and the mdx mouse model. We recently found that satellite cells in MSVski transgenic mice, a muscle hypertrophy model showing progressive muscle degeneration, display a severe ageing-related differentiation defect in vitro. We tested the hypothesis that similar changes contribute to the gradual loss of muscle function with age in mdx and PMP22 mice, a model of human motor and sensory neuropathy type 1A (HMSN1A. Methods Single extensor digitorum longus muscle fibres were cultured from mdx and PMP22 mice and age- and genetic background-matched controls. Mice at several ages were compared with regard to the differentiation of satellite cells, assayed as the proportion of desmin-expressing cells that accumulated sarcomeric myosin heavy chain. Results Satellite cells of 2 month, 6 month, and 12 month old mdx mice were capable of differentiating to a similar extent to age-matched wild type control animals in an in vitro proliferation/differentiation model. Strikingly, differentiation efficiency in individual 6 month and 12 month old mdx animals varies to a much higher extent than in age-matched controls, younger mdx animals, or PMP22 mice. In contrast, differentiation of myoblasts from all myoD null mice assayed was severely impaired in this assay system. The defect in satellite cell differentiation that occurs in some mdx animals arises from a delay in differentiation that is not overcome by IGF-1 treatment at any phase of cultivation. Conclusion Overall, a defect in satellite cell differentiation above that arising through normal ageing does not occur in mdx or PMP22 mouse models of human disease. Nonetheless, the impaired differentiation of satellite cells from some mdx animals

  1. Activation of Pax7-positive cells in a non-contractile tissue contributes to regeneration of myogenic tissues in the electric fish S. macrurus.

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    Christopher M Weber

    Full Text Available The ability to regenerate tissues is shared across many metazoan taxa, yet the type and extent to which multiple cellular mechanisms come into play can differ across species. For example, urodele amphibians can completely regenerate all lost tissues, including skeletal muscles after limb amputation. This remarkable ability of urodeles to restore entire limbs has been largely linked to a dedifferentiation-dependent mechanism of regeneration. However, whether cell dedifferentiation is the fundamental factor that triggers a robust regeneration capacity, and whether the loss or inhibition of this process explains the limited regeneration potential in other vertebrates is not known. Here, we studied the cellular mechanisms underlying the repetitive regeneration of myogenic tissues in the electric fish S. macrurus. Our in vivo microinjection studies of high molecular weight cell lineage tracers into single identified adult myogenic cells (muscle or noncontractile muscle-derived electrocytes revealed no fragmentation or cellularization proximal to the amputation plane. In contrast, ultrastructural and immunolabeling studies verified the presence of myogenic stem cells that express the satellite cell marker Pax7 in mature muscle fibers and electrocytes of S. macrurus. These data provide the first example of Pax-7 positive muscle stem cells localized within a non-contractile electrogenic tissue. Moreover, upon amputation, Pax-7 positive cells underwent a robust replication and were detected exclusively in regions that give rise to myogenic cells and dorsal spinal cord components revealing a regeneration process in S. macrurus that is dependent on the activation of myogenic stem cells for the renewal of both skeletal muscle and the muscle-derived electric organ. These data are consistent with the emergent concept in vertebrate regeneration that different tissues provide a distinct progenitor cell population to the regeneration blastema, and these

  2. Activation of Pax7-positive cells in a non-contractile tissue contributes to regeneration of myogenic tissues in the electric fish S. macrurus.

    Science.gov (United States)

    Weber, Christopher M; Martindale, Mark Q; Tapscott, Stephen J; Unguez, Graciela A

    2012-01-01

    The ability to regenerate tissues is shared across many metazoan taxa, yet the type and extent to which multiple cellular mechanisms come into play can differ across species. For example, urodele amphibians can completely regenerate all lost tissues, including skeletal muscles after limb amputation. This remarkable ability of urodeles to restore entire limbs has been largely linked to a dedifferentiation-dependent mechanism of regeneration. However, whether cell dedifferentiation is the fundamental factor that triggers a robust regeneration capacity, and whether the loss or inhibition of this process explains the limited regeneration potential in other vertebrates is not known. Here, we studied the cellular mechanisms underlying the repetitive regeneration of myogenic tissues in the electric fish S. macrurus. Our in vivo microinjection studies of high molecular weight cell lineage tracers into single identified adult myogenic cells (muscle or noncontractile muscle-derived electrocytes) revealed no fragmentation or cellularization proximal to the amputation plane. In contrast, ultrastructural and immunolabeling studies verified the presence of myogenic stem cells that express the satellite cell marker Pax7 in mature muscle fibers and electrocytes of S. macrurus. These data provide the first example of Pax-7 positive muscle stem cells localized within a non-contractile electrogenic tissue. Moreover, upon amputation, Pax-7 positive cells underwent a robust replication and were detected exclusively in regions that give rise to myogenic cells and dorsal spinal cord components revealing a regeneration process in S. macrurus that is dependent on the activation of myogenic stem cells for the renewal of both skeletal muscle and the muscle-derived electric organ. These data are consistent with the emergent concept in vertebrate regeneration that different tissues provide a distinct progenitor cell population to the regeneration blastema, and these progenitor cells

  3. Vestibular evoked myogenic potential: its use in Sudden Sensorineural Hearing Loss.

    Science.gov (United States)

    Ledesma, Alleluia Lima Losno; Barreto, Monique Antunes de Souza Chelminski; Bahmad, Fayez

    2014-01-01

    Sudden sensorineural hearing loss (SHL) has uncertain origin and evolution. Vestibular evoked myogenic potential (VEMP) estimates of the vestibular pathway that can not be analyzed by the other entrance examinations, which can be helpful in diagnosing the extent of hearing damage in these patients. To investigate the clinical applicability of VEMP in patients with SHL. This is a systematic review. Searches were conducted in the databases PubMed/Medline, SciELO and LILACS. Data were tabulated. We found 45 articles, 15 of these made up the study by fitting either the inclusion factors. The objective of 60% of the studies was to determine whether the VEMP can be used as predictive hearing recovery Conclusion: VEMP may be useful as hearing recovery predictor in patients with sudden sensorineural hearing loss, justifying the implementation of such examination in this population.

  4. A simple model for the generation of the vestibular evoked myogenic potential (VEMP)

    NARCIS (Netherlands)

    Wit, HP; Kingma, CM

    2006-01-01

    Objective: To describe the mechanism by which the vestibular evoked myogenic potential is generated. Methods: Vestibular evoked myogenic potential generation is modeled by adding a large number of muscle motor unit action potentials. These action potentials occur randomly in time along a 100 ms long

  5. Ocular vestibular evoked myogenic potentials in normal-hearing adults

    OpenAIRE

    Mohammad Kamali; Homa Zarinkoub; Akram Pourbakht; Abdoreza Sheibanizade; Maryam Ramezani; Seyede Nazanin Hajari

    2012-01-01

    Background and Aim: Ocular vestibular-evoked myogenic potential (oVEMP) is a novel vestibular function test. This short-latency response can be recorded through contracting extraocular muscles by high-intensity acoustic stimulation and can be used to evaluate contralateral ocular-vestibular reflex. The aim of this study was to record and compare the amplitude, latency, asymmetry ratio and occurrence percentage of oVEMP (n10) and cervical VEMP (p13) responses in a group of normal adult subject...

  6. Clinical application of vestibular evoked myogenic potential (VEMP).

    Science.gov (United States)

    Murofushi, Toshihisa

    2016-08-01

    The author reviewed clinical aspects of vestibular evoked myogenic potentials (VEMPs). Now two types of VEMPs are available. The first one is cervical VEMP, which is recorded in the sternocleidomastoid muscle and predominantly reflects sacculo-collic reflex. The other is ocular VEMP, which is usually recorded below the lower eye lid and predominantly reflects utriculo-ocular reflex. VEMPs play important roles not only for assessment of common vestibular diseases but also for establishment of new clinical entities. Clinical application in Meniere's disease, vestibular neuritis, benign paroxysmal positional vertigo, vestibular migraine, idiopathic otolithic vertigo, and central vertigo/dizziness was reviewed. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  7. Population sex-ratio affecting behavior and physiology of overwintering bank voles (Myodes glareolus).

    Science.gov (United States)

    Sipari, Saana; Haapakoski, Marko; Klemme, Ines; Palme, Rupert; Sundell, Janne; Ylönen, Hannu

    2016-05-15

    Many boreal rodents are territorial during the breeding season but during winter become social and aggregate for more energy efficient thermoregulation. Communal winter nesting and social interactions are considered to play an important role for the winter survival of these species, yet the topic is relatively little explored. Females are suggested to be the initiators of winter aggregations and sometimes reported to survive better than males. This could be due to the higher social tolerance observed in overwintering females than males. Hormonal status could also affect winter behavior and survival. For instance, chronic stress can have a negative effect on survival, whereas high gonadal hormone levels, such as testosterone, often induce aggressive behavior. To test if the winter survival of females in a boreal rodent is better than that of males, and to assess the role of females in the winter aggregations, we generated bank vole (Myodes glareolus) populations of three different sex ratios (male-biased, female-biased and even density) under semi-natural conditions. We monitored survival, spatial behavior and hormonal status (stress and testosterone) during two winter months. We observed no significant differences in survival between the sexes or among populations with differing sex-ratios. The degree of movement area overlap was used as an indicator of social tolerance and potential communal nesting. Individuals in male biased populations showed a tendency to be solitary, whereas in female biased populations there was an indication of winter aggregation. Females living in male-biased populations had higher stress levels than the females from the other populations. The female-biased sex-ratio induced winter breeding and elevated testosterone levels in males. Thus, our results suggest that the sex-ratio of the overwintering population can lead to divergent overwintering strategies in bank voles. Copyright © 2016 Elsevier Inc. All rights reserved.

  8. BAMBI Promotes C2C12 Myogenic Differentiation by Enhancing Wnt/β-Catenin Signaling

    Directory of Open Access Journals (Sweden)

    Qiangling Zhang

    2015-08-01

    Full Text Available Bone morphogenic protein and activin membrane-bound inhibitor (BAMBI is regarded as an essential regulator of cell proliferation and differentiation that represses transforming growth factor-β and enhances Wnt/β-catenin signaling in various cell types. However, its role in skeletal muscle remains largely unknown. In the current study, we found that the expression level of BAMBI peaked in the early differentiation phase of the C2C12 rodent myoblast cell line. Knockdown of BAMBI via siRNA inhibited C2C12 differentiation, indicated by repressed MyoD, MyoG, and MyHC expression as well as reductions in the differentiation and fusion indices. BAMBI knockdown reduced the activity of Wnt/β-catenin signaling, as characterized by the decreased nuclear translocation of β-catenin and the lowered transcription of Axin2, which is a well-documented target gene of the Wnt/β-catenin signaling pathway. Furthermore, treatment with LiCl, an activator of Wnt/β-catenin signaling, rescued the reduction in C2C12 differentiation caused by BAMBI siRNA. Taken together, our data suggest that BAMBI is required for normal C2C12 differentiation, and that its role in myogenesis is mediated by the Wnt/β-catenin pathway.

  9. Vestibular-evoked myogenic potentials in miniature pigs

    Institute of Scientific and Technical Information of China (English)

    Xi Shi; Yan Zhang; Ya Li; Shiwei Qiu; Shili Zhang; Yaohan Li; Na Yuan; Yuehua Qiao; Shiming Yang

    2016-01-01

    Objective:To report detection of vestibular-evoked myogenic potentials (VEMPs) in the miniature pig. Methods:Potentials evoked by 1000 Hz tone bursts were recorded from neck extensor muscles and the masseter muscles in normal adult Bama miniature pigs anesthetized with 3%pentobarbital sodium and Carbachol II. Results:The latency of the first positive wave P from neck extensor muscles was 7.65 ± 0.64 ms, with an amplitude of 1.66 ± 0.34 uv and a rate of successful induction of 75%at 80 dB SPL. The latency of potentials evoked from the masseter muscles was 7.60 ± 0.78 ms, with an amplitude of 1.31 ± 0.28 uv and a rate successful induction of 66%at 80 dB SPL. Conclusion:The latencies and thresholds of VEMPs recorded from the neck extensor muscle and the masseter muscle appear to be comparable in normal adult Bama miniature pigs, although the amplitude recorded from the neck extensor muscle seems to be higher than that from the masseter muscle. However, because of their usually relatively superficial and easily accessible location, as well as their large volume and strong contractions, masseter muscles may be better target muscles for recording myogenic potentials.

  10. Myogenic cytodifferentiation of the precardiac mesoderm in the rat.

    Science.gov (United States)

    Baldwin, H S; Jensen, K L; Solursh, M

    1991-08-01

    The contractile cells of the primitive heart are derived from a subpopulation of the lateral plate splanchnic mesoderm. While the formation of the cardiac primordia has been studied in the avian embryo, little is known about this cell population in the mammal. To investigate the distribution and cellular differentiation of the myocardial precursors in the early mammalian embryo, we studied the sequential immunohistochemical appearance of desmin and myosin in whole mounts of rat embryos from the presomite (gestational day 9) through the 6-8 somite, straight heart tube (gestational day 10) stages of early cardiac morphogenesis. In contrast to the chicken, and previous reports in the mouse, our results show that myogenic differentiation of the muscle precursor cells of the heart begins in the presomite embryo prior to formation of the anterior intestinal portal or foregut. In addition, this cell population of the precardiac mesoderm appears as a single crescent-shaped population of cells in continuity across the midline which extends caudally during development and then fuses in the midline to form the primitive heart tube. Unlike skeletal myogenesis, desmin and myosin appear simultaneously and are codistributed throughout this initial period of heart development. These results suggest that myocardial differentiation in the rat is precocious when compared to the chicken and precedes the morphogenetic processes involved in formation of the primitive heart tube. Furthermore, this study provides the first description in the mammal of the spatial distribution of the myogenic precardiac mesoderm.

  11. Vestibular evoked myogenic potentials (VEMPs) in central neurological disorders.

    Science.gov (United States)

    Venhovens, J; Meulstee, J; Verhagen, W I M

    2016-01-01

    Several types of acoustic stimulation (i.e. tone bursts or clicks), bone-conducted vibration, forehead taps, and galvanic stimulation elicit myogenic potentials. These can be recorded in cervical and ocular muscles, the so called vestibular evoked myogenic potentials (VEMPs). The cervical VEMP (cVEMP) resembles the vestibulo-collic reflex and the responses can be recorded from the ipsilateral sternocleidomastoid muscle. The ocular VEMP resembles the vestibulo-ocular reflex and can be recorded from extra-ocular muscles by a surface electrode beneath the contralateral infraorbital margin. Initially, the literature concerning VEMPs was limited to peripheral vestibular disorders, however, the field of VEMP testing is rapidly expanding, with an increasing focus on central neurological disorders. The current literature concerning VEMP abnormalities in central neurological disorders is critically reviewed, especially regarding the methodological aspects in relation to quality as well as the clinical interpretation of the VEMP results. Suggestions for further research are proposed as well as some clinically useful indications. Copyright © 2015 International Federation of Clinical Neurophysiology. Published by Elsevier Ireland Ltd. All rights reserved.

  12. Arterial Myogenic Activation through Smooth Muscle Filamin A

    Directory of Open Access Journals (Sweden)

    Kevin Retailleau

    2016-03-01

    Full Text Available Mutations in the filamin A (FlnA gene are frequently associated with severe arterial abnormalities, although the physiological role for this cytoskeletal element remains poorly understood in vascular cells. We used a conditional mouse model to selectively delete FlnA in smooth muscle (sm cells at the adult stage, thus avoiding the developmental effects of the knockout. Basal blood pressure was significantly reduced in conscious smFlnA knockout mice. Remarkably, pressure-dependent tone of the resistance caudal artery was lost, whereas reactivity to vasoconstrictors was preserved. Impairment of the myogenic behavior was correlated with a lack of calcium influx in arterial myocytes upon an increase in intraluminal pressure. Notably, the stretch activation of CaV1.2 was blunted in the absence of smFlnA. In conclusion, FlnA is a critical upstream element of the signaling cascade underlying the myogenic tone. These findings allow a better understanding of the molecular basis of arterial autoregulation and associated disease states.

  13. The SNPs Analysis of the Exons of Three Genes of MyoD Gene Family in Seven Swine Breeds (Line)

    Institute of Scientific and Technical Information of China (English)

    LI Jing-fen; LIU Di; YU Hao

    2005-01-01

    The study objects includes seven swine breeds: Minzhu, Sanjiangbaizhu, Yorkshire, Landrace, Junmuyihao, Duroc and Double muscle Yorkshire. According to the sequences of MyoG, MyoD and Myf5 of swine in GenBank, seventeen pairs of primers for MyoG, MyoD and Myf5 were designed. PCR-SSCP technology was applied to detect SNPs of the exons of the three genes. The results showed that no polymorphism was in MyoG and MyoD, and some SNPs were in three exons of Myf5. There was one mutant site in the first exon of Myf5 (G → C), three mutant sites in the second exon of Myf5 (C → A, A → G and G → A); in the third exon of Myf5, there was one base A deficiency at 3 387 bp, three bases T deficiency at 3 417 bp, one mutant site at 3 443 bp (T → C).This study obtained a tendency conclusion that gene frequency of allele M of Myf5 on the one hand is positively correlated with lean meat percentage, on the other hand is correlated with the orientation of selective breeding; it also deduced that allele F is possibly correlated with high lean meat percentage. Through statistical analysis, allele A, B, C of Myf5 have no obvious correlation with lean meat percentage of different swine breeds, In addition, the high polymorphism of Myf5 showed that seven swine breeds are rich in genetic variation, and have high selective competency.

  14. Distinct populations of adipogenic and myogenic Myf5-lineage progenitors in white adipose tissues.

    Science.gov (United States)

    Shan, Tizhong; Liang, Xinrong; Bi, Pengpeng; Zhang, Pengpeng; Liu, Weiyi; Kuang, Shihuan

    2013-08-01

    Brown adipose tissues (BAT) are derived from a myogenic factor 5 (Myf5)-expressing cell lineage and white adipose tissues (WAT) predominantly arise from non-Myf5 lineages, although a subpopulation of adipocytes in some WAT depots can be derived from the Myf5 lineage. However, the functional implication of the Myf5- and non-Myf5-lineage cells in WAT is unclear. We found that the Myf5-lineage constitution in subcutaneous WAT depots is negatively correlated to the expression of classical BAT and newly defined beige/brite adipocyte-specific genes. Consistently, fluorescent-activated cell sorting (FACS)-purified Myf5-lineage adipo-progenitors give rise to adipocytes expressing lower levels of BAT-specific Ucp1, Prdm16, Cidea, and Ppargc1a genes and beige adipocyte-specific CD137, Tmem26, and Tbx1 genes compared with the non-Myf5-lineage adipocytes from the same depots. Ablation of the Myf5-lineage progenitors in WAT stromal vascular cell (SVC) cultures leads to increased expression of BAT and beige cell signature genes. Strikingly, the Myf5-lineage cells in WAT are heterogeneous and contain distinct adipogenic [stem cell antigen 1(Sca1)-positive] and myogenic (Sca1-negative) progenitors. The latter differentiate robustly into myofibers in vitro and in vivo, and they restore dystrophin expression after transplantation into mdx mouse, a model for Duchenne muscular dystrophy. These results demonstrate the heterogeneity and functional differences of the Myf5- and non-Myf5-lineage cells in the white adipose tissue.

  15. Abnormality of cervical vestibular-evoked myogenic potentials and ocular vestibular-evoked myogenic potentials in patients with recurrent benign paroxysmal postitional vertigo.

    Science.gov (United States)

    Lee, Jong Dae; Park, Moo Kyun; Lee, Byung Don; Lee, Tae Kyeong; Sung, Ki-Bum; Park, Ji Yun

    2013-02-01

    Our results show that cervical vestibular-evoked myogenic potential (cVEMP) or ocular VEMP (oVEMP) abnormalities in the recurrent benign paroxysmal positional vertigo (BPPV) group were significantly higher than those in the non-recurrent BPPV group. Therefore, we can infer that VEMP abnormality is one of risk factors for BPPV recurrence. This prospective study aimed to test the hypothesis that otolith dysfunction using the VEMP test is a cause of recurrence of BPPV. cVEMP and oVEMP tests using 500 Hz tone-burst stimuli were performed on 16 patients with recurrent BPPV between March 2010 and December 2011. Both VEMP tests were performed in 20 patients with non-recurrent BPPV. The differences in age, sex, and involved canal between the recurrent and non-recurrent BPPV groups were not significant. Abnormal cVEMP responses were detected in 5 of 16 (31.3%) subjects in the recurrent BPPV group and abnormal oVMEP responses were detected in 4 of 16 (25%) subjects in the recurrent BPPV group. When we defined VEMP abnormality as an abnormal cVEMP or abnormal oVEMP, VEMP abnormalities were detected in eight (50%) subjects in the recurrent BPPV group and in three (15%) subjects in the non-recurrent BPPV group; the difference between groups was significant.

  16. Vitamin D(3) enhances the expression of I-mfa, an inhibitor of the MyoD family, in osteoblasts.

    Science.gov (United States)

    Tsuji, K; Kraut, N; Groudine, M; Noda, M

    2001-05-28

    I-mfa (inhibitor of the MyoD family) is a transcription modulator that binds to and suppresses the transcriptional activity of MyoD family members. I-mfa transcripts are expressed in sclerotome, suggesting a role of I-mfa in skeletogenesis. The aim of this study was to examine the expression and regulation of I-mfa in osteoblasts. We found that I-mfa is expressed at a low level in an osteoblast-like cell line, MC3T3E1, and a pluripotent differentiation modulator, 1,25-dihydroxyvitamin D(3), specifically enhanced I-mfa mRNA expression. This effect was completely blocked by the presence of an RNA polymerase inhibitor, but not by a protein synthesis inhibitor, suggesting that 1,25-dihydroxyvitamin D(3) upregulates transcription of the I-mfa gene without requirement for new protein synthesis. Western blot analysis indicated that 1,25-dihydroxyvitamin D(3) increased the I-mfa protein levels severalfold in MC3T3E1 cells. I-mfa expression was also observed in primary mouse calvaria cells and ROS17/2.8 cells and 1,25-dihydroxyvitamin D(3) enhanced I-mfa expression in these cells. These data indicate that I-mfa is a novel transcriptional regulator gene expressed in osteoblasts and that its level is under the control of 1,25-dihydroxyvitamin D(3).

  17. Myogenic differentiation of laryngeal mucosal mesenchymal stem cells%喉黏膜间充质干细胞肌向分化的实验研究

    Institute of Scientific and Technical Information of China (English)

    杨润琴; 明磊国; 邓志宏; 金岩

    2014-01-01

    Objective To investigate the myogenic differentiation of laryngeal mucosal mesenchymal stem cells (LM-MSCs) and the possibility of LM-MSCs as new alternative seed cells for laryngeal tissue engineering.Methods LM-MSCs were separated from normal epiglottis mucosa and the cell surface markers including CD44,CD105,CD90,CD29,CD34 and CD45 were analyzed through flow cytometry.The osteogenesis and adipogenesis differentiation of LM-MSCs were investigated by oil red staining and alizarin red S staining.Immunofluorescence staining and RT-PCR were used to detect the expressions of myogenic differentiation markers including Myod1,Myogenin and myosin heavy chain (MyHc).Results The separated LM-MSCs were in a fibrocyte-like form with long fusiform shape and grew adherently.The expression rates of cell surface markers LM-MSCs were CD44 (100.0%),CD105 (90.4%),CD90 (99.9%),CD29 (93.0%),CD34 (0.4%) and CD45 (1.3%) respectively.A number of beaded lipid drops and mineral deposition were observed after 14 days of adipogenesis differentiation and 21 days of osteogenesis differentiation.Myod1,Myogenin and MyHc genes appeared after 1 week and 3 weeks of myogenesis differentiation respectively.Conclusions The LM-MSCs have the properties of mesenchymal stem cells and could be differentiated into myoblasts,providing with the possibility to repair the damaged vocal cords with LM-MSCs through tissue engineering techniques.%目的 将喉黏膜间充质干细胞(laryngeal mucosal mesenchymal stem cells,LM-MSC)向成肌细胞诱导分化,为声带部分或全部缺失的组织工程修复寻求优势种子细胞.方法 从正常会厌舌面黏膜分离培养获得LM-MSC,流式细胞仪检测细胞表面分子标志CD44、CD105、CD90、CD29、CD34和CD45;向成脂、成骨细胞分化鉴定细胞多向分化潜能,油红O、茜素红染色鉴定成脂、成骨分化情况;加入成肌诱导液向成肌细胞方向分化,于成肌诱导分化后1周和3周时行免

  18. Changes in calsequestrin, TNF-α, TGF-β and MyoD levels during the progression of skeletal muscle dystrophy in mdx mice: a comparative analysis of the quadriceps, diaphragm and intrinsic laryngeal muscles.

    Science.gov (United States)

    Barros Maranhão, Juliana; de Oliveira Moreira, Drielen; Maurício, Adriana Fogagnolo; de Carvalho, Samara Camaçari; Ferretti, Renato; Pereira, Juliano Alves; Santo Neto, Humberto; Marques, Maria Julia

    2015-10-01

    In Duchenne muscular dystrophy (DMD), the search for new biomarkers to follow the evolution of the disease is of fundamental importance in the light of the evolving gene and pharmacological therapies. In addition to the lack of dystrophin, secondary events including changes in calcium levels, inflammation and fibrosis greatly contribute to DMD progression and the molecules involved in these events may represent potential biomarkers. In this study, we performed a comparative evaluation of the progression of dystrophy within muscles that are differently affected by dystrophy (diaphragm; DIA and quadriceps; QDR) or spared (intrinsic laryngeal muscles) using the mdx mice model of DMD. We assessed muscle levels of calsequestrin (calcium-related protein), tumour necrosis factor (TNF-α; pro-inflammatory cytokine), tumour growth factor (TGF-β; pro-fibrotic factor) and MyoD (muscle proliferation) vs. histopathology at early (1 and 4 months of age) and late (9 months of age) stages of dystrophy. Fibrosis was the primary feature in the DIA of mdx mice (9 months: 32% fibrosis), which was greater than in the QDR (9 months: 0.6% fibrosis). Muscle regeneration was the primary feature in the QDR (9 months: 90% of centrally nucleated fibres areas vs. 33% in the DIA). The QDR expressed higher levels of calsequestrin than the DIA. Laryngeal muscles showed normal levels of TNF-α, TGF-β and MyoD. A positive correlation between histopathology and cytokine levels was observed only in the diaphragm, suggesting that TNF-α and TGF-β serve as markers of dystrophy primarily for the diaphragm.

  19. Analysis of interaction between TGF and the myogenic response in renal blood flow autoregulation

    DEFF Research Database (Denmark)

    Feldberg, R; Colding-Jørgensen, M; Holstein-Rathlou, N H

    1995-01-01

    The present study investigates the interaction between the tubuloglomerular feedback (TGF) response and the myogenic mechanism by use of a mathematical model. The two control mechanisms are implemented in a spatially distributed model of the rat renal juxtamedullary afferent arteriole. The model....... The contribution of TGF to smooth muscle activity is assumed to be a linear function of the glomerular capillary pressure. The results show that the myogenic response plays an important role in renal blood flow autoregulation. Without a myogenic response, mechanisms such as TGF that are localized in the distal...... located in the distal part of the afferent arteriole. An ascending myogenic response could enhance the regulatory efficiency of the TGF mechanism by increasing the open-loop gain of the system. However, such a synergistic interaction will only be observed when the two mechanisms operate on more or less...

  20. Vestibular Dysfunctions in Cochlear Implant Patients; A Vestibular Evoked Myogenic Potential Study

    Directory of Open Access Journals (Sweden)

    Masoud Motasaddi Zarandy

    2011-12-01

    Full Text Available Background and Aim: Vestibular evoked myogenic potential in response to click or short tone burst stimuli have been used as a clinical test for distinguish saccule and inferior vestibular nerve diseases. Different studies show that cochlear implant could have inverse effects on vestibular structures. We aimed to investigate vestibular evoked myogenic potential in unilateral cochlear implanted individuals in compare to normal individuals.Methods: Thirty-three unilateral cochlear implanted patients (mean age 19.96 years and 30 normal hearing individuals (mean age 24-27 years as control group were enrolled in this cross- sectional study. Absolute latencies and amplitudes of myogenic potential responses were measured and compared in both groups.Results: Myogenic potential recorded in both ears of all controls were normal. No response could be recorded in 16 patients (48.48% from both ears. In three patients, responses were recorded in both ears though the amplitude of waves was reduced in implanted ear. Unilateral response could be recorded in 14 patients only in their non-implanted ear.Conclusion: Vestibular evoked myogenic potential test is a useful tool for assessing saccular function in cochlear implant patients. Damages of osseous spiral lamina and basilar membrane after cochlear implantation could result in dysfunctions of vestibular organs specially saccule. It seems that saccule could be easily damaged after cochlear implantation. This would cause absence or reduced amplitudes in myogenic potential.

  1. RNA/DNA ratio and LPL and MyoD mRNA expressions in muscle of Oreochromis niloticus fed with elevated levels of palm oil

    Science.gov (United States)

    Ayisi, Christian Larbi; Zhao, Jinliang

    2016-02-01

    Palm oil is of great potential as one of the sustainable alternatives to fish oil (FO) in aquafeeds. In this present study, five isonitrogenous diets (32% crude protein) with elevated palm oil levels of 0%, 2%, 4%, 6% and 8% were used during an 8-week feeding trial to evaluate its effects on RNA/DNA ratio and lipoprotein lipase (LPL) and MyoD mRNA expressions in muscle of Oreochromis niloticus. The results showed that RNA, DNA content as well as ratio of RNA to DNA were significantly affected ( P palm oil level. There was a strong positive correlation between nucleic acid concentration (RNA concentration and RNA: DNA ratio) and specific growth rate (SGR), protein efficiency ratio (PER), while a negative correlation existed between nucleic acid concentration (RNA concentration and RNA: DNA ratio) and feed conversion ratio (FCR). The mRNA expressions of LPL and MyoD in muscle were not significantly affected by the different palm oil levels, although the highest expression was observed in fish fed with 6% palm oil level. There also existed a strong positive correlation between the mRNA expression of LPL, MyoD and SGR, PER, while their correlation with FCR was negative. In conclusion, elevated palm oil affected the RNA, DNA concentration as well as RNA/DNA ratio significantly, although the mRNA expression of LPL and MyoD were not affected significantly by elevated palm oil levels.

  2. Platelet-rich plasma, especially when combined with a TGF-β inhibitor promotes proliferation, viability and myogenic differentiation of myoblasts in vitro.

    Directory of Open Access Journals (Sweden)

    Robi Kelc

    Full Text Available Regeneration of skeletal muscle after injury is limited by scar formation, slow healing time and a high recurrence rate. A therapy based on platelet-rich plasma (PRP has become a promising lead for tendon and ligament injuries in recent years, however concerns have been raised that PRP-derived TGF-β could contribute to fibrotic remodelling in skeletal muscle after injury. Due to the lack of scientific grounds for a PRP -based muscle regeneration therapy, we have designed a study using human myogenic progenitors and evaluated the potential of PRP alone and in combination with decorin (a TGF-β inhibitor, to alter myoblast proliferation, metabolic activity, cytokine profile and expression of myogenic regulatory factors (MRFs. Advanced imaging multicolor single-cell analysis enabled us to create a valuable picture on the ratio of quiescent, activated and terminally committed myoblasts in treated versus control cell populations. Finally high-resolution confocal microscopy validated the potential of PRP and decorin to stimulate the formation of polynucleated myotubules. PRP was shown to down-regulate fibrotic cytokines, increase cell viability and proliferation, enhance the expression of MRFs, and contribute to a significant myogenic shift during differentiation. When combined with decorin further synergistc effects were identified. These results suggest that PRP could not only prevent fibrosis but could also stimulate muscle commitment, especially when combined with a TGF-β inhibitor.

  3. Effects of Dietary Supplementation of β-hydroxy-β-methylbutyrate on Sow Performance and mRNA Expression of Myogenic Markers in Skeletal Muscle of Neonatal Piglets.

    Science.gov (United States)

    Wan, H F; Zhu, J T; Shen, Y; Xiang, X; Yin, H J; Fang, Z F; Che, L Q; Lin, Y; Xu, S Y; Feng, B; Wu, D

    2016-02-01

    The effects of dietary β-hydroxy-β-methylbutyrate (HMB) supplementation during gestation on reproductive performance of sows and the mRNA expression of myogenic markers in skeletal muscle of neonatal pigs were determined. At day 35 of gestation, a total of 20 sows (Landrace × Yorkshire, at third parity) were randomly assigned to two groups, with each group receiving either a basal diet or the same diet supplemented with 4 g/day β-hydroxy-β-methylbutyrate calcium (HMB-Ca) until parturition. At parturition, the total and live litter size were not markedly different between treatments, however, the sows fed HMB diet had a decreased rate of stillborn piglets compared with the sows fed the control (CON) diets (p sows fed HMB diet tended to have an increased birth weight (p = 0.08), and a reduced rate of low birth weight piglets (p = 0.05) compared with piglets from the CON sows. Nevertheless, lower feed intake during lactation was observed in the sows fed the HMB diet compared with those on the CON diet (p sows. Furthermore, maternal HMB treatment increased the mRNA levels of the myogenic genes, including muscle regulatory factor-4 (MRF4, p sows at 4 g/day from day 35 of gestation to term significantly improves pregnancy outcomes and increases the expression of myogenic genes in skeletal muscle of neonatal piglets, but reduces feed intake of sows during lactation.

  4. Influences of dietary vitamin D restriction on bone strength, body composition and muscle in rats fed a high-fat diet: involvement of mRNA expression of MyoD in skeletal muscle.

    Science.gov (United States)

    Oku, Yuno; Tanabe, Rieko; Nakaoka, Kanae; Yamada, Asako; Noda, Seiko; Hoshino, Ayumi; Haraikawa, Mayu; Goseki-Sone, Masae

    2016-06-01

    Vitamin D insufficiency is associated with a greater risk of osteoporosis and also influences skeletal muscle functions, differentiation and development. The present study investigated the influences of vitamin D restriction on the body composition, bone and skeletal muscle in rats fed a high-fat diet. Sprague-Dawley strain male rats (11weeks old) were divided into four groups and fed experimental diets: a basic control diet (Cont.), a basic control diet with vitamin D restriction (DR), a high-fat diet (F) and a high-fat diet with vitamin D restriction (FDR). At 28days after starting the experimental diets, the visceral fat mass was significantly increased in the F group compared with Cont. group, and the muscle mass tended to decrease in the DR group compared with Cont. group. The total volume of the femur was significantly lower in the DR group compared with Cont. group, and the bone mineral density (BMD) of the femur was significantly lower in the FDR group compared with F group. MyoD is one of the muscle-specific transcription factors. The levels of mRNA expression of MyoD of the gastrocnemius and soleus muscles from the DR group were reduced markedly compared with those from the Cont. group. In conclusion, our findings revealed the influences of a vitamin D-restricted high-fat diet on the bone strength, body composition and muscle. Further studies on vitamin D insufficiency in the regulation of muscle as well as fat and bone metabolism would provide valuable data for the prevention of lifestyle-related disorders, including osteoporosis and sarcopenia.

  5. Selective predation of tawny owls (Strix aluco) on yellow-necked mice (Apodemus flavicollis) and bank voles (Myodes glareolus)

    DEFF Research Database (Denmark)

    Sunde, Peter; Forsom, Heidi Malene; Al-Sabi, Mohammad Nafi Solaiman

    2012-01-01

    Differential predation on certain classes of individuals within prey populations might make owls strong selective agents on their prey. We investigated selective predation of tawny owls (Strix aluco) on yellow-necked mice (Apodemus flavicollis, A.f.) and bank voles (Myodes glareolus, M.g.) for two...... years by comparing prey from owl nests with live-trapped individuals. The owls killed significantly more male M.g. (73%) than females, but not more than expected from traps (57%). For A.f., owls selected adults in favour of subadults, and for adults, individuals with longer femurs. Adult males of A.......f. killed by owls had significantly heavier testes in relation their size than the trapped males. Prey selection did not correlate with size-adjusted body or spleen mass. Owl-killed A.f. had higher prevalences of the intestinal helminth Heligmosomoides sp. than trapped individuals, but hosted similar...

  6. Selective Predation of Tawny Owls (Strix aluco) on Yellow-Necked Mice (Apodemus flavicollis) and Bank Voles (Myodes glareolus)

    DEFF Research Database (Denmark)

    Sunde, Peter; Forsom, Heidi Malene; Al-Sabi, Mohammad Nafi Solaiman

    2012-01-01

    Differential predation on certain classes of individuals within prey populations might make owls strong selective agents on their prey. We investigated selective predation of tawny owls (Strix aluco) on yellow-necked mice (Apodemus flavicollis, A.f.) and bank voles (Myodes glareolus, M.g.) for two...... years by comparing prey from owl nests with live-trapped individuals. The owls killed significantly more male M.g. (73%) than females, but not more than expected from traps (57%). For A.f., owls selected adults in favour of subadults, and for adults, individuals with longer femurs. Adult males of A.......f. killed by owls had significantly heavier testes in relation their size than the trapped males. Prey selection did not correlate with size-adjusted body or spleen mass. Owl-killed A.f. had higher prevalences of the intestinal helminth Heligmosomoides sp. than trapped individuals, but hosted similar...

  7. Schisandrae fructus enhances myogenic differentiation and inhibits atrophy through protein synthesis in human myotubes

    Science.gov (United States)

    Kim, Cy Hyun; Shin, Jin-Hong; Hwang, Sung Jun; Choi, Yung Hyun; Kim, Dae-Seong; Kim, Cheol Min

    2016-01-01

    Schisandrae fructus (SF) has recently been reported to increase skeletal muscle mass and inhibit atrophy in mice. We investigated the effect of SF extract on human myotube differentiation and its acting pathway. Various concentrations (0.1–10 μg/mL) of SF extract were applied on human skeletal muscle cells in vitro. Myotube area and fusion index were measured to quantify myotube differentiation. The maximum effect was observed at 0.5 μg/mL of SF extract, enhancing differentiation up to 1.4-fold in fusion index and 1.6-fold in myotube area at 8 days after induction of differentiation compared to control. Phosphorylation of eukaryotic translation initiation factor 4E-binding protein 1 and 70 kDa ribosomal protein S6 kinase, which initiate translation as downstream of mammalian target of rapamycin pathway, was upregulated in early phases of differentiation after SF treatment. SF also attenuated dexamethasone-induced atrophy. In conclusion, we show that SF augments myogenic differentiation and attenuates atrophy by increasing protein synthesis through mammalian target of rapamycin/70 kDa ribosomal protein S6 kinase and eukaryotic translation initiation factor 4E-binding protein 1 signaling pathway in human myotubes. SF can be a useful natural dietary supplement in increasing skeletal muscle mass, especially in the aged with sarcopenia and the patients with disuse atrophy. PMID:27330287

  8. Mesenchymal Stromal Cells for Sphincter Regeneration: Role of Laminin Isoforms upon Myogenic Differentiation.

    Science.gov (United States)

    Seeger, Tanja; Hart, Melanie; Patarroyo, Manuel; Rolauffs, Bernd; Aicher, Wilhelm K; Klein, Gerd

    2015-01-01

    Multipotent mesenchymal stromal cells (MSCs) are well known for their tri-lineage potential and ability to differentiate in vitro into osteogenic, chondrogenic or adipogenic lineages. By selecting appropriate conditions MSCs can also be differentiated in vitro into the myogenic lineage and are therefore a promising option for cell-based regeneration of muscle tissue such as an aged or damaged sphincter muscle. For the differentiation into the myogenic lineage there is still a need to evaluate the effects of extracellular matrix proteins such as laminins (LM) which are crucial for different stem cell types and for normal muscle function. The laminin family consists of 16 functionally different isoforms with LM-211 being the most abundant isoform of adult muscle tissues. In the sphincter tissue a strong expression of the isoforms LM-211/221, LM-411/421 and LM-511/521 can be detected in the different cell layers. Bone marrow-derived MSCs in culture, however, mainly express the isoforms LM-411 and LM-511, but not LM-211. Even after myogenic differentiation, LM-211 can hardly be detected. All laminin isoforms tested (LM-211, LM-411, LM-511 and LM-521) showed a significant inhibition of the proliferation of undifferentiated MSCs but, with the exception of LM-521, they had no influence on the proliferation of MSCs cultivated in myogenic medium. The strongest cellular adhesion of MSCs was to LM-511 and LM-521, whereas LM-211 was only a weakly-adhesive substrate for MSCs. Myogenic differentiation of MSCs even reduced the interaction with LM-211, but it did not affect the interaction with LM-511 and LM-521. Since during normal myogenesis the latter two isoforms are the major laminins surrounding developing myogenic progenitors, α5 chain-containing laminins are recommended for further improvements of myogenic differentiation protocols of MSCs into smooth muscle cells.

  9. Mesenchymal Stromal Cells for Sphincter Regeneration: Role of Laminin Isoforms upon Myogenic Differentiation.

    Directory of Open Access Journals (Sweden)

    Tanja Seeger

    Full Text Available Multipotent mesenchymal stromal cells (MSCs are well known for their tri-lineage potential and ability to differentiate in vitro into osteogenic, chondrogenic or adipogenic lineages. By selecting appropriate conditions MSCs can also be differentiated in vitro into the myogenic lineage and are therefore a promising option for cell-based regeneration of muscle tissue such as an aged or damaged sphincter muscle. For the differentiation into the myogenic lineage there is still a need to evaluate the effects of extracellular matrix proteins such as laminins (LM which are crucial for different stem cell types and for normal muscle function. The laminin family consists of 16 functionally different isoforms with LM-211 being the most abundant isoform of adult muscle tissues. In the sphincter tissue a strong expression of the isoforms LM-211/221, LM-411/421 and LM-511/521 can be detected in the different cell layers. Bone marrow-derived MSCs in culture, however, mainly express the isoforms LM-411 and LM-511, but not LM-211. Even after myogenic differentiation, LM-211 can hardly be detected. All laminin isoforms tested (LM-211, LM-411, LM-511 and LM-521 showed a significant inhibition of the proliferation of undifferentiated MSCs but, with the exception of LM-521, they had no influence on the proliferation of MSCs cultivated in myogenic medium. The strongest cellular adhesion of MSCs was to LM-511 and LM-521, whereas LM-211 was only a weakly-adhesive substrate for MSCs. Myogenic differentiation of MSCs even reduced the interaction with LM-211, but it did not affect the interaction with LM-511 and LM-521. Since during normal myogenesis the latter two isoforms are the major laminins surrounding developing myogenic progenitors, α5 chain-containing laminins are recommended for further improvements of myogenic differentiation protocols of MSCs into smooth muscle cells.

  10. Mesenchymal Stromal Cells for Sphincter Regeneration: Role of Laminin Isoforms upon Myogenic Differentiation

    Science.gov (United States)

    Seeger, Tanja; Hart, Melanie; Patarroyo, Manuel; Rolauffs, Bernd; Aicher, Wilhelm K.; Klein, Gerd

    2015-01-01

    Multipotent mesenchymal stromal cells (MSCs) are well known for their tri-lineage potential and ability to differentiate in vitro into osteogenic, chondrogenic or adipogenic lineages. By selecting appropriate conditions MSCs can also be differentiated in vitro into the myogenic lineage and are therefore a promising option for cell-based regeneration of muscle tissue such as an aged or damaged sphincter muscle. For the differentiation into the myogenic lineage there is still a need to evaluate the effects of extracellular matrix proteins such as laminins (LM) which are crucial for different stem cell types and for normal muscle function. The laminin family consists of 16 functionally different isoforms with LM-211 being the most abundant isoform of adult muscle tissues. In the sphincter tissue a strong expression of the isoforms LM-211/221, LM-411/421 and LM-511/521 can be detected in the different cell layers. Bone marrow-derived MSCs in culture, however, mainly express the isoforms LM-411 and LM-511, but not LM-211. Even after myogenic differentiation, LM-211 can hardly be detected. All laminin isoforms tested (LM-211, LM-411, LM-511 and LM-521) showed a significant inhibition of the proliferation of undifferentiated MSCs but, with the exception of LM-521, they had no influence on the proliferation of MSCs cultivated in myogenic medium. The strongest cellular adhesion of MSCs was to LM-511 and LM-521, whereas LM-211 was only a weakly-adhesive substrate for MSCs. Myogenic differentiation of MSCs even reduced the interaction with LM-211, but it did not affect the interaction with LM-511 and LM-521. Since during normal myogenesis the latter two isoforms are the major laminins surrounding developing myogenic progenitors, α5 chain-containing laminins are recommended for further improvements of myogenic differentiation protocols of MSCs into smooth muscle cells. PMID:26406476

  11. Juvenile growth reduces the influence of epithelial sodium channels on myogenic tone in skeletal muscle arterioles.

    Science.gov (United States)

    Kang, Lori S; Masilamani, Shyama; Boegehold, Matthew A

    2016-12-01

    Previous studies have documented that rapid juvenile growth is accompanied by functional changes in the arteriolar endothelium, but much less is known about functional changes in arteriolar smooth muscle over this period. In this study, we investigate the possible contribution of epithelial sodium channels (ENaC) to the myogenic behaviour of arterioles at two stages of juvenile growth. The effects of the ENaC inhibitor benzamil on different levels of myogenic tone were studied in isolated gracilis muscle arterioles from rats aged 21-28 days ("weanlings") and 42-49 days ("juveniles"). ENaC subunit expression in the arteriolar wall was also determined, and the interaction between ENaC and nitric oxide (NO) in regulating vascular tone was explored by combined use of benzamil and N(G) -monomethyl-l-arginine (l-NMMA). At physiological pressures, both steady-state myogenic tone and the dynamic adjustments in this tone triggered by acute pressure changes were less in juvenile arterioles than in weanling arterioles. α, β and γ ENaC protein was present in arterioles at both ages, but benzamil only had an effect on myogenic tone in weanling arterioles. In these vessels, benzamil increased, rather than decreased, myogenic tone, and this effect was prevented by l-NMMA or endothelial removal. These findings suggest that although ENaC is present in gracilis muscle arterioles of both weanling and juvenile rats, it is not obligatory for the genesis of myogenic activity in these vessels at either age. However, ENaC activity can significantly modulate the level of myogenic tone through stimulation of endothelial NO release at an early stage of growth. © 2016 John Wiley & Sons Australia, Ltd.

  12. DNA methylation analysis of human myoblasts during in vitro myogenic differentiation: de novo methylation of promoters of muscle-related genes and its involvement in transcriptional down-regulation.

    Science.gov (United States)

    Miyata, Kohei; Miyata, Tomoko; Nakabayashi, Kazuhiko; Okamura, Kohji; Naito, Masashi; Kawai, Tomoko; Takada, Shuji; Kato, Kiyoko; Miyamoto, Shingo; Hata, Kenichiro; Asahara, Hiroshi

    2015-01-15

    Although DNA methylation is considered to play an important role during myogenic differentiation, chronological alterations in DNA methylation and gene expression patterns in this process have been poorly understood. Using the Infinium HumanMethylation450 BeadChip array, we obtained a chronological profile of the genome-wide DNA methylation status in a human myoblast differentiation model, where myoblasts were cultured in low-serum medium to stimulate myogenic differentiation. As the differentiation of the myoblasts proceeded, their global DNA methylation level increased and their methylation patterns became more distinct from those of mesenchymal stem cells. Gene ontology analysis revealed that genes whose promoter region was hypermethylated upon myoblast differentiation were highly significantly enriched with muscle-related terms such as 'muscle contraction' and 'muscle system process'. Sequence motif analysis identified 8-bp motifs somewhat similar to the binding motifs of ID4 and ZNF238 to be most significantly enriched in hypermethylated promoter regions. ID4 and ZNF238 have been shown to be critical transcriptional regulators of muscle-related genes during myogenic differentiation. An integrated analysis of DNA methylation and gene expression profiles revealed that de novo DNA methylation of non-CpG island (CGI) promoters was more often associated with transcriptional down-regulation than that of CGI promoters. These results strongly suggest the existence of an epigenetic mechanism in which DNA methylation modulates the functions of key transcriptional factors to coordinately regulate muscle-related genes during myogenic differentiation.

  13. Vestibular evoked myogenic potentials in patients with ankylosing spondylitis.

    Science.gov (United States)

    Özgür, Abdulkadir; Serdaroğlu Beyazal, Münevver; Terzi, Suat; Coşkun, Zerrin Özergin; Dursun, Engin

    2016-10-01

    Ankylosing spondylitis (AS) is a chronic systemic inflammatory disease with unknown etiology. Although sacroiliac joint involvement is the classic sign along with the formed immune mediators, it may result in immune-mediated inner ear disease and may cause damage to the audiovestibular system. Vestibular evoked myogenic potentials (VEMP) is a clinical reflex test used in the diagnosis of vestibular diseases and is performed by recording and evaluating the muscle potentials resulting from the stimulation of the vestibular system with different stimuli. The aim of this study is to evaluate the cervical VEMP test results in AS patients without vestibular symptoms. Thirty-three patients with AS and a control group of 30 healthy volunteers with similar demographic characteristics were evaluated in the study. VEMP wave latency, P13-N23 wave amplitude, and VEMP asymmetry ratio (VAR) values were compared between the groups. The relationship between clinical and laboratory findings of the AS patients and VEMP data were also investigated. Compared with healthy people, this study shows the response rate of patients with ankylosing spondylitis was reduced in the VEMP test, and P13-N23 wave amplitude showed a decrease in AS patients who had VEMP response (p ankylosing spondylitis. The data obtained from this study suggest that AS may lead to decreased sensitivity of the vestibular system.

  14. Vestibular evoked myogenic potential in sudden sensorineural hearing loss

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    Feroze Kancharu Khan

    2013-01-01

    Full Text Available Aim and Objective: To investigate saccular damage in patients with sudden sensorineural hearing loss (SSNHL with or without vertigo and to evaluate the saccular damage according to the hearing loss and presence or absence of vertigo. Materials and Methods: All tests done in this study were performed in the audio vestibular unit of ENT department from September 2009 to November 2010. Statistical Analysis Used: The association between the severity of hearing loss and changes in the vestibular evoked myogenic potential (VEMP recordings were assessed using descriptive statistics. The pattern of VEMP in different diseases and also the behavior of VEMP in presence or absence of vertigo were evaluated using SPSS 15. Results: Among 27 patients there were 11 cases of idiopathic SSNHL. Out of nine unaffected ears, 88% had normal and 12% had absent VEMP. Whereas out of 13 affected ears, only 53.9% had normal VEMP. Among all the 54 ears, 17 ears had normal hearing. In this group 76.47% had normal VEMP. The group with hearing loss > 90 dB had 61.53% absent VEMP. Conclusions: In patients with unilateral SSNHL, there was a tendency for the affected ear to have absent VEMP indicating the saccular involvement. The extent of saccular damage did not correspond to the amount of hearing loss or presence or absence of vertigo.

  15. Ocular vestibular evoked myogenic potentials are abnormal in internuclear ophthalmoplegia.

    Science.gov (United States)

    Rosengren, S M; Colebatch, J G

    2011-06-01

    The cervical vestibular evoked myogenic potential (cVEMP) is sensitive to lower brainstem lesions affecting the vestibulo-collic pathway. We wished to determine whether the ocular VEMP (oVEMP), a recently-described otolith-ocular reflex, is also abnormal in patients with brainstem lesions. We tested patients with internuclear ophthalmoplegia (INO), caused by a brainstem lesion in the medial longitudinal fasciculus (MLF), to investigate whether the oVEMP is abnormal in patients with a lesion of the otolith-ocular pathway. We describe a patient who developed a right INO during his first episode of demyelination, and report results from 12 additional patients, most of whom had multiple sclerosis. All subjects were stimulated with air-conducted tone bursts. cVEMPs and oVEMPs were measured using surface electrodes placed over the neck and beneath the eyes. Overall, oVEMPs showed significantly more abnormalities (69%) than cVEMPs (8%). Ocular VEMPs were absent with stimulation of 13/26 ears, significantly delayed in 5/26 cases and normal in only 8/26 cases. Ocular VEMPs are often abnormal in patients with multiple sclerosis who have an INO, while cVEMPs are usually normal. Ocular VEMPs provide a new, non-invasive method for examining central vestibular pathways in humans and are sensitive to lesions of the MLF. Copyright © 2010 International Federation of Clinical Neurophysiology. Published by Elsevier Ireland Ltd. All rights reserved.

  16. Vestibular evoked myogenic potential findings in multiple sclerosis.

    Science.gov (United States)

    Escorihuela García, Vicente; Llópez Carratalá, Ignacio; Orts Alborch, Miguel; Marco Algarra, Jaime

    2013-01-01

    Multiple sclerosis is an inflammatory disease involving the occurrence of demyelinating, chronic neurodegenerative lesions in the central nervous system. We studied vestibular evoked myogenic potentials (VEMPs) in this pathology, to allow us to evaluate the saccule, inferior vestibular nerve and vestibular-spinal pathway non-invasively. There were 23 patients diagnosed with multiple sclerosis who underwent VEMP recordings, comparing our results with a control group consisting of 35 healthy subjects. We registered p13 and n23 wave latencies, interaural amplitude difference and asymmetry ratio between both ears. Subjects also underwent an otoscopy and audiometric examination. The prolongation of p13 and n23 wave latencies was the most notable characteristic, with a mean p13 wave latency of 19.53 milliseconds and a mean latency of 30.06 milliseconds for n23. In contrast, the asymmetry index showed no significant differences with our control group. In case of multiple sclerosis, the prolongation of the p13 and n23 VEMP wave latencies is a feature that has been attributed to slowing of conduction by demyelination of the vestibular-spinal pathway. In this regard, alteration of the response or lack thereof in these potentials has a locator value of injury to the lower brainstem. Copyright © 2013 Elsevier España, S.L. All rights reserved.

  17. Non-myogenic Contribution to Muscle Development and Homeostasis: The Role of Connective Tissues.

    Science.gov (United States)

    Nassari, Sonya; Duprez, Delphine; Fournier-Thibault, Claire

    2017-01-01

    Skeletal muscles belong to the musculoskeletal system, which is composed of bone, tendon, ligament and irregular connective tissue, and closely associated with motor nerves and blood vessels. The intrinsic molecular signals regulating myogenesis have been extensively investigated. However, muscle development, homeostasis and regeneration require interactions with surrounding tissues and the cellular and molecular aspects of this dialogue have not been completely elucidated. During development and adult life, myogenic cells are closely associated with the different types of connective tissue. Connective tissues are defined as specialized (bone and cartilage), dense regular (tendon and ligament) and dense irregular connective tissue. The role of connective tissue in muscle morphogenesis has been investigated, thanks to the identification of transcription factors that characterize the different types of connective tissues. Here, we review the development of the various connective tissues in the context of the musculoskeletal system and highlight their important role in delivering information necessary for correct muscle morphogenesis, from the early step of myoblast differentiation to the late stage of muscle maturation. Interactions between muscle and connective tissue are also critical in the adult during muscle regeneration, as impairment of the regenerative potential after injury or in neuromuscular diseases results in the progressive replacement of the muscle mass by fibrotic tissue. We conclude that bi-directional communication between muscle and connective tissue is critical for a correct assembly of the musculoskeletal system during development as well as to maintain its homeostasis in the adult.

  18. Dynamic clustering and dispersion of lipid rafts contribute to fusion competence of myogenic cells

    Energy Technology Data Exchange (ETDEWEB)

    Mukai, Atsushi [Department of Regenerative Medicine, National Institute for Longevity Sciences, National Center for Geriatrics and Gerontology, 36-3 Gengo, Morioka, Oobu, Aichi 474-8522 (Japan); Kurisaki, Tomohiro [Department of Growth Regulation, Institute for Frontier Medical Sciences, Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto 606-8507 (Japan); Sato, Satoshi B. [Research Center for Low Temperature and Material Sciences, Kyoto University, Yoshida-honmachi, Kyoto 606-8501 (Japan); Kobayashi, Toshihide [Lipid Biology Laboratory, Discovery Research Institute, RIKEN, Wako, Saitama 351-0198 (Japan); Kondoh, Gen [Laboratory of Animal Experiments for Regeneration, Institute for Frontier Medical Sciences, Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto 606-8507 (Japan); Hashimoto, Naohiro, E-mail: nao@nils.go.jp [Department of Regenerative Medicine, National Institute for Longevity Sciences, National Center for Geriatrics and Gerontology, 36-3 Gengo, Morioka, Oobu, Aichi 474-8522 (Japan)

    2009-10-15

    Recent research indicates that the leading edge of lamellipodia of myogenic cells (myoblasts and myotubes) contains presumptive fusion sites, yet the mechanisms that render the plasma membrane fusion-competent remain largely unknown. Here we show that dynamic clustering and dispersion of lipid rafts contribute to both cell adhesion and plasma membrane union during myogenic cell fusion. Adhesion-complex proteins including M-cadherin, {beta}-catenin, and p120-catenin accumulated at the leading edge of lamellipodia, which contains the presumptive fusion sites of the plasma membrane, in a lipid raft-dependent fashion prior to cell contact. In addition, disruption of lipid rafts by cholesterol depletion directly prevented the membrane union of myogenic cell fusion. Time-lapse recording showed that lipid rafts were laterally dispersed from the center of the lamellipodia prior to membrane fusion. Adhesion proteins that had accumulated at lipid rafts were also removed from the presumptive fusion sites when lipid rafts were laterally dispersed. The resultant lipid raft- and adhesion complex-free area at the leading edge fused with the opposing plasma membrane. These results demonstrate a key role for dynamic clustering/dispersion of lipid rafts in establishing fusion-competent sites of the myogenic cell membrane, providing a novel mechanistic insight into the regulation of myogenic cell fusion.

  19. Arecoline inhibits myogenic differentiation of C2C12 myoblasts by reducing STAT3 phosphorylation.

    Science.gov (United States)

    Chang, Yung-Fu; Liu, Ting-Yuan; Liu, Shao-Tung; Tseng, Chao-Neng

    2012-10-01

    Areca nut (Areca catechu) is chewed regularly as a medical and psychoactive food by about 10% of the world population, in countries including India, Taiwan and parts of Southern Asia. Areca nut chewing during pregnancy has been associated with both lower birth weight and premature birth. Animals of low birth weights showed retardation of muscle development. Our previous study showed that arecoline, the major areca alkaloid, decreased the number of implanted embryos. Here we sought to determine the effects of arecoline in myogenic differentiation by in vitro assays using C2C12 myoblast cells. The results showed that arecoline higher than 0.4mM significantly increased apoptosis and decreased viability of C2C12 cells. Morphometric measurements of myotube formation and analyses of myogenic markers, myosin heavy chain and myogenin, revealed that myogenic differentiation was inhibited by 0.04-0.08 mM arecoline. Moreover, phosphorylated but not total STAT3 was significantly inhibited by arecoline during myotube formation. These results indicate that arecoline inhibits the myogenic differentiation of C2C12 cells by reducing the activation of STAT3, an upstream regulator of myogenesis. Improved understanding of the effects of arecoline during myogenic differentiation may help to establish public health policies and to develop potential treatments for such patients. Copyright © 2012 Elsevier Ltd. All rights reserved.

  20. YB-1 gene expression is kept constant during myocyte differentiation through replacement of different transcription factors and then falls gradually under the control of neural activity.

    Science.gov (United States)

    Kobayashi, Shunsuke; Tanaka, Toru; Moue, Masamitsu; Ohashi, Sachiyo; Nishikawa, Taishi

    2015-11-01

    We have previously reported that translation of acetylcholine receptor α-subunit (AChR α) mRNA in skeletal muscle cells is regulated by Y-box binding protein 1 (YB-1) in response to neural activity, and that in the postnatal mouse developmental changes in the amount of YB-1 mRNA are similar to those of AChR α mRNA, which is known to be regulated by myogenic transcription factors. Here, we examined transcriptional regulation of the YB-1 gene in mouse skeletal muscle and differentiating C2C12 myocytes. Although neither YB-1 nor AChR α was detected at either the mRNA or protein level in adult hind limb muscle, YB-1 expression was transiently activated in response to denervation of the sciatic nerve and completely paralleled that of AChR α, suggesting that these genes are regulated by the same transcription factors. However, during differentiation of C2C12 cells to myotubes, the level of YB-1 remained constant even though the level of AChR α increased markedly. Reporter gene, gel mobility shift and ChIP assays revealed that in the initial stage of myocyte differentiation, transcription of the YB-1 gene was regulated by E2F1 and Sp1, and was then gradually replaced under the control of both MyoD and myogenin through an E-box sequence in the proximal region of the YB-1 gene promoter. These results suggest that transcription factors for the YB-1 gene are exchanged during skeletal muscle cell differentiation, perhaps playing a role in translational control of mRNAs by YB-1 in both myotube formation and the response of skeletal muscle tissues to neural stimulation.

  1. Hormonal induction of an immediate-early gene response in myogenic cell lines--a paradigm for heart growth.

    Science.gov (United States)

    Maass, A; Grohé, C; Kubisch, C; Wollnik, B; Vetter, H; Neyses, L

    1995-05-01

    Cardiac hypertrophy is characterized by growth of myocardial cells without proliferation. Many endo- paracrine stimuli such as angiotensin II, endothelin, alpha 1-adrenergic agonists, and insulin have been shown to be able to induce cardiac hypertrophy either in vivo or in vitro. We have used the myoblast model of differentiation and proliferation to determine nuclear signal transduction mechanisms in muscle and (by analogy) cardiac growth. The first nuclear event known to occur when a growth stimulus acts upon a cell is induction of a family of immediate-early genes. Our group focused on the role of one of these genes, the early growth response gene-1 (Egr-1). We have shown that this gene is induced in isolated adult cardiac myocytes in the presence of endothelin. An anti-sense oligonucleotide complementary to the first six codons of the Egr-1 mRNA abolishes the stimulation of protein synthesis induced by endothelin. In the present study we further characterized paracrine growth stimuli in the myogenic cell line Sol8, which was used as a paradigm to further investigate mechanisms of paracrine growth induction. We demonstrated that a variety of candidate endo- paracrine stimuli for the induction of cardiac hypertrophy induced the Egr-1 messenger RNA in the myogenic cell line Sol8. Among these are endothelin, insulin, basic fibroblast growth factor, and platelet-derived growth factor BB (PDGF BB). We conclude: (1) In analogy to the myocardium, these growth factors act upon myoblasts. (2) This line appears to be a suitable model for the molecular characterization of Egr-1 target genes.

  2. Adipose-derived stem cells enhance myogenic differentiation in the mdx mouse model of muscular dystrophy via paracrine signaling

    Directory of Open Access Journals (Sweden)

    Ji-qing Cao

    2016-01-01

    Full Text Available Adipose-derived stem cells have been shown to promote peripheral nerve regeneration through the paracrine secretion of neurotrophic factors. However, it is unclear whether these cells can promote myogenic differentiation in muscular dystrophy. Adipose-derived stem cells (6 × 10 6 were injected into the gastrocnemius muscle of mdx mice at various sites. Dystrophin expression was found in the muscle fibers. Phosphorylation levels of Akt, mammalian target of rapamycin (mTOR, eIF-4E binding protein 1 and S6 kinase 1 were increased, and the Akt/mTOR pathway was activated. Simultaneously, myogenin levels were increased, whereas cleaved caspase 3 and vimentin levels were decreased. Necrosis and fibrosis were reduced in the muscle fibers. These findings suggest that adipose-derived stem cells promote the regeneration and survival of muscle cells by inhibiting apoptosis and fibrosis, thereby alleviating muscle damage in muscular dystrophy.

  3. Adipose-derived stem cells enhance myogenic differentiation in the mdx mouse model of muscular dystrophyvia paracrine signaling

    Institute of Scientific and Technical Information of China (English)

    Ji-qing Cao; Jie Kong; Cheng Zhang; Ying-yin Liang; Ya-qin Li; Hui-li Zhang; Yu-ling Zhu; Jia Geng; Li-qing Yang; Shan-wei Feng; Juan Yang

    2016-01-01

    Adipose-derived stem cells have been shown to promote peripheral nerve regeneration through the paracrine secretion of neurotrophic factors. However, it is unclear whether these cells can promote myogenic differentiation in muscular dystrophy. Adipose-derived stem cells (6 × 106) were injected into the gastrocnemius muscle of mdx mice at various sites. Dystrophin expression was found in the muscle ifbers. Phosphorylation levels of Akt, mammalian target of rapamycin (mTOR), eIF-4E binding protein 1 and S6 kinase 1 were increased, and the Akt/mTOR pathway was activated. Simultaneously, myogenin levels were increased, whereas cleaved caspase 3 and vimentin levels were decreased. Necrosis and ifbrosis were reduced in the muscle ifbers. hTese ifndings suggest that adipose-derived stem cells promote the re-generation and survival of muscle cells by inhibiting apoptosis and ifbrosis, thereby alleviating muscle damage in muscular dystrophy.

  4. Vestibular-evoked myogenic potentials in central vestibular disorders.

    Science.gov (United States)

    Oh, Sun-Young; Kim, Hyo-Jeong; Kim, Ji-Soo

    2016-02-01

    Vestibular-evoked myogenic potentials (VEMPs) are short latency manifestations of vestibulo-ocular and vestibulocollic reflexes that originate from the utricle and saccule. Although cervical and ocular VEMPs have mostly been applied to peripheral vestibular disorders, the characteristics and the diagnostic values of VEMPs have been expanded to assess the function of the central otolithic pathways. In the central nervous system, the cervical VEMPs (cVEMPs) are mediated by the vestibular nuclei and uncrossed medial vestibulospinal tract descending in the lower brainstem and spinal cord. In contrast, the ocular VEMPs (oVEMPs) reflect the function of the vestibular nuclei and the crossed vestibulo-ocular reflex (VOR) pathways, mostly contained in the medial longitudinal fasciculus (MLF). Therefore, lesions involving the vestibular nuclei can present abnormalities of both cVEMPs and oVEMPs. The medullary lesions involving the descending MLF or the spinal accessory nucleus impair cVEMPs. In contrast, the lesions involving the MLF, the crossed ventral tegmental tract, oculomotor nuclei and the interstitial nucleus of Cajal can impair oVEMPs. Patients with unilateral cerebellar infarctions may show abnormal VEMPs especially when they have the ocular tilt reaction. Delayed responses of VEMPs are characteristic of multiple sclerosis (MS). Reduced VEMP responses can be observed in patients with vestibular migraine. VEMPs are useful in evaluating central as well as peripheral otolithic function that are not readily defined by conventional vestibular function tests, and can aid in detecting and localizing central lesions, especially silent brainstem lesions such as tiny infarctions or MS plaques.

  5. Ocular vestibular evoked myogenic potentials in normal-hearing adults

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    Mohammad Kamali

    2012-06-01

    Full Text Available Background and Aim: Ocular vestibular-evoked myogenic potential (oVEMP is a novel vestibular function test. This short-latency response can be recorded through contracting extraocular muscles by high-intensity acoustic stimulation and can be used to evaluate contralateral ocular-vestibular reflex. The aim of this study was to record and compare the amplitude, latency, asymmetry ratio and occurrence percentage of oVEMP (n10 and cervical VEMP (p13 responses in a group of normal adult subjects.Methods: We carried out a cross-sectional study on 20 adult subjects' mean age 22.18 years, SD=2.19 with normal hearing sensitivity and no history of vestibular diseases. oVEMP and cVEMP responses in both ears were recorded using air conducted stimuli 500 Hz short tone burst, 95 dB nHL via insert earphone and compared.Results: cVEMP was recorded in all subjects but oVEMP was absent in two subjects. Mean amplitude and latency were 140.77 μv and 15.56 ms in p13; and 3.18 μv and 9.32 ms in n10. There were statistically significant differences between p13 and n10 amplitudes (p<0.001.Conclusion: This study showed that occurrence percentage and amplitude of oVEMP were less than those of cVEMP. Since these two tests originate from different sections of vestibular nerve, we can consider them as parallel vestibular function tests and utilize them for evaluation of vestibular disorders.

  6. Promontory electrical stimulation to elicit vestibular evoked myogenic potentials (VEMPs).

    Science.gov (United States)

    Park, Jonas J-H; Shen, Anmin; Westhofen, Martin

    2015-03-01

    Vestibular evoked myogenic potentials (VEMPs) provoked electrically at the promontory provide a feasible method to record vestibular responses in awake patients. Electrically evoked VEMP testing has been performed by galvanic stimulation at the mastoid so far. The present study examined an electrical stimulation mode close to the otolith organs at the promontory. Fourteen cochlear implant candidates who were planned for clinical routine promontory stimulation testing (PST) to assess auditory nerve function underwent promontory VEMP testing. After testing the cochlear nerve function during PST promontory cervical VEMPs (p-c-VEMPs) and promontory ocular VEMPs (p-o-VEMPs) were recorded during subsequent transtympanic electrical stimulation at the promontory. Promontory VEMP testing was well tolerated by the patients. Mean latencies for p-c-VEMPs were 10.30 ± 2.23 ms (p1) and 17.86 ± 3.83 ms (n1). Mean latencies for p-o-VEMPs were 7.64 ± 1.24 ms (n1) and 11.2 ± 1.81 ms (p1). The stimulation threshold level was measured at 0.15 ± 0.07 mA for p-c-VEMPs and at 0.19 ± 0.11 mA for p-o-VEMPs. The discomfort level was found to be at 0.78 ± 0.29 mA for p-c-VEMPs and at 0.69 ± 0.25 mA for p-oVEMPs. Mean p1-n1 amplitude in p-c-VEMPs was 124.78 ± 56.55 µV and p-o-VEMPs showed a mean n1-p1 amplitude of 30.94 ± 18.98 µV.

  7. Community effect triggers terminal differentiation of myogenic cells derived from muscle satellite cells by quenching Smad signaling

    Energy Technology Data Exchange (ETDEWEB)

    Yanagisawa, Michiko [Department of Regenerative Medicine, National Institute for Longevity Sciences, National Center for Geriatrics and Gerontology, 35 Gengo, Morioka, Oobu, Aichi 474-8522 (Japan); Aging Research, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, Aichi 466-8550 (Japan); Mukai, Atsushi; Shiomi, Kosuke [Department of Regenerative Medicine, National Institute for Longevity Sciences, National Center for Geriatrics and Gerontology, 35 Gengo, Morioka, Oobu, Aichi 474-8522 (Japan); Song, Si-Yong [Institute of Neuroscience, Faculty of Pharmaceutical Sciences at Kagawa, Tokushima Bunri University, 1314-1 Shido, Sanuki-shi, Kagawa 769-2193 (Japan); Hashimoto, Naohiro, E-mail: nao@ncgg.go.jp [Department of Regenerative Medicine, National Institute for Longevity Sciences, National Center for Geriatrics and Gerontology, 35 Gengo, Morioka, Oobu, Aichi 474-8522 (Japan)

    2011-01-15

    A high concentration of bone morphogenetic proteins (BMPs) stimulates myogenic progenitor cells to undergo heterotopic osteogenic differentiation. However, the physiological role of the Smad signaling pathway during terminal muscle differentiation has not been resolved. We report here that Smad1/5/8 was phosphorylated and activated in undifferentiated growing mouse myogenic progenitor Ric10 cells without exposure to any exogenous BMPs. The amount of phosphorylated Smad1/5/8 was severely reduced during precocious myogenic differentiation under the high cell density culture condition even in growth medium supplemented with a high concentration of serum. Inhibition of the Smad signaling pathway by dorsomorphin, an inhibitor of Smad activation, or noggin, a specific antagonist of BMP, induced precocious terminal differentiation of myogenic progenitor cells in a cell density-dependent fashion even in growth medium. In addition, Smad1/5/8 was transiently activated in proliferating myogenic progenitor cells during muscle regeneration in rats. The present results indicate that the Smad signaling pathway is involved in a critical switch between growth and differentiation of myogenic progenitor cells both in vitro and in vivo. Furthermore, precocious cell density-dependent myogenic differentiation suggests that a community effect triggers the terminal muscle differentiation of myogenic cells by quenching the Smad signaling.

  8. [Recording cervical and ocular vestibular evoked myogenic potentials: part 1: anatomy, physiology, methods and normal findings].

    Science.gov (United States)

    Walther, L E; Hörmann, K; Pfaar, O

    2010-10-01

    Vestibular evoked myogenic potentials (VEMP) have gained in clinical significance in recent years, now forming an integral part of neurootological examinations to establish the functional status of the otolith organs. They are sensitive to low-frequency acoustic stimuli. When stimulated, receptors in the sacculus and utriculous are activated. By means of reflexive connections, myogenic potentials can be recorded when the relevant muscles are tonically activated. The vestibulocolic (sacculocollic) reflex travels from the otolith organs over the central circuitry to the ipsilateral sternocleidomastoid muscle. Myogenic potentials can be recorded by means of cervical VEMP (cVEMP). The vestibuloocular reflex crosses contralaterally to the extraocular eye muscle. Ocular VEMP (oVEMP) are recorded periocularly, preferably from the inferior oblique muscle. Various stimulation methods are used including air conduction and bone conduction.

  9. Refractory episodic vertigo: role of intratympanic gentamicin and vestibular evoked myogenic potentials.

    Science.gov (United States)

    Celis-Aguilar, Erika; Hinojosa-González, Ramon; Vales-Hidalgo, Olivia; Coutinho-Toledo, Heloisa

    Even today, the treatment of intractable vertigo remains a challenge. Vestibular ablation with intratympanic gentamicin stands as a good alternative in the management of refractory vertigo patients. To control intractable vertigo through complete saccular and horizontal canal vestibular ablation with intratympanic gentamicin treatment. Patients with refractory episodic vertigo were included. The inclusion criteria were: unilateral ear disease, moderate to profound sensorineural hearing loss, and failure to other treatments. Included patients underwent 0.5-0.8mL of gentamicin intratympanic application at a 30mg/mL concentration. Vestibular ablation was confirmed by the absence of response on cervical vestibular evoked myogenic potentials and no response on caloric tests. Audiometry, electronystagmography with iced water, and vestibular evoked myogenic potentials were performed in all patients. Ten patients were included; nine patients with Meniere's disease and one patient with (late onset) delayed hydrops. Nine patients showed an absent response on vestibular evoked myogenic potentials and no response on caloric tests. The only patient with low amplitude on cervical vestibular evoked myogenic potentials had vertigo recurrence. Vertigo control was achieved in 90% of the patients. One patient developed hearing loss >30dB. Cervical vestibular evoked myogenic potentials confirmed vestibular ablation in patients treated with intratympanic gentamicin. High-grade vertigo control was due to complete saccular and horizontal canal ablation (no response to iced water in electronystagmography and no response on cervical vestibular evoked myogenic potentials). Copyright © 2016 Associação Brasileira de Otorrinolaringologia e Cirurgia Cérvico-Facial. Published by Elsevier Editora Ltda. All rights reserved.

  10. Comparison of cervical and ocular vestibular evoked myogenic potentials in dancers and non-dancers

    OpenAIRE

    Sujeet Kumar Sinha; Vaishnavi Bohra; Himanshu Kumar Sanju

    2013-01-01

    The objective of the study was to assess the sacculocollic and otolith ocular pathway function using cervical vestibular evoked myogenic potentials (cVEMP) and ocular vestibular myogenic potentials (oVEMP) in dancers and non dancers. Total 16 subjects participated in the study. Out of 16 participants, 8 were trained in Indian classical form of dance (dancers) and other 8 participants who were not trained in any dance form (non dancers). cVEMP and oVEMP responses were recorded for all the subj...

  11. A new permanent cell line derived from the bank vole (Myodes glareolus as cell culture model for zoonotic viruses

    Directory of Open Access Journals (Sweden)

    Herzog Sibylle

    2011-07-01

    Full Text Available Abstract Background Approximately 60% of emerging viruses are of zoonotic origin, with three-fourths derived from wild animals. Many of these zoonotic diseases are transmitted by rodents with important information about their reservoir dynamics and pathogenesis missing. One main reason for the gap in our knowledge is the lack of adequate cell culture systems as models for the investigation of rodent-borne (robo viruses in vitro. Therefore we established and characterized a new cell line, BVK168, using the kidney of a bank vole, Myodes glareolus, the most abundant member of the Arvicolinae trapped in Germany. Results BVK168 proved to be of epithelial morphology expressing tight junctions as well as adherence junction proteins. The BVK168 cells were analyzed for their infectability by several arbo- and robo-viruses: Vesicular stomatitis virus, vaccinia virus, cowpox virus, Sindbis virus, Pixuna virus, Usutu virus, Inkoo virus, Puumalavirus, and Borna disease virus (BDV. The cell line was susceptible for all tested viruses, and most interestingly also for the difficult to propagate BDV. Conclusion In conclusion, the newly established cell line from wildlife rodents seems to be an excellent tool for the isolation and characterization of new rodent-associated viruses and may be used as in vitro-model to study properties and pathogenesis of these agents.

  12. Increased radiation from Chernobyl decreases the expression of red colouration in natural populations of bank voles (Myodes glareolus).

    Science.gov (United States)

    Boratyński, Zbyszek; Lehmann, Philipp; Mappes, Tapio; Mousseau, Timothy A; Møller, Anders Pape

    2014-11-21

    Pheomelanin is a pink to red version of melanin pigment deposited in skin and hair. Due to its bright colour, pheomelanin plays a crucial function in signalling, in particular sexual signalling. However, production of pheomelanin, as opposed to its dark alternative, eumelanin, bears costs in terms of consumption of antioxidants important for protection of DNA against naturally produced reactive oxidative species. Therefore, decreased expression of pheomelanin is expected in organisms exposed to severe oxidative stress such as that caused by exposure to chronic ionizing radiation. We tested if variable exposure to radiation among natural populations of bank voles Myodes glareolus in Chernobyl affected expression of red colouration in their dorsal fur. The relative redness of dorsal fur was positively correlated with weight, but also negatively correlated with the level of background radiation. These results suggest that the development of the natural red colouration in adult bank voles is affected by ionizing background radiation, and potentially causing elevated levels of oxidative stress. Reduced production of pheomelanin allows more antioxidants to mitigate the oxidative stress caused by radiation. However, changing natural animal colouration for physiological reasons can have ecological costs, if e.g. it causes mismatch with habitat colouration and conspicuousness for predators.

  13. Microevolution of Puumala hantavirus during a complete population cycle of its host, the bank vole (Myodes glareolus.

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    Maria Razzauti

    Full Text Available Microevolution of Puumala hantavirus (PUUV was studied throughout a population cycle of its host, the bank vole (Myodes glareolus. We monitored PUUV variants circulating in the host population in Central Finland over a five-year period that included two peak-phases and two population declines. Of 1369 bank voles examined, 360 (26.3% were found infected with PUUV. Partial sequences of each of the three genome segments were recovered (approx. 12% of PUUV genome from 356 bank voles. Analyses of these sequences disclosed the following features of PUUV evolution: 1 nucleotide substitutions are mostly silent and deduced amino acid changes are mainly conservative, suggesting stabilizing selection at the protein level; 2 the three genome segments accumulate mutations at a different rate; 3 some of the circulating PUUV variants are frequently observed while others are transient; 4 frequently occurring PUUV variants are composed of the most abundant segment genotypes (copious and new transient variants are continually generated; 5 reassortment of PUUV genome segments occurs regularly and follows a specific pattern of segments association; 6 prevalence of reassortant variants oscillates with season and is higher in the autumn than in the spring; and 7 reassortants are transient, i.e., they are not competitively superior to their parental variants. Collectively, these observations support a quasi-neutral mode of PUUV microevolution with a steady generation of transient variants, including reassortants, and preservation of a few preferred genotypes.

  14. Band limited chirp stimulation in vestibular evoked myogenic potentials.

    Science.gov (United States)

    Walther, Leif Erik; Cebulla, Mario

    2016-10-01

    Air conducted vestibular evoked myogenic potentials (VEMP) can be elicited by various low frequency and intense sound stimuli, mainly clicks or short tone bursts (STB). Chirp stimuli are increasingly used in diagnostic audiological evaluations as an effective means to obtain acoustically evoked responses in narrowed or extended frequency ranges. We hypothesized in this study that band limited chirp stimulation, which covers the main sensitivity range of sound sensitive otolithic afferents (around 500 Hz), might be useful for application in cervical and ocular VEMP to air conduction. For this purpose we designed a chirp stimulus ranging 250-1000 Hz (up chirp). The chirp stimulus was delivered with a stimulus intensity of 100 dB nHL in normal subjects (n = 10) and patients with otolith involvement (vestibular neuritis) (n = 6). Amplitudes of the designed chirp ("CW-VEMP-chirp, 250-1000 Hz") were compared with amplitudes of VEMPs evoked by click stimuli (0.1 ms) and a short tone burst (STB, 1-2-1, 8 ms, 500 Hz). CVEMPs and oVEMPs were detectable in 9 of 10 normal individuals. Statistical evaluation in healthy patients revealed significantly larger cVEMP and oVEMP amplitudes for CW-VEMP-chirp (250-1000 Hz) stimuli. CVEMP amplitudes evoked by CW-VEMP-chirp (250-1000 Hz) showed a high stability in comparison with click and STB stimulation. CW-VEMP-chirp (250-1000 Hz) showed abnormal cVEMP and oVEMP amplitudes in patients with vestibular neuritis, with the same properties as click and STB stimulated VEMPs. We conclude that the designed CW-VEMP-chirp (250-1000 Hz) is an effective stimulus which can be further used in VEMP diagnostic. Since a chirp stimulus can be easily varied in its properties, in particular with regard to frequency, this might be a promising tool for further investigations.

  15. G-protein mediated signaling pathways in myogenic responsiveness of mouse mesenteric artery

    DEFF Research Database (Denmark)

    Jensen, Lars Jørn; Joseph, Philomeena Daphne; Haanes, Kristian Agmund

    2015-01-01

    Myogenic responsiveness (MR) is the ability of a small artery to constrict to an increase in intraluminal pressure and dilate to a decrease in pressure. The mechanisms linking an increase in pressure to VSMC contraction are suggested to involve mechanical activation of AT1-receptors. Our aim...

  16. The methyl-CpG-binding protein CIBZ suppresses myogenic differentiation by directly inhibiting myogenin expression

    Institute of Scientific and Technical Information of China (English)

    Yu Oikawa; Reiko Omori; Tomonori Nishii; Yasumasa Ishida; Masashi Kawaichi; Eishou Matsuda

    2011-01-01

    Postnatal growth and regeneration of skeletal muscle are carried out mainly by satellite cells,which,upon stimulation,begin to express myogenin (Myog),the critical determinant of myogenic differentiation.DNA methylation status has been associated with the expression of Myog,but the causative mechanism remains almost unknown.Here,we report that the level of CIBZ,a methyI-CpG-binding protein,decreases upon myogenic differentiation of satellitederived C2C12 cells,and during skeletal muscle regeneration in mice.We present data showing that the loss of CIBZ promotes myogenic differentiation,whereas exogenous expression of CIBZ impairs it,in cultured cells.CIBZ binds to a Myog promoter-proximal region and inhibits Myog transcription in a methylation-dependent manner.These data suggest that the suppression of myogenic differentiation by CIBZ is dependent,at least in part,on the regulation of Myog.Our data show that the methylation status of this proximal Myog promoter inversely correlates with Myog transcription in cells and tissues,and during postnatal growth of skeletal muscle.Notably,induction of Myog transcription by CIBZ suppression is independent of the demethylation of CpG sites in the Myog promoter.These observations provide the first reported molecular mechanism illustrating how Myog transcription is coordinately regulated by a methyI-CpG-binding protein and the methylation status of the proximal Myog promoter.

  17. Chronic hypoxia during gestation enhances uterine arterial myogenic tone via heightened oxidative stress.

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    Daliao Xiao

    Full Text Available Chronic hypoxia during gestation has profound adverse effects on the adaptation of uteroplacental circulation in pregnancy. Yet, the underlying mechanisms are not fully understood. The present study tested the hypothesis that enhanced production of reactive oxygen species (ROS in uterine arteries plays a critical role in the maladaptation of uterine circulation associated with chronic hypoxia. Uterine arteries were isolated from nonpregnant and near-term pregnant sheep maintained at sea level (~300 m or exposed to high-altitude (3801 m hypoxia for 110 days. Hypoxia significantly increased ROS production in uterine arteries of pregnant, but not nonpregnant, sheep. This was associated with a significant increase in NADPH oxidase (Nox 2, but not Nox1 or Nox4, protein abundance and total Nox activity in uterine arteries of pregnant animals. Chronic hypoxia significantly increased pressure-dependent uterine arterial myogenic tone in pregnant sheep, which was abrogated by a Nox inhibitor apocynin. Additionally, the hypoxia-induced increase in myogenic reactivity of uterine arteries to phorbol 12,13-dibutyrate in pregnant sheep was blocked by apocynin and tempol. In consistence with the myogenic responses, the hypoxia-mediated down-regulation of BKCa channel activity in uterine arteries of pregnant animals was reversed by apocynin. The findings suggest that heightened oxidative stress in uterine arteries plays a key role in suppressing the BKCa channel activity, resulting in increased myogenic reactivity and maladaptation of uteroplacental circulation caused by chronic hypoxia during gestation.

  18. In vivo myogenic potential of human CD133+ muscle-derived stem cells: a quantitative study.

    Science.gov (United States)

    Negroni, Elisa; Riederer, Ingo; Chaouch, Soraya; Belicchi, Marzia; Razini, Paola; Di Santo, James; Torrente, Yvan; Butler-Browne, Gillian S; Mouly, Vincent

    2009-10-01

    In recent years, numerous reports have identified in mouse different sources of myogenic cells distinct from satellite cells that exhibited a variable myogenic potential in vivo. Myogenic stem cells have also been described in humans, although their regenerative potential has rarely been quantified. In this study, we have investigated the myogenic potential of human muscle-derived cells based on the expression of the stem cell marker CD133 as compared to bona fide satellite cells already used in clinical trials. The efficiency of these cells to participate in muscle regeneration and contribute to the renewal of the satellite cell pool, when injected intramuscularly, has been evaluated in the Rag2(-/-) gammaC(-/-) C5(-/-) mouse in which muscle degeneration is induced by cryoinjury. We demonstrate that human muscle-derived CD133+ cells showed a much greater regenerative capacity when compared to human myoblasts. The number of fibers expressing human proteins and the number of human cells in a satellite cell position are all dramatically increased when compared to those observed after injection of human myoblasts. In addition, CD133+/CD34+ cells exhibited a better dispersion in the host muscle when compared to human myoblasts. We propose that muscle-derived CD133+ cells could be an attractive candidate for cellular therapy.

  19. In Vivo Myogenic Potential of Human CD133+ Muscle-derived Stem Cells: A Quantitative Study

    Science.gov (United States)

    Negroni, Elisa; Riederer, Ingo; Chaouch, Soraya; Belicchi, Marzia; Razini, Paola; Di Santo, James; Torrente, Yvan; Butler-Browne, Gillian S; Mouly, Vincent

    2009-01-01

    In recent years, numerous reports have identified in mouse different sources of myogenic cells distinct from satellite cells that exhibited a variable myogenic potential in vivo. Myogenic stem cells have also been described in humans, although their regenerative potential has rarely been quantified. In this study, we have investigated the myogenic potential of human muscle–derived cells based on the expression of the stem cell marker CD133 as compared to bona fide satellite cells already used in clinical trials. The efficiency of these cells to participate in muscle regeneration and contribute to the renewal of the satellite cell pool, when injected intramuscularly, has been evaluated in the Rag2−/− γC−/− C5−/− mouse in which muscle degeneration is induced by cryoinjury. We demonstrate that human muscle–derived CD133+ cells showed a much greater regenerative capacity when compared to human myoblasts. The number of fibers expressing human proteins and the number of human cells in a satellite cell position are all dramatically increased when compared to those observed after injection of human myoblasts. In addition, CD133+/CD34+ cells exhibited a better dispersion in the host muscle when compared to human myoblasts. We propose that muscle-derived CD133+ cells could be an attractive candidate for cellular therapy. PMID:19623164

  20. Ocular vestibular evoked myogenic potential in patients with benign paroxysmal positional vertigo

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    Mozhgan Masoom

    2014-06-01

    Full Text Available Background and Aim: Since utricle is the main damaged organ in benign paroxysmal positional vertigo (BPPV, ocular vestibular evoked myogenic potential (oVEMP may be an appropriate method to evaluate the utricule dysfunction and the effect of disease recurrence rate on it. This study aimed to record myogenic potential in patients with benign paroxysmal positional vertigo.Methods: In a cross-sectional study, ocular myogenic potential was recorded in 25 healthy subjects and 20 patients with benign paroxysmal positional vertigo using 500 Hz-tone bursts (95 dB nHL.Results: In the affected ear, mean amplitude was lower and mean threshold was higher than those in the unaffected ear and in the normal group (p<0.05. Mean amplitude asymmetry ratio of patients was more than the healthy subjects (p0.05. Frequencies of abnormal responses in the affected ears were higher than in unaffected ears and in the normal group (p<0.05. Furthermore, the patients with recurrent vertigo showed more abnormalities than the patients with non-recurrent (p=0.030.Conclusion: In the recurrent benign paroxysmal positional vertigo, ocular vestibular evoked myogenic potential showed more damage in the utricle, suggesting this response could be used to evaluate the patients with benign paroxysmal positional vertigo.

  1. INFLUENCE OF DANCE TRAINING ON SACCULOCOLLIC PATHWAY: VESTIBULAR EVOKED MYOGENIC POTENTIALS (VEMP) AS AN OBJECTIVE TOOL

    OpenAIRE

    Swathi; Sathish Kumar

    2013-01-01

    ABSTRACT : Auditory system is shaped by experience and training. Training (s ensory experience) induces neurophysiologic changes & plasticity in normal hearing individuals, hearing loss patients, hearing aid users and cochlear implanted subjects. Not only speech stimulus, but music also brings about functional and structural organi zation of the brain in musician compared to non - musicians. The Vestibular evoked myogenic potentials (VEMP) are a biphasic in...

  2. Commentary on the Special Issue on the Impact of Myogenic Tone in Health and Disease Preface

    NARCIS (Netherlands)

    Roman, Richard J.; Van Dokkum, Richard P. E.

    2014-01-01

    Autoregulation is a vital homeostatic mechanism that helps maintain constant delivery of oxygen to organs despite fluctuations in arteriolar pressure. Autoregulation of blood flow to elevations in pressure is largely mediated by the myogenic response of small arteries and arterioles which constrict

  3. Sphingosine-1-Phosphate Signaling Regulates Myogenic Responsiveness in Human Resistance Arteries.

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    Sonya Hui

    Full Text Available We recently identified sphingosine-1-phosphate (S1P signaling and the cystic fibrosis transmembrane conductance regulator (CFTR as prominent regulators of myogenic responsiveness in rodent resistance arteries. However, since rodent models frequently exhibit limitations with respect to human applicability, translation is necessary to validate the relevance of this signaling network for clinical application. We therefore investigated the significance of these regulatory elements in human mesenteric and skeletal muscle resistance arteries. Mesenteric and skeletal muscle resistance arteries were isolated from patient tissue specimens collected during colonic or cardiac bypass surgery. Pressure myography assessments confirmed endothelial integrity, as well as stable phenylephrine and myogenic responses. Both human mesenteric and skeletal muscle resistance arteries (i express critical S1P signaling elements, (ii constrict in response to S1P and (iii lose myogenic responsiveness following S1P receptor antagonism (JTE013. However, while human mesenteric arteries express CFTR, human skeletal muscle resistance arteries do not express detectable levels of CFTR protein. Consequently, modulating CFTR activity enhances myogenic responsiveness only in human mesenteric resistance arteries. We conclude that human mesenteric and skeletal muscle resistance arteries are a reliable and consistent model for translational studies. We demonstrate that the core elements of an S1P-dependent signaling network translate to human mesenteric resistance arteries. Clear species and vascular bed variations are evident, reinforcing the critical need for further translational study.

  4. Differential effects of superoxide and hydrogen peroxide on myogenic signaling, membrane potential, and contractions of mouse renal afferent arterioles.

    Science.gov (United States)

    Li, Lingli; Lai, En Yin; Wellstein, Anton; Welch, William J; Wilcox, Christopher S

    2016-06-01

    Myogenic contraction is the principal component of renal autoregulation that protects the kidney from hypertensive barotrauma. Contractions are initiated by a rise in perfusion pressure that signals a reduction in membrane potential (Em) of vascular smooth muscle cells to activate voltage-operated Ca(2+) channels. Since ROS have variable effects on myogenic tone, we investigated the hypothesis that superoxide (O2 (·-)) and H2O2 differentially impact myogenic contractions. The myogenic contractions of mouse isolated and perfused single afferent arterioles were assessed from changes in luminal diameter with increasing perfusion pressure (40-80 mmHg). O2 (·-), H2O2, and Em were assessed by fluorescence microscopy during incubation with paraquat to increase O2 (·-) or with H2O2 Paraquat enhanced O2 (·-) generation and myogenic contractions (-42 ± 4% vs. -19 ± 4%, P contractions (-10 ± 1% vs. -19 ± 2%, P contractions with paraquat without preventing the reduction in Em Myogenic contractions were independent of the endothelium and largely independent of nitric oxide. We conclude that O2 (·-) and H2O2 activate different signaling pathways in vascular smooth muscle cells linked to discreet membrane channels with opposite effects on Em and voltage-operated Ca(2+) channels and therefore have opposite effects on myogenic contractions.

  5. Epidemiology of vestibular evoked myogenic potentials: Data from the Baltimore Longitudinal Study of Aging.

    Science.gov (United States)

    Li, Carol; Layman, Andrew J; Carey, John P; Agrawal, Yuri

    2015-11-01

    To evaluate whether age-related changes in vestibular evoked myogenic potentials (VEMPs) differ by demographic and cardiovascular risk groups. Participants in the Baltimore Longitudinal Study of Aging underwent cervical and ocular VEMP testing. VEMP latency, amplitude, asymmetry ratios, and prevalence of absent responses were compared across demographic and cardiovascular risk groups. In 257 participants (mean age 72.9, 57% female), ocular VEMP (oVEMP) n10 latency increased by 0.12ms/decade while amplitude decreased by 2.9μV/decade. Black participants had better oVEMP function (shorter latency, increased amplitude, and decreased odds of absent responses) relative to white participants. In 250 participants (mean age 72.6, 54% female), EMG-corrected cervical VEMP (cVEMP) amplitude decreased by 0.14μV/decade and p13 latency was 0.38ms longer in males. The odds of absent responses were significantly higher in individuals age ⩾80 for oVEMPs, and age ⩾70 for cVEMPs. Cardiovascular risk factors had no association with VEMP parameters. We confirmed age-related declines in otolith function, and observed a protective effect of black race on oVEMP latency and amplitude. These results illustrate how measures of otolith function change with age in community-dwelling adults. Further investigations are needed to ascertain whether better otolith function in blacks might contribute to a lower risk of mobility disability and falls. Copyright © 2015 International Federation of Clinical Neurophysiology. Published by Elsevier Ireland Ltd. All rights reserved.

  6. PC-PLC/sphingomyelin synthase activity plays a central role in the development of myogenic tone in murine resistance arteries.

    Science.gov (United States)

    Mauban, Joseph R H; Zacharia, Joseph; Fairfax, Seth; Wier, Withrow Gil

    2015-06-15

    Myogenic tone is an intrinsic property of the vasculature that contributes to blood pressure control and tissue perfusion. Earlier investigations assigned a key role in myogenic tone to phospholipase C (PLC) and its products, inositol 1,4,5-trisphosphate (IP3) and diacylglycerol (DAG). Here, we used the PLC inhibitor, U-73122, and two other, specific inhibitors of PLC subtypes (PI-PLC and PC-PLC) to delineate the role of PLC in myogenic tone of pressurized murine mesenteric arteries. U-73122 inhibited depolarization-induced contractions (high external K(+) concentration), thus confirming reports of nonspecific actions of U-73122 and its limited utility for studies of myogenic tone. Edelfosine, a specific inhibitor of PI-PLC, did not affect depolarization-induced contractions but modulated myogenic tone. Because PI-PLC produces IP3, we investigated the effect of blocking IP3 receptor-mediated Ca(2+) release on myogenic tone. Incubation of arteries with xestospongin C did not affect tone, consistent with the virtual absence of Ca(2+) waves in arteries with myogenic tone. D-609, an inhibitor of PC-PLC and sphingomyelin synthase, strongly inhibited myogenic tone and had no effect on depolarization-induced contraction. D-609 appeared to act by lowering cytoplasmic Ca(2+) concentration to levels below those that activate contraction. Importantly, incubation of pressurized arteries with a membrane-permeable analog of DAG induced vasoconstriction. The results therefore mandate a reexamination of the signaling pathways activated by the Bayliss mechanism. Our results suggest that PI-PLC and IP3 are not required in maintaining myogenic tone, but DAG, produced by PC-PLC and/or SM synthase, is likely through multiple mechanisms to increase Ca(2+) entry and promote vasoconstriction.

  7. Neurogenic and myogenic properties of pan-colonic motor patterns and their spatiotemporal organization in rats.

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    Ji-Hong Chen

    Full Text Available BACKGROUND AND AIMS: Better understanding of intrinsic control mechanisms of colonic motility will lead to better treatment options for colonic dysmotility. The aim was to investigate neurogenic and myogenic control mechanisms underlying pan-colonic motor patterns. METHODS: Analysis of in vitro video recordings of whole rat colon motility was used to explore motor patterns and their spatiotemporal organizations and to identify mechanisms of neurogenic and myogenic control using pharmacological tools. RESULTS: Study of the pan-colonic spatiotemporal organization of motor patterns revealed: fluid-induced or spontaneous rhythmic propulsive long distance contractions (LDCs, 0.4-1.5/min, involving the whole colon, rhythmic propulsive motor complexes (RPMCs (0.8-2.5/min, dominant in distal colon, ripples (10-14/min, dominant in proximal colon, segmentation and retrograde contractions (0.1-0.8/min, prominent in distal and mid colon. Spontaneous rhythmic LDCs were the dominant pattern, blocked by tetrodotoxin, lidocaine or blockers of cholinergic, nitrergic or serotonergic pathways. Change from propulsion to segmentation and distal retrograde contractions was most prominent after blocking 5-HT3 receptors. In the presence of all neural blockers, bethanechol consistently evoked rhythmic LDC-like propulsive contractions in the same frequency range as the LDCs, indicating the existence of myogenic mechanisms of initiation and propulsion. CONCLUSIONS: Neurogenic and myogenic control systems orchestrate distinct and variable motor patterns at different regions of the pan-colon. Cholinergic, nitrergic and serotonergic pathways are essential for rhythmic LDCs to develop. Rhythmic motor patterns in presence of neural blockade indicate the involvement of myogenic control systems and suggest a role for the networks of interstitial cells of Cajal as pacemakers.

  8. Sodium pump α2 subunits control myogenic tone and blood pressure in mice

    Science.gov (United States)

    Zhang, Jin; Lee, Moo Yeol; Cavalli, Maurizio; Chen, Ling; Berra-Romani, Roberto; Balke, C William; Bianchi, Giuseppe; Ferrari, Patrizia; Hamlyn, John M; Iwamoto, Takahiro; Lingrel, Jerry B; Matteson, Donald R; Wier, W Gil; Blaustein, Mordecai P

    2005-01-01

    A key question in hypertension is: How is long-term blood pressure controlled? A clue is that chronic salt retention elevates an endogenous ouabain-like compound (EOLC) and induces salt-dependent hypertension mediated by Na+/Ca2+ exchange (NCX). The precise mechanism, however, is unresolved. Here we study blood pressure and isolated small arteries of mice with reduced expression of Na+ pump α1 (α1+/−) or α2 (α2+/−) catalytic subunits. Both low-dose ouabain (1–100 nm; inhibits only α2) and high-dose ouabain (≥1 μm; inhibits α1) elevate myocyte Ca2+ and constrict arteries from α1+/−, as well as α2+/− and wild-type mice. Nevertheless, only mice with reduced α2 Na+ pump activity (α2+/−), and not α1 (α1+/−), have elevated blood pressure. Also, isolated, pressurized arteries from α2+/−, but not α1+/−, have increased myogenic tone. Ouabain antagonists (PST 2238 and canrenone) and NCX blockers (SEA0400 and KB-R7943) normalize myogenic tone in ouabain-treated arteries. Only the NCX blockers normalize the elevated myogenic tone in α2+/− arteries because this tone is ouabain independent. All four agents are known to lower blood pressure in salt-dependent and ouabain-induced hypertension. Thus, chronically reduced α2 activity (α2+/− or chronic ouabain) apparently regulates myogenic tone and long-term blood pressure whereas reduced α1 activity (α1+/−) plays no persistent role: the in vivo changes in blood pressure reflect the in vitro changes in myogenic tone. Accordingly, in salt-dependent hypertension, EOLC probably increases vascular resistance and blood pressure by reducing α2 Na+ pump activity and promoting Ca2+ entry via NCX in myocytes. PMID:16166162

  9. MAPK signaling pathways and HDAC3 activity are disrupted during emerin-null myogenic progenitor differentiation.

    Science.gov (United States)

    Collins, Carol M; Ellis, Joseph; Holaska, James M

    2017-02-10

    Mutations in the gene encoding emerin cause Emery-Dreifuss muscular dystrophy (EDMD). Emerin is an integral inner nuclear membrane protein and a component of the nuclear lamina. EDMD is characterized by skeletal muscle wasting, cardiac conduction defects and tendon contractures. The failure to regenerate skeletal muscle is predicted to contribute to the skeletal muscle pathology of EDMD. We hypothesize muscle regeneration defects are caused by impaired muscle stem cell differentiation. Myogenic progenitors derived from emerin-null mice were used to confirm their impaired differentiation and analyze selected myogenic molecular pathways. Emerin-null progenitors were delayed in their cell cycle exit, had decreased myosin heavy chain (MyHC) expression and formed fewer myotubes. Emerin binds to and activates histone deacetylase 3 (HDAC3). Here we show theophylline, an HDAC3-specific activator, improved myotube formation in emerin-null cells. Addition of the HDAC3-specific inhibitor RGFP966 blocked myotube formation and MyHC expression in wildtype and emerin-null myogenic progenitors, but did not affect cell cycle exit. Downregulation of emerin was previously shown to affect the p38 and ERK MAPK pathways in C2C12 myoblast differentiation. Using a pure population of myogenic progenitors completely lacking emerin expression we show these pathways are also disrupted. ERK inhibition improved MyHC expression in emerin-null cells, but failed to rescue myotube formation or cell cycle exit. p38 MAPK inhibition prevented differentiation in both wildtype and emerin-null progenitors. These results show each of these molecular pathways specifically regulate particular stages of myogenic differentiation in an emerin-dependent manner. Thus, pharmacological targeting of multiple pathways acting at specific differentiation stages may be a better therapeutic approach in the future to rescue muscle regeneration in vivo.

  10. Woodland recovery after suppression of deer: cascade effects for small mammals, wood mice (Apodemus sylvaticus and bank voles (Myodes glareolus.

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    Emma R Bush

    Full Text Available Over the past century, increases in both density and distribution of deer species in the Northern Hemisphere have resulted in major changes in ground flora and undergrowth vegetation of woodland habitats, and consequentially the animal communities that inhabit them. In this study, we tested whether recovery in the vegetative habitat of a woodland due to effective deer management (from a peak of 0.4-1.5 to <0.17 deer per ha had translated to the small mammal community as an example of a higher order cascade effect. We compared deer-free exclosures with neighboring open woodland using capture-mark-recapture (CMR methods to see if the significant difference in bank vole (Myodes glareolus and wood mouse (Apodemus sylvaticus numbers between these environments from 2001-2003 persisted in 2010. Using the multi-state Robust Design method in program MARK we found survival and abundance of both voles and mice to be equivalent between the open woodland and the experimental exclosures with no differences in various metrics of population structure (age structure, sex composition, reproductive activity and individual fitness (weight, although the vole population showed variation both locally and temporally. This suggests that the vegetative habitat--having passed some threshold of complexity due to lowered deer density--has allowed recovery of the small mammal community, although patch dynamics associated with vegetation complexity still remain. We conclude that the response of small mammal communities to environmental disturbance such as intense browsing pressure can be rapidly reversed once the disturbing agent has been removed and the vegetative habitat is allowed to increase in density and complexity, although we encourage caution, as a source/sink dynamic may emerge between old growth patches and the recently disturbed habitat under harsh conditions.

  11. Diversity in the utilization of glucose and lactate in synthetic mammalian myotubes generated by engineered configurations of MyoD and E12 in otherwise non-differentiation growth conditions.

    Science.gov (United States)

    Grubišić, Vladimir; Parpura, Vladimir

    2015-03-01

    We previously used the expression of various combinations and configurations of MyoD and E12, two basic helix-loop-helix transcription factors (TF), to produce populations of myotubes assuming distinct morphology, myofibrillar development and Ca2+ dynamics, from mammalian C2C12 myoblasts in non-differentiation growth conditions. Here, we assessed the synthetically generated myotubes in terms of energetics, otherwise necessary to sustain their mechanical output as bio-actuators. We found that the myotubes exhibit changed expression of key regulators for the uptake and utilization of two major cellular fuels, glucose and lactate. Furthermore, while lactate transport was uniformly slowed in all the populations of myotubes, glucose uptake and utilization were modified by particular TF configuration. Our approach allows the production of a class of biomaterials with predetermined energetics that could be applied in biorobotics, where fuel of choice could be used, and also in reparative medicine where, for example, particular population of myotubes could be additionally employed as glucose sinks to mitigate effects of secondary metabolic syndrome.

  12. Losartan protects mesenteric arteries from ROS-associated decrease in myogenic constriction following 5/6 nephrectomy

    NARCIS (Netherlands)

    Vavrinec, Peter; van Dokkum, Richard P. E.; Goris, Maaike; Buikema, Hendrik; Henning, Robert H.

    2011-01-01

    Background: Chronic renal failure (CRF) is associated with hypertension, proteinuria, loss of myogenic constriction (MC) of mesenteric arteries and increased production of reactive oxygen species (ROS) under experimental conditions. Previous results showed that ACE (angiotensin-converting enzyme act

  13. Investigation of LDHA and COPB1 as candidate genes for muscle development in the MYOD1 region of pig chromosome 2.

    Science.gov (United States)

    Qiu, Haifeng; Xu, Xuewen; Fan, Bing; Rothschild, Max F; Martin, Yerle; Liu, Bang

    2010-01-01

    Porcine MYOD1 gene has been mapped to swine chromosome (SSC) 2p14-p17, which is involved in the regulation of the proliferation and differentiation of skeletal muscle cells. The LDHA (lactate dehydrogenase A) and COPB1 (coatomer protein complex, subunit beta 1) genes, which map close to MYOD1, are involved in energy metabolism and protein transport processes. Both genes might play important roles in muscle development. However, little is known about the porcine LDHA and COPB1 genes. In the present study, the full-length cDNA of these two genes were cloned. The mapping results demonstrated that porcine LDHA and COPB1 were all mapped to SSC 2p14-p17. In this region, there are several QTL for growth and carcass traits, including average backfat thickness, lean and fat percentage. The RT-PCR results revealed that both LDHA and COPB1 were highly expressed in porcine skeletal muscle tissues, implying their potential regulatory function of muscle development. LDHA and COPB1 were then mapped to the region and multipoint analyses generated a best sex-averaged map order of each gene between linked markers: MYOD1_75.2 cM _LDHA_79 cM _CSRP3_83.8 cM _TEF-1_86.5 cM _COPB1_90 cM. Association analyses revealed that the substitution of c.423A>G had a significant effect on average daily gain on test, average backfat thickness (BFT), loin muscle area, lumbar BFT, marbling score, tenth rib BFT, average drip loss and fiber type II ratio. The substitution of c.3096C>T had a significant effect on average BFT, lumbar BFT, tenth rib BFT, carcass weight and last rib BFT. Interestingly, both SNPs were all associated with average BFT, lumbar BFT and tenth rib BFT.

  14. Concomitant influence of helminth infection and landscape on the distribution of Puumala hantavirus in its reservoir, Myodes glareolus

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    Henttonen Heikki

    2011-02-01

    Full Text Available Abstract Background Puumala virus, the agent of nephropathia epidemica (NE, is the most prevalent hantavirus in Europe. The risk for human infection seems to be strongly correlated with the prevalence of Puumala virus (PUUV in populations of its reservoir host species, the bank vole Myodes glareolus. In humans, the infection risks of major viral diseases are affected by the presence of helminth infections. We therefore proposed to analyse the influence of both helminth community and landscape on the prevalence of PUUV among bank vole populations in the Ardennes, a PUUV endemic area in France. Results Among the 313 voles analysed, 37 had anti-PUUV antibodies. Twelve gastro-intestinal helminth species were recorded among all voles sampled. We showed that PUUV seroprevalence strongly increased with age or sexual maturity, especially in the northern forests (massif des Ardennes. The helminth community structure significantly differed between this part and the woods or hedgerows of the southern cretes pre-ardennaises. Using PUUV RNA quantification, we identified significant coinfections between PUUV and gastro-intestinal helminths in the northern forests only. More specifically, PUUV infection was positively associated with the presence of Heligmosomum mixtum, and in a lesser extent, Aonchotheca muris-sylvatici. The viral load of PUUV infected individuals tended to be higher in voles coinfected with H. mixtum. It was significantly lower in voles coinfected with A. muris-sylvatici, reflecting the influence of age on these latter infections. Conclusions This is the first study to emphasize hantavirus - helminth coinfections in natural populations. It also highlights the importance to consider landscape when searching for such associations. We have shown that landscape characteristics strongly influence helminth community structure as well as PUUV distribution. False associations might therefore be evidenced if geographic patterns of helminths or PUUV

  15. Intraluminal pressure stimulates MAPK phosphorylation in arterioles: temporal dissociation from myogenic contractile response.

    Science.gov (United States)

    Spurrell, Brian E; Murphy, Timothy V; Hill, Michael A

    2003-10-01

    Members of the MAPK family of enzymes, p42/44 and p38, have been implicated in both the regulation of contractile function and growth responses in vascular smooth muscle. We determined whether such kinases are activated during the arteriolar myogenic response after increases in intraluminal pressure. Particular emphasis was placed on temporal aspects of activation to determine whether such phosphorylation events parallel the known time course for myogenic contraction. Experiments used single cannulated arterioles isolated from the cremaster muscle of rats with some vessels loaded with the fluorescent Ca2+-sensitive dye fura 2 (2 microM). The p42/44 inhibitor PD-98059 (50 microM) caused vasodilation but did not prevent pressure-induced myogenic constriction. The vasodilator response was accompanied by decreased smooth muscle intracellular Ca2+. Western blotting revealed a significant increase in the level of phosphorylation of p42/44 15 min after the application of a 30- to 100-mmHg pressure step. Phosphorylation of p42/44 was a late event that appeared to be temporally dissociated from contraction, which was complete within 1-5 min. EGF (80 nM) caused marked phosphorylation of p42/44 but only acted as a weak vasoconstrictor. The p38 inhibitor SB-203580 (10 microM) did not alter baseline diameter, nor did it prevent myogenic vasoconstriction. Consistent with these observations, SB-203580 did not cause a measurable change in intracellular Ca2+. The results demonstrate activation of the p42/44 class of MAPK resulting from increased transmural pressure. Such activation is, however, dissociated from the acute pressure-induced vasoconstrictor response in terms of time course and may represent the activation of compensatory, but parallel, pathways, including those related to growth and remodeling.

  16. Myogenic temporomandibular disorders: Clinical systemic comorbidities in a female population sample

    OpenAIRE

    de-Pedro-Herráez, Miguel; Mesa-Jiménez, Juan; Fernández-de-las-Peñas, César; de-la-Hoz-Aizpurua, José-Luis

    2016-01-01

    Background Myogenic temporomandibular disorders (MTMD) frequently coexist with other clinical conditions in the same individual. In the last decades, several authors have analyzed these comorbidities looking for the origin of this overlapping. Objetives The aim of this study was to perform a comparative anaylisis between a group of patients with MTMD and a control group of dental patients without dysfunctional pathology to assess whether there are significant differences in the presence of sy...

  17. Asynchronous Inflammation and Myogenic Cell Migration Limit Muscle Tissue Regeneration Mediated by a Cellular Scaffolds

    Science.gov (United States)

    2015-02-11

    over two-times that observed with muscle grafts, but they appeared to be less active, as gene expression of pro- and anti- inflammatory cytokines ( TNF -α...injury) the inflammatory and myogenic response to the muscle scaffold [16], which relies solely on host cell migration for regeneration [18]. Vital...cells [37] to induce myogenesis. Following injury, the type of the inflammatory response and the significance of transition from pro- to an anti

  18. Cervical vestibular evoked myogenic potentials and caloric test results in individuals with auditory neuropathy spectrum disorders.

    Science.gov (United States)

    Sujeet, Kumar Sinha; Niraj, Kumar Singh; Animesh, Barman; Rajeshwari, G; Sharanya, R

    2014-01-01

    Auditory neuropathy spectrum disorder is a type of hearing loss where outer hair cell function are normal (as evidenced by the preservation of OAEs and cochlear microphonics), whereas auditory nerve functions are abnormal (as evidenced by abnormal auditory brainstem evoked potentials beginning with wave I of the ABR) and acoustic reflexes to ipsilateral and contralateral tones are absent. It is likely that in cases with auditory neuropathy spectrum disorder not only the cochlear nerve, but also the vestibular nerves might get involved. The present study was conducted with an aim of finding out the inferior and superior vestibular nerve involvement through cervical vestibular evoked myogenic potentials and Caloric test results respectively in individuals with Auditory Neuropathy Spectrum Disorders. Total 26 participants who fulfilled the criteria of auditory neuropathy spectrum disorder participated for the study. Vestibular evoked myogenic potentials results showed absence of responses from most of the subjects also caloric responses showed bilateral hypofunctional responses in most of the participants, which is suggestive of involvement of both the inferior as well as superior vestibular nerve in individuals with auditory neuropathy spectrum disorders. Additionally there was no association between the pattern and degree of hearing loss to caloric test results and vestibular evoked myogenic potentials results findings.

  19. FSHD myoblasts fail to downregulate intermediate filament protein vimentin during myogenic differentiation.

    Directory of Open Access Journals (Sweden)

    Lipinski M.

    2011-10-01

    Full Text Available Facioscapulohumeral muscular dystrophy (FSHD is an autosomal dominant hereditary neuromuscular disorder. The clinical features of FSHD include weakness of the facial and shoulder girdle muscles followed by wasting of skeletal muscles of the pelvic girdle and lower extremities. Although FSHD myoblasts grown in vitro can be induced to differentiate into myotubes by serum starvation, the resulting FSHD myotubes have been shown previously to be morphologically abnormal. Aim. In order to find the cause of morphological anomalies of FSHD myotubes we compared in vitro myogenic differentiation of normal and FSHD myoblasts at the protein level. Methods. We induced myogenic differentiation of normal and FSHD myoblasts by serum starvation. We then compared protein extracts from proliferating myoblasts and differentiated myotubes using SDS-PAGE followed by mass spectrometry identification of differentially expressed proteins. Results. We demonstrated that the expression of vimentin was elevated at the protein and mRNA levels in FSHD myotubes as compared to normal myotubes. Conclusions. We demonstrate for the first time that in contrast to normal myoblasts, FSHD myoblasts fail to downregulate vimentin after induction of in vitro myogenic differentiation. We suggest that vimentin could be an easily detectable marker of FSHD myotubes

  20. Myogenic Precursors from iPS Cells for Skeletal Muscle Cell Replacement Therapy

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    Isart Roca

    2015-01-01

    Full Text Available The use of adult myogenic stem cells as a cell therapy for skeletal muscle regeneration has been attempted for decades, with only moderate success. Myogenic progenitors (MP made from induced pluripotent stem cells (iPSCs are promising candidates for stem cell therapy to regenerate skeletal muscle since they allow allogenic transplantation, can be produced in large quantities, and, as compared to adult myoblasts, present more embryonic-like features and more proliferative capacity in vitro, which indicates a potential for more self-renewal and regenerative capacity in vivo. Different approaches have been described to make myogenic progenitors either by gene overexpression or by directed differentiation through culture conditions, and several myopathies have already been modeled using iPSC-MP. However, even though results in animal models have shown improvement from previous work with isolated adult myoblasts, major challenges regarding host response have to be addressed and clinically relevant transplantation protocols are lacking. Despite these challenges we are closer than we think to bringing iPSC-MP towards clinical use for treating human muscle disease and sporting injuries.

  1. Functional expression of gap junction gene Cx43 and the myogenic differentiation of rhabdomyosarcoma cells

    Institute of Scientific and Technical Information of China (English)

    林仲翔; 张志谦; 韩亚玲; C.C.G.Naus; K.R.Yu; H.Holtzer

    1995-01-01

    Rhabdomyosarcoma (RD) cells express low levels of the gap junction protein connexin 43 (Cx43), and its mRNA, and display very weak gap junctional intercellular communication (GJIC) as detected by Cx43 immunofluorescence, slot-blot and dye-transfer methods. These cells grow rapidly and show aberrant and incomplete myogenic differentiation. To investigate the role of gap junctions in these cells, the expression of Cx43 with relation to cell growth and myogenic differentiation in RD single-cell subclones-RDL3 and RDL6 is studied. The subclone RDL3 grows slowly and displays better myogenic differentiation. The expression of Cx43, its mRNA and the GJIC in RDL3 is comparable to that in normal myoblasts. Another subclone RDL6 which grows rapidly, but is poorly differentiated, expresses very low levels of Cx43 and its mRNA, and very weak GJIC. By using the calcium phosphate precipitate transfection technique, a full-length cDNA-encoding Cx43 and a pSV2neo have been introduced into the RDL6 cells. Several stably

  2. Conditional Cripto overexpression in satellite cells promotes myogenic commitment and enhances early regeneration.

    Science.gov (United States)

    Prezioso, Carolina; Iaconis, Salvatore; Andolfi, Gennaro; Zentilin, Lorena; Iavarone, Francescopaolo; Guardiola, Ombretta; Minchiotti, Gabriella

    2015-01-01

    Skeletal muscle regeneration mainly depends on satellite cells, a population of resident muscle stem cells. Despite extensive studies, knowledge of the molecular mechanisms underlying the early events associated with satellite cell activation and myogenic commitment in muscle regeneration remains still incomplete. Cripto is a novel regulator of postnatal skeletal muscle regeneration and a promising target for future therapy. Indeed, Cripto is expressed both in myogenic and inflammatory cells in skeletal muscle after acute injury and it is required in the satellite cell compartment to achieve effective muscle regeneration. A critical requirement to further explore the in vivo cellular contribution of Cripto in regulating skeletal muscle regeneration is the possibility to overexpress Cripto in its endogenous configuration and in a cell and time-specific manner. Here we report the generation and the functional characterization of a novel mouse model for conditional expression of Cripto, i.e., the Tg:DsRed (loxP/loxP) Cripto-eGFP mice. Moreover, by using a satellite cell specific Cre-driver line we investigated the biological effect of Cripto overexpression in vivo, and provided evidence that overexpression of Cripto in the adult satellite cell compartment promotes myogenic commitment and differentiation, and enhances early regeneration in a mouse model of acute injury.

  3. Ccndbp1 is a new positive regulator of skeletal myogenesis.

    Science.gov (United States)

    Huang, Yan; Chen, Bohong; Ye, Miaoman; Liang, Puping; Zhangfang, Yingnan; Huang, Junjiu; Liu, Mingyao; Songyang, Zhou; Ma, Wenbin

    2016-07-15

    Skeletal myogenesis is a multistep process in which basic helix-loop-helix (bHLH) transcription factors, such as MyoD (also known as MyoD1), bind to E-boxes and activate downstream genes. Ccndbp1 is a HLH protein that lacks a DNA-binding region, and its function in skeletal myogenesis is currently unknown. We generated Ccndbp1-null mice by using CRISPR-Cas9. Notably, in Ccndbp1-null mice, the cross sectional area of the skeletal tibialis anterior muscle was smaller, and muscle regeneration ability and grip strength were impaired, compared with those of wild type. This phenotype resembled that of myofiber hypotrophy in some human myopathies or amyoplasia. Ccndbp1 expression was upregulated during C2C12 myogenesis. Ccndbp1 overexpression promoted myogenesis, whereas knockdown of Ccndbp1 inhibited myogenic differentiation. Co-transfection of Ccndbp1 with MyoD and/or E47 (encoded by TCF3) significantly enhanced E-box-dependent transcription. Furthermore, Ccndbp1 physically associated with MyoD but not E47. These data suggest that Ccndbp1 regulates muscle differentiation by interacting with MyoD and enhancing its binding to target genes. Our study newly identifies Ccndbp1 as a positive modulator of skeletal myogenic differentiation in vivo and in vitro, providing new insights in order to decipher the complex network involved in skeletal myogenic development and related diseases. © 2016. Published by The Company of Biologists Ltd.

  4. INFLUENCE OF DANCE TRAINING ON SACCULOCOLLIC PATHWAY: VESTIBULAR EVOKED MYOGENIC POTENTIALS (VEMP AS AN OBJECTIVE TOOL

    Directory of Open Access Journals (Sweden)

    Swathi

    2013-10-01

    Full Text Available ABSTRACT : Auditory system is shaped by experience and training. Training (s ensory experience induces neurophysiologic changes & plasticity in normal hearing individuals, hearing loss patients, hearing aid users and cochlear implanted subjects. Not only speech stimulus, but music also brings about functional and structural organi zation of the brain in musician compared to non - musicians. The Vestibular evoked myogenic potentials (VEMP are a biphasic inhibitory response elicited by loud clicks or tone bursts recorded from the tonically contracted sternocleidomastoid muscle (SCM, b eing the only resource available to assess the function of the saccule and the inferior vestibular nerve ( sacculocollic pathway DESIGN: prospective study. AIM : The present study was conducted with an aim of studying plasticity of the sacculocollic pathway in professional dancers who are receiving dance training. METHOD : Two groups of subjects participated for the study a experimental group; b control group, experimental group was further divided in to two subgroups - Professional dancers who have received training in salsa as well as Bharath natyam. Experimental group consisted of total 40 subjects (80 ears, 20 (40 ears in each subgroup. Control group consisted of 40 individuals who have not received any professional training in dance (80 ears . RESULT: Results showed that there was statistically increase in amplitude of P13, N23 and P13 - N23 as well as early latency of P13, and N23 in professional dancers compared to the control group. The difference in amplitude and latency between the two groups was att ributed to plasticity of sacculocollic pathway in dancers. CONCLUSION: during aging process there is considerable deterioration of balance capability, loss of balance is a major risk factor for falls in middle aged and elderly people , to slow this deterior ation of balance one should gradually and continuously stimulate balance through motor activity, People

  5. Divergent and conserved roles of Dll1 signaling in development of craniofacial and trunk muscle.

    Science.gov (United States)

    Czajkowski, Maciej T; Rassek, Claudia; Lenhard, Diana C; Bröhl, Dominique; Birchmeier, Carmen

    2014-11-15

    Craniofacial and trunk skeletal muscles are evolutionarily distinct and derive from cranial and somitic mesoderm, respectively. Different regulatory hierarchies act upstream of myogenic regulatory factors in cranial and somitic mesoderm, but the same core regulatory network - MyoD, Myf5 and Mrf4 - executes the myogenic differentiation program. Notch signaling controls self-renewal of myogenic progenitors as well as satellite cell homing during formation of trunk muscle, but its role in craniofacial muscles has been little investigated. We show here that the pool of myogenic progenitor cells in craniofacial muscle of Dll1(LacZ/Ki) mutant mice is depleted in early fetal development, which is accompanied by a major deficit in muscle growth. At the expense of progenitor cells, supernumerary differentiating myoblasts appear transiently and these express MyoD. The progenitor pool in craniofacial muscle of Dll1(LacZ/Ki) mutants is largely rescued by an additional mutation of MyoD. We conclude from this that Notch exerts its decisive role in craniofacial myogenesis by repression of MyoD. This function is similar to the one previously observed in trunk myogenesis, and is thus conserved in cranial and trunk muscle. However, in cranial mesoderm-derived progenitors, Notch signaling is not required for Pax7 expression and impinges little on the homing of satellite cells. Thus, Dll1 functions in satellite cell homing and Pax7 expression diverge in cranial- and somite-derived muscle. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

  6. Mitogen-activated protein kinase inhibition and cardioplegia-cardiopulmonary bypass reduce coronary myogenic tone.

    Science.gov (United States)

    Khan, Tanveer A; Bianchi, Cesario; Ruel, Marc; Voisine, Pierre; Li, Jianyi; Liddicoat, John R; Sellke, Frank W

    2003-09-09

    Cardioplegia-cardiopulmonary bypass (C/CPB) is associated with coronary microcirculatory dysfunction. Regulation of the microcirculation includes myogenic tone. Mitogen-activated protein kinases (MAPK) have been implicated in coronary vasomotor function. We hypothesized that vasomotor dysfunction of the coronary microcirculation is mediated in part by alterations in extracellular signal regulated kinase 1/2 (ERK1/2) activity following C/CPB in humans. Atrial myocardium was harvested from patients (n=15) before and after blood cardioplegia and short-term reperfusion under conditions of CPB. Myogenic tone of coronary arterioles was measured by videomicroscopy. Microvessel tone was determined post-C/CPB and after PD98059, a MAPK/ERK kinase 1/2 (MEK1/2) inhibitor. MAPK phosphatase-1 (MKP-1) and activated ERK1/2 were measured by Western blot. MKP-1 gene expression was determined by Northern blot. In situ hybridization and immunohistochemistry were used to localize myocardial MKP-1 and activated ERK1/2, respectively. Myogenic tone was reduced in coronary arterioles post-C/CPB (-10.5+/-0.9%, P<0.01 versus control/pre-C/CPB, n=5). Myogenic tone was decreased in coronary microvessels after 30 micromol/L (n=5) and 50 micromol/L (n=5) PD98059 treatment (-11.0+/-0.8% and -14.6+/-2.0%, respectively, both P<0.01 versus control/pre-C/CPB). Myocardial levels of activated ERK1/2 were reduced post-C/CPB (0.6+/-0.1, post/pre-C/CPB ratio, P<0.05, n=5) while MKP-1 levels increased (4.2+/-0.6, post/pre-C/CPB ratio, P<0.05, n=5). Myocardial MKP-1 gene expression increased post-C/CPB (3.0+/-0.8, post/pre-C/CPB ratio, P<0.05, n=5). MKP-1 and activated ERK1/2 localized to coronary arterioles in myocardial sections. Coronary myogenic tone is dependent on ERK1/2 and decreased after C/CPB. C/CPB reduces levels of activated ERK1/2, potentially by increased levels of MKP-1. The ERK1/2 signal transduction pathway in part mediates coronary microvascular dysfunction after C/CPB in humans.

  7. N-cadherin and integrin blockade inhibit arteriolar myogenic reactivity but not pressure-induced increases in intracellular Ca2+

    Directory of Open Access Journals (Sweden)

    Teresa Y. Jackson

    2010-12-01

    Full Text Available The vascular myogenic response is characterized by arterial constriction in response to an increase in intraluminal pressure and dilatation to a decrease in pressure. This mechanism is important for the regulation of blood flow, capillary pressure and arterial pressure. The identity of the mechanosensory mechanism(s for this response is incompletely understood but has been shown to include the integrins as cell-extracellular matrix receptors. The possibility that a cell-cell adhesion receptor is involved has not been studied. Thus, we tested the hypothesis that N-cadherin, a cell-cell adhesion molecule in vascular smooth muscle cells (VSMCs, was important for myogenic responsiveness. The purpose of this study was to investigate:
    1. whether cadherin inhibition blocks myogenic responses to increases in intraluminal pressure and 2. the effect of the cadherin or integrin blockade on pressure-induced changes in [Ca2+]i. Cadherin blockade was tested in isolated rat cremaster arterioles on myogenic responses to acute pressure steps from 60 – 100 mmHg and changes in VSMC Ca2+ were measured using fura-2. In the presence of a synthetic cadherin inhibitory peptide or a function blocking antibody, myogenic responses were inhibited. In contrast, during N-cadherin blockade, pressure-induced changes in [Ca2+]i were not altered. Similarly, vessels treated with function-blocking β1- or β3-integrin antibodies maintained pressure-induced [Ca2+]i responses despite inhibition of myogenic constriction. Collectively, these data suggest that both cadherins and integrins play a fundamental role in mediating myogenic constriction but argue against their direct involvement in mediating pressure-induced [Ca2+]i increases.

  8. Myogenic-induced mesenchymal stem cells are capable of modulating the immune response by regulatory T cells

    Directory of Open Access Journals (Sweden)

    Sunyoung Joo

    2014-02-01

    Full Text Available Cell therapy for patients who have intractable muscle disorders may require highly regenerative cells from young, healthy allogeneic donors. Mesenchymal stem cells are currently under clinical investigation because they are known to induce muscle regeneration and believed to be immune privileged, thus making them suitable for allogeneic applications. However, it is unclear whether allogeneic and myogenic-induced mesenchymal stem cells retain their immunomodulatory characteristics. Therefore, our aim was to evaluate the effects of mesenchymal stem cell differentiation on the immune characteristics of cells in vitro. We investigated the immunologic properties of mesenchymal stem cells after myogenic induction. Mesenchymal stem cells were obtained from C57BL/6 mice and the C3H/10T1/2 murine mesenchymal stem cell line. Two different 5-aza-2′-deoxycytidine doses (0.5 and 3 µM were evaluated for their effects on mesenchymal stem cell skeletal myogenic differentiation potential, immune antigen expression, and mixed lymphocytic reactions. Using a mixed lymphocytic reaction, we determined the optimal splenocyte proliferation inhibition dose. The induction of regulatory T cells was markedly increased by the addition of 3 µM 5-aza-2′-deoxycytidine–treated mesenchymal stem cells. Myogenic-induced mesenchymal stem cells do not elicit alloreactive lymphocyte proliferative responses and are able to modulate immune responses. These findings support the hypothesis that myogenic-induced mesenchymal stem cells may be transplantable across allogeneic barriers.

  9. Redox signaling via oxidative inactivation of PTEN modulates pressure-dependent myogenic tone in rat middle cerebral arteries.

    Directory of Open Access Journals (Sweden)

    Debebe Gebremedhin

    Full Text Available The present study examined the level of generation of reactive oxygen species (ROS and roles of inactivation of the phosphatase PTEN and the PI3K/Akt signaling pathway in response to an increase in intramural pressure-induced myogenic cerebral arterial constriction. Step increases in intraluminal pressure of cannulated cerebral arteries induced myogenic constriction and concomitant formation of superoxide (O2 (.- and its dismutation product hydrogen peroxide (H2O2 as determined by fluorescent HPLC analysis, microscopic analysis of intensity of dihydroethidium fluorescence and attenuation of pressure-induced myogenic constriction by pretreatment with the ROS scavenger 4,hydroxyl-2,2,6,6-tetramethylpiperidine1-oxyl (tempol or Mito-tempol or MitoQ in the presence or absence of PEG-catalase. An increase in intraluminal pressure induced oxidation of PTEN and activation of Akt. Pharmacological inhibition of endogenous PTEN activity potentiated pressure-dependent myogenic constriction and caused a reduction in NPo of a 238 pS arterial KCa channel current and an increase in [Ca(2+]i level in freshly isolated cerebral arterial muscle cells (CAMCs, responses that were attenuated by Inhibition of the PI3K/Akt pathway. These findings demonstrate an increase in intraluminal pressure induced increase in ROS production triggered redox-sensitive signaling mechanism emanating from the cross-talk between oxidative inactivation of PTEN and activation of the PI3K/Akt signaling pathway that involves in the regulation of pressure-dependent myogenic cerebral arterial constriction.

  10. Peroral Echinococcus multilocularis egg inoculation in Myodes glareolus, Mesocricetus auratus and Mus musculus (CD-1 IGS and C57BL/6j)

    DEFF Research Database (Denmark)

    Woolsey, Ian David; Jensen, Per Moestrup; Deplazes, Peter

    2016-01-01

    Echinococcus multilocularis transmission predominantly occurs in Europe between the red fox (Vulpes vulpes) and various species of rodent intermediate hosts. We infected 3 species of rodent, Myodes glareolus (n = 47), Mesocricetus auratus (n = 11) and outbred Mus musculus (CD-1 IGS) (n = 9...

  11. Treatment of myogenic temporomandibular disorder: a prospective randomized clinical trial, comparing a mechanical stretching device (TheraBite®) with standard physical therapy exercise

    NARCIS (Netherlands)

    Kraaijenga, S.; van der Molen, L.; van Tinteren, H.; Hilgers, F.; Smeele, L.

    2014-01-01

    Aims: To compare in a randomized controlled clinical trial (RCT) the application of the TheraBite® (TB) Jaw Motion Rehabilitation System with a standard physical therapy (PT) exercise regimen for the treatment of myogenic temporomandibular disorder (TMD). Methodology: Myogenic TMD patients were

  12. Bank voles (Myodes glareolus) and house mice (Mus musculus musculus; M. m. domesticus) in Europe are each parasitized by their own distinct species of Aspiculuris (Nematoda, Oxyurida).

    Science.gov (United States)

    Behnke, J M; Stewart, A; Bajer, A; Grzybek, M; Harris, P D; Lowe, A; Ribas, A; Smales, L; Vandegrift, K J

    2015-10-01

    The molecular phylogeny and morphology of the oxyuroid nematode genus Aspiculuris from voles and house mice has been examined. Worms collected from Myodes glareolus in Poland, Eire and the UK are identified as Aspiculuris tianjinensis, previously known only from China, while worms from Mus musculus from a range of locations in Europe and from laboratory mice, all conformed to the description of Aspiculuris tetraptera. Worms from voles and house mice are not closely related and are not derived from each other, with A. tianjinensis being most closely related to Aspiculuris dinniki from snow voles and to an isolate from Microtus longicaudus in the Nearctic. Both A. tianjinensis and A. tetraptera appear to represent recent radiations within their host groups; in voles, this radiation cannot be more than 2 million years old, while in commensal house mice it is likely to be less than 10,000 years old. The potential of Aspiculuris spp. as markers of host evolution is highlighted.

  13. Gender differences in myogenic regulation along the vascular tree of the gerbil cochlea.

    Directory of Open Access Journals (Sweden)

    Katrin Reimann

    Full Text Available Regulation of cochlear blood flow is critical for hearing due to its exquisite sensitivity to ischemia and oxidative stress. Many forms of hearing loss such as sensorineural hearing loss and presbyacusis may involve or be aggravated by blood flow disorders. Animal experiments and clinical outcomes further suggest that there is a gender preference in hearing loss, with males being more susceptible. Autoregulation of cochlear blood flow has been demonstrated in some animal models in vivo, suggesting that similar to the brain, blood vessels supplying the cochlea have the ability to control flow within normal limits, despite variations in systemic blood pressure. Here, we investigated myogenic regulation in the cochlear blood supply of the Mongolian gerbil, a widely used animal model in hearing research. The cochlear blood supply originates at the basilar artery, followed by the anterior inferior cerebellar artery, and inside the inner ear, by the spiral modiolar artery and the radiating arterioles that supply the capillary beds of the spiral ligament and stria vascularis. Arteries from male and female gerbils were isolated and pressurized using a concentric pipette system. Diameter changes in response to increasing luminal pressures were recorded by laser scanning microscopy. Our results show that cochlear vessels from male and female gerbils exhibit myogenic regulation but with important differences. Whereas in male gerbils, both spiral modiolar arteries and radiating arterioles exhibited pressure-dependent tone, in females, only radiating arterioles had this property. Male spiral modiolar arteries responded more to L-NNA than female spiral modiolar arteries, suggesting that NO-dependent mechanisms play a bigger role in the myogenic regulation of male than female gerbil cochlear vessels.

  14. Effects of Chronic Blood-Flow Restriction Exercise on Skeletal Muscle Size and Myogenic Satellite Cell Expression

    DEFF Research Database (Denmark)

    Aagaard, Per; Jacobsen, Mikkel; Jensen, Kasper Yde

    2016-01-01

    of continued sports activity, resulting in visible hypertrophy of his left leg. AIM: To study the effect of chronic blood-flow restricted (BFR) exercise conditions on skeletal muscle size and myogenic satellite cell (SC) expression in an arterio-venous shunt patient. METHODS: Muscle biopsies were obtained from......-regulation in myogenic satellite cell activity within all stages of the cell cycle, which was accompanied by substantial muscle hypertrophy. Specifically, muscle fiber cross-sectional area (40%) and myonuclei number (15%) were elevated in the affected leg, together with an elevated myonuclear domain (20%). This single......-case study confirms previous result from our Lab demonstrating that blood-flow restricted muscle exercise leads to a marked activation of myogenic SCs, upregulated myonuclei number and marked myofiber hypertrophy....

  15. Myogenic-specific ablation of Fgfr1 impairs FGF2-mediated proliferation of satellite cells at the myofiber niche but does not abolish the capacity for muscle regeneration

    Directory of Open Access Journals (Sweden)

    Zipora eYablonka-Reuveni

    2015-05-01

    Full Text Available Skeletal muscle satellite cells (SCs are Pax7+ myogenic stem cells that reside between the basal lamina and the plasmalemma of the myofiber. In mature muscles, SCs are typically quiescent, but can be activated in response to muscle injury. Depending on the magnitude of tissue trauma, SCs may divide minimally to repair subtle damage within individual myofibers or produce a larger progeny pool that forms new myofibers in cases of overt muscle injury. SC transition through proliferation, differentiation and renewal is governed by the molecular blueprint of the cells as well as by the extracellular milieu at the SC niche. In particular, the role of the fibroblast growth factor (FGF family in regulating SCs during growth and aging is well recognized. Of the several FGFs shown to affect SCs, FGF1, FGF2 and FGF6 proteins have been documented in adult skeletal muscle. These prototypic paracrine FGFs transmit their mitogenic effect through the FGFRs, which are transmembrane tyrosine kinase receptors. Using the mouse model, we show here that of the four FGFRs, only Fgfr1 and Fgfr4 are expressed at relatively high levels in quiescent SCs and their proliferating progeny. To further investigate the role of FGFR1 in adult myogenesis, we have employed a genetic (Cre/loxP approach for myogenic-specific (MyoDCre-driven ablation of Fgfr1. Neither muscle histology nor muscle regeneration following cardiotoxin-induced injury were overtly affected in Fgfr1-ablated mice. This suggests that FGFR1 is not obligatory for SC performance in this acute muscle trauma model, where compensatory growth factor/cytokine regulatory cascades may exist. However, the SC mitogenic response to FGF2 is drastically repressed in isolated myofibers prepared from Fgfr1-ablated mice. Collectively, our study indicates that FGFR1 is important for FGF-mediated proliferation of SCs and its mitogenic role is not compensated by FGFR4 that is also highly expressed in SCs.

  16. The effect of alcohol on cervical and ocular vestibular evoked myogenic potentials in healthy volunteers

    OpenAIRE

    Rosengren, S M; Weber, K P; Hegemann, S C A; Roth, T N

    2014-01-01

    OBJECTIVE: We investigated the effect of alcohol on the cervical and ocular vestibular evoked myogenic potentials (cVEMPs and oVEMPs). As alcohol produces gaze-evoked nystagmus (GEN), we also tested the effect of nystagmus independent of alcohol by recording oVEMPs during optokinetic stimulation (OKS). METHODS: The effect of alcohol was tested in 14 subjects over multiple rounds of alcohol consumption up to a maximum breath alcohol concentration (BrAC) of 1.5‰ (mean 0.97‰). The effect of O...

  17. ADAM12 and alpha9beta1 integrin are instrumental in human myogenic cell differentiation

    DEFF Research Database (Denmark)

    Lafuste, Peggy; Sonnet, Corinne; Chazaud, Bénédicte

    2005-01-01

    Knowledge on molecular systems involved in myogenic precursor cell (mpc) fusion into myotubes is fragmentary. Previous studies have implicated the a disintegrin and metalloproteinase (ADAM) family in most mammalian cell fusion processes. ADAM12 is likely involved in fusion of murine mpc and human...... extracellular matrix, suggesting specific involvement of ADAM12-alpha9beta1 interaction in the fusion process. Evaluation of the fusion rate with regard to the size of myotubes showed that both ADAM12 antisense oligonucleotides and alpha9beta1 blockade inhibited more importantly formation of large (> or =5...

  18. Protein nitration impairs the myogenic tone of rat middle cerebral arteries in both ischemic and nonischemic hemispheres after ischemic stroke.

    Science.gov (United States)

    Coucha, Maha; Li, Weiguo; Johnson, Maribeth H; Fagan, Susan C; Ergul, Adviye

    2013-12-01

    The myogenic response is crucial for maintaining vascular resistance to achieve constant perfusion during pressure fluctuations. Reduced cerebral blood flow has been reported in ischemic and nonischemic hemispheres after stroke. Ischemia-reperfusion injury and the resulting oxidative stress impair myogenic responses in the ischemic hemisphere. Yet, the mechanism by which ischemia-reperfusion affects the nonischemic side is still undetermined. The goal of the present study was to determine the effect of ischemia-reperfusion injury on the myogenic reactivity of cerebral vessels from both hemispheres and whether protein nitration due to excess peroxynitrite production is the underlying mechanism of loss of tone. Male Wistar rats were subjected to sham operation or 30-min middle cerebral artery occlusion/45-min reperfusion. Rats were administered saline, the peroxynitrite decomposition catalyst 5,10,15,20-tetrakis(4-sulfonatophenyl)prophyrinato iron (III), or the nitration inhibitor epicatechin at reperfusion. Middle cerebral arteries isolated from another set of control rats were exposed to ex vivo oxygen-glucose deprivation with and without glycoprotein 91 tat (NADPH oxidase inhibitor) or N(ω)-nitro-l-arginine methyl ester. Myogenic tone and nitrotyrosine levels were determined. Ischemia-reperfusion injury impaired the myogenic tone of vessels in both hemispheres compared with the sham group (P nitration improved the myogenic tone of vessels from ischemic (P Nitration was significantly increased in both hemispheres versus the sham group and was normalized with epicatechin treatment. In conclusion, ischemia-reperfusion injury impairs vessel reactivity in both hemispheres via nitration. We suggest that sham operation rather than the nonischemic side should be used as a control in preclinical stroke studies.

  19. Bone morphogenic protein-2 regulates the myogenic differentiation of PMVECs in CBDL rat serum-induced pulmonary microvascular remodeling

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Chang; Chen, Lin; Zeng, Jing; Cui, Jian; Ning, Jiao-nin [Department of Anesthesia, Southwest Hospital, The Third Military Medical University, Chongqing 400038 (China); Wang, Guan-song [Institute of Respiratory Disease, Xinqiao Hospital, The Third Military Medical University, Chongqing 400037 (China); Belguise, Karine; Wang, Xiaobo [Université P. Sabatier Toulouse III and CNRS, LBCMCP, 31062 Toulouse Cedex 9 (France); Qian, Gui-sheng [Institute of Respiratory Disease, Xinqiao Hospital, The Third Military Medical University, Chongqing 400037 (China); Lu, Kai-zhi [Department of Anesthesia, Southwest Hospital, The Third Military Medical University, Chongqing 400038 (China); Yi, Bin, E-mail: yibin1974@163.com [Department of Anesthesia, Southwest Hospital, The Third Military Medical University, Chongqing 400038 (China)

    2015-08-01

    Hepatopulmonary syndrome (HPS) is characterized by an arterial oxygenation defect induced by intrapulmonary vasodilation (IPVD) that increases morbidity and mortality. In our previous study, it was determined that both the proliferation and the myogenic differentiation of pulmonary microvascular endothelial cells (PMVECs) play a key role in the development of IPVD. However, the molecular mechanism underlying the relationship between IPVD and the myogenic differentiation of PMVECs remains unknown. Additionally, it has been shown that bone morphogenic protein-2 (BMP2), via the control of protein expression, may regulate cell differentiation including cardiomyocyte differentiation, neuronal differentiation and odontoblastic differentiation. In this study, we observed that common bile duct ligation (CBDL)-rat serum induced the upregulation of the expression of several myogenic proteins (SM-α-actin, calponin, SM-MHC) and enhanced the expression levels of BMP2 mRNA and protein in PMVECs. We also observed that both the expression levels of Smad1/5 and the activation of phosphorylated Smad1/5 were significantly elevated in PMVECs following exposure to CBDL-rat serum, which was accompanied by the down-regulation of Smurf1. The blockage of the BMP2/Smad signaling pathway with Noggin inhibited the myogenic differentiation of PMVECs, a process that was associated with relatively low expression levels of both SM-α-actin and calponin in the setting of CBDL-rat serum exposure, although SM-MHC expression was not affected. These findings suggested that the BMP2/Smad signaling pathway is involved in the myogenic differentiation of the PMVECs. In conclusion, our data highlight the pivotal role of BMP2 in the CBDL-rat serum-induced myogenic differentiation of PMVECs via the activation of both Smad1 and Smad5 and the down-regulation of Smurf1, which may represent a potential therapy for HPS-induced pulmonary vascular remodeling. - Highlights: • CBDL-rat serum promotes the myogenic

  20. The migration of myogenic cells from the somites at the wing level in avian embryos.

    Science.gov (United States)

    Solursh, M; Drake, C; Meier, S

    1987-06-01

    This study is concerned with establishing a morphological basis for the initiation of migration of putative myogenic cells from the somites into the presumptive wing bud in avian embryos. At the 22 somite stage (stage 14) vasculogenesis is a prevalent activity. By use of a quail specific monoclonal antibody to vascular endothelial cells, vascular cells are recognized in the lateral plate, on the intermediate mesoderm, and on somite surfaces. Cells that are found between the lateral plate mesoderm and somites are shown to be vascular endothelial cells. The lateral body folds progressively bring the lateral plate mesoderm close to the lateral margin of the somites and vascular elements disappear from surface view. It is not until the 24 somite stage (stage 15) that some cells in the ventral lateral margin of somites at the wing level can be seen in scanning electron micrographs to extend basal cell processes toward adjacent vascular tubes. These results provide a morphological basis for the early migratory behavior of myogenic cells and demonstrate their close proximity to the prepatterned vascular network.

  1. Molecular Basis for the Regulation of Transcriptional Coactivator p300 in Myogenic Differentiation.

    Science.gov (United States)

    Chen, Jihong; Wang, Yingjian; Hamed, Munerah; Lacroix, Natascha; Li, Qiao

    2015-09-10

    Skeletal myogenesis is a highly ordered process which specifically depends on the function of transcriptional coactivator p300. Previous studies have established that Akt/protein kinase B (PKB), a positive regulator of p300 in proliferating cells, is also important for proper skeletal muscle development. Nevertheless, it is not clear as to how the p300 is regulated by myogenic signaling events given that both p300 and Akt are involved in many cellular processes. Our studies revealed that the levels of p300 protein are temporally maintained in ligand-enhanced skeletal myocyte development. Interestingly, this maintenance of p300 protein is observed at the stage of myoblast differentiation, which coincides with an increase in Akt phosphorylation. Moreover, regulation of p300 during myoblast differentiation appears to be mediated by Akt signaling. Blunting of p300 impairs myogenic expression and myoblast differentiation. Thus, our data suggests a particular role for Akt in myoblast differentiation through interaction with p300. Our studies also establish the potential of exploiting p300 regulation and Akt activation to decipher the complex signaling cascades involved in skeletal muscle development.

  2. Fibromodulin: a master regulator of myostatin controlling progression of satellite cells through a myogenic program.

    Science.gov (United States)

    Lee, Eun Ju; Jan, Arif Tasleem; Baig, Mohammad Hassan; Ashraf, Jalaluddin Mohammad; Nahm, Sang-Soep; Kim, Yong-Woon; Park, So-Young; Choi, Inho

    2016-08-01

    Differentiation of muscle satellite cells (MSCs) involves interaction of the proteins present in the extracellular matrix (ECM) with MSCs to regulate their activity, and therefore phenotype. Herein, we report fibromodulin (FMOD), a member of the proteoglycan family participating in the assembly of ECM, as a novel regulator of myostatin (MSTN) during myoblast differentiation. In addition to having a pronounced effect on the expression of myogenic marker genes [myogenin (MYOG) and myosin light chain 2 (MYL2)], FMOD was found to maintain the transcriptional activity of MSTN Moreover, coimmunoprecipitation and in silico studies performed to investigate the interaction of FMOD helped confirm that it antagonizes MSTN function by distorting its folding and preventing its binding to activin receptor type IIB. Furthermore, in vivo studies revealed that FMOD plays an active role in healing by increasing satellite cell recruitment to sites of injury. Together, these findings disclose a hitherto unrecognized regulatory role for FMOD in MSCs and highlight new mechanisms whereby FMOD circumvents the inhibitory effects of MSTN and triggers myoblast differentiation. These findings offer a basis for the design of novel MSTN inhibitors that promote muscle regeneration after injury or for the development of pharmaceutical agents for the treatment of different muscle atrophies.-Lee, E. J., Jan, A. T., Baig, M. H., Ashraf, J. M., Nahm, S.-S., Kim, Y.-W., Park, S.-Y., Choi, I. Fibromodulin: a master regulator of myostatin controlling progression of satellite cells through a myogenic program.

  3. Accuracy of the surface electromyography RMS processing for the diagnosis of myogenous temporomandibular disorder.

    Science.gov (United States)

    Berni, Kelly Cristina dos Santos; Dibai-Filho, Almir Vieira; Pires, Paulo Fernandes; Rodrigues-Bigaton, Delaine

    2015-08-01

    Due to the multifactor etiology of temporomandibular disorder (TMD), the precise diagnosis remains a matter of debate and validated diagnostic tools are needed. The aim was to determine the accuracy of surface electromyography (sEMG) activity, assessed in the amplitude domain by the root mean square (RMS), in the diagnosis of TMD. One hundred twenty-three volunteers were evaluated using the Research Diagnostic Criteria for Temporomandibular Disorders and distributed into two groups: women with myogenous TMD (n=80) and women without TMD (n=43). The volunteers were then submitted to sEMG evaluation of the anterior temporalis, masseter and suprahyoid muscles at rest and during maximum voluntary teeth clenching (MVC) on parafilm. The accuracy, sensitivity and specificity of the muscle activity were analyzed. Differences between groups were found in all muscles analyzed at rest as well as in the masseter and suprahyoid muscles during MVC on parafilm. Moderate accuracy (AUC: 0.74-0.84) of the RMS sEMG was found in all muscles regarding the diagnosis of TMD at rest and in the suprahyoid muscles during MVC on parafilm. Moreover, sensitivity ranging from 71.3% to 80% and specificity from 60.5% to 76.6%. In contrast, RMS sEMG did not exhibit acceptable degrees of accuracy in the other masticatory muscles during MVC on parafilm. It was concluded that the RMS sEMG is a complementary tool for clinical diagnosis of the myogenous TMD.

  4. Myogenic differentiation of mesenchymal stem cells for muscle regeneration in urinary tract

    Institute of Scientific and Technical Information of China (English)

    YANG Bin; ZHENG Jun-hua; ZHANG Yuan-yuan

    2013-01-01

    Objective This article was to review the current status of adult mesenchymal stem cells transplantation for muscle regeneration in urinary tract and propose the future prospect in this field.Data sources The data used in this review were mainly obtained from articles listed in Medline and PubMed (2000-2013).The search terms were "mesenchymal stem cells","bladder","stress urinary incontinence" and "tissue engineering".Study selection Articles regarding the adult mesenchymal stem cells for tissue engineering of bladder and stress urinary incontinence were selected and reviewed.Results Adult mesenchymal stem cells had been identified and well characterized in human bone marrow,adipose tissue,skeletal muscle and urine,and demonstrated the capability of differentiating into smooth muscle cells and skeletal muscle cells under myogenic differentiation conditions in vitro.Multiple preclinical and clinical studies indicated that adult mesenchymal stem cells could restore and maintain the structure and function of urinary muscle tissues after transplanted,and potentially improve the quality of life in patients.Conclusions Smooth or skeletal myogenic differentiation of mesenchymal stem cells with regenerative medicine technology may provide a novel approach for muscle regeneration and tissue repair in urinary tract.The long-term effect and safety of mesenchymal stem cell transplantation should be further evaluated before this approach becomes widely used in patients.

  5. Intrinsic Transient Tracheal Occlusion Training and Myogenic Remodeling of Rodent Parasternal Intercostal Fibers

    Science.gov (United States)

    Smith, Barbara K.; Mathur, Sunita; Ye, Fan; Martin, A. Daniel; Truelson, Sara Attia; Vandenborne, Krista; Davenport, Paul W.

    2014-01-01

    It is recognized that diaphragm muscle plasticity occurs with mechanical overloads, yet less is known regarding synergistic parasternal intercostal muscle fiber remodeling. We conducted overload training with intrinsic transient tracheal occlusion (ITTO) exercises in conscious animals. We hypothesized ITTO would yield significant fiber hypertrophy and myogenic activation that would parallel diaphragm fiber remodeling. Sprague-Dawley rats underwent placement of a tracheal cuff and were randomly assigned to receive daily ten-minute sessions of conscious ITTO or observation (SHAM) over two weeks. After training, fiber morphology, myosin heavy chain isoform composition, cross-sectional area, proportion of Pax7-positive nuclei, and presence of embryonic myosin (eMHC) were quantified. Type IIx/b fibers were 20% larger after ITTO training than with SHAM training (ITTO: 4431±676 μm2, SHAM: 3689±400 μm2, p<0.05), and type I fibers were more prevalent after ITTO (p<0.01). Expression of Pax7 was increased in ITTO parasternals and diaphragm (p<0.05). In contrast, the proportion of eMHC-positive fibers was increased only in ITTO parasternals (1.2 (3.4-0.6)%, SHAM: 0 (0.6-0%, p<0.05). Although diaphragm and parasternal type II fibers hypertrophy to a similar degree, myogenic remodeling appears to differ between the two muscles. PMID:25509059

  6. Can a finding of cervical vestibular evoked myogenic potentials contribute to vestibular migraine diagnostics?

    Directory of Open Access Journals (Sweden)

    Tihana Vešligaj

    2016-02-01

    Full Text Available Aim To investigate differences in vestibular evoked myogenic potentials (VEMP results with patients suffering from vestibular migraine and healthy people, taking into consideration values of threshold and latency of occurrence of the characteristic wave complex, size of amplitude, and interaural amplitude ratio. According to the results, determine the importance and usefulness of VEMP in vestibular migraine diagnostics. Methods A total number of 62 subjects were included in the study, 32 of them belonging to a group of patients suffering from vestibular migraine (VM, while other 30 were in a control group of healthy subjects. Information was collected during the diagnostic evaluation. General and otoneurological history of patients and bedside tests, audiological results, videonystagmography and cervical vestibular evoked myogenic potentials (cVEMP were made. Results There was a difference in an interaural ratio of amplitudes in the experimental and control groups, but it was not found to be clinically significant. By ToneBurst 500 Hz method, the interaural amplitude ratio higher than 35% was measured in 46.97% subjects, while the response was totally unilaterally missing in 28.8% patients. Conclusion Even the sophisticated method as cVEMP does not give the ultimate result confirming the vestibular migraine diagnosis, and neither do other diagnostic methods. cVEMP result can contribute to the completion of full mosaic of vestibular migraine diagnostics.

  7. Nociception contributes to the formation of myogenic contracture in the early phase of adjuvant-induced arthritis in a rat knee.

    Science.gov (United States)

    Kaneguchi, Akinori; Ozawa, Junya; Moriyama, Hideki; Yamaoka, Kaoru

    2017-07-01

    It is unknown how joint contracture is generated in inflamed joints. This study aimed to clarify the role of nociception on the formation of joint contracture secondary to arthritis. Monoarthritis was induced by intra-articular injections of complete Freund's adjuvant (CFA) into rat knees. On day 5 after CFA injection, the passive extension range of motion (ROM) of knee joints were measured, both before and after myotomy of knee flexors, to evaluate the extent of muscular contribution to CFA-induced joint contracture. The steroidal anti-inflammatory drug dexamethasone could prevent ROM restrictions completely, both before and after myotomy. On the other hand, the opioid analgesic drug morphine did not prevent the development of restricted ROM observed after myotomy, while it did before myotomy. This indicates that nociception contributes to joint contracture through alterations in muscular structure (myogenic factors). Next, we tested the hypothesis that nociception-induced reflexive flexor muscle contractions cause myogenic contracture in arthritic joints. To do this, chemical denervation was performed by Botulinum toxin type A (BTX-A) injections into knee flexor muscles, simultaneously with CFA injections into the knee. As expected, BTX-A could alleviate ROM restrictions observed before myotomy. These findings suggest that nociceptive-related muscle contractions play an essential role in the formation of joint contracture. Thus, our study indicates that analgesic management during an early stage of joint arthritis is an essential mean to prevent the formation of joint contracture. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:1404-1413, 2017. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

  8. The myogenic electric organ of Sternopygus macrurus: a non-contractile tissue with a skeletal muscle transcriptome

    Science.gov (United States)

    Samanta, Manoj P.; Chaidez, Alexander

    2016-01-01

    In most electric fish species, the electric organ (EO) derives from striated muscle cells that suppress many muscle properties. In the gymnotiform Sternopygus macrurus, mature electrocytes, the current-producing cells of the EO, do not contain sarcomeres, yet they continue to make some cytoskeletal and sarcomeric proteins and the muscle transcription factors (MTFs) that induce their expression. In order to more comprehensively examine the transcriptional regulation of genes associated with the formation and maintenance of the contractile sarcomere complex, results from expression analysis using qRT-PCR were informed by deep RNA sequencing of transcriptomes and miRNA compositions of muscle and EO tissues from adult S. macrurus. Our data show that: (1) components associated with the homeostasis of the sarcomere and sarcomere-sarcolemma linkage were transcribed in EO at levels similar to those in muscle; (2) MTF families associated with activation of the skeletal muscle program were not differentially expressed between these tissues; and (3) a set of microRNAs that are implicated in regulation of the muscle phenotype are enriched in EO. These data support the development of a unique and highly specialized non-contractile electrogenic cell that emerges from a striated phenotype and further differentiates with little modification in its transcript composition. This comprehensive analysis of parallel mRNA and miRNA profiles is not only a foundation for functional studies aimed at identifying mechanisms underlying the transcription-independent myogenic program in S. macrurus EO, but also has important implications to many vertebrate cell types that independently activate or suppress specific features of the skeletal muscle program. PMID:27114860

  9. MAPK signaling pathways and HDAC3 activity are disrupted during differentiation of emerin-null myogenic progenitor cells

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    Carol M. Collins

    2017-04-01

    Full Text Available Mutations in the gene encoding emerin cause Emery–Dreifuss muscular dystrophy (EDMD. Emerin is an integral inner nuclear membrane protein and a component of the nuclear lamina. EDMD is characterized by skeletal muscle wasting, cardiac conduction defects and tendon contractures. The failure to regenerate skeletal muscle is predicted to contribute to the skeletal muscle pathology of EDMD. We hypothesize that muscle regeneration defects are caused by impaired muscle stem cell differentiation. Myogenic progenitors derived from emerin-null mice were used to confirm their impaired differentiation and analyze selected myogenic molecular pathways. Emerin-null progenitors were delayed in their cell cycle exit, had decreased myosin heavy chain (MyHC expression and formed fewer myotubes. Emerin binds to and activates histone deacetylase 3 (HDAC3. Here, we show that theophylline, an HDAC3-specific activator, improved myotube formation in emerin-null cells. Addition of the HDAC3-specific inhibitor RGFP966 blocked myotube formation and MyHC expression in wild-type and emerin-null myogenic progenitors, but did not affect cell cycle exit. Downregulation of emerin was previously shown to affect the p38 MAPK and ERK/MAPK pathways in C2C12 myoblast differentiation. Using a pure population of myogenic progenitors completely lacking emerin expression, we show that these pathways are also disrupted. ERK inhibition improved MyHC expression in emerin-null cells, but failed to rescue myotube formation or cell cycle exit. Inhibition of p38 MAPK prevented differentiation in both wild-type and emerin-null progenitors. These results show that each of these molecular pathways specifically regulates a particular stage of myogenic differentiation in an emerin-dependent manner. Thus, pharmacological targeting of multiple pathways acting at specific differentiation stages may be a better therapeutic approach in the future to rescue muscle regeneration in vivo.

  10. Zinc promotes proliferation and activation of myogenic cells via the PI3K/Akt and ERK signaling cascade

    Energy Technology Data Exchange (ETDEWEB)

    Ohashi, Kazuya, E-mail: asuno10k@yahoo.co.jp [Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo (Japan); Nagata, Yosuke, E-mail: cynagata@mail.ecc.u-tokyo.ac.jp [Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo (Japan); Wada, Eiji, E-mail: gacchu1@yahoo.co.jp [Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo (Japan); Zammit, Peter S., E-mail: peter.zammit@kcl.ac.uk [Randall Division of Cell and Molecular Biophysics, King' s College London, London SE1 1UL (United Kingdom); Shiozuka, Masataka, E-mail: cmuscle@mail.ecc.u-tokyo.ac.jp [Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo (Japan); Matsuda, Ryoichi, E-mail: cmatsuda@mail.ecc.u-tokyo.ac.jp [Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo (Japan)

    2015-05-01

    Skeletal muscle stem cells named muscle satellite cells are normally quiescent but are activated in response to various stimuli, such as injury and overload. Activated satellite cells enter the cell cycle and proliferate to produce a large number of myogenic progenitor cells, and these cells then differentiate and fuse to form myofibers. Zinc is one of the essential elements in the human body, and has multiple roles, including cell growth and DNA synthesis. However, the role of zinc in myogenic cells is not well understood, and is the focus of this study. We first examined the effects of zinc on differentiation of murine C2C12 myoblasts and found that zinc promoted proliferation, with an increased number of cells incorporating EdU, but inhibited differentiation with reduced myogenin expression and myotube formation. Furthermore, we used the C2C12 reserve cell model of myogenic quiescence to investigate the role of zinc on activation of myogenic cells. The number of reserve cells incorporating BrdU was increased by zinc in a dose dependent manner, with the number dramatically further increased using a combination of zinc and insulin. Akt and extracellular signal-regulated kinase (ERK) are downstream of insulin signaling, and both were phosphorylated after zinc treatment. The zinc/insulin combination-induced activation involved the phosphoinositide 3-kinase (PI3K)/Akt and ERK cascade. We conclude that zinc promotes activation and proliferation of myogenic cells, and this activation requires phosphorylation of PI3K/Akt and ERK as part of the signaling cascade. - Highlights: • Zinc has roles for promoting proliferation and inhibition differentiation of C2C12. • Zinc promotes activation of reserve cells. • Insulin and zinc synergize activation of reserve cells. • PI3K/Akt and ERK cascade affect zinc/insulin-mediated activation of reserve cells.

  11. Role of T-type calcium channels in myogenic tone of skeletal muscle resistance arteries

    DEFF Research Database (Denmark)

    VanBavel, Ed; Sorop, Oana; Andreasen, Ditte

    2002-01-01

    T-type calcium channels may be involved in the maintenance of myogenic tone. We tested their role in isolated rat cremaster arterioles obtained after CO(2) anesthesia and decapitation. Total RNA was analyzed by RT-PCR and Southern blotting for calcium channel expression. We observed expression...... of voltage-operated calcium (Ca(V)) channels Ca(V)3.1 (T-type), Ca(V)3.2 (T-type), and Ca(V)1.2 (L-type) in cremaster arterioles (n = 3 rats). Amplification products were observed only in the presence of reverse transcriptase and cDNA. Concentration-response curves of the relatively specific L-type blocker......); K(+) -5.4 +/- 0.3 (n = 4); all log(IC(50)) P cremaster muscle arterioles....

  12. Augmented ocular vestibular evoked myogenic potentials to air-conducted sound in large vestibular aqueduct syndrome.

    Science.gov (United States)

    Taylor, Rachael L; Bradshaw, Andrew P; Magnussen, John S; Gibson, William P R; Halmagyi, G Michael; Welgampola, Miriam S

    2012-01-01

    To demonstrate the value of recording air-conducted ocular Vestibular Evoked Myogenic Potentials (oVEMP) in a patient with bilaterally enlarged vestibular aqueducts. Cervical VEMP and oVEMP were recorded from a patient presenting with bilateral hearing loss and imbalance, attributable to large vestibular aqueduct syndrome. The stimuli were air-conducted tone bursts at octave frequencies from 250 to 2000 Hz. Amplitudes and thresholds were measured and compared with the normal response range of 32 healthy control subjects. oVEMP reflexes demonstrated pathologically increased amplitudes and reduced thresholds for low-frequency tone bursts. Cervical VEMP amplitudes and thresholds were within normal limits for both ears across all frequencies of stimulation. This study is the first to describe the augmentation of AC oVEMPs in an adult with large vestibular aqueduct syndrome.

  13. Development of Bipotent Cardiac/Skeletal Myogenic Progenitors from MESP1+ Mesoderm

    Directory of Open Access Journals (Sweden)

    Sunny Sun-Kin Chan

    2016-01-01

    Full Text Available The branchiomeric skeletal muscles co-evolved with new chambers of the heart to enable predatory feeding in chordates. These co-evolved tissues develop from a common population in anterior splanchnic mesoderm, referred to as cardiopharyngeal mesoderm (CPM. The regulation and development of CPM are poorly understood. We describe an embryonic stem cell-based system in which MESP1 drives a PDGFRA+ population with dual cardiac and skeletal muscle differentiation potential, and gene expression resembling CPM. Using this system, we investigate the regulation of these bipotent progenitors, and find that cardiac specification is governed by an antagonistic TGFβ-BMP axis, while skeletal muscle specification is enhanced by Rho kinase inhibition. We define transcriptional signatures of the first committed CPM-derived cardiac and skeletal myogenic progenitors, and discover surface markers to distinguish cardiac (PODXL+ from the skeletal muscle (CDH4+ CPM derivatives. These tools open an accessible window on this developmentally and evolutionarily important population.

  14. Ageing effect on air-conducted ocular vestibular evoked myogenic potential

    Directory of Open Access Journals (Sweden)

    Kaushlendra Kumar

    2015-08-01

    Full Text Available One of the recent diagnostic tests to assess the function of otolithic organs is through vestibular evoked myogenic potential (VEMP testing. There are equivocal findings on effect of aging on ocular VEMP (oVEMP parameters with reference to latencies. Hence this study was taken up to investigate the age related changes in oVEMP parameters. This present study considered 30 participants in each age group i.e., young adults, middle-aged adults and older adults. oVEMP were recorded using insert earphone at 100dBnHL at 500hZ short duration tone burst. The results showed in older adult significant difference in response rate, latencies and amplitude as compared to young and middle adult. Hence age should be taken into consideration when interpreting oVEMP results.

  15. Role of RhoA/Rho kinase signaling pathway in microgroove induced stem cell myogenic differentiation.

    Science.gov (United States)

    Li, Huaqiong; Wen, Feng; Wang, Xincai; Tan, Lay Poh

    2015-06-01

    In our previous report, the authors have demonstrated that direct laser machined microchannels would trigger upregulation of myogenic markers in human mesenchymal stem cells (hMSCs) through promotion of cell elongation. However, the molecular basis signaling pathways behind this observation remains unclear. In this work, three types of microchannels generated by femtosecond laser were utilized to investigate possible mechanisms behind the induction of hMSCs myogenesis by microchannels. The authors hypothesized that small G-proteins RhoA and Rac1 play a vital role on myogenesis of hMSCs through regulating cytoskeleton rearrangement, via cell tension signaling cascades. The RhoA and Rac1 activities were evaluated for cells cultured on the micropatterned substrates, using a flat unpatterned substrate as control. It was found that significant activation of RhoA GTPase was exhibited for cells cultured on narrow microchannels (20-20-20 and 30-30-20), while no obvious differences were obtained on wide ones (80-30-20). Meanwhile, no significant difference was found for Rac1 activities on all tested groups. To further deduce the role of RhoA signaling pathway in microchannel directed stem cell myogenesis, the effectors of Rho, Rho kinase (ROCK) was chosen to explore how cell shape regulate myogenesis of hMSCs cultured on laser micropatterned substrate. A pharmacological ROCK inhibitor, Y-27632, was used to treat the cells and the effect on RhoA activation was investigated. Our data on the role of RhoA/ROCK in regulating cell myogenic differentiation on lasered microchannels substrates may provide a mechanistic insight on hMSCs fate directed by substrate topography.

  16. An experimental method to identify neurogenic and myogenic active mechanical states of intestinal motility

    Directory of Open Access Journals (Sweden)

    Marcello eCosta

    2013-04-01

    Full Text Available Excitatory and inhibitory enteric neural input to intestinal muscle acting on ongoing myogenic activity determines the rich repertoire of motor patterns involved in digestive function. The enteric neural activity cannot yet be established during movement of intact intestine in vivo or in vitro. We propose the hypothesis that is possible to deduce indirectly, but reliably, the state of activation of the enteric neural input to the muscle from measurements of the mechanical state of the intestinal muscle. The fundamental biomechanical model on which our hypothesis is based is the ‘three-element model’ proposed by Hill. Our strategy is based on simultaneous video recording of changes in diameters and intraluminal pressure with a fibre-optic manometry in isolated segments of rabbit colon. We created a composite spatiotemporal map (DPMap from diameter (DMap and pressure changes (PMaps. In this composite map rhythmic myogenic motor patterns can readily be distinguished from the distension induced neural peristaltic contractions. Plotting the diameter changes against corresponding pressure changes at each location of the segment, generates ‘orbits’ that represent the state of the muscle according to its ability to contract or relax actively or undergoing passive changes. With a software developed in MatLab, we identified twelve possible discrete mechanical states and plotted them showing where the intestine actively contracted and relaxed isometrically, auxotonically or isotonically, as well as where passive changes occurred or was quiescent. Clustering all discrete active contractions and relaxations states generated for the first time a spatio-temporal map of where enteric excitatory and inhibitory neural input to the muscle occurs during physiological movements. Recording internal diameter by an impedance probe proved equivalent to measuring external diameter, making possible to further develop similar strategy in vivo and humans.

  17. Peroral Echinococcus multilocularis egg inoculation in Myodes glareolus, Mesocricetus auratus and Mus musculus (CD-1 IGS and C57BL/6j

    Directory of Open Access Journals (Sweden)

    Ian David Woolsey

    2016-08-01

    Full Text Available Echinococcus multilocularis transmission predominantly occurs in Europe between the red fox (Vulpes vulpes and various species of rodent intermediate hosts. We infected 3 species of rodent, Myodes glareolus (n = 47, Mesocricetus auratus (n = 11 and outbred Mus musculus (CD-1 IGS (n = 9 with an E. multilocularis egg suspension that contained 100 eggs with viable oncospheres and performed post mortem examination 6, 8 (M. glareolus and 10 weeks post inoculation (wpi. C57BL/6j mice (n = 4 were used as positive controls as they have been shown to exhibit macroscopic liver lesions 4 wpi. To the best of our knowledge, this is the first study to experimentally assess susceptibility in the ostensibly competent host M. glareolus. Lesions were only detected in 2 of 47 M. glareolus (4.3% at 8 and 10 wpi and although both contained protoscolices (1675 at 8 wpi and 88 at 12 wpi the low percentage of infected animals brings into question their role as transmitters of the parasite. Significant differences were observed between inbred and outbred mice with E. multilocularis infection in the former demonstrating increased establishment (p ≤ 0.0001 and growth (p ≤ 0.0001. No lesions were found in all 11 M. auratus.

  18. Effects of an MHC-DRB genotype and allele number on the load of gut parasites in the bank vole Myodes glareolus.

    Science.gov (United States)

    Kloch, Agnieszka; Babik, Wiesław; Bajer, Anna; Siński, Edward; Radwan, Jacek

    2010-03-01

    The major histocompatibility complex (MHC) genes code for the proteins responsible for pathogen recognition. The MHC class II DRB gene is multiplicated in the bank vole, Myodes glareolus, with different numbers of loci found in different individuals. Possessing large numbers of loci should increase the probability of pathogen recognition, but according to the optimality hypothesis, there is a cost of possessing too many MHC alleles. Using 454 technology, we determined the individual DRB allelic diversity and related it to the load of intestinal parasites in voles collected from three sites separated by a distance of 12 to 27 km. The analysis of six microsatellite loci revealed significant population structure (F(ST) = 0.07). The sites differed significantly in the prevalence and abundance of nematode species as well. We found two significant associations between MHC alleles and the intensity of the infection with the most prevalent nematode, Aspiculuris tetraptera. One of these associations was population-specific. This result suggests that the directions of selection can differ between populations connected by a low level of gene flow, which may contribute to the maintenance of high DRB allele diversity. In accordance with the optimality hypothesis, individuals with an intermediate number of alleles carried the lowest number of nematode species and had the lowest prevalence of A. tetraptera. However, the intensity of infection with A. tetraptera was linearly and negatively associated with the number of alleles.

  19. Yellow-necked mice (Apodemus flavicollis and bank voles (Myodes glareolus as zoomonitors of environmental contamination at a polluted area in Slovakia

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    Jančová Alena

    2010-11-01

    Full Text Available Abstract Background Free-living wild rodents are often used as zoomonitors of environmental contamination. In the present study, accumulation of cadmium (Cd, copper (Cu, iron (Fe, and zinc (Zn in critical organs of yellow-necked mice (Apodemus flavicollis and bank voles (Myodes glareolus trapped in a polluted area in Nováky, Slovakia was investigated. Methods Yellow-necked mice (n = 8 and bank voles (n = 10 were collected using standard theriological methods for wood ecosystems. All animals were adult males in good physical condition. The concentrations of Cd, Cu, Fe, and Zn in the liver, kidney, and bone were determined by atomic absorption spectrophotometry. Results The highest concentrations of Cd and Zn were found in the bone of both species while Cu and Fe accumulated mainly in kidney or liver. Significant higher concentrations of Cd and Cu were detected in the liver of bank voles than in yellow-necked mice. Similar significant higher levels of Cd and Zn were found in the bone of bank voles. In contrast, significant higher concentrations of Cu and Fe were present in the kidney of yellow-necked mice. Conclusions In the yellow-necked mouse and bank vole, bone seems to accumulate Cd and Zn following prolonged exposure. On the contrary, kidney and liver store Cu and Fe after a long-term environmental exposure. In the present study, bank voles seemed to be more heavy metal loaded zoomonitors than yellow-necked mice.

  20. Wnt3a signaling promotes proliferation,myogenic differentiation,and migration of rat bone marrow mesenchymal stem cells

    Institute of Scientific and Technical Information of China (English)

    Yan-chang SHANG; Shu-hui WANG; Fu XIONG; Cui-ping ZHAO; Fu-ning PENG; Shan-wei FENG; Mei-shan LI; Yong LI; Cheng ZHANG

    2007-01-01

    Aim:To investigate the effects of the wingless-related MMTV integration site 3A (Wnt3a) signaling on the proliferation,migration,and the myogenic and adipogenic differentiation of rat bone marrow mesenchymal stem cells (rMSC). Methods:Primary MSC were isolated and cultured from Spragne-Dawley rats and characterized by flow cytometry. Mouse L cells were transfected with Wnt3a cDNA,and conditioned media containing active Wnt3a proteins were prepared. Cell proliferation was evaluated by cell count and 5-bromodeoxyuridine incorporation assay.The migration of rMSC was performed by using a transwell migration and wound healing assay. The myogenic and adipogenic differentiation in rMSC were examined by light microscopy,immunofluorescence,and RT-PCR at different time points after myogenic or adipogenic introduction. Results:Wnt3a signaling induced β-catenin nuclear translocation and activated the Writ pathway in rMSC.In the presence of Wnt3a,rMSC proliferated more rapidly than the control cells,keeping their differentiation potential. Moreover,Wnt3a signaling induced 2.62%and 3.76% of rMSC-expressed desmin and myosin heavy chain after being cultured in myogenic medium. The myogenic differentiation genes,including Pax7,MyoD,Myf5,Myf4,and myogenin,were activated after Wnt3a treatment. On the other hand,Wnt3a inhibited the adipogenic differentiation in rMSC through the downregulated expression of CCAAT/enhancer-binding protein alpha (C/EBPalpha)and peroxisome proliferator-activaled receptor gamma (PPARgamma). Furthermore,Wnt3a promoted the migration capacity of rMSC. Conclusion:The results indicate that Wnt3a signaling can induce myogenic differentiation in rMSC. Wnt3a signaling is also involved in the regulation of the proliferation and migration of rMSC. These results could provide a rational foundation for cell-based tissue repair in humans.

  1. Combined ocular and cervical vestibular evoked myogenic potential in individuals with vestibular hyporeflexia and in patients with Ménière's disease.

    Science.gov (United States)

    Silva, Tatiana Rocha; de Resende, Luciana Macedo; Santos, Marco Aurélio Rocha

    The vestibular evoked myogenic potential is a potential of mean latency that measures the muscle response to auditory stimulation. This potential can be generated from the contraction of the sternocleidomastoid muscle and also from the contraction of extraocular muscles in response to high-intensity sounds. This study presents a combined or simultaneous technique of cervical and ocular vestibular evoked myogenic potential in individuals with changes in the vestibular system, for use in otoneurologic diagnosis. To characterize the records and analyze the results of combined cervical and ocular VEMP in individuals with vestibular hyporeflexia and in those with Ménière's disease. The study included 120 subjects: 30 subjects with vestibular hyporeflexia, 30 with Ménière's disease, and 60 individuals with normal hearing. Data collection was performed by simultaneously recording the cervical and ocular vestibular evoked myogenic potential. There were differences between the study groups (individuals with vestibular hyporeflexia and individuals with Ménière's disease) and the control group for most of wave parameters in combined cervical and ocular vestibular evoked myogenic potential. For cervical vestibular evoked myogenic potential, it was observed that the prolongation of latency of the P13 and N23 waves was the most frequent finding in the group with vestibular hyporeflexia and in the group with Ménière's disease. For ocular vestibular evoked myogenic potential, prolonged latency of N10 and P15 waves was the most frequent finding in the study groups. Combined cervical and ocular vestibular evoked myogenic potential presented relevant results for individuals with vestibular hyporeflexia and for those with Ménière's disease. There were differences between the study groups and the control group for most of the wave parameters in combined cervical and ocular vestibular evoked myogenic potential. Copyright © 2016 Associação Brasileira de Otorrinolaringologia

  2. Immortalization of mouse myogenic cells can occur without loss of p16INK4a, p19ARF, or p53 and is accelerated by inactivation of Bax

    Directory of Open Access Journals (Sweden)

    Kravetz Amanda J

    2004-01-01

    Full Text Available Abstract Background Upon serial passaging of mouse skeletal muscle cells, a small number of cells will spontaneously develop the ability to proliferate indefinitely while retaining the ability to differentiate into multinucleate myotubes. Possible gene changes that could underlie myogenic cell immortalization and their possible effects on myogenesis had not been examined. Results We found that immortalization occurred earlier and more frequently when the myogenic cells lacked the pro-apoptotic protein Bax. Furthermore, myogenesis was altered by Bax inactivation as Bax-null cells produced muscle colonies with more nuclei than wild-type cells, though a lower percentage of the Bax-null nuclei were incorporated into multinucleate myotubes. In vivo, both the fast and slow myofibers in Bax-null muscles had smaller cross-sectional areas than those in wild-type muscles. After immortalization, both Bax-null and Bax-positive myogenic cells expressed desmin, retained the capacity to form multinucleate myotubes, expressed p19ARF protein, and retained p53 functions. Expression of p16INK4a, however, was found in only about half of the immortalized myogenic cell lines. Conclusions Mouse myogenic cells can undergo spontaneous immortalization via a mechanism that can include, but does not require, loss of p16INK4a, and also does not require inactivation of p19ARF or p53. Furthermore, loss of Bax, which appears to be a downstream effector of p53, accelerates immortalization of myogenic cells and alters myogenesis.

  3. Genome-wide association analysis of tolerance to methylmercury toxicity in Drosophila implicates myogenic and neuromuscular developmental pathways.

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    Sara L Montgomery

    Full Text Available Methylmercury (MeHg is a persistent environmental toxin present in seafood that can compromise the developing nervous system in humans. The effects of MeHg toxicity varies among individuals, despite similar levels of exposure, indicating that genetic differences contribute to MeHg susceptibility. To examine how genetic variation impacts MeHg tolerance, we assessed developmental tolerance to MeHg using the sequenced, inbred lines of the Drosophila melanogaster Genetic Reference Panel (DGRP. We found significant genetic variation in the effects of MeHg on development, measured by eclosion rate, giving a broad sense heritability of 0.86. To investigate the influence of dietary factors, we measured MeHg toxicity with caffeine supplementation in the DGRP lines. We found that caffeine counteracts the deleterious effects of MeHg in the majority of lines, and there is significant genetic variance in the magnitude of this effect, with a broad sense heritability of 0.80. We performed genome-wide association (GWA analysis for both traits, and identified candidate genes that fall into several gene ontology categories, with enrichment for genes involved in muscle and neuromuscular development. Overexpression of glutamate-cysteine ligase, a MeHg protective enzyme, in a muscle-specific manner leads to a robust rescue of eclosion of flies reared on MeHg food. Conversely, mutations in kirre, a pivotal myogenic gene identified in our GWA analyses, modulate tolerance to MeHg during development in accordance with kirre expression levels. Finally, we observe disruptions of indirect flight muscle morphogenesis in MeHg-exposed pupae. Since the pathways for muscle development are evolutionarily conserved, it is likely that the effects of MeHg observed in Drosophila can be generalized across phyla, implicating muscle as an additional hitherto unrecognized target for MeHg toxicity. Furthermore, our observations that caffeine can ameliorate the toxic effects of MeHg show

  4. Subclinical vestibular dysfunction in migraine patients: a preliminary study of ocular and rectified cervical vestibular evoked myogenic potentials

    OpenAIRE

    Kim, Chul-Ho; Jang, Min-Uk; Choi, Hui-Chul; Sohn, Jong-Hee

    2015-01-01

    Background Many studies have identified various vestibular symptoms and laboratory abnormalities in migraineurs. Although the vestibular tests may be abnormal, the changes may exist without vestibular symptoms. To date, vestibular-evoked myogenic potential (VEMP) has been the easiest and simplest test for measuring vestibular function in clinical practice. Cervical VEMP (cVEMP) represents a vestibulo-collic reflex, whereas ocular VEMP (oVEMP) reflects a vestibulo-ocular pathway. Therefore, we...

  5. Ocular vestibular evoked myogenic potentials induced by bone-conducted vibration in patients with unilateral inner ear disease

    OpenAIRE

    Nagai, Noriko; Ogawa, Yasuo; Hagiwara, Akira; Otsuka, Koji; Inagaki, Taro; Shimizu, Shigetaka; Suzuki, Mamoru

    2013-01-01

    Conclusion Patients with vestibular neuritis (VN) with complete canal paresis (CP) showed a higher rate of abnormal ocular vestibular evoked myogenic potential (oVEMP) than those with partial CP. From these results, it is speculated that the superior vestibular nerve function mainly affects oVEMP. Significant correlation was found between the grades of the hearing outcome and oVEMP in sudden sensorineural hearing loss (SSHL). Objective We attempted to correlate the results of oVEMP with the r...

  6. Vestibular evoked myogenic potentials: an overview Potencial evocado miogênico vestibular: uma visão geral

    OpenAIRE

    Renato Cal; Fayez Bahmad Jr

    2009-01-01

    The vestibular evoked myogenic potential (VEMP) test is a relatively new diagnostic tool that is in the process of being investigated in patients with specific vestibular disorders. Briefly, the VEMP is a biphasic response elicited by loud clicks or tone bursts recorded from the tonically contracted sternocleidomastoid muscle, being the only resource available to assess the function of the saccule and the lower portion of the vestibular nerve. AIM: In this review, we shall highlight the histo...

  7. Comparison Acoustically Evoked Short Latency Negative Response with Vestibular Evoked Myogenic Potential in Adults with Profound Hearing Loss

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    Maryam Ramezani

    2012-03-01

    Full Text Available Background and Aim: A negative deflection with a 3-4 ms latency period has been reported to exist within the auditory brainstem response of some patients with profound hearing loss following a strong acoustic stimulus. This deflection, namingly the n3 or the acoustically evoked short latency negative response is assumed to be a vestibular-evoked potential, especially of saccular origin. Since the myogenic potential is also saccular in origin, the purpose of the present study was to investigate the relationship between these two tests in adults with profound hearing loss.Methods: The present cross sectional study was performed on 20 profoundly deaf volunteers(39 ears who aged between 18-40 years old, randomly selected from available deaf adults in Tehran. The auditory brainstem response of all subjects was recorded following a 1000 Hz tone burst in 70-100dB nHL. Subjects were also tested for vestibular evoked myogenic potential.Results: Only 34 of 39 ears recorded myogenic potential that negative response was recorded in 27 of 34 ears with normal p13 and n23. In seven ears with normal p13 and n23, the negative response was absent. In 3 ears with no p13 and n23, the negative response was observed, and two none.Conclusion: In view of the high prevalance of the negative response in profoundly deaf ears with normal p13 and n23, it could be concluded that the negative response can be used when for any reason, it is not possible to record myogenic potential and be considered as a new test in vestibular test battery.

  8. Role of miR-181a-5p and endoplasmic reticulum stress in the regulation of myogenic differentiation.

    Science.gov (United States)

    Wei, Yingying; Tao, Xuelian; Xu, Huaming; Chen, Yan; Zhu, Li; Tang, Guoqing; Li, Mingzhou; Jiang, Anan; Shuai, Surong; Ma, Jideng; Jin, Long; Wen, Anxiang; Wang, Qin; Zhu, Guangxiang; Xie, Meng; Wu, Jiayun; He, Tao; Jiang, Yanzhi; Li, Xuewei

    2016-10-30

    Accumulating evidence has indicated that microRNAs (miRNAs) and endoplasmic reticulum (ER) stress play critical roles in myoblast differentiation. However, the regulation roles of miRNAs and ER stress in myogenic differentiation have not been fully revealed and need to be further studied. Here, we discovered that the expression levels of miR-181a-5p were strongly upregulated during C2C12 cell differentiation. miR-181a-5p overexpression promoted ER stress and differentiation of C2C12 cells, which was accompanied by increasing expression levels of marker genes related to ER stress-mediated apoptosis and myogenic differentiation. Opposite results were observed after inhibition of the miR-181a-5p expression. The gain- and loss-of-function experiments on C2C12 cells showed that miR-181a-5p affected the development of muscle fiber type, but had no significant influence on C2C12 cell proliferation. In the ER-stressed C2C12 cells induced by thapsigargin (Tg), the expression levels of both miR-181a-5p and marker genes related to ER stress and myogenesis were upregulated. In the ER-stressed C2C12 cells and porcine muscle fibroblast (PMF) cells pretreated with Tg, we found that miR-181a-5p targeted glucose-regulated protein, 78kDa/binding immunoglobulin protein (GRP78/BIP), and influenced cell apoptosis. In conclusion, these results indicate that miR-181a-5p and ER stress have positive synergistic effects on myogenic differentiation by increasing the expression levels of myogenic differentiation key genes and activating the ER stress-mediated apoptosis signaling pathway.

  9. Vestibular-evoked myogenic potentials and subjective visual vertical testing in patients with vitamin D deficiency/insufficiency.

    Science.gov (United States)

    Sanyelbhaa, Hossam; Sanyelbhaa, Ahmed

    2015-11-01

    Otolith function in subjects with vitamin D deficiency/insufficiency is investigated through vestibular-evoked myogenic potentials (VEMP) and subjective visual vertical (SVV) testing. The study group included 62 patients with vitamin D deficiency/insufficiency (30 females, 32 males), with age range 24-56 years (40.6 ± 9.1). The control group included 44 healthy volunteers of similar age and gender distribution. The entire study group had: (1) serum level of 25-hydroxyvitamin D -1; (3) normal middle ear function; (4) Age is ≤60 years. All subjects enrolled in the current study underwent audiovestibular evaluation consisting of pure-tone audiometry, immittancemetry, cervical VEMP (cVEMP), ocular VEMP (oVEMP), and SSV. The entire control group had normal cVEMP, two subjects had abnormal oVEMP. Thirty-three subjects (53%) in the study group had abnormal oVEMP and 31 subjects (50%) had abnormal cVEMP. Forty-one (66%) had abnormal VEMP when abnormal VEMP was considered as either abnormal oVEMP or cVEMP. The entire control and study groups had normal SSV test results. Vitamin D deficiency may be associated with development of otolith dysfunction affecting both the utricle and saccule. This was suggested by the high prevalence of abnormal ocular vestibular-evoked myogenic potentials (oVEMP) and cervical vestibular-evoked myogenic potentials (cVEMP) in the study group.

  10. Effect of Low Power Laser Irradiation on the Ability of Cell Growth and Myogenic Differentiation of Myoblasts Cultured In Vitro

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    Cui-Ping Zhang

    2014-01-01

    Full Text Available As a therapeutic modality, low power laser irradiation (LPLI has been used clinically in the treatment of skeletal muscle injuries and other myopathic conditions, but the cellular and molecular mechanisms attributed to this therapy were still unclear. Myoblasts are a type of myogenic stem cells quiescence in mature skeletal muscle fibers and are considered as the source cells during the regenerating process. The purpose of this paper was to investigate the effects of LPLI on the proliferation and myogenic differentiation of the cultured myoblasts and to find out the major candidates responsible for LPLI-induced muscle regeneration in vivo. In this study, primary rat myoblasts were exposed to helium-neon (He-Ne laser. Cell proliferation, differentiation, and the cellular responses to LPLI were monitored by using morphological observation and molecular biological methods. It was found that LPLI at a certain fluence could increase the cell growth potential for myoblasts and further induce more cells entering into S phase of the mitotic cycle as indicated by high levels of bromodeoxyuridine (BrdU incorporation, while at the same time inhibiting their in vitro differentiation and decreasing the expression of myogenic regulatory genes to a certain extent. Taken together, these results provide experimental evidence for the clinical applications of LPLI in regenerating skeletal muscle.

  11. Sparing of extraocular muscle in aging and muscular dystrophies: A myogenic precursor cell hypothesis

    Energy Technology Data Exchange (ETDEWEB)

    Kallestad, Kristen M.; Hebert, Sadie L.; McDonald, Abby A.; Daniel, Mark L.; Cu, Sharon R.; McLoon, Linda K., E-mail: mcloo001@tc.umn.edu

    2011-04-01

    The extraocular muscles (EOM) are spared from pathology in aging and many forms of muscular dystrophy. Despite many studies, this sparing remains an enigma. The EOM have a distinct embryonic lineage compared to somite-derived muscles, and we have shown that they continuously remodel throughout life, maintaining a population of activated satellite cells even in aging. These data suggested the hypothesis that there is a population of myogenic precursor cells (mpcs) in EOM that is different from those in limb, with either elevated numbers of stem cells and/or mpcs with superior proliferative capacity compared to mpcs in limb. Using flow cytometry, EOM and limb muscle mononuclear cells were compared, and a number of differences were seen. Using two different cell isolation methods, EOM have significantly more mpcs per mg muscle than limb skeletal muscle. One specific subpopulation significantly increased in EOM compared to limb was positive for CD34 and negative for Sca-1, M-cadherin, CD31, and CD45. We named these the EOMCD34 cells. Similar percentages of EOMCD34 cells were present in both newborn EOM and limb muscle. They were retained in aged EOM, whereas the population decreased significantly in adult limb muscle and were extremely scarce in aged limb muscle. Most importantly, the percentage of EOMCD34 cells was elevated in the EOM from both the mdx and the mdx/utrophin{sup -/-} (DKO) mouse models of DMD and extremely scarce in the limb muscles of these mice. In vitro, the EOMCD34 cells had myogenic potential, forming myotubes in differentiation media. After determining a media better able to induce proliferation in these cells, a fusion index was calculated. The cells isolated from EOM had a 40% higher fusion index compared to the same cells isolated from limb muscle. The EOMCD34 cells were resistant to both oxidative stress and mechanical injury. These data support our hypothesis that the EOM may be spared in aging and in muscular dystrophies due to a

  12. Sparing of extraocular muscle in aging and muscular dystrophies: a myogenic precursor cell hypothesis.

    Science.gov (United States)

    Kallestad, Kristen M; Hebert, Sadie L; McDonald, Abby A; Daniel, Mark L; Cu, Sharon R; McLoon, Linda K

    2011-04-01

    The extraocular muscles (EOM) are spared from pathology in aging and many forms of muscular dystrophy. Despite many studies, this sparing remains an enigma. The EOM have a distinct embryonic lineage compared to somite-derived muscles, and we have shown that they continuously remodel throughout life, maintaining a population of activated satellite cells even in aging. These data suggested the hypothesis that there is a population of myogenic precursor cells (mpcs) in EOM that is different from those in limb, with either elevated numbers of stem cells and/or mpcs with superior proliferative capacity compared to mpcs in limb. Using flow cytometry, EOM and limb muscle mononuclear cells were compared, and a number of differences were seen. Using two different cell isolation methods, EOM have significantly more mpcs per mg muscle than limb skeletal muscle. One specific subpopulation significantly increased in EOM compared to limb was positive for CD34 and negative for Sca-1, M-cadherin, CD31, and CD45. We named these the EOMCD34 cells. Similar percentages of EOMCD34 cells were present in both newborn EOM and limb muscle. They were retained in aged EOM, whereas the population decreased significantly in adult limb muscle and were extremely scarce in aged limb muscle. Most importantly, the percentage of EOMCD34 cells was elevated in the EOM from both the mdx and the mdx/utrophin(-/-) (DKO) mouse models of DMD and extremely scarce in the limb muscles of these mice. In vitro, the EOMCD34 cells had myogenic potential, forming myotubes in differentiation media. After determining a media better able to induce proliferation in these cells, a fusion index was calculated. The cells isolated from EOM had a 40% higher fusion index compared to the same cells isolated from limb muscle. The EOMCD34 cells were resistant to both oxidative stress and mechanical injury. These data support our hypothesis that the EOM may be spared in aging and in muscular dystrophies due to a subpopulation

  13. Accumulation of point mutations and reassortment of genomic RNA segments are involved in the microevolution of Puumala hantavirus in a bank vole (Myodes glareolus) population.

    Science.gov (United States)

    Razzauti, Maria; Plyusnina, Angelina; Henttonen, Heikki; Plyusnin, Alexander

    2008-07-01

    The genetic diversity of Puumala hantavirus (PUUV) was studied in a local population of its natural host, the bank vole (Myodes glareolus). The trapping area (2.5 x 2.5 km) at Konnevesi, Central Finland, included 14 trapping sites, at least 500 m apart; altogether, 147 voles were captured during May and October 2005. Partial sequences of the S, M and L viral genome segments were recovered from 40 animals. Seven, 12 and 17 variants were detected for the S, M and L sequences, respectively; these represent new wild-type PUUV strains that belong to the Finnish genetic lineage. The genetic diversity of PUUV strains from Konnevesi was 0.2-4.9 % for the S segment, 0.2-4.8 % for the M segment and 0.2-9.7 % for the L segment. Most nucleotide substitutions were synonymous and most deduced amino acid substitutions were conservative, probably due to strong stabilizing selection operating at the protein level. Based on both sequence markers and phylogenetic clustering, the S, M and L sequences could be assigned to two groups, 'A' and 'B'. Notably, not all bank voles carried S, M and L sequences belonging to the same group, i.e. S(A)M(A)L(A) or S(B)M(B)L(B). A substantial proportion (8/40, 20 %) of the newly characterized PUUV strains possessed reassortant genomes such as S(B)M(A)L(A), S(A)M(B)L(B) or S(B)M(A)L(B). These results suggest that at least some of the PUUV reassortants are viable and can survive in the presence of their parental strains.

  14. Mercury in soil, earthworms and organs of voles Myodes glareolus and shrew Sorex araneus in the vicinity of an industrial complex in Northwest Russia (Cherepovets).

    Science.gov (United States)

    Komov, V T; Ivanova, E S; Poddubnaya, N Y; Gremyachikh, V A

    2017-03-01

    The characteristic properties of uptake and distribution of mercury in terrestrial ecosystems have received much lesser attention compared to aquatic particularly in Russia. Terrestrial ecosystems adjacent to large industrial manufactures-potential sources of mercury inflow into the environment frequently remain unstudied. This is the first report on mercury (Hg) levels in the basic elements of terrestrial ecosystems situated close to a large metallurgical complex.Mean values of mercury concentration (mg Hg/kg dry weight) in the vicinity of city of Cherepovets were the following: 0.056 ± 0.033-in the humus layer of soil; 0.556 ± 0.159-in earthworms; in the organs of voles Myodes glareolus (kidneys-0.021 ± 0.001; liver-0.014 ± 0.003; muscle-0.014 ± 0.001; brain-0.008 ± 0.002); in the organs of shrew Sorex araneus (kidneys-0.191 ± 0.016; liver-0.124 ± 0.011; muscle-0.108 ± 0.009; brain-0.065 ± 0.000). Correlation dependences between Hg content in soil and earthworms (r s  = 0.85, p < 0.01) as well as soil and all studied shrews' organs (rs = 0.44-0.58; p ≤ 0.01) were found.The results obtained evidence for a strong trophic link in the bioaccumulation of Hg in terrestrial food webs. Despite the vicinity to a large metallurgical complex, mercury content in the studied objects was significantly lower than values of corresponding parameters in the soils and biota from industrial (polluted) areas of Great Britain, the USA, and China.

  15. 眼肌前庭诱发肌源性电位的研究现状及展望%Present situation and development of ocular vestibular-evoked myogenic potential

    Institute of Scientific and Technical Information of China (English)

    胡娟; 张青

    2013-01-01

    Myogenic potentials evoked by air conducted sound (ACS) , bone conducted vibration (BCV) or galvanic pulses can be recorded with surface electrodes over contracted muscles. These myogenic potentials are of vestibular origin (utricle and saccule) and so these potentials are called vestibular evoked myogenic potentials (VEMPs). Since the vestibular system has projections to many muscle systems, there are many such VEMPs. In this review, we discuss the generated origin, response pathway, waveform characteristics and clinical application of ocular vestibular-evoked myogenic potential(oVEMP).

  16. Potenciais miogênicos evocados vestibulares: metodologias de registro em homens e cobaias Vestibular evoked myogenic potential: recording methods in humans and guinea pigs

    Directory of Open Access Journals (Sweden)

    Aline Cabral de Oliveira

    2008-10-01

    Full Text Available O potencial miogênico evocado vestibular (VEMP é um teste clínico que avalia a função vestibular através de um reflexo vestíbulo-cervical inibitório captado nos músculos do corpo em resposta à estimulação acústica de alta intensidade. OBJETIVO: Verificar e analisar os diversos métodos de registro dos potenciais miogênicos evocados vestibulares no homem e em cobaias. MATERIAL E MÉTODO: Realizou-se busca eletrônica nas bases de dados MEDLINE, LILACS, SCIELO e COCHRANE. RESULTADOS: Foram verificadas divergências quanto às formas de registro dos potenciais miogênicos evocados vestibulares, relacionadas com os seguintes fatores: posição do paciente no momento do registro, tipo de estímulo sonoro utilizado (clicks ou tone bursts, parâmetros para a promediação dos estímulos (intensidade, freqüência, tempo de apresentação, filtros, ganho de amplificação das respostas e janelas para captação dos estímulos, tipo de fone utilizado e forma de apresentação dos estímulos (monoaural ou binaural, ipsi ou contralateral. CONCLUSÃO: Não existe consenso na literatura quanto ao melhor método de registro dos potenciais evocados miogênicos vestibulares, havendo necessidade de pesquisas mais específicas para comparação entre estes registros e a definição de um modelo padrão para a utilização na prática clínica.The vestibular evoked myogenic potential (VEMP is a clinical test that assess the vestibular function by means of an inhibitory vestibulo-neck reflex, recorded in body muscles in response to high intensity acoustic stimuli. AIM: To check and analyze the different methods used to record VEMPs in humans and in guinea pigs. MATERIALS AND METHODS: We researched the following databases: MEDLINE, LILACS, SCIELO and COCHRANE. RESULTS: we noticed discrepancies in relation to the ways used to record the vestibular evoked myogenic potentials in relation to the following factors: patient position at the time of recording

  17. miR-206 represses hypertrophy of myogenic cells but not muscle fibers via inhibition of HDAC4.

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    Catherine E Winbanks

    Full Text Available microRNAs regulate the development of myogenic progenitors, and the formation of skeletal muscle fibers. However, the role miRNAs play in controlling the growth and adaptation of post-mitotic musculature is less clear. Here, we show that inhibition of the established pro-myogenic regulator miR-206 can promote hypertrophy and increased protein synthesis in post-mitotic cells of the myogenic lineage. We have previously demonstrated that histone deacetylase 4 (HDAC4 is a target of miR-206 in the regulation of myogenic differentiation. We confirmed that inhibition of miR-206 de-repressed HDAC4 accumulation in cultured myotubes. Importantly, inhibition of HDAC4 activity by valproic acid or sodium butyrate prevented hypertrophy of myogenic cells otherwise induced by inhibition of miR-206. To test the significance of miRNA-206 as a regulator of skeletal muscle mass in vivo, we designed recombinant adeno-associated viral vectors (rAAV6 vectors expressing miR-206, or a miR-206 "sponge," featuring repeats of a validated miR-206 target sequence. We observed that over-expression or inhibition of miR-206 in the muscles of mice decreased or increased endogenous HDAC4 levels respectively, but did not alter muscle mass or myofiber size. We subsequently manipulated miR-206 levels in muscles undergoing follistatin-induced hypertrophy or denervation-induced atrophy (models of muscle adaptation where endogenous miR-206 expression is altered. Vector-mediated manipulation of miR-206 activity in these models of cell growth and wasting did not alter gain or loss of muscle mass respectively. Our data demonstrate that although the miR-206/HDAC4 axis operates in skeletal muscle, the post-natal expression of miR-206 is not a key regulator of basal skeletal muscle mass or specific modes of muscle growth and wasting. These studies support a context-dependent role of miR-206 in regulating hypertrophy that may be dispensable for maintaining or modifying the adult skeletal

  18. miR-206 represses hypertrophy of myogenic cells but not muscle fibers via inhibition of HDAC4.

    Science.gov (United States)

    Winbanks, Catherine E; Beyer, Claudia; Hagg, Adam; Qian, Hongwei; Sepulveda, Patricio V; Gregorevic, Paul

    2013-01-01

    microRNAs regulate the development of myogenic progenitors, and the formation of skeletal muscle fibers. However, the role miRNAs play in controlling the growth and adaptation of post-mitotic musculature is less clear. Here, we show that inhibition of the established pro-myogenic regulator miR-206 can promote hypertrophy and increased protein synthesis in post-mitotic cells of the myogenic lineage. We have previously demonstrated that histone deacetylase 4 (HDAC4) is a target of miR-206 in the regulation of myogenic differentiation. We confirmed that inhibition of miR-206 de-repressed HDAC4 accumulation in cultured myotubes. Importantly, inhibition of HDAC4 activity by valproic acid or sodium butyrate prevented hypertrophy of myogenic cells otherwise induced by inhibition of miR-206. To test the significance of miRNA-206 as a regulator of skeletal muscle mass in vivo, we designed recombinant adeno-associated viral vectors (rAAV6 vectors) expressing miR-206, or a miR-206 "sponge," featuring repeats of a validated miR-206 target sequence. We observed that over-expression or inhibition of miR-206 in the muscles of mice decreased or increased endogenous HDAC4 levels respectively, but did not alter muscle mass or myofiber size. We subsequently manipulated miR-206 levels in muscles undergoing follistatin-induced hypertrophy or denervation-induced atrophy (models of muscle adaptation where endogenous miR-206 expression is altered). Vector-mediated manipulation of miR-206 activity in these models of cell growth and wasting did not alter gain or loss of muscle mass respectively. Our data demonstrate that although the miR-206/HDAC4 axis operates in skeletal muscle, the post-natal expression of miR-206 is not a key regulator of basal skeletal muscle mass or specific modes of muscle growth and wasting. These studies support a context-dependent role of miR-206 in regulating hypertrophy that may be dispensable for maintaining or modifying the adult skeletal muscle phenotype

  19. Cervical vestibular evoked myogenic potentials in children with spastic cerebral palsy and normal children with 7-12 years of age

    Directory of Open Access Journals (Sweden)

    Nazila Akbarfahimi

    2014-10-01

    Full Text Available Background and Aim: Cervical vestibular-evoked myogenic potential (cVEMP is one of the diagnostic tests used in assessing vestibular function. Two aims of this study were to investigate implications of cervical vestibular evoked myogenic potential in children with spastic cerebral palsy (7-12 years, and to compare vestibular function in these children and normal children.Methods: In this cross-sectional study, myogenic potential was recorded in 31 children with spastic cerebral palsy (8 girls, 23 boys,7-12 years of age, with mean age of 8.77 years old and standard deviation of 1.52 years and 31 normal children (13 girls, 18 boys with mean age of 8.77 years and standard deviation of 1.52 years. Cervical vestibular evoked myogenic potential was recorded with 500 Hz tone burst. The recorded parameters included p13 and n23 latency, p13-n23 pick to pick amplitude, and threshold.Results: Myogenic Potential was recorded in 31 normal children. They had bilateral responses. In children with cerebral palsy, 21 children showed bilateral responses, 3 children had only right-sided responses, 8 children had only left-sided responses, and two children did not show any responses. The statistical significant differences were shown between the two groups in n23, p13-n23 pick to pick amplitude, and threshold (p<0.05.Conclusion: These findings showed that cervical vestibular evoked myogenic potential can be used in children with cerebral palsy. There were significant differences in myogenic potential parameters between the two groups. More studies are needed to investigate the causes of these differences.

  20. Effect of caffeine on cervical vestibular-evoked myogenic potential in healthy individuals

    Directory of Open Access Journals (Sweden)

    Ana Maria Almeida de Sousa

    2014-06-01

    Full Text Available INTRODUCTION: Caffeine is the most common psychoactive drug in use around the world and is found at different concentrations in a variety of common food items. Clinically, a strong association between caffeine consumption and diseases of the vestibular system has been established. Cervical vestibular-evoked myogenic potential (cVEMP is an electrophysiological test that is used to assess the sacculocollic pathway by measuring changes in the vestialibulocollic reflex. AIM: The present study aimed to evaluate the effect of an acute dose of caffeine on the vestibulocollic reflex by using cVEMP. METHOD: A prospective experimental study was performed in which healthy volunteers were submitted to the test before and after the intake of 420 mg of caffeine. The following parameters were compared: p13 and n23 latencies and p13-n23 amplitude. RESULT: No statistically significant difference was found in the test results before and after caffeine use. CONCLUSION: The vestibulocollic reflex is not altered by caffeine intake.

  1. Stem Cell Differentiation Toward the Myogenic Lineage for Muscle Tissue Regeneration: A Focus on Muscular Dystrophy.

    Science.gov (United States)

    Ostrovidov, Serge; Shi, Xuetao; Sadeghian, Ramin Banan; Salehi, Sahar; Fujie, Toshinori; Bae, Hojae; Ramalingam, Murugan; Khademhosseini, Ali

    2015-12-01

    Skeletal muscle tissue engineering is one of the important ways for regenerating functionally defective muscles. Among the myopathies, the Duchenne muscular dystrophy (DMD) is a progressive disease due to mutations of the dystrophin gene leading to progressive myofiber degeneration with severe symptoms. Although current therapies in muscular dystrophy are still very challenging, important progress has been made in materials science and in cellular technologies with the use of stem cells. It is therefore useful to review these advances and the results obtained in a clinical point of view. This article focuses on the differentiation of stem cells into myoblasts, and their application in muscular dystrophy. After an overview of the different stem cells that can be induced to differentiate into the myogenic lineage, we introduce scaffolding materials used for muscular tissue engineering. We then described some widely used methods to differentiate different types of stem cell into myoblasts. We highlight recent insights obtained in therapies for muscular dystrophy. Finally, we conclude with a discussion on stem cell technology. We discussed in parallel the benefits brought by the evolution of the materials and by the expansion of cell sources which can differentiate into myoblasts. We also discussed on future challenges for clinical applications and how to accelerate the translation from the research to the clinic in the frame of DMD.

  2. Vestibular evoked myogenic potential eliciting in normal subjects: comparison of four different methods.

    Science.gov (United States)

    Eleftheriadou, Anna; Deftereos, Spyros N; Zarikas, Vasilios; Panagopoulos, Grigoris; Korres, Stavros; Sfetsos, Sotirios; Karageorgiou, Klimentini L; Ferekidou, Elisa; Kandiloros, Dimitrios

    2008-10-01

    Vestibular evoked myogenic potential (VEMP) recording is a new method for testing the otolith receptors and vestibulospinal pathways. The aim of this study was to evaluate the characteristics of VEMP using four different techniques to find reasons to prefer one type of recording over the others. Twenty healthy persons, 10 males and 10 females with ages ranging from 20 to 57 years (mean age 41 years), were enrolled in this study. Eliciting of VEMPs by using monaural or binaural acoustic stimulation and unilateral or bilateral SCM contraction was evaluated; 105 dB NHL acoustic stimulation consisting of 145 dB rarefaction clicks was applied. Latencies of p13, n23, n34, p44 peaks; amplitudes p13-n23 and n34-p44; and interaural amplitude differences (IADs) were assessed. All four methods elicited constant and evident waveforms. The reliability coefficients of amplitudes were high for all four methods and for both waves. However, the higher scores of reliability appeared for the monaural-ipsilateral recording. The results indicated no statistically significant difference between the right and left sides for all four types of VEMP eliciting. No correlation was found between IAD13-23 and IAD34-44 for all four methods. Statistically significant differences were found only for n23 latency among the four methods. Although no evidence to reject or strongly favour a specific method was found, the monaural-ipsilateral recording was associated with some advantages.

  3. Expression of myogenes in longissimus dorsi muscle during prenatal development in commercial and local Piau pigs

    Science.gov (United States)

    dos Reis, Evelyze Pinheiro; Paixão, Débora Martins; Brustolini, Otávio José Bernardes; Silva, Fabyano Fonseca e; Silva, Walmir; de Araújo, Flávio Marcos Gomes; Salim, Anna Christina de Matos; Oliveira, Guilherme; Guimarães, Simone Eliza Facioni

    2016-01-01

    Abstract This study used qRT-PCR to examine variation in the expression of 13 myogenes during muscle development in four prenatal periods (21, 40, 70 and 90 days post-insemination) in commercial (the three-way Duroc, Landrace and Large-White cross) and local Piau pig breeds that differ in muscle mass. There was no variation in the expression of the CHD8, EID2B, HIF1AN, IKBKB, RSPO3, SOX7 and SUFU genes at the various prenatal ages or between breeds. The MAP2K1 and RBM24 genes showed similar expression between commercial and Piau pigs but greater expression (p < 0.05) in at least one prenatal period. Pair-wise comparisons of prenatal periods in each breed showed that only the CSRP3, LEF1, MRAS and MYOG genes had higher expression (p < 0.05) in at least one prenatal period in commercial and Piau pigs. Overall, these results identified the LEF1 gene as a primary candidate to account for differences in muscle mass between the pig breeds since activation of this gene may lead to greater myoblast fusion in the commercial breed compared to Piau pigs. Such fusion could explain the different muscularity between breeds in the postnatal periods. PMID:27801482

  4. Electrical events underlying organized myogenic contractions of the guinea pig stomach.

    Science.gov (United States)

    Hirst, G David S; Edwards, Frank R

    2006-11-01

    The stomach generates a characteristic pattern of coordinated activity whereby rings of contraction regularly start in the corpus and migrate slowly down the stomach to the duodenum. This behaviour persists after isolating the stomach and after blocking nervous activity; hence the response is myogenic, resulting from organized contractions of smooth muscle cells lying in the stomach wall. Each ring of contraction is triggered by a long lasting wave of depolarization, termed a slow wave. Slow waves are now known to be generated by sets of interstitial cells of Cajal (ICC), which intermingle with gastric smooth muscle cells. This article describes some studies which identify the roles played by ICC in the on-going generation of coordinated gastric movements. Intramuscular ICC in the corpus generate slow waves and these provide the dominant pacemaker frequency in the stomach. Corporal slow waves, in turn, activate a network of myenteric ICC, which starts in the antrum and slowly conducts waves of depolarization down the stomach. As these waves pass over bundles of circularly orientated muscle cells, they activate a set of intramuscular ICC which lie in the circular muscle layer: these generate slow waves that rapidly spread radially, so triggering each ring of contraction.

  5. Correlation between caloric and ocular vestibular evoked myogenic potential test results.

    Science.gov (United States)

    Huang, Chi-Hsuan; Wang, Shou-Jen; Young, Yi-Ho

    2012-02-01

    The ocular vestibular evoked myogenic potential (o-VEMP) test results correlate significantly with caloric test results for patients with acoustic neuroma (AN), but not for patients with Meniere's disease (MD), indicating that the o-VEMP test may replace the caloric test for evaluating the vestibular nerve from which the AN arises. Conversely, the caloric, o-VEMP, and cervical VEMP (c-VEMP) tests should be performed to map lesion sites in the vestibular labyrinth. This study performed caloric, o-VEMP, and c-VEMP tests on patients with central and peripheral vestibular disorders to investigate their relationships. In all, 66 patients comprising 16 with unilateral AN and 50 with unilateral definite MD were enrolled. All patients underwent caloric, o-VEMP, and c-VEMP tests. In the AN group, the caloric test identified canal paresis and caloric areflexia in 10 ears, while the o-VEMP and c-VEMP tests identified abnormal (absent or delayed) responses in 12 and 11 ears, respectively. A significant correlation existed between caloric and o-VEMP test results, but not between caloric and c-VEMP test results, or between o-VEMP and c-VEMP test results. For the MD group, abnormal caloric, o-VEMP, and c-VEMP test results were obtained for 24%, 44%, and 38% of hydropic ears, respectively. No correlation existed between any two test results.

  6. Ocular vestibular evoked myogenic potentials to air conduction (AC oVEMP): useful in clinical practice?

    Science.gov (United States)

    Walther, L E; Rogowski, M; Hörmann, K; Schaaf, H; Löhler, J

    2011-01-01

    Cervical vestibular-evoked myogenic potential (cVEMP) and ocular VEMP (oVEMP) stimuli can be used to measure otolith function using air (AC) and bone conducted (BC) stimuli. Cervical VEMPs reflect saccular function and can be recorded using air conduction (AC), whereas oVEMPs reflect probably predominantly utricular function. Air- and bone-conducted vibration can be used, because AC oVEMP methodology seems to be fast and simple in clinical practice to measure otolith function. In this study we discuss the advantages and problems of AC oVEMP stimulation. AC oVEMP can be easily and quickly obtained within a few seconds. N10 (first negative peak) and p15 (first positive peak) latencies may be used as parameters for clinical interpretation but amplitude fluctuations are relatively large. For daily clinical use of VEMP visualization in a normogram seems feasible. Especially the AC oVEMP methodology (100 dB nHL, tone burst 500 Hz) is fast and efficient in clinical practice to measure otolith function, predominantly utricular function. Copyright © 2011 Polish Otolaryngology Society. Published by Elsevier Urban & Partner (Poland). All rights reserved.

  7. Development of ocular vestibular-evoked myogenic potentials in small children.

    Science.gov (United States)

    Wang, Shou-Jen; Hsieh, Wu-Shiun; Young, Yi-Ho

    2013-02-01

    This study investigated the development of otolithic-ocular reflex in small children (ocular vestibular-evoked myogenic potential (oVEMP) test. Prospective study. Twenty full-term newborns (group A), 15 children aged 1 to 3 years (group B), and 15 children aged 4 to 13 years (group C) were enrolled in this study. All children underwent auditory brainstem response testing or audiometry, and the oVEMP test. All subjects had normal hearing. Typical biphasic oVEMP waveforms were not observed in the 20 newborns, but were present in six (40%) of 15 children aged 1 to 3 years and all (100%) children aged 4 to 13 years, exhibiting a significant difference. In group B, except for the nine children aged 12 to 24 months, the remaining six children, aged 25 to 47 months, had clear oVEMPs, with the mean nI latency and nI-pI amplitude resembling those in children aged 4 to 13 years, indicating that the otolithic-ocular reflex is mature in children aged >2 years. Despite the well-developed caloric and cervical VEMP responses in early life, oVEMPs are not present in newborns, but are present in children aged >2 years who can walk with a gait resembling an adult. Maturation of the otolithic-ocular reflex is important to balance control, which is necessary in small children for independent gait. Copyright © 2012 The American Laryngological, Rhinological, and Otological Society, Inc.

  8. An indicator of probable semicircular canal dehiscence: ocular vestibular evoked myogenic potentials to high frequencies.

    Science.gov (United States)

    Manzari, Leonardo; Burgess, Ann M; McGarvie, Leigh A; Curthoys, Ian S

    2013-07-01

    The n10 component of the ocular vestibular evoked myogenic potential (oVEMP) to sound and vibration stimuli is a crossed response that has enhanced amplitude and decreased threshold in patients with CT-verified superior semicircular canal dehiscence (SSCD). However, demonstrating enhanced VEMP amplitude and reduced VEMP thresholds requires multiple trials and can be very time consuming and tiring for patients, so a specific indicator of probable SCD that is fast and not tiring would be preferred. Here we report a 1-trial indicator: that the oVEMP n10 in response to a very high frequency stimulus (4000 Hz), either air-conducted sound (ACS) or bone conducted vibration (BCV), is such a fast indicator of probable SCD. In 22 healthy subjects, oVEMP n10 at 4000 Hz was not detectable; however, in all 22 CT-verified SSCD patients tested, oVEMP n10 responses were clearly present to 4000 Hz to either ACS or BCV stimuli.

  9. Downbeat nystagmus: evidence for enhancement of utriculo-ocular pathways by ocular vestibular evoked myogenic potentials?

    Science.gov (United States)

    Bremova, Tatiana; Glasauer, Stefan; Strupp, Michael

    2015-11-01

    Downbeat nystagmus (DBN) is caused by an impairment of Purkinje cells in the flocculus. The decreased cerebellar inhibitory input affects otolith pathways. Since ocular and cervical vestibular evoked myogenic potentials (o-/cVEMP) test the otoliths, the VEMP were measured in DBN patients and in controls. Sixteen patients with DBN, 14 cerebellar oculomotor disorder patients without DBN (COMD), and 16 healthy controls were examined with o-/cVEMP. Computational modeling was used to predict VEMP differences between groups. DBN patients had significantly higher oVEMP peak-to-peak (PP) amplitudes than COMD patients without DBN and controls. Cervical VEMP did not differ. The computational model of DBN predicted a twofold oVEMP increase for DBN patients. These findings suggest an enhancement of the utriculo-ocular response. The unchanged cVEMP indicate no effect on the otolith-cervical reflex in DBN. Computational modeling suggests that the utriculo-ocular enhancement is caused by an impaired vertical neural integrator resulting in the increased influence of utricular signals. This also explains the gravitational dependence of DBN.

  10. Pathological eye movements influence on the recordings of ocular vestibular-evoked myogenic potential.

    Science.gov (United States)

    Yang, Ting-Hua; Chen, Hsin-Lin; Young, Yi-Ho

    2017-08-01

    This study investigated the influence of pathological eye movements on the recordings of ocular vestibulo-evoked myogenic potential (oVEMP). Ten patients with pathological eye movements of non-vestibular origin (nine congenital nystagmus and one opsoclonus) who had negative MRI result were assigned to Group A. Another 20 vestibular neuritis (VN) patients with spontaneous nystagmus were assigned to Group B. Both groups underwent audiometry, and caloric, oVEMP and cervical VEMP (cVEMP) tests. In Group A, the caloric, oVEMP and cVEMP tests showed 40%, 55% and 50% abnormalities, respectively. In Group B (VN), caloric, oVEMP and cVEMP tests revealed 100%, 80% and 45% abnormalities on the lesion ears, and 0%, 40% and 0% abnormalities on the healthy (opposite) ears, respectively. The 40% oVEMP abnormality on the healthy ears may be due to recording failure from the presence of spontaneous nystagmus, since five of five VN patients showed normal oVEMPs on the healthy ears, one year after presentation. Presence of pathological eye movements may affect the recordings of oVEMP. Thus, the oVEMP test is recommended to perform after acute vertiginous episode to exclude the influence of pathological eye movements on the oVEMP recordings.

  11. Potential application of ocular and cervical vestibular-evoked myogenic potentials in Meniere's disease: a review.

    Science.gov (United States)

    Young, Yi-Ho

    2013-02-01

    By stimulating the ear with air-conducted sound or bone-conducted vibration stimuli, vestibular-evoked myogenic potential (VEMP) can be recorded on the contracted neck muscles, termed cervical VEMP (cVEMP), and on the extraocular muscles, termed ocular VEMP (oVEMP). These two electrophysiological tests expand the test battery for clinicians to explore the dynamic otolithic function, adding a potential usefulness to the sacculocollic reflex and vestibulo-ocular reflex, respectively. The inner ear test battery, including audiometry, and cVEMP, oVEMP and caloric tests, is designed for complete evaluation of the inner ear function, namely, the cochlea, saccule, utricle, and semicircular canals, respectively. Using this test battery to study the localization and prevalence of hydrops formation reveals that the declining function in the cochlea, saccule, utricle, and semicircular canals mimics the declining sequence of hydrops formation in temporal bone studies. This study reviewed the physiological results in Meniere's patients via the inner ear test battery, especially the potential application of the oVEMP and cVEMP tests, to correlate with the histopathological findings of Meniere's disease. Copyright © 2012 The American Laryngological, Rhinological, and Otological Society, Inc.

  12. Correlation between subjective visual horizontal test and ocular vestibular-evoked myogenic potential test.

    Science.gov (United States)

    Lin, Kuei-You; Young, Yi-Ho

    2011-02-01

    The static subjective visual horizontal (SVH) test correlates with the dynamic ocular vestibular-evoked myogenic potential (oVEMP) test in healthy and pathological ears, and further confirms that both tests may, at least in part, share the same utricular reflex pathway. This study correlated the SVH test results with those of the oVEMP and cervical VEMP (cVEMP) tests to investigate their relationships. Twenty healthy subjects underwent the SVH test at a view pattern angle of 30° or 70° horizontal tilt under various background distractions to establish the optimal stimulation mode for SVH test. Thereafter, 20 patients with unilateral Meniere's disease underwent the SVH test using the optimal mode. In addition, oVEMP and cVEMP tests were performed in all subjects. The preliminary study in 20 healthy subjects at a view pattern angle of 70° under counterclockwise square background distraction revealed that the mean deviation degree of the SVH test was -0.61 ± 1.17°. Based on the criteria, abnormal percentages of SVH in 20 Meniere's patients were 40%. All healthy subjects had normal oVEMPs and cVEMPs. In contrast, eight patients (40%) showed abnormal oVEMPs and nine (45%) revealed abnormal cVEMPs. The SVH test results correlated significantly with oVEMP results, but not with cVEMP results.

  13. Sudden Bilateral Hearing Loss After Cervical and Ocular Vestibular Evoked Myogenic Potentials.

    Science.gov (United States)

    Mattingly, Jameson K; Portnuff, Cory D F; Hondorp, Brian M; Cass, Stephen P

    2015-07-01

    Cervical and ocular vestibular evoked myogenic potentials (cVEMPs and oVEMPs) are commonly used in evaluation of neurotologic disorders. We present a case of sudden bilateral hearing loss immediately after oVEMP and cVEMP testing. The hearing loss did not recover. To our knowledge, no previous case reports discuss sudden hearing loss, especially bilateral, associated with VEMP testing. A single patient with sudden bilateral hearing loss that has persisted after cVEMP and oVEMP. The patient had a history of chronic daily dizziness. She underwent vestibular function testing that included cVEMP and oVEMP testing. A significant bilateral sensorineural hearing loss was noted immediately after cVEMP and oVEMP testing and confirmed with audiometric testing. Despite the use of oral steroids, her hearing loss did not recover. Serial audiograms, calculated maximum total sound energies to each ear. Pre-VEMP versus post-VEMP audiograms show increased thresholds and decreased word recognition scores; total sound energy delivered to each ear shows significant sound exposure. Although VEMP testing is thought to be safe and well tolerated, a significant amount of sound can be delivered to the cochlea, and certain individuals may be susceptible to acoustic trauma at these levels. We recommend limits for VEMP stimuli levels and attention to total sound exposure when multiple trials are used.

  14. Vestibular evoked myogenic potential testing as an objective measure of vestibular stimulation with cochlear implants.

    Science.gov (United States)

    Parkes, William J; Gnanasegaram, Joshua J; Cushing, Sharon L; McKnight, Carmen L; Papsin, Blake C; Gordon, Karen A

    2017-02-01

    To determine if vestibular potentials could be elicited with electrical stimulation from cochlear implants. Prospective cohort study. Vestibular responsiveness to electrical stimulation from cochlear implants was assessed via vestibular evoked myogenic potential (VEMP) testing in 53 pediatric and young adult patients. Thirty-one participants (58%) showed at least one vestibular potential in response to acoustic stimulation; 33 (62%) had an electrically evoked vestibular response. A cervical VEMP (cVEMP) was present in 45 of the 96 tested ears (47%) in response to acoustic stimulation, and in 34 ears (35%) with electrical stimulation. An ocular VEMP (oVEMP) was elicited acoustically in 25 ears (26%) and electrically in 34 (35%) ears. In the ears with absent responses to acoustic stimuli, electrically evoked cVEMPs and oVEMPs were present in 14 (27%) and 18 (25%) ears, respectively. Electric VEMPs demonstrated shorter latencies than acoustic VEMPs (P VEMPs was seen at high stimulation levels (P .05). VEMPs can be elicited with electrical stimulation in a proportion of children with cochlear implants, demonstrating current spread from the cochlea to the vestibular system. The presence of electric VEMPs in acoustically nonresponsive ears, along with the shorter latencies of electrically driven VEMPs, suggests that electrical current can bypass the otoliths and directly stimulate vestibular neural elements. 4. Laryngoscope, 2016 127:E75-E81, 2017. © 2016 The American Laryngological, Rhinological and Otological Society, Inc.

  15. Ocular and cervical vestibular evoked myogenic potentials in multiple sclerosis patients.

    Science.gov (United States)

    Gazioglu, Sibel; Boz, Cavit

    2012-09-01

    Vestibular evoked myogenic potentials (VEMPs) are thought to provide useful information about brainstem functions, as the neural pathways of both ocular and cervical VEMPs pass through the brainstem. The aim of this study was to investigate the clinical value of ocular and cervical VEMP tests in the evaluation of brainstem involvement in multiple sclerosis (MS) patients and to assess their relation with clinical and cranial MRI findings. Ocular and cervical VEMPs were recorded in 62 MS patients and 35 age and sex matched healthy volunteers. The latencies, amplitude asymmetry ratios of both VEMP responses and abnormality ratios (prolonged latencies and absent responses) were compared between the MS patients and the control group and among the groups of MS patients. oVEMP mean n1 and p1 latencies and cVEMP mean p13 latency were significantly prolonged in MS patients. Although the abnormality ratios of both VEMPs were higher in patients with brainstem clinical or MRI lesions, the correlation was not statistically significant. Both ocular and cervical VEMP latencies were significantly correlated with expanded disability status scale. Although there is no significant correlation with clinical or MRI findings, MS patients show high frequency of abnormality in VEMP tests, especially in oVEMP tests. VEMP tests may be useful as an adjunct test in the evaluation of brainstem dysfunction in MS patients. Copyright © 2012 International Federation of Clinical Neurophysiology. Published by Elsevier Ireland Ltd. All rights reserved.

  16. Electromotive Triggering and Single Sweep Analysis of Vestibular Evoked Myogenic Potentials (VEMPs).

    Science.gov (United States)

    Hecker, Dietmar J; Lohscheller, Joerg; Schorn, Bianca; Koch, Klaus Peter; Schick, Bernhard; Dlugaiczyk, Julia

    2014-01-01

    Cervical (c) and ocular (o) vestibular evoked myogenic potentials (VEMPs) provide important tools for measuring otolith function. However, two major drawbacks of this method are encountered in clinical practice. First, recording of oVEMPs is compromised by small n10 amplitudes. Second, VEMP analysis is currently based on the averaging technique, resulting in a loss of information compared to single sweep analysis. Here, we: 1) developed a novel electromotive trigger mechanism for evoking VEMPs by bone-conducted vibration to the forehead and 2) established maximum entropy extraction of complex wavelet transforms for calculation of phase synchronization between VEMP single sweeps. Both c- and oVEMPs were recorded for n=10 healthy individuals. The oVEMP n10 amplitude was consistently higher (right: 24.84±9.71 μV; left: 27.40±14.55 μV) than previously described. Stable VEMP signals were reached after a smaller number of head taps (oVEMPs 6; cVEMPs 11) compared to current recommendations. Phase synchronization vectors and phase shift values were successfully determined for simulated and clinically recorded VEMPs, providing information about the impact of noise and phase jitter on the VEMP signal. Thus, the proposed method constitutes an easy-to-use approach for the fast detection and analysis of VEMPs in clinical practice.

  17. Differentiating cerebellar and brainstem lesions with ocular vestibular-evoked myogenic potential test.

    Science.gov (United States)

    Su, Chia-Hung; Young, Yi-Ho

    2011-06-01

    This study applied both ocular vestibular-evoked myogenic potential (oVEMP) and cervical VEMP (cVEMP) tests in patients with cerebellar disorders to determine whether VEMP test can differentiate between cerebellar and brainstem lesions. A total of 12 patients with cerebellar disorder, including extended cerebellar lesion (involving the brainstem) in 8 and localized cerebellar lesion (excluding the brainstem) in 4, were enrolled in this study. All patients underwent caloric, visual suppression, and oVEMP and cVEMP tests via bone-conducted vibration stimuli. The abnormal rates for the caloric, visual suppression, and oVEMP and cVEMP tests were 62, 83, 88 and 75% in patients with extended cerebellar lesion and 0, 25, 0 and 0% in those with localized cerebellar lesion, respectively. The rate of abnormal oVEMP results significantly differed between the two groups, but caloric, visual suppression and cVEMP test results did not differ. In another ten healthy subjects, characteristic parameters of oVEMPs obtained under light and dark conditions did not significantly differ. In conclusion, ocular VEMP test can differentiate between cerebellar and brainstem lesions. Abnormal oVEMPs in patients with cerebellar disorder may indicate adjacent brainstem involvement.

  18. Auditory evoked potentials and vestibular evoked myogenic potentials in evaluation of brainstem lesions in multiple sclerosis.

    Science.gov (United States)

    Ivanković, Anita; Nesek Mađarić, Vesna; Starčević, Katarina; Krbot Skorić, Magdalena; Gabelić, Tereza; Adamec, Ivan; Habek, Mario

    2013-05-15

    The aim of this study was to determine the roles of magnetic resonance imaging (MRI), auditory evoked potentials (AEP) and vestibular evoked myogenic potentials (VEMP) in the evaluation of brainstem involvement in multiple sclerosis (MS). Altogether 32 patients with the diagnosis of MS participated in the study. The following data was collected from all patients: age, gender, Expanded Disability Status Scale (EDSS) score, brainstem functional system score (BSFS) (part of the EDSS evaluating brainstem symptomatology), and involvement of the brainstem on the brain MRI. AEP and ocular VEMP (oVEMP) and cervical VEMP (cVEMP) were studied in all patients. BSFS, MRI, AEP, oVEMP and cVEMP involvement of the brainstem was evident in 9 (28.1%), 14 (43.8%), 7 (21.9%), 12 (37.5%) and 10 (31.0%) patients, respectively. None of the tests used showed statistically significant advantage in the detection of brainstem lesions. When combining oVEMP and cVEMP 18 (56.3%) patients showed brainstem involvement. This combination showed brainstem involvement in greater percentage than BSFS or AEP, with statistical significance (p=0.035 and p=0.007, respectively). VEMP is a reliable method in detection of brainstem involvement in MS. It is comparable with MRI, but superior to clinical examination or AEP. Copyright © 2013 Elsevier B.V. All rights reserved.

  19. The effect of increased intracranial pressure on vestibular evoked myogenic potentials in superior canal dehiscence syndrome.

    Science.gov (United States)

    Janky, Kristen L; Zuniga, M Geraldine; Schubert, Michael C; Carey, John P

    2015-04-01

    To determine if vestibular evoked myogenic potential (VEMP) responses change during inversion in patients with superior canal dehiscence syndrome (SCDS) compared to controls. Sixteen subjects with SCDS (mean: 43, range 30-57 years) and 15 age-matched, healthy subjects (mean: 41, range 22-57 years) completed cervical VEMP (cVEMP) in response to air conduction click stimuli and ocular VEMP (oVEMP) in response to air conduction 500 Hz tone burst stimuli and midline tap stimulation. All VEMP testing was completed in semi-recumbent and inverted conditions. SCDS ears demonstrated significantly larger oVEMP peak-to-peak amplitudes in comparison to normal ears in semi-recumbency. While corrected cVEMP peak-to-peak amplitudes were larger in SCDS ears; this did not reach significance in our sample. Overall, there was not a differential change in o- or cVEMP amplitude with inversion between SCDS and normal subjects. Postural-induced changes in o- and cVEMP responses were measured in the steady state regardless of whether the labyrinth was intact or dehiscent. VEMP responses are blunted during inversion. Although steady-state measurements of VEMPs during inversion do not increase diagnostic accuracy for SCDS, the findings suggest that inversion may provide more general insights into the equilibration of pressures between intracranial and intralabyrinthine fluids. Copyright © 2014 International Federation of Clinical Neurophysiology. Published by Elsevier Ireland Ltd. All rights reserved.

  20. A dual mode pulsed electro-magnetic cell stimulator produces acceleration of myogenic differentiation.

    Science.gov (United States)

    Leon-Salas, Walter D; Rizk, Hatem; Mo, Chenglin; Weisleder, Noah; Brotto, Leticia; Abreu, Eduardo; Brotto, Marco

    2013-04-01

    This paper presents the design and test of a dual-mode electric and magnetic biological stimulator (EM-Stim). The stimulator generates pulsing electric and magnetic fields at programmable rates and intensities. While electric and magnetic stimulators have been reported before, this is the first device that combines both modalities. The ability of the dual stimulation to target bone and muscle tissue simultaneously has the potential to improve the therapeutic treatment of osteoporosis and sarcopenia. The device is fully programmable, portable and easy to use, and can run from a battery or a power supply. The device can generate magnetic fields of up to 1.6 mT and output voltages of +/- 40 V. The EM-Stim accelerated myogenic differentiation of myoblasts into myotubes as evidenced by morphometric, gene expression, and protein content analyses. Currently, there are many patents concerned with the application of single electrical or magnetic stimulation, but none that combine both simultaneously. However, we applied for and obtained a provisional patent for new device to fully explore its therapeutic potential in pre-clinical models.

  1. Analysis of Saccular Function With Vestibular Evoked Myogenic Potential Test in Meniere's Disease.

    Science.gov (United States)

    Dabiri, Sasan; Yazdani, Nasrin; Esfahani, Mahdis; Tari, Niloufar; Adil, Susan; Mahvi, Zahra; Rezazadeh, Nima

    2017-02-01

    Meniere's disease is the disorder of inner ear characterized by vertigo, tinnitus and sensorineural hearing loss. The vestibular evoked myogenic potential (VEMP) test could be useful in the analysis of saccular function, and diagnosis of Meniere's disease. In this study, we've analyzed the saccular function, using VEMP test in different groups of Meniere's disease. Patients were categorized as possible, probable or definite Meniere's disease groups according to the guideline of American Academy of Otolaryngology-Head and Neck Surgery. The exclusion criteria were neuromuscular system diseases, diseases of central nervous system, inner ear disorders, conductive hearing loss, a history of ototoxic drug consumption, being a drug abuser and a positive history of inner ear surgery or manipulations. The VEMP test is the recording of positive and negative waves from sternocleidomastoid muscle that is made by an auditory click to the ear. From the total of 100 patients, the waves of VEMP test was seen in 59 patients which 19 patients had abnormal amplitude, and latency and 40 patients were with normally recorded waves. There was a significant relationship between the severity of hearing loss and a VEMP test without any recorded waves. Most of the cases with 'no wave recorded' VEMP test, were patients with severe hearing loss. However, there wasn't any relation between the pattern of hearing loss and 'no wave recorded' VEMP test. VEMP test could be a valuable diagnostic clue especially in patients with definite Meniere's disease.

  2. Myogenic response of human skeletal muscle to 12 weeks of resistance training at light loading intensity.

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    Mackey, A L; Holm, L; Reitelseder, S; Pedersen, T G; Doessing, S; Kadi, F; Kjaer, M

    2011-12-01

    There is strong evidence for enhanced numbers of satellite cells with heavy resistance training. The satellite cell response to very light muscle loading is, however, unknown. We, therefore, designed a 12-week training protocol where volunteers trained one leg with a high load (H) and the other leg with a light load (L). Twelve young healthy men [mean age 25 ± 3 standard deviation (SD) years] volunteered for the study. Muscle biopsies were collected from the m. vastus lateralis of both legs before and after the training period and satellite cells were visualized by CD56 immunohistochemistry. A significant main effect of time was observed (Pfiber (L: from 0.11 ± 0.02 to 0.13 ± 0.03; H: from 0.12 ± 0.03 to 0.15 ± 0.05, mean ± SD). The finding that 12 weeks of training skeletal muscle even with very light loads can induce an increase in the number of satellite cells reveals a new aspect of myogenic precursor cell activation and suggests that satellite cells may play a role in skeletal muscle adaptation over a broad physiological range.

  3. Myogenic Progenitor Cells Control Extracellular Matrix Production by Fibroblasts during Skeletal Muscle Hypertrophy.

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    Fry, Christopher S; Kirby, Tyler J; Kosmac, Kate; McCarthy, John J; Peterson, Charlotte A

    2017-01-05

    Satellite cells, the predominant stem cell population in adult skeletal muscle, are activated in response to hypertrophic stimuli and give rise to myogenic progenitor cells (MPCs) within the extracellular matrix (ECM) that surrounds myofibers. This ECM is composed largely of collagens secreted by interstitial fibrogenic cells, which influence satellite cell activity and muscle repair during hypertrophy and aging. Here we show that MPCs interact with interstitial fibrogenic cells to ensure proper ECM deposition and optimal muscle remodeling in response to hypertrophic stimuli. MPC-dependent ECM remodeling during the first week of a growth stimulus is sufficient to ensure long-term myofiber hypertrophy. MPCs secrete exosomes containing miR-206, which represses Rrbp1, a master regulator of collagen biosynthesis, in fibrogenic cells to prevent excessive ECM deposition. These findings provide insights into how skeletal stem and progenitor cells interact with other cell types to actively regulate their extracellular environments for tissue maintenance and adaptation. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. Female host sex-biased parasitism with the rodent stomach nematode Mastophorus muris in wild bank voles (Myodes glareolus).

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    Grzybek, Maciej; Bajer, Anna; Behnke-Borowczyk, Jolanta; Al-Sarraf, Mohammed; Behnke, Jerzy M

    2015-02-01

    Abundance and prevalence of helminth infections often differ between host sexes, and are usually biased in favor of males. Relatively few cases of female-biased parasitism have been reported. We sampled bank voles in three woodland sites in N.E. Poland over 11 years at 3-4-year intervals, and assessed their parasite burdens. Prevalence and abundance of the stomach nematode Mastophorus muris were consistently higher among females. Among adult female bank voles from the two sites that showed the highest prevalence with M. muris, both prevalence and abundance were significantly higher in lactating bank voles, but not pregnant animals, and the effect of lactation was evident in both sites, in all four surveys, and in both age classes. Although the magnitude of the effect of lactation varied between years, it was not confounded by any significant interactions with other factors. We hypothesize that mature and reproductively active female bank voles are subject to higher exposure compared with males of similar age, as a consequence of the increased content of invertebrates in their diet, including the intermediate hosts of M. muris, required to meet the higher increased energy and protein demands of nursing litters throughout the summer months.

  5. Long-term spatiotemporal stability and dynamic changes in the haemoparasite community of bank voles (Myodes glareolus) in NE Poland.

    Science.gov (United States)

    Bajer, Anna; Welc-Falęciak, Renata; Bednarska, Małgorzata; Alsarraf, Mohammed; Behnke-Borowczyk, Jolanta; Siński, Edward; Behnke, Jerzy M

    2014-08-01

    Long-term field studies on parasite communities are rare but provide a powerful insight into the ecological and evolutionary processes shaping host-parasite interactions. The aim of our study was to identify the principal factors regulating long-term trends in the haemoparasite communities of bank voles, and to this end, we sampled three semi-isolated populations of bank voles (n = 880) in 1999, 2002, 2006 and 2010 in the Mazury lake district region of NE Poland. Overall, 90.8 % of the bank voles harboured at least one of the species of haemoparasites studied. Whilst overall prevalence (all species combined) did not vary significantly between the surveys, different temporal changes were detected among voles in each of the three sites. In voles from Urwitałt, prevalence increased consistently with successive surveys, whereas in Tałty, the peak years were 2002 and 2006, and in Pilchy, prevalence oscillated without a clear pattern. Across the study, bank voles harboured a mean of 1.75 ± 0.034 haemoparasite species, and species richness remained stable with no significant between-year fluctuations or trends. However, each of the five constituent species/genera showed a different pattern of spatio-temporal changes. The overall prevalence of Babesia microti was 4.9 %, but this varied significantly between years peaking in 2006 and declining again by 2010. For Bartonella spp., overall prevalence was 38.7 %, and this varied with year of study, but the temporal pattern of changes differed among the three sites. The overall prevalence of Haemobartonella (Mycoplasma) was 68.3 % with an increase in prevalence with year of study in all three sites. Hepatozoon erhardovae had an overall prevalence of 46.8 % but showed a marked reduction with each successive year of the study, and this was consistent in all three sites. The overall prevalence of Trypanosoma evotomys was 15.4 % varying significantly between sites, but showing temporal stability. While overall prevalence

  6. Clone-derived human AF-amniotic fluid stem cells are capable of skeletal myogenic differentiation in vitro and in vivo.

    Science.gov (United States)

    Ma, Xiaorong; Zhang, Shengli; Zhou, Junmei; Chen, Baisong; Shang, Yafeng; Gao, Tongbing; Wang, Xue; Xie, Hua; Chen, Fang

    2012-08-01

    Stem cell-based therapy may be the most promising method to cure skeletal muscle degenerative diseases such as Duchenne muscular dystrophy (DMD) and trauma in the future. Human amniotic fluid is enriched with early-stage stem cells from developing fetuses and these cells have cardiomyogenic potential both in vitro and in vivo. In the present study, we investigated the characteristics of human amniotic fluid-derived AF-type stem (HAF-AFS) cells by flow cytometry, immunofluorescence staining, reverse-transcription polymerase chain reaction, and osteogenic and adipogenic differentiation analysis. After confirming the stemness of HAF-AFS cells, we tested whether HAF-AFS cells could differentiate into skeletal myogenic cells in vitro and incorporate into regenerating skeletal muscle in vivo. By temporary exposure to the DNA demethylation agent 5-aza-2'-deoxycytidine (5-Aza dC) or co-cultured with C2C12 myoblasts, HAF-AFS cells differentiated into skeletal myogenic cells, expressing skeletal myogenic cell-specific markers such as Desmin, Troponin I (Tn I) and α-Actinin. Four weeks after transplantation into cardiotoxin-injured and X-ray-irradiated tibialis anterior (TA) muscles of NOD/SCID mice, HAF-AFS cells survived, differentiated into myogenic precursor cells and fused with host myofibres. The findings that HAF-AFS cells differentiate into myogenic cells in vitro and incorporate in skeletal muscle regeneration in vivo hold the promise of HAF-AFS cell-based therapy for skeletal muscle degenerative diseases.

  7. A CRISPR/Cas9-Based System for Reprogramming Cell Lineage Specification

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    Syandan Chakraborty

    2014-12-01

    Full Text Available Gene activation by the CRISPR/Cas9 system has the potential to enable new approaches to science and medicine, but the technology must be enhanced to robustly control cell behavior. We show that the fusion of two transactivation domains to Cas9 dramatically enhances gene activation to a level that is necessary to reprogram cell phenotype. Targeted activation of the endogenous Myod1 gene locus with this system led to stable and sustained reprogramming of mouse embryonic fibroblasts into skeletal myocytes. The levels of myogenic marker expression obtained by the activation of endogenous Myod1 gene were comparable to that achieved by overexpression of lentivirally delivered MYOD1 transcription factor.

  8. Vestibular evoked myogenic potentials and video head impulse test in patients with vertigo, dizziness and imbalance.

    Science.gov (United States)

    Skorić, Magdalena Krbot; Adamec, Ivan; Pavičić, Tin; Pavlović, Ivan; Ruška, Berislav; Crnošija, Luka; Habek, Mario

    2017-05-01

    The aim of this study was to compare vestibular evoked myogenic potentials (VEMP) and video head impulse test (vHIT) results in patients presenting with vertigo and dizziness. We retrospectively analyzed data of all patients with the chief complaint of vertigo, dizziness, or imbalance that underwent VEMP and vHIT from January 2015 to January 2016. A total of 117 patients (73 females, mean age 53.92±16.76) fulfilled inclusion criteria: group 1 included patients with the final diagnosis of vestibular neuritis (VN) (N=31 (16 right and 15 left VN)), group 2 included patients with the final diagnosis of vertigo of central origin (N=23) and group 3 included patients with the final diagnosis of unspecified dizziness (N=63). There was significant correlation between oVEMP asymmetry and asymmetry of the lateral canals 60ms gains on vHIT (r=0.225, p=0.026). Significant correlation between oVEMP and vHIT asymmetry was present in VN patients (r=0.749, p<0.001), while no correlation was found in the groups 2 and 3. oVEMP and vHIT lateral canals asymmetries were significantly greater in patients with vestibular neuritis. Furthermore, positive correlations of oVEMP amplitudes with 60ms gain of the lateral semicircular canal and slope of the anterior semicircular canal on vHIT, and cVEMP with slope of the posterior semicircular canal on the vHIT were found. These changes were significantly more pronounced in patients with vestibular neuritis. In conclusion, VEMPs and vHIT data should be used complementarily; asymmetry on both tests strongly supports peripheral vestibular system involvement. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Splenius capitis is a reliable target for measuring cervical vestibular evoked myogenic potentials in adults.

    Science.gov (United States)

    Camp, Aaron J; Gu, Chao; Cushing, Sharon L; Gordon, Karen A; Corneil, Brian D

    2017-02-08

    The cervical vestibular evoked myogenic potential (cVEMP) is a common and simple test of vestibulospinal reflex patency. In the clinic, cVEMPs are measured in response to loud sounds from the sternocleidomastoid (SCM) on the ventral neck, as subjects maintain an uncomfortable head posture needed to recruit SCM. Here we characterize the cVEMP in a dorsal neck turner (splenius capitis; SPL), and compare it with the SCM cVEMP. cVEMPs were recorded simultaneously via surface electromyography from SCM and SPL from 17 healthy subjects in a variety of postures, including head-turned postures adopted while either seated or standing, and the clinical posture. Like the SCM cVEMP recorded ipsilateral to the side of sound stimulation, the cVEMP on the contralateral SPL (synergistic with ipsilateral SCM) was characterized by a biphasic wave of muscle activity that began at ~ 13 ms. cVEMP reliability was higher on SPL vs. SCM in standing postures (chi-squared; P < 0.05), and equivalent results were obtained from SPL in a standing or seated posture. In 9 of the 17 subjects, we also obtained bilateral intramuscular (IM) recordings from SPL at the same time as the surface recordings. In these subjects, the initial surface response in SPL was associated with a consistent decrease in multi-unit IM SPL activity. Overall, these results demonstrate that SPL recordings offer a complimentary target for cVEMP assessments. The expression of SPL cVEMPs in simple head-turned postures may also improve the utility of cVEMP testing for vestibular assessment in children, the elderly, or non-compliant.

  10. Saccular function in otosclerosis patients: bone conducted-vestibular evoked myogenic potential analysis.

    Directory of Open Access Journals (Sweden)

    Amin Amali

    2014-02-01

    Full Text Available Vestibular involvements have long been observed in otosclerotic patients. Among vestibular structures saccule has the closest anatomical proximity to the sclerotic foci, so it is the most prone vestibular structure to be affected during the otosclerosis process. The aim of this study was to investigate the saccular function in patients suffering from otosclerosis, by means of Vestibular Evoked Myogenic Potential (VEMP. The material consisted of 30 otosclerosis patients and 20 control subjects. All participants underwent audiometric and VEMP testing. Analysis of tests results revealed that the mean values of Air-Conducted Pure Tone Average (AC-PTA and Bone-Conducted Pure Tone Average (BC-PTA in patients were 45.28 ± 15.57 and 19.68 ± 10.91, respectively and calculated 4 frequencies Air Bone Gap (ABG was 25.64 ± 9.95. The VEMP response was absent in 14 (28.57% otosclerotic ears. A statistically significant increase in latency of the p13 was found in the affected ears (P=0.004, differences in n23 latency did not reach a statistically significant level (P=0.112. Disparities in amplitude of p13-n23 in between two study groups was statistically meaningful (P=0.009, indicating that the patients with otosclerosis had lower amplitudes. This study tends to suggest that due to the direct biotoxic effect of the materials released from the otosclerosis foci on saccular receptors, there might be a possibility of vestibular dysfunction in otosclerotic patients.

  11. The Clinical Utility of Vestibular Evoked Myogenic Potentials in Patients of Benign Paroxysmal Positional Vertigo

    Science.gov (United States)

    Sreenivasan, Anuprasad; Parida, Pradiptata Kumar; Alexander, Arun; Saxena, Sunil Kumar; Suria, Gopalakrishnan

    2015-01-01

    Context Vestibular Evoked Myogenic Potentials (VEMP) is an emerging tool to diagnose Benign Paroxysmal Positional Vertigo (BPPV). The clinical utility of VEMP has been reported only in small accord in Indian literature. Aim To study the latency and amplitude of VEMP in patients with BPPV and compare it with that of normal subjects. Settings and Design The study included two groups. Group one (control group) were 18 normal subjects. Group two (test group) were 15 subjects with unilateral BPPV. Materials and Methods Those subjects who fulfilled the selection criteria based on case history and audiological assessment were taken for the VEMP recording. The VEMP response consist of positive and negative successive waves (pI-nI), with latency values in adults about 13 and 23 milliseconds respectively. Statistical Analysis Data was analysed using Statistical Package for Social Sciences (SPSS) version 12 (Chicago, IL, USA). Unpaired t-test was employed to measure the statistical difference between control group and test group. Results The difference in n23 and the peak to peak amplitude between the ipsilateral and contralateral ears of the test group were statistically significant, whereas such a difference in the p13 latency turned out to be statistically insignificant. It should be noted that, out of 15 patients in the test group, five patients showed only artifact tracer recordings in both the ears which is considered as no response. The heterogeneity of the results extended from absence of VEMP to prolongation of both p13, n23; prolongation of p13 alone; and even side to side variations. Conclusion Absent response from the ipsilateral ear, prolonged latency of n23 and decreased peak to peak amplitude (p13, n23), indicates the disease pathology. However, large sample size is required to draw further conclusions and to consolidate the usage of VEMP in the diagnosis of BPPV. PMID:26266140

  12. Vestibular-evoked myogenic potentials in three patients with large vestibular aqueduct.

    Science.gov (United States)

    Sheykholeslami, Kianoush; Schmerber, Sébastien; Habiby Kermany, Mohammad; Kaga, Kimitaka

    2004-04-01

    An enlarged vestibular aqueduct (LVA) is a common congenital inner ear anomaly responsible for some unusual vestibular and audiological symptoms. Most of the cases show bilateral early onset and progressive hearing loss in children. The gross appearance on CT scan of the inner ear is generally normal. However, precise measurements of the inner ear components reveal abnormal dimensions, which may account for the accompanying auditory and vestibular dysfunction. Despite extensive studies on hearing and the vestibular apparatus, saccular function is not studied. To our knowledge this is the first report of saccular malfunction in three patients with LVA by means of vestibular evoked myogenic potentials. Conventional audiograms revealed bilateral severe sensorineural hearing loss in two patients and mixed type hearing loss in one patient. Two of the patients complained about vertigo and dizziness but vestibular assessments of the patients showed normal results. The diagnosis had been made by high-resolution CT scans and MR images of the skull that showed LVA in the absence of other anomalies. The VEMP threshold measured from the ear with LVA in two patients with unilateral enlargement of the vestibular aqueduct was 75-80 dB nHL whereas the threshold from normal ears was 95 dB nHL. The third patient with mixed type hearing loss and bilateral LVA had VEMP responses despite a big air-bone gap in the low frequency range. The VEMP in this patient was greater in amplitude and lower in threshold in the operated ear (the patient had a tympanoplasty which did not improve her hearing). These findings and results of other patients with Tullio phenomenon and superior semicircular canal dehiscence, who also showed lower VEMP threshold, confirmed the theory of a 'third window' that allows volume and pressure displacements, and thus larger deflection of the vestibular sensors, which would cause the vestibular organ to be more responsive to sound and pressure changes.

  13. Feasibility of the simultaneous ocular and cervical vestibular-evoked myogenic potentials in unilateral vestibular hypofunction.

    Science.gov (United States)

    Chou, Chen-Han; Wang, Shou-Jen; Young, Yi-Ho

    2009-09-01

    This study compared the results of combined and individual ocular vestibular-evoked myogenic potential (oVEMP) and cervical VEMP (cVEMP) tests in healthy subjects and patients with unilateral vestibular hypofunction to confirm the effectiveness of the combined oVEMP and cVEMP test. Twenty healthy volunteers and 12 patients with unilateral vestibular hypofunction underwent individual oVEMP and cVEMP tests in a random order, and combined oVEMP and cVEMP test on another day. Twenty healthy subjects had 100% response rates for oVEMPs and cVEMPs in both individual and combined stimulation modes. Significant positive correlations exist between individual and combined oVEMPs/cVEMPs in terms of latencies and amplitudes. In 12 patients with unilateral vestibular hypofunction, differences in abnormal percentages of oVEMPs or cVEMPs were non-significant. Additionally, the characteristic parameters of oVEMP/cVEMP among ears of healthy subjects, good and pathological ears of patients with unilateral vestibular hypofunction did not differ significantly, regardless of whether the individual or combined mode was used. The combined oVEMP and cVEMP test generates similar information to that obtained by individual mode, regardless of whether subjects are healthy or have unilateral vestibular hypofunction. Simultaneous oVEMP and cVEMP tests may be a convenient screening tool for assessing crossed vestibulo-ocular reflex and ipsilateral sacculo-collic reflex, which definitely shortens the diagnostic test time. Thus, it may favor the large diffusion of these techniques.

  14. Ocular and cervical vestibular-evoked myogenic potentials in Tumarkin falls.

    Science.gov (United States)

    Huang, Chi-Hsuan; Young, Yi-Ho

    2012-09-01

    This study applied ocular vestibular-evoked myogenic potential (oVEMP) and cervical VEMP (cVEMP) tests in Ménière's patients with Tumarkin falls to investigate the etiologic role of the saccule/utricle in the event of Tumarkin falls. Retrospective study. University hospital. Twenty unilateral definite Ménière's patients were divided into 2 age- and sex-matched groups. Ten patients had a history of drop attacks, and 10 had no history of drop attacks. All patients underwent audiometry and caloric, oVEMP, and cVEMP tests. No significant differences were observed between the fall and nonfall groups in terms of Ménière staging, and caloric and cVEMP test results. In the fall group, oVEMP test via Fz tapping showed augmented, reduced, and absent responses in 1, 1, and 8 patients, respectively. In 3 of the latter 8 patients, however, oVEMPs could be obtained by lateral mastoid tapping. Thus, 100% of patients in the fall group had abnormal oVEMPs, which significantly differed from 50% oVEMP abnormalities in the nonfall group. Comparison of the abnormal results for the caloric, oVEMP, and cVEMP tests revealed that poorer residual vestibular function in the fall group than the nonfall group. Absence of oVEMP by Fz tapping and presence of oVEMPs via lateral tapping indicates a residual utricular function. With residual canal function (canal paresis) and absent cVEMPs, subsequently residual utricular function may trigger the Tumarkin falls in Ménière's patients.

  15. Myogenic temporomandibular disorders: Clinical systemic comorbidities in a female population sample

    Science.gov (United States)

    Mesa-Jiménez, Juan; Fernández-de-las-Peñas, César; de-la-Hoz-Aizpurua, José-Luis

    2016-01-01

    Background Myogenic temporomandibular disorders (MTMD) frequently coexist with other clinical conditions in the same individual. In the last decades, several authors have analyzed these comorbidities looking for the origin of this overlapping. Objetives The aim of this study was to perform a comparative anaylisis between a group of patients with MTMD and a control group of dental patients without dysfunctional pathology to assess whether there are significant differences in the presence of systemic medical comorbidities between the two groups. Material and Methods Restrospective epidemiological analysis, based on medical questionnaires in a group of 31 patients, women, aged from 24 to 58 (average 39.96 years), diagnosed with MTMD (Masticatory Myofascial Pain), with a control group with the same number of individuals, gender and age range to evaluate if there is a significant statistical difference in the presence of medical comorbidities in this group of patients with MTMD and if they are in a higher risk of suffering different pathological conditions. Results It was found that the group affected by MTMD presented many more associated medical conditions than the control group: health changes during the last year, medical evaluations and treatments, presence of pain, sinus disease, tinnitus, headache, joint pain, ocular disorders, fatigue, dizziness, genitourinary disorders and xerostomia among others; and they were also in a higher risk to suffer other pathological entities as headaches and articular pain. Conclusions These results reinforce our hypothesis that MTMD belong to a group of medical conditions triggered by a loss of equilibrium of the individual’s Psycho-Neuro-Endocrine-Immune (PNEI) Axis that produces alterations in the response against external stimuli in some genetically predisposed individuals. It is, therefore, necessary to change the way of diagnosing and managing these individual’s medical conditions, being mandatory to look from a more

  16. Effects of deoxycholylglycine, a conjugated secondary bile acid, on myogenic tone and agonist-induced contraction in rat resistance arteries.

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    Sandeep Khurana

    Full Text Available BACKGROUND: Bile acids (BAs regulate cardiovascular function via diverse mechanisms. Although in both health and disease serum glycine-conjugated BAs are more abundant than taurine-conjugated BAs, their effects on myogenic tone (MT, a key determinant of systemic vascular resistance (SVR, have not been examined. METHODOLOGY/PRINCIPAL FINDINGS: Fourth-order mesenteric arteries (170-250 µm isolated from Sprague-Dawley rats were pressurized at 70 mmHg and allowed to develop spontaneous constriction, i.e., MT. Deoxycholylglycine (DCG; 0.1-100 µM, a glycine-conjugated major secondary BA, induced reversible, concentration-dependent reduction of MT that was similar in endothelium-intact and -denuded arteries. DCG reduced the myogenic response to stepwise increase in pressure (20 to 100 mmHg. Neither atropine nor the combination of L-NAME (a NOS inhibitor plus indomethacin altered DCG-mediated reduction of MT. K(+ channel blockade with glibenclamide (K(ATP, 4-aminopyradine (K(V, BaCl(2 (K(IR or tetraethylammonium (TEA, K(Ca were also ineffective. In Fluo-2-loaded arteries, DCG markedly reduced vascular smooth muscle cell (VSM Ca(2+ fluorescence (∼50%. In arteries incubated with DCG, physiological salt solution (PSS with high Ca(2+ (4 mM restored myogenic response. DCG reduced vascular tone and VSM cytoplasmic Ca(2+ responses (∼50% of phenylephrine (PE- and Ang II-treated arteries, but did not affect KCl-induced vasoconstriction. CONCLUSION: In rat mesenteric resistance arteries DCG reduces pressure- and agonist-induced vasoconstriction and VSM cytoplasmic Ca(2+ responses, independent of muscarinic receptor, NO or K(+ channel activation. We conclude that BAs alter vasomotor responses, an effect favoring reduced SVR. These findings are likely pertinent to vascular dysfunction in cirrhosis and other conditions associated with elevated serum BAs.

  17. Regulation of turkey myogenic satellite cell migration by MicroRNAs miR-128 and miR-24.

    Science.gov (United States)

    Velleman, S G; Harding, R L

    2016-12-05

    Myogenic satellite cells are an adult stem cell responsible for all post-hatch muscle growth in poultry. As a stem cell population, satellite cells are highly heterogeneous, but the origin of this heterogeneity remains unclear. Heterogeneity is, in part, regulated by gene expression. One method of endogenous gene regulation that may contribute to heterogeneity is microRNAs (miRNAs). Two miRNAs previously shown to regulate poultry myogenic satellite cell proliferation and differentiation, miR-128 and miR-24, were studied to determine if they also affected satellite cell migration. Satellite cell migration is an essential step for both proliferation and differentiation. During proliferation, satellite cells will migrate and align to form new myofibers or donate their nuclei to existing myofibers leading to muscle fiber hypertrophy or regeneration. Transient transfection of miRNA specific mimics to each miRNA reduced migration of satellite cells following a cell culture scratch at 72 h of proliferation when the cultures were 90 to 100% confluent. However, only the migration in cells transfected with miR-24 mimics at 24 and 30 h following the scratch was significantly reduced (P ≤ 0.05) to around 70% of the distance migrated by controls. Alternately, transfection with inhibitors specific to miR-128 or miR-24 significantly (P ≤ 0.05) increased migration between 147 and 252% compared to their controls between 24 and 48 h following the scratch. These data demonstrate that miR-128 and miR-24 play a role in myogenic satellite cell migration, which will impact muscle development and growth.

  18. Effect of pregnancy and nitric oxide on the myogenic vasodilation of posterior cerebral arteries and the lower limit of cerebral blood flow autoregulation.

    Science.gov (United States)

    Chapman, Abbie C; Cipolla, Marilyn J; Chan, Siu-Lung

    2013-09-01

    Hemorrhage during parturition can lower blood pressure beyond the lower limit of cerebral blood flow (CBF) autoregulation that can cause ischemic brain injury. However, the impact of pregnancy on the lower limit of CBF autoregulation is unknown. We measured myogenic vasodilation, a major contributor of CBF autoregulation, in isolated posterior cerebral arteries (PCAs) from nonpregnant and late-pregnant rats (n = 10/group) while the effect of pregnancy on the lower limit of CBF autoregulation was studied in the posterior cerebral cortex during controlled hemorrhage (n = 8). Pregnancy enhanced myogenic vasodilation in PCA and shifted the lower limit of CBF autoregulation to lower pressures. Inhibition of nitric oxide synthase (NOS) prevented the enhanced myogenic vasodilation during pregnancy but did not affect the lower limit of CBF autoregulation. The shift in the autoregulatory curve to lower pressures during pregnancy is likely protective of ischemic injury during hemorrhage and appears to be independent of NOS.

  19. VASCULAR: Inhibition of myogenic tone in rat cremaster and cerebral arteries by SKA-31, an activator of endothelial KCa2.3 and KCa3.1 channels

    OpenAIRE

    Mishra, Ramesh C.; Wulff, Heike; Hill, Michael A.; Braun, Andrew P.

    2015-01-01

    Endothelial KCa2.3 and KCa3.1 channels contribute to the regulation of myogenic tone in resistance arteries by Ca2+-mobilizing, vasodilatory hormones. To define further the functional role of these channels in distinct vascular beds, we have examined the vasodilatory actions of the KCa channel activator SKA-31 in myogenically active, rat cremaster and middle cerebral arteries. Vessels pressurized to 70 mmHg constricted by 80–100 microns (i.e. 25–45% of maximal diameter). SKA-31 (10 μM) inhibi...

  20. Body position effects on EMG activity of sternocleidomastoid and masseter muscles in patients with myogenic cranio-cervical-mandibular dysfunction.

    Science.gov (United States)

    Palazzi, C; Miralles, R; Soto, M A; Santander, H; Zuñiga, C; Moya, H

    1996-07-01

    This study was conducted in order to determine the effects of body position on integrated electromyographic (IEMG) activity of sternocleidomastoid and masseter muscles in 17 patients with myogenic cranio-cervical-mandibular dysfunction. EMG recordings at rest and during swallowing of saliva and maximal voluntary clenching were performed by placing surface electrodes on the sternocleidomastoid and masseter muscles (contralateral to the habitual side of sleeping of each patient), in the following body positions: standing, seated, supine, and lateral decubitus position. Significant higher EMG activities were recorded in the sternocleidomastoid muscle in the lateral decubitus position and in the supine position (except during swallowing), whereas a significant higher EMG activity was recorded in the masseter muscle during maximal voluntary clenching in standing and seated positions. The EMG pattern observed suggests that the presence of parafunctional habits and body position could be closely correlated with the clinical symptomatology in the sternocleidomastoid and masseter muscles at wakening and during waking hours, respectively, in patients with myogenic cranio-cervical-mandibular dysfunction.

  1. Doxycycline Inhibits IL-17-Stimulated MMP-9 Expression by Downregulating ERK1/2 Activation: Implications in Myogenic Differentiation

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    Hristina Obradović

    2016-01-01

    Full Text Available Interleukin 17 (IL-17 is a cytokine with pleiotropic effects associated with several inflammatory diseases. Although elevated levels of IL-17 have been described in inflammatory myopathies, its role in muscle remodeling and regeneration is still unknown. Excessive extracellular matrix degradation in skeletal muscle is an important pathological consequence of many diseases involving muscle wasting. In this study, the role of IL-17 on the expression of matrix metalloproteinase- (MMP- 9 in myoblast cells was investigated. The expression of MMP-9 after IL-17 treatment was analyzed in mouse myoblasts C2C12 cell line. The increase in MMP-9 production by IL-17 was concomitant with its capacity to inhibit myogenic differentiation of C2C12 cells. Doxycycline (Doxy treatment protected the myogenic capacity of myoblasts from IL-17 inhibition and, moreover, increased myotubes hypertrophy. Doxy blocked the capacity of IL-17 to stimulate MMP-9 production by regulating IL-17-induced ERK1/2 MAPK activation. Our results imply that MMP-9 mediates IL-17’s capacity to inhibit myoblast differentiation during inflammatory diseases and indicate that Doxy can modulate myoblast response to inflammatory induction by IL-17.

  2. Doxycycline Inhibits IL-17-Stimulated MMP-9 Expression by Downregulating ERK1/2 Activation: Implications in Myogenic Differentiation.

    Science.gov (United States)

    Obradović, Hristina; Krstić, Jelena; Kukolj, Tamara; Trivanović, Drenka; Đorđević, Ivana Okić; Mojsilović, Slavko; Jauković, Aleksandra; Jovčić, Gordana; Bugarski, Diana; Santibañez, Juan Francisco

    2016-01-01

    Interleukin 17 (IL-17) is a cytokine with pleiotropic effects associated with several inflammatory diseases. Although elevated levels of IL-17 have been described in inflammatory myopathies, its role in muscle remodeling and regeneration is still unknown. Excessive extracellular matrix degradation in skeletal muscle is an important pathological consequence of many diseases involving muscle wasting. In this study, the role of IL-17 on the expression of matrix metalloproteinase- (MMP-) 9 in myoblast cells was investigated. The expression of MMP-9 after IL-17 treatment was analyzed in mouse myoblasts C2C12 cell line. The increase in MMP-9 production by IL-17 was concomitant with its capacity to inhibit myogenic differentiation of C2C12 cells. Doxycycline (Doxy) treatment protected the myogenic capacity of myoblasts from IL-17 inhibition and, moreover, increased myotubes hypertrophy. Doxy blocked the capacity of IL-17 to stimulate MMP-9 production by regulating IL-17-induced ERK1/2 MAPK activation. Our results imply that MMP-9 mediates IL-17's capacity to inhibit myoblast differentiation during inflammatory diseases and indicate that Doxy can modulate myoblast response to inflammatory induction by IL-17.

  3. Vestibular evoked myogenic potentials and digital vectoelectronystagmography's study in patients with benign paroxysmal positional vertigo

    Directory of Open Access Journals (Sweden)

    Lira-Batista, Marta Maria da Silva

    2013-04-01

    Full Text Available Introduction: Benign Paroxysmal Positional Vertigo (BPPV is a very common vestibular disorder characterized by brief but intense attacks of rotatory vertigo triggered by simple rapid movement of the head. The integrity of the vestibular pathways can be assessed using tests such as digital vectoelectronystagmography (VENG and vestibular evoked myogenic potentials (VEMP. Aim: This study aimed to determine the VEMP findings with respect to latency, amplitude, and waveform peak to peak and the results of the oculomotor and vestibular components of VENG in patients with BPPV. Method: Although this otoneurological condition is quite common, little is known of the associated VEMP and VENG changes, making it important to research and describe these results. Results: We examined the records of 4438 patients and selected 35 charts after applying the inclusion and exclusion criteria. Of these, 26 patients were women and 9 men. The average age at diagnosis was 52.7 years, and the most prevalent physiological cause, accounting for 97.3% of cases, was ductolithiasis. There was a statistically significant association between normal hearing and mild contralateral sensorineural hearing loss. The results of the oculomotor tests were within the normal reference ranges for all subjects. Patients with BPPV exhibited symmetrical function of the semicircular canals in their synergistic pairs (p < 0.001. The caloric test showed statistically normal responses from the lateral canals. The waveforms of all patients were adequate, but the VEMP results for the data-crossing maneuver with positive positioning showed a trend toward a relationship for the left ear Lp13. There was also a trend towards an association between normal reflexes in the caloric test and the inter-peak VEMP of the left ear. It can be concluded that although there are some differences between the average levels of the VENG and VEMP results, these differences were not statistically significant

  4. Effects of stimulus and recording parameters on the air conduction ocular vestibular evoked myogenic potential.

    Science.gov (United States)

    Murnane, Owen D; Akin, Faith W; Kelly, Kip J; Byrd, Stephanie

    2011-01-01

    Vestibular evoked myogenic potentials (VEMPs) have been recorded from the sternocleidomastoid muscle (cervical VEMP or cVEMP) and more recently from the eye muscles (ocular VEMP or oVEMP) in response to air conduction and bone conduction stimuli. Both cVEMPs and oVEMPs are mediated by the otoliths and thereby provide diagnostic information that is complementary to videonystagmography and rotational chair tests. In contrast to the air conduction cVEMP, which originates from the saccule/inferior vestibular nerve, recent evidence suggests the possibility that the air conduction oVEMP may be mediated by the utricle/superior vestibular nerve. The oVEMP, therefore, may provide complementary diagnostic information relative to the cVEMP. There are relatively few studies, however, that have quantified the effects of stimulus and recording parameters on the air conduction oVEMP, and there is a paucity of normative data. To evaluate the effects of several stimulus and recording parameters on the air conduction oVEMP and to establish normative data for clinical use. A prospective repeated measures design was utilized. Forty-seven young adults with no history of neurologic disease, hearing loss, middle ear pathology, open or closed head injury, cervical injury, or audiovestibular disorder participated in the study. The effects of stimulus frequency, stimulus level, gaze elevation, and recording electrode location on the amplitude and latency of the oVEMP for monaural air conduction stimuli were assessed using repeated measures analyses of variance in an initial group of 17 participants. The optimal stimulus and recording parameters obtained in the initial group were used subsequently to obtain oVEMPs from 30 additional participants. The effects of stimulus frequency, stimulus level, gaze elevation, and electrode location on the response prevalence, amplitude, and latency of the oVEMP for monaural air conduction stimuli were significant. The maximum N1-P1 amplitude and response

  5. Influence of gender on the vestibular evoked myogenic potential Influência do gênero no potencial miogênico evocado vestibular

    Directory of Open Access Journals (Sweden)

    Aline Tenório Lins Carnaúba

    2011-04-01

    Full Text Available There is no consensus on the relevance of factors that influence gender differences in the behavior of muscles. Some studies have reported a relationship between muscle tension and amplitude of the vestibular evoked myogenic potential; others, that results depend on which muscles are studied or on how much load is applied. AIMS: This study aims to compare vestibular evoked myogenic potential parameters between genders in young individuals. METHODS: Eighty young adults were selected - 40 men and 40 women. Stimuli were averaged tone-bursts at 500 Hz, 90 dBHL intensity, and a 10-1000 Hz bandpass filter with amplification of 10-25 microvolts per division. The recordings were made in 80 ms windows. STUDY TYPE: An experimental and prospective study. RESULTS: No significant gender differences were found in wave latency - p = 0.19 and p = 0.50 for waves P13 and N23, respectively. No differences were found in amplitude values - p = 0.28 p = 0.40 for waves P13 and N23, respectively. CONCLUSION: There were no gender differences in latency and amplitude factors; the sternocleidomastoid muscle strain was monitored during the examination.Não existe consenso sobre a relevância dos fatores que influenciam as diferenças entre gêneros no comportamento dos músculos. Alguns estudos relatam existir uma relação entre tensão muscular e amplitude do potencial miogênico evocado vestibular, outros apenas que os resultados dependem dos músculos estudados ou do aumento da carga imposta. OBJETIVOS: Este estudo tem como objetivo comparar os parâmetros do potencial miogênico evocado vestibular, entre os gêneros, em indivíduos jovens. MATERIAL E MÉTODO: Selecionaram-se 80 adultos jovens, sendo 40 homens e 40 mulheres. Foram promediados estímulos tone burts na frequência de 500Hz, na intensidade de 90 dBNA, utilizando-se um filtro passa banda de 10 a 1000 Hz, com amplificação de 10 a 25 microvolts por divisão. Os registros foram realizados em janelas de 80

  6. The effect of some α-adrenoceptor antagonists on spontaneous myogenic activity in the rat portal vein and the putative involvement of ATP-sensitive K+channels

    NARCIS (Netherlands)

    Schwietert, R.; Wilhelm, D.; Wilffert, B.; Van Zwieten, P.A.

    1992-01-01

    In the present study we showed that the α-adrenoceptor antagonists phentolamine, yohimbine, prazosin, corynanthine and idazoxan, when cumulatively applied in high concentrations (1-100 μmol/l), can increase spontaneous myogenic activity in the rat portal vein. 5-Methyl-urapidil and rauwolscine were

  7. THE EFFECT OF SOME ALPHA-ADRENOCEPTOR ANTAGONISTS ON SPONTANEOUS MYOGENIC ACTIVITY IN THE RAT PORTAL-VEIN AND THE PUTATIVE INVOLVEMENT OF ATP-SENSITIVE K+ CHANNELS

    NARCIS (Netherlands)

    SCHWIETERT, R; WILHELM, D; WILFFERT, B; VANZWIETEN, PA

    1992-01-01

    In the present study we showed that the alpha-adrenoceptor antagonists phentolamine, yohimbine, prazosin, corynanthine and idazoxan, when cumulatively applied in high concentrations (1-100-mu-mol/l), can increase spontaneous myogenic activity in the rat portal vein. 5-Methyl-urapidil and rauwolscine

  8. Responses to noradrenaline of tail arteries in hypertensive, hypotensive and normotensive rats under different regimens of perfusion: role of the myogenic response

    DEFF Research Database (Denmark)

    Matchkov, Vladimir; Vlasova, Maria A; Tarasova, Olga S;

    1998-01-01

    The vasoconstrictor effects of noradrenaline were studied in spontaneously hypertensive rats (SHR) compared with Wistar Kyoto rats (WKY), and in Wistar rats with regional hypotension (WH) compared to control Wistar rats (WC). The abdominal aorta was ligated in WH distal to the renal arteries...... thickness causing different degrees of activation of the myogenic response to distension....

  9. Myogenic constriction is increased in mesenteric resistance arteries from rats with chronic heart failure : instantaneous counteraction by acute AT(1) receptor blockade

    NARCIS (Netherlands)

    Gschwend, S; Henning, RH; Pinto, YM; de Zeeuw, D; van Gilst, WH; Buikema, H

    2003-01-01

    1 Increased vascular resistance in chronic heart failure (CHF) has been attributed to stimulated neurohumoral systems. However, local mechanisms may also importantly contribute to set arterial tone. Our aim, therefore, was to test whether pressure-induced myogenic constriction of resistance arteries

  10. Vildagliptin restores renal myogenic function and attenuates renal sclerosis independently of effects on blood glucose or proteinuria in Zucker Diabetic Fatty rat

    NARCIS (Netherlands)

    Vavrinec, Peter; Henning, Robert H.; Landheer, Sjoerd W.; Wang, Yumei; Deelman, Leo E.; van Dokkum, Richard P. E.; Buikema, Hendrik

    2014-01-01

    Type 2 diabetes mellitus (T2DM) is associated with risk for chronic kidney disease (CKD), which is associated with a decrease in renal myogenic tone - part of renal autoregulatory mechanisms. Novel class of drugs used for the treatment of T2DM, dipeptidyl peptidase-4 (DPP-4) inhibitors, have protect

  11. Transcriptome analysis of post-hatch breast muscle in legacy and modern broiler chickens reveals enrichment of several regulators of myogenic growth.

    Science.gov (United States)

    Davis, Richard V N; Lamont, Susan J; Rothschild, Max F; Persia, Michael E; Ashwell, Chris M; Schmidt, Carl J

    2015-01-01

    Agriculture provides excellent model systems for understanding how selective pressure, as applied by humans, can affect the genomes of plants and animals. One such system is modern poultry breeding in which intensive genetic selection has been applied for meat production in the domesticated chicken. As a result, modern meat-type chickens (broilers) exhibit enhanced growth, especially of the skeletal muscle, relative to their legacy counterparts. Comparative studies of modern and legacy broiler chickens provide an opportunity to identify genes and pathways affected by this human-directed evolution. This study used RNA-seq to compare the transcriptomes of a modern and a legacy broiler line to identify differentially enriched genes in the breast muscle at days 6 and 21 post-hatch. Among the 15,945 genes analyzed, 10,841 were expressed at greater than 0.1 RPKM. At day 6 post-hatch 189 genes, including several regulators of myogenic growth and development, were differentially enriched between the two lines. The transcriptional profiles between lines at day 21 post-hatch identify 193 genes differentially enriched and still include genes associated with myogenic growth. This study identified differentially enriched genes that regulate myogenic growth and differentiation between the modern and legacy broiler lines. Specifically, differences in the ratios of several positive (IGF1, IGF1R, WFIKKN2) and negative (MSTN, ACE) myogenic growth regulators may help explain the differences underlying the enhanced growth characteristics of the modern broilers.

  12. Transcriptome analysis of post-hatch breast muscle in legacy and modern broiler chickens reveals enrichment of several regulators of myogenic growth.

    Directory of Open Access Journals (Sweden)

    Richard V N Davis

    Full Text Available Agriculture provides excellent model systems for understanding how selective pressure, as applied by humans, can affect the genomes of plants and animals. One such system is modern poultry breeding in which intensive genetic selection has been applied for meat production in the domesticated chicken. As a result, modern meat-type chickens (broilers exhibit enhanced growth, especially of the skeletal muscle, relative to their legacy counterparts. Comparative studies of modern and legacy broiler chickens provide an opportunity to identify genes and pathways affected by this human-directed evolution. This study used RNA-seq to compare the transcriptomes of a modern and a legacy broiler line to identify differentially enriched genes in the breast muscle at days 6 and 21 post-hatch. Among the 15,945 genes analyzed, 10,841 were expressed at greater than 0.1 RPKM. At day 6 post-hatch 189 genes, including several regulators of myogenic growth and development, were differentially enriched between the two lines. The transcriptional profiles between lines at day 21 post-hatch identify 193 genes differentially enriched and still include genes associated with myogenic growth. This study identified differentially enriched genes that regulate myogenic growth and differentiation between the modern and legacy broiler lines. Specifically, differences in the ratios of several positive (IGF1, IGF1R, WFIKKN2 and negative (MSTN, ACE myogenic growth regulators may help explain the differences underlying the enhanced growth characteristics of the modern broilers.

  13. Pbx and Prdm1a transcription factors differentially regulate subsets of the fast skeletal muscle program in zebrafish

    Directory of Open Access Journals (Sweden)

    Zizhen Yao

    2013-04-01

    The basic helix–loop–helix factor Myod initiates skeletal muscle differentiation by directly and sequentially activating sets of muscle differentiation genes, including those encoding muscle contractile proteins. We hypothesize that Pbx homeodomain proteins direct Myod to a subset of its transcriptional targets, in particular fast-twitch muscle differentiation genes, thereby regulating the competence of muscle precursor cells to differentiate. We have previously shown that Pbx proteins bind with Myod on the promoter of the zebrafish fast muscle gene mylpfa and that Pbx proteins are required for Myod to activate mylpfa expression and the fast-twitch muscle-specific differentiation program in zebrafish embryos. Here we have investigated the interactions of Pbx with another muscle fiber-type regulator, Prdm1a, a SET-domain DNA-binding factor that directly represses mylpfa expression and fast muscle differentiation. The prdm1a mutant phenotype, early and increased fast muscle differentiation, is the opposite of the Pbx-null phenotype, delayed and reduced fast muscle differentiation. To determine whether Pbx and Prdm1a have opposing activities on a common set of genes, we used RNA-seq analysis to globally assess gene expression in zebrafish embryos with single- and double-losses-of-function for Pbx and Prdm1a. We find that the levels of expression of certain fast muscle genes are increased or approximately wild type in pbx2/4-MO;prdm1a−/− embryos, suggesting that Pbx activity normally counters the repressive action of Prdm1a for a subset of the fast muscle program. However, other fast muscle genes require Pbx but are not regulated by Prdm1a. Thus, our findings reveal that subsets of the fast muscle program are differentially regulated by Pbx and Prdm1a. Our findings provide an example of how Pbx homeodomain proteins act in a balance with other transcription factors to regulate subsets of a cellular differentiation program.

  14. Big Stimulus, Little Ears: Safety in Administering Vestibular-Evoked Myogenic Potentials in Children.

    Science.gov (United States)

    Thomas, Megan L A; Fitzpatrick, Denis; McCreery, Ryan; Janky, Kristen L

    2017-05-01

    Cervical and ocular vestibular-evoked myogenic potentials (VEMPs) have become common clinical vestibular assessments. However, VEMP testing requires high intensity stimuli, raising concerns regarding safety with children, where sound pressure levels may be higher due to their smaller ear canal volumes. The purpose of this study was to estimate the range of peak-to-peak equivalent sound pressure levels (peSPLs) in child and adult ears in response to high intensity stimuli (i.e., 100 dB normal hearing level [nHL]) commonly used for VEMP testing and make a determination of whether acoustic stimuli levels with VEMP testing are safe for use in children. Prospective experimental. Ten children (4-6 years) and ten young adults (24-35 years) with normal hearing sensitivity and middle ear function participated in the study. Probe microphone peSPL measurements of clicks and 500 Hz tonebursts (TBs) were recorded in tubes of small, medium, and large diameter, and in a Brüel & Kjær Ear Simulator Type 4157 to assess for linearity of the stimulus at high levels. The different diameter tubes were used to approximate the range of cross-sectional areas in infant, child, and adult ears, respectively. Equivalent ear canal volume and peSPL measurements were then recorded in child and adult ears. Lower intensity levels were used in the participant's ears to limit exposure to high intensity sound. The peSPL measurements in participant ears were extrapolated using predictions from linear mixed models to determine if equivalent ear canal volume significantly contributed to overall peSPL and to estimate the mean and 95% confidence intervals of peSPLs in child and adult ears when high intensity stimulus levels (100 dB nHL) are used for VEMP testing without exposing subjects to high-intensity stimuli. Measurements from the coupler and tubes suggested: 1) each stimuli was linear, 2) there were no distortions or nonlinearities at high levels, and 3) peSPL increased with decreased tube diameter

  15. Contributions of biarticular myogenic components to the limitation of the range of motion after immobilization of rat knee joint

    Science.gov (United States)

    2014-01-01

    Background Muscle atrophy caused by immobilization in the shortened position is characterized by a decrease in the size or cross-sectional area (CSA) of myofibers and decreased muscle length. Few studies have addressed the relationship between limitation of the range of motion (ROM) and the changes in CSA specifically in biarticular muscles after atrophy because of immobilization. We aimed to determine the contribution of 2 distinct muscle groups, the biarticular muscles of the post thigh (PT) and those of the post leg (PL), to the limitation of ROM as well as changes in the myofiber CSAs after joint immobilization surgery. Methods Male Wistar rats (n = 40) were randomly divided into experimental and control groups. In the experimental group, the left knee was surgically immobilized by external fixation for 1, 2, 4, 8, or 16 weeks (n = 5 each) and sham surgery was performed on the right knee. The rats in the control groups (n = 3 per time point) did not undergo surgery. After the indicated immobilization periods, myotomy of the PT or PL biarticular muscles was performed and the ROM was measured. The hamstrings and gastrocnemius muscles from the animals operated for 1 or 16 weeks were subjected to morphological analysis. Results In immobilized knees, the relative contribution of the PT biarticular myogenic components to the total restriction reached 80% throughout the first 4 weeks and decreased thereafter. The relative contribution of the PL biarticular myogenic components remained contracture did not significantly change during the experimental period. However, the ratio of hamstrings CSAs to the sham side was larger than the ratio of medial gastrocnemius CSAs to the sham side after complete atrophy because of immobilization. PMID:25001065

  16. The Cervical Vestibular-Evoked Myogenic Potentials (cVEMPs) Recorded Along the Sternocleidomastoid Muscles During Head Rotation and Flexion in Normal Human Subjects.

    Science.gov (United States)

    Ashford, Alexander; Huang, Jun; Zhang, Chunming; Wei, Wei; Mustain, William; Eby, Thomas; Zhu, Hong; Zhou, Wu

    2016-08-01

    Tone burst-evoked myogenic potentials recorded from tonically contracted sternocleidomastoid muscles (SCM) (cervical VEMP or cVEMP) are widely used to assess the vestibular function. Since the cVEMP response is mediated by the vestibulo-collic reflex (VCR) pathways, it is important to understand how the cVEMPs are determined by factors related to either the sensory components (vestibular end organs) or the motor components (SCM) of the VCR pathways. Compared to the numerous studies that have investigated effects of sound parameters on the cVEMPs, there are few studies that have examined effects of SCM-related factors on the cVEMPs. The goal of the present study is to fill this knowledge gap by testing three SCM-related hypotheses. The first hypothesis is that contrary to the current view, the cVEMP response is only present in the SCM ipsilateral to the stimulated ear. The second hypothesis is that the cVEMP response is not only dependent on tonic level of the SCM, but also on how the tonic level is achieved, i.e., by head rotation or head flexion. The third hypothesis is that the SCM is compartmented and the polarity of the cVEMP response is dependent on the recording site. Seven surface electrodes were positioned along the left SCMs in 12 healthy adult subjects, and tone bursts were delivered to the ipsilateral or contralateral ear (8 ms plateau, 1 ms rise/fall, 130 dB SPL, 50-4000 Hz) while subjects activated their SCMs by head rotation (HR condition) or chin downward head flexion (CD condition). The first hypothesis was confirmed by the finding that the contralateral cVEMPs were minimal at all recording sites for all the tested tones during both HR and CD conditions. The second hypothesis was confirmed by the finding that the ipsilateral cVEMPs were larger in HR condition than in CD condition at recording sites above and below the SCM midpoint. Finally, the third hypothesis was confirmed by the finding that the cVEMPs exhibit reversed polarities at the sites

  17. 电针血清对多裂肌卫星细胞增殖及Pax-7、成肌分化抗原、磷酸化蛋白激酶B表达的影响%Effect of Electroacupuncture Serum on Proliferation of Cultured Multifidus Muscle Satellite Cells and Expression of Pax-7,MyoD and p-Akt

    Institute of Scientific and Technical Information of China (English)

    刘通; 于佳妮; 邹德辉; 陈玉佩; 卢宗孝; 晏珺; 张莉; 霍则军

    2016-01-01

    目的::观察电针血清对大鼠多裂肌卫星细胞增殖及 Pax-7、成肌分化抗原(MyoD)、磷酸化蛋白激酶 B(p-Akt)蛋白表达的影响,从血清学角度探讨电针促进多裂肌损伤修复的部分作用机制。方法:SD 大鼠随机分为正常组、模型组、电针委中组、电针肾俞组,每组各8只,采用0.5%布比卡因肌肉注射复制多裂肌损伤模型,各电针组分别选取“委中”穴、“肾俞”穴进行电针治疗,每日1次,每次20 min,4 d 后收集血清。原代培养大鼠腰多裂肌卫星细胞并进行鉴定,随机分为正常血清组、模型血清组、电针委中血清组、电针委中血清+LY 294002组、电针肾俞血清组、电针肾俞血清+LY 294002组,分别以各组血清培养24 h,CCK-8、EdU 法观察肌卫星细胞的增殖,Western blot 法检测肌卫星细胞 Pax-7、MyoD、p-Akt 蛋白表达。结果:与正常血清组相比,模型血清组多裂肌卫星细胞增殖速度明显加快(P 0.05)。结论:电针“委中”血清和电针“肾俞”血清均可通过磷脂酰肌醇3-激酶-蛋白激酶 B 通路促进肌卫星细胞的增殖,且有促进成肌分化的趋势。%Objective To observe the effect of electroacupuncture (EA)serum on proliferation of multifidus muscle sa-tellite cells (SCs)and expression of paired box transcription factor Pax-7,MyoD and protein kinase B (PKB or Akt)proteins of SCs,so as to explore its underlying mechanism in promoting repair of multifidus muscles.Methods Thirty-two SD rats were randomly assigned to control,model,EA-Weizhong (BL 40 )and EA-Shenshu (BL 23 )groups.The multifidus muscle injury (MFMI)model was established by injection of 0.5% bupivacaine hydrochloride (400 μL)into the bilateral L4 -L5 paravertebral muscles (4 points,1 00 μL for each point).EA stimulation was separately applied to bilateral BL 40 and BL 23 for 20 min,once daily,4 days altogether.Blood samples of the abdominal artery of rats in the above mentioned 4

  18. Skeletal Muscle Cell Induction from Pluripotent Stem Cells

    Directory of Open Access Journals (Sweden)

    Yusaku Kodaka

    2017-01-01

    Full Text Available Embryonic stem cells (ESCs and induced pluripotent stem cells (iPSCs have the potential to differentiate into various types of cells including skeletal muscle cells. The approach of converting ESCs/iPSCs into skeletal muscle cells offers hope for patients afflicted with the skeletal muscle diseases such as the Duchenne muscular dystrophy (DMD. Patient-derived iPSCs are an especially ideal cell source to obtain an unlimited number of myogenic cells that escape immune rejection after engraftment. Currently, there are several approaches to induce differentiation of ESCs and iPSCs to skeletal muscle. A key to the generation of skeletal muscle cells from ESCs/iPSCs is the mimicking of embryonic mesodermal induction followed by myogenic induction. Thus, current approaches of skeletal muscle cell induction of ESCs/iPSCs utilize techniques including overexpression of myogenic transcription factors such as MyoD or Pax3, using small molecules to induce mesodermal cells followed by myogenic progenitor cells, and utilizing epigenetic myogenic memory existing in muscle cell-derived iPSCs. This review summarizes the current methods used in myogenic differentiation and highlights areas of recent improvement.

  19. Myogenic, matrix and growth factor mRNA expression in human skeletal muscle: effect of contraction intensity and feeding

    DEFF Research Database (Denmark)

    Agergaard, Jakob; Reitelseder, Søren; Pedersen, T.G.

    2013-01-01

    exercise. No major differences were seen in atrophy-related genes between HL and LL resistance exercise. No changes were seen over 12-week training for any of the targets. CONCLUSIONS: Resistance exercise at LL and HL elevated the expression of genes involved in skeletal muscle hypertrophy, although......INTRODUCTION: We examined short-term (3-hour) and long-term (12-week) training effects after heavy load [HL; 70% 1RM] and light load (LL; 16% 1RM) exercise. METHODS: mRNA expression of genes involved in skeletal muscle remodeling were analyzed and muscle activity (EMG measurements) was measured....... RESULTS: Relative muscle activity differed between HL and LL resistance exercise, whereas median power frequency was even, suggesting an equal muscle-fiber-type recruitment distribution. mRNA expression of Myf6, myogenin, and p21 was mostly increased, and myostatin was mostly depressed by HL resistance...

  20. Cancer-associated myositis associated with oesophageal adenocarcinoma arising in Barrett's oesophagus without serum myogenic enzymes elevation: an example suggesting the importance of MRI.

    Science.gov (United States)

    Sasaki, Yosuke; Shimizu, Hiroshige; Nemoto, Tetsuo; Urita, Yoshihisa

    2016-04-21

    The strong association between myositis and malignancy has been well recognised. Cancer-associated myositis (CAM) is thought to be a cross-reaction to regenerating muscle tissue similar to tumour antigen. We report a case of CAM due to oesophageal adenocarcinoma arising in Barrett's oesophagus without elevation of myogenic enzymes, diagnosed by MRI and repeated endoscopy. Elderly onset, prominent symptoms, lack of interstitial pneumonia, poorer response to immunosuppressive therapies, and the combination of negative conventional myositis-related antibodies and positive anti-p155/140 antibody may help to distinguish CAM from idiopathic inflammatory myopathy. As the prognosis of patients with CAM depends on the malignancy, aggressive diagnosis of CAM and the causative malignancy is required. Our experience underscores the importance of avoiding the over-reliance on serum myogenic enzymes for excluding CAM and recognising MRI as a useful diagnostic tool of myositis.

  1. Ectopic expression of a polyalanine expansion mutant of poly(A)-binding protein N1 in muscle cells in culture inhibits myogenesis.

    Science.gov (United States)

    Wang, Qishan; Bag, Jnanankur

    2006-02-17

    Oculopharyngeal muscular dystrophy (OPMD) is an adult-onset dominant genetic disease caused by the expansion of a GCG trinucleotide repeat that encodes the polyalanine tract at the N-terminus of the nuclear poly(A)-binding protein (PABPN1). Presence of intranuclear inclusions (INIs) containing PABPN1 aggregates in the skeletal muscles is the hallmark of OPMD. Here, we show that ectopic expression of the mutant PABPN1 produced INIs in a muscle cell culture model and reduced expression of several muscle-specific proteins including alpha-actin, slow troponin C, muscle creatine kinase, and two myogenic transcription factors, myogenin and MyoD. However, the levels of two upstream regulators of the MyoD gene, the Myf-5 and Pax3/7, were not affected, but both proteins co-localized with the PABPN1 aggregates in the mutant PABPN1 overexpressing cells. In these cells, although myogenin and MyoD levels were reduced, these two transcription factors did not co-localize with the mutant PABPN1 aggregates. Therefore, sequestration of Myf5 and Pax3/7 by the mutant PABPN1 aggregates was a specific effect on these factors. Our results suggest that trapping of these two important myogenic determinants may interfere with an early step in myogenesis.

  2. Association of 17-β Estradiol with Adipose-Derived Stem Cells: New Strategy to Produce Functional Myogenic Differentiated Cells with a Nano-Scaffold for Tissue Engineering.

    Science.gov (United States)

    Feng, Chunxiang; Hu, Jinqian; Liu, Chang; Liu, Shiliang; Liao, Guiying; Song, Linjie; Zeng, Xiaoyong

    2016-01-01

    The increased incidence of stress urinary incontinence (SUI) in postmenopausal women has been proposed to be associated with a reduction in the level of 17-β estradiol (E2). E2 has also been shown to enhance the multi-differentiation ability of adipose-derived stem cells (ASCs) in vitro. However, studies on the potential value of E2 for tissue engineering in SUI treatment are rare. In the present study, we successfully fabricated myogenically differentiated ASCs (MD-ASCs), which were seeded onto a Poly(l-lactide)/Poly(e-caprolactone) electrospinning nano-scaffold, and incorporated E2 into the system, with the aim of improving the proliferation and myogenic differentiation of ASCs. ASCs were collected from the inguinal subcutaneous fat of rats. The proliferation and myogenic differentiation of ASCs, as well as the nano-scaffold biocompatibility of MD-ASCs, with or without E2 supplementation, were investigated. We demonstrated that E2 incorporation enhanced the proliferation of ASCs in vitro, and the most optimal concentration was 10-9 M. E2 also led to modulation of the MD-ASCs phenotype toward a concentrated type with smooth muscle-inductive medium. The expression of early (alpha-smooth muscle actin), mid (calponin), and late-stage (myosin heavy chain) contractile markers in MD-ASCs was enhanced by E2 during the different differentiation stages. Furthermore, the nano-scaffold was biocompatible with MD-ASCs, and cell proliferation was significantly enhanced by E2. Taken together, these results demonstrate that E2 can enhance the proliferation and myogenic differentiation of ASCs and can be used to construct a biocompatible cell/nano-scaffold. These scaffolds with desirable differentiation cells show promising applications for tissue engineering.

  3. PAX7 Targets, CD54, Integrin α9β1, and SDC2, Allow Isolation of Human ESC/iPSC-Derived Myogenic Progenitors

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    Alessandro Magli

    2017-06-01

    Full Text Available Pluripotent stem (PS-cell-derived cell types hold promise for treating degenerative diseases. However, PS cell differentiation is intrinsically heterogeneous; therefore, clinical translation requires the development of practical methods for isolating progenitors from unwanted and potentially teratogenic cells. Muscle-regenerating progenitors can be derived through transient PAX7 expression. To better understand the biology, and to discover potential markers for these cells, here we investigate PAX7 genomic targets and transcriptional changes in human cells undergoing PAX7-mediated myogenic commitment. We identify CD54, integrin α9β1, and Syndecan2 (SDC2 as surface markers on PAX7-induced myogenic progenitors. We show that these markers allow for the isolation of myogenic progenitors using both fluorescent- and CGMP-compatible magnetic-based sorting technologies and that CD54+α9β1+SDC2+ cells contribute to long-term muscle regeneration in vivo. These findings represent a critical step toward enabling the translation of PS-cell-based therapies for muscle diseases.

  4. Magnetization Transfer MR Imaging to Monitor Muscle Tissue Formation during Myogenic in Vivo Differentiation of Muscle Precursor Cells.

    Science.gov (United States)

    Rottmar, Markus; Haralampieva, Deana; Salemi, Souzan; Eberhardt, Christian; Wurnig, Moritz C; Boss, Andreas; Eberli, Daniel

    2016-11-01

    Purpose To determine whether magnetization transfer (MT) magnetic resonance (MR) imaging may serve as a quantitative measure of the degree of fiber formation during differentiation of muscle precursor cells into engineered muscle tissue as a potential noninvasive monitoring tool in mice. Materials and Methods The study was approved by the local ethics committee (no. StV 01/2008) and the local Veterinary Office (license no. 99/2013). Human muscle progenitor cells (hMPCs) derived from rectus abdominis muscles were subcutaneously injected into CD-1 nude mice (CD-1 nude mice, Crl:CD1-Foxn1(nu); Charles River Laboratories, Wilmington, Mass) for development of muscle tissue. The mice underwent MR imaging examinations at 4.7 T at days 1, 3, 7, 14, 21, and 28 after cell transplantation by using a gradient-echo sequence with an MT prepulse and systematic variation of the off-resonance frequency (50-37 500 Hz) at an amplitude of 800°. Direct saturation was estimated from a Bloch equation simulation. The MT ratio (MTR) was correlated to immunohistochemistry findings, Western blot results, and results of myography. Data were analyzed by using one-way or two-way analysis of variance with the Sidak or Tukey multiple comparisons test. Results In the reference skeletal muscle, highest MT was found for 2500 Hz off-resonance frequency with an MTR ± standard deviation of 57.5% ± 3.5. The developing muscle tissue exhibited increasing MT values during the 28 days of myogenic in vivo differentiation and did not reach the values of native skeletal muscle. Mean values of MTR (2500 Hz) for hMPCs were 27.6% ± 6.3 (day 1), 24.7% ± 8.7 (day 3), 28.2% ± 5.7 (day 7), 35.9% ± 5.0 (day 14), 37.0% ± 7.9 (day 21), and 39.9% ± 8.1 (day 28). The results from MT MR imaging correlated qualitatively well with muscle tissue expression of specific skeletal markers, as well as muscle contractility. Conclusion MT MR imaging may be used to noninvasively monitor the process of myogenic in vivo

  5. Long lasting pain hypersensitivity following ligation of the tendon of the masseter muscle in rats: A model of myogenic orofacial pain

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    Dubner Ronald

    2010-07-01

    Full Text Available Abstract Background A major subgroup of patients with temporomandibular joint (TMJ disorders have masticatory muscle hypersensitivity. To study myofacial temporomandibular pain, a number of preclinical models have been developed to induce myogenic pain of the masseter muscle, one of the four muscles involved in mastication. The currently used models, however, generate pain that decreases over time and only lasts from hours to weeks and hence are not suitable for studying chronicity of the myogenic pain in TMJ disorders. Here we report a model of constant myogenic orofacial pain that lasts for months. Results The model involves unilateral ligation of the tendon of the anterior superficial part of the rat masseter muscle (TASM. The ligation of the TASM was achieved with two chromic gut (4.0 ligatures via an intraoral approach. Nocifensive behavior of the rat was assessed by probing the skin site above the TASM with a series of von Frey filaments. The response frequencies were determined and an EF50 value, defined as the von Frey filament force that produces a 50% response frequency, was derived and used as a measure of mechanical sensitivity. Following TASM ligation, the EF50 of the injured side was significantly reduced and maintained throughout the 8-week observation period, suggesting the presence of mechanical hyperalgesia/allodynia. In sham-operated rats, the EF50 of the injured side was transiently reduced for about a week, likely due to injury produced by the surgery. Somatotopically relevant Fos protein expression was indentified in the subnucleus caudalis of the spinal trigeminal sensory complex. In the same region, persistent upregulation of NMDA receptor NR1 phosphorylation and protein expression and increased expression of glial markers glial fibrillary acidic protein (astroglia and CD11b (microglia were found. Morphine (0.4-8 mg/kg, s.c. and duloxetine (0.4-20 mg/kg, i.p., a selective serotonin-norepinephrine reuptake inhibitor

  6. Pulsed low-level infrared laser alters mRNA levels from muscle repair genes dependent on power output in Wistar rats

    Science.gov (United States)

    Trajano, L. A. S. N.; Trajano, E. T. L.; Thomé, A. M. C.; Sergio, L. P. S.; Mencalha, A. L.; Stumbo, A. C.; Fonseca, A. S.

    2017-10-01

    Satellite cells are present in skeletal muscle functioning in the repair and regeneration of muscle injury. Activation of these cells depends on the expression of myogenic factor 5 (Myf5), myogenic determination factor 1(MyoD), myogenic regulatory factor 4 (MRF4), myogenin (MyoG), paired box transcription factors 3 (Pax3), and 7 (Pax7). Low-level laser irradiation accelerates the repair of muscle injuries. However, data from the expression of myogenic factors have been controversial. Furthermore, the effects of different laser beam powers on the repair of muscle injuries have been not evaluated. The aim of this study was to evaluate the effects of low-level infrared laser at different powers and in pulsed emission mode on the expression of myogenic regulatory factors and on Pax3 and Pax7 in injured skeletal muscle from Wistar rats. Animals that underwent cryoinjury were divided into three groups: injury, injury laser 25 Mw, and injury laser 75 mW. Low-level infrared laser irradiation (904 nm, 3 J cm‑2, 5 kHz) was carried out at 25 and 75 mW. After euthanasia, skeletal muscle samples were withdrawn and the total RNA was extracted for the evaluation of mRNA levels from the MyoD, MyoG, MRF4, Myf5, Pax3, and Pax7 gene. Pax 7 mRNA levels did not alter, but Pax3 mRNA levels increased in the injured and laser-irradiated group at 25 mW. MyoD, MyoG, and MYf5 mRNA levels increased in the injured and laser-irradiated animals at both powers, and MRF4 mRNA levels decreased in the injured and laser-irradiated group at 75 mW. In conclusion, exposure to pulsed low-level infrared laser, by power-dependent effect, could accelerate the muscle repair process altering mRNA levels from paired box transcription factors and myogenic regulatory factors.

  7. Experiment list: SRX029147 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available growth protocol=forced to undergo myogenic differentiation by expression of MyoD || sample type=ChIP || flag immunoprecipitation...=anti-FLAG M2 agarose resin || 6xhis immunoprecipitation=His-Select nickel beads http://

  8. Is Alteration of Tuning Property in Cervical Vestibular-Evoked Myogenic Potential Specific for Ménière's Disease?

    Science.gov (United States)

    Murofushi, Toshihisa; Tsubota, Masahito; Suizu, Ryota; Yoshimura, Eriko

    2017-01-01

    The aim of this study is to show sensitivity and specificity of cervical vestibular-evoked myogenic potential (cVEMP) tuning property test to Ménière's disease (MD) in comparison with healthy controls (HC) and patients with other vestibular diseases. Totally 92 subjects (50 women and 42 men, 20-77 years of age) were enrolled in this study. Subjects were composed of 38 definite unilateral MD patients, 11 unilateral benign paroxysmal positional vertigo patients, 14 vestibular migraine patients, 19 unilateral vestibular neuritis patients, and 10 HC. The subjects underwent cVEMP testing to 500 and 1,000 Hz short tone bursts (125 dBSPL). The corrected amplitudes of the first biphasic responses (p13-n23) (cVEMP) were measured. Then, a tuning property index (the 500-1,000 Hz cVEMP slope) was calculated. The area of under the ROC curve (AUC) was 0.75 in comparison with other vestibular disease patients, while AUC was 0.77 in comparison with other vestibular disease patients plus HC. The best cutoff point of the 500-1,000 Hz cVEMP slope was -19.9. Sensitivity of the tuning property test to MD was 0.74, while specificity was 0.76 to other vestibular disease patients. The tuning property test of cVEMP is useful as a screening test of MD.

  9. Comparison of Vestibular Evoked Myogenic Potential and Caloric Tests Findings in Noise Induced Hearing Loss-Affected and Healthy Individuals

    Directory of Open Access Journals (Sweden)

    Farinoosh Fakharnia

    2010-06-01

    Full Text Available Background and Aim: Balance disturbance is one of the non-auditory effects of noisy industrial environments that is usually neglected. The aim of the current study was to investigate the effect of occupational noise on vestibular system among workers with noise-induced hearing loss (NIHL, based on both vestibular evoked myogenic potentials (VEMP and caloric tests.Methods: Thirty male workers with noise-induced hearing loss and thirty male matched controls were examined by VEMP and caloric tests. Study parameters included unilateral weakness, p13 and n23 latencies, and p13-n23 amplitude. Caloric test was performed only for 20 patients.Results: No significant difference was observed in unilateral weakness between the two groups. On the other hand, the difference in mean latencies of p13 in the right ear (p=0.003 and left ear (p=0.01 was significant between the two groups. However, the difference in n23 latency was significant only in the right ear (p=0.03. There was no significant difference between groups in p13-n23 amplitude.Conclusion: It seems that pars inferior of vestibule is the susceptible part in individuals with NIHL. In general, abnormal findings in both VEMP and caloric tests were more common compared to functional symptoms such as vertigo, which may be due to central compensation and the symmetry of the disorder.

  10. Multivariate modeling of cognitive-motor stimulation on neurovascular coupling: transcranial Doppler used to characterize myogenic and metabolic influences.

    Science.gov (United States)

    Panerai, Ronney B; Eyre, Michelle; Potter, John F

    2012-08-15

    Neural activation induces changes in cerebral blood flow velocity (CBFV) with separate contributions from resistance-area product (V(RAP)) and critical closing pressure (V(CrCP)). We modeled the dependence of V(RAP) and V(CrCP) on arterial blood pressure (ABP), end-tidal CO(2) (EtCO(2)), and cognitive stimulation to test the hypothesis that V(RAP) reflects myogenic activity while V(CrCP) reflects metabolic pathways. In 14 healthy subjects, CBFV was measured with transcranial Doppler ultrasound, ABP with the Finapres device and EtCO(2) with infrared capnography. Two different paradigms (word or puzzle) were repeated 10 times (30 s on-off), and the corresponding square-wave signal was used, together with ABP and EtCO(2), as inputs to autoregressive-moving average (ARMA) models, which allowed identification of the separate contributions of the three inputs to either V(RAP) or V(CrCP). For both paradigms, the contribution of ABP was mainly manifested through V(RAP) (P modeling of V(RAP) and V(CrCP) allows the separation of the effects of cerebral autoregulation and CO(2) reactivity from the main effects of cognitive-motor stimulation and have the potential to improve the diagnostic value of neurovascular coupling testing in physiological and clinical studies.

  11. Can Vestibular-Evoked Myogenic Potentials Help Differentiate Ménière Disease from Vestibular Migraine?

    Science.gov (United States)

    Zuniga, M. Geraldine; Janky, Kristen L.; Schubert, Michael C.; Carey, John P.

    2013-01-01

    Objectives To characterize both cervical and ocular vestibular-evoked myogenic potential (cVEMP, oVEMP) responses to air-conducted sound (ACS) and midline taps in Ménière disease (MD), vestibular migraine (VM), and controls, as well as to determine if cVEMP or oVEMP responses can differentiate MD from VM. Study Design Prospective cohort study. Setting Tertiary referral center. Subjects and Methods Unilateral definite MD patients (n = 20), VM patients (n = 21) by modified Neuhauser criteria, and age-matched controls (n = 28). cVEMP testing used ACS (clicks), and oVEMP testing used ACS (clicks and 500-Hz tone bursts) and midline tap stimuli (reflex hammer and Mini-Shaker). Outcome parameters were cVEMP peak-to-peak amplitudes and oVEMP n10 amplitudes. Results Relative to controls, MD and VM groups both showed reduced click-evoked cVEMP (P .210). Conclusions Using these techniques, VM and MD behaved similarly on most of the VEMP test battery. A link in their pathophysiology may be responsible for these responses. The data suggest a difference in 500-Hz tone burst–evoked oVEMP responses between MD and MV as a group. However, no VEMP test that was investigated segregated individuals with MD from those with VM. PMID:22267492

  12. Frequency-tuning characteristics of cervical and ocular vestibular evoked myogenic potentials induced by air-conducted tone bursts.

    Science.gov (United States)

    Park, Hong Ju; Lee, In-Sik; Shin, Jung Eun; Lee, Yeo Jin; Park, Mun Su

    2010-01-01

    To better characterize both ocular and cervical vestibular evoked myogenic potentials (VEMP) responses at different frequencies of sound in 20 normal subjects. Cervical and ocular VEMPs were recorded. The intensities of sound stimulation decreased from the maximal intensity, until no responses were evoked. Thresholds, amplitudes, latencies and interaural amplitude difference ratio (IADR) at the maximal stimulation were calculated. Both tests showed the similar frequency tuning, with the lowest threshold and highest amplitude for 500-Hz tone-burst stimuli. Sound stimulation at 500Hz showed the response rates of 100% in both tests. Cervical VEMPs showed higher incidence than ocular VEMPs. Ocular VEMP thresholds were significantly higher than those of cervical VEMP. Cervical VEMP amplitudes were significantly higher than ocular VEMP amplitudes. IADRs of ocular and cervical VEMPs did not differ significantly. Ocular VEMP showed the similar frequency tuning to cervical VEMP. Cervical VEMP responses showed higher incidence, lower thresholds and larger amplitudes than ocular VEMP. Cervical VEMP is a more reliable measure than ocular VEMP, though the results of both tests will be complementary. Five hundred Hertz is the optimal frequency to use. Copyright 2009 International Federation of Clinical Neurophysiology. Published by Elsevier Ireland Ltd. All rights reserved.

  13. Subclinical vestibular dysfunction in migraine patients: a preliminary study of ocular and rectified cervical vestibular evoked myogenic potentials.

    Science.gov (United States)

    Kim, Chul-Ho; Jang, Min-Uk; Choi, Hui-Chul; Sohn, Jong-Hee

    2015-01-01

    Many studies have identified various vestibular symptoms and laboratory abnormalities in migraineurs. Although the vestibular tests may be abnormal, the changes may exist without vestibular symptoms. To date, vestibular-evoked myogenic potential (VEMP) has been the easiest and simplest test for measuring vestibular function in clinical practice. Cervical VEMP (cVEMP) represents a vestibulo-collic reflex, whereas ocular VEMP (oVEMP) reflects a vestibulo-ocular pathway. Therefore, we determined whether ocular and rectified cervical VEMPs differed in patients with migraine or tension type headache (TTH) and compared the results to controls with no accompanying vestibular symptoms. The present study included 38 females with migraine without aura, 30 with episodic TTH, and 50 healthy controls without vestibular symptoms. oVEMP and cVEMP using a blood pressure manometer were recorded during a headache-free period. From the VEMP graphs, latency and amplitude parameters were analyzed, especially following EMG rectification in cVEMP. With respect to oVEMP, the migraine group exhibited significantly longer mean latencies of bilateral n1 and left p1 than the other groups (p ocular reflex. oVEMP is a more reliable measure than cVEMP to evaluate vestibular function in migraineurs, although results from the two tests in patients with migraine are complementary.

  14. The vestibular evoked myogenic potentials (VEMP) score: a promising tool for evaluation of brainstem involvement in multiple sclerosis.

    Science.gov (United States)

    Gabelić, T; Krbot Skorić, M; Adamec, I; Barun, B; Zadro, I; Habek, M

    2015-02-01

    Concerning the great importance of brainstem involvement in multiple sclerosis (MS), the aim of this study was to explore the role of the newly developed vestibular evoked myogenic potentials (VEMP) score as a possible marker of brainstem involvement in MS patients. This was a prospective case-control study which included 100 MS patients divided into two groups (without and with clinical signs of brainstem involvement) and 50 healthy controls. Ocular VEMP (oVEMP) and cervical VEMP (cVEMP) measurements were performed in all participants and analyzed for latencies, conduction block and amplitude asymmetry ratio. Based on this the VEMP score was calculated and compared with Expanded Disability Status Scale (EDSS), disease duration and magnetic resonance imaging data. Multiple sclerosis patients with clinical signs of brainstem involvement (group 2) had a statistically significant higher percentage of VEMP conduction blocks compared with patients without clinical signs of brainstem involvement (group 1) and healthy controls (P = 0.027 and P VEMP score was significantly higher in group 2 compared with group 1 (P = 0.018) and correlated with EDSS and disease duration (P = 0.011 and P = 0.032, respectively). Multivariate linear regression analysis showed that the VEMP score has a statistically significant influence on the EDSS score (P VEMP score enables better evaluation of brainstem involvement than either of these evoked potentials alone and correlates well with disability. © 2014 EAN.

  15. Ocular vestibular evoked myogenic potentials induced by air-conducted sound in patients with acute brainstem lesions.

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    Oh, Sun-Young; Kim, Ji Soo; Lee, Jong-Min; Shin, Byoung-Soo; Hwang, Seung-Bae; Kwak, Ki-Chang; Kim, Chanmi; Jeong, Seul-Ki; Kim, Tae-Woo

    2013-04-01

    The ocular vestibular-evoked myogenic potential (oVEMP), a recently documented otolith-ocular reflex, is considered to reflect the central projections of the primary otolithic afferent fibers to the oculomotor nuclei. The aim of our study is to define air-conducted sound oVEMP abnormality in patients with acute brainstem lesions and to determine the brainstem structures involved in the generation of oVEMPs. In response to air-conducted tone burst sounds (ACS), oVEMP was measured in 52 patients with acute brainstem lesions. Individualized brainstem lesions were analyzed by means of MRI-based voxel-wise lesion-behavior mapping, and the probabilistic lesion maps were constructed. More than half (n=28, 53.8%) of the patients with acute brainstem lesions showed abnormal oVEMP in response to ACS. The majority of patients with abnormal oVEMPs had lesions in the dorsomedial brainstem that contains the medial longitudinal fasciculus (MLF), the crossed ventral tegmental tract (CVTT), and the oculomotor nuclei and nerves. MLF, CVTT, and the oculomotor nuclei and nerves appear to be responsible for otolith-ocular responses in the brainstem. Complemented to cervical VEMP for the uncrossed otolith-spinal function, oVEMP to ACS may be applied to evaluate the crossed otolith-ocular function in central vestibulopathies. Copyright © 2012 International Federation of Clinical Neurophysiology. Published by Elsevier Ireland Ltd. All rights reserved.

  16. Assessment of the otolith-ocular reflex using ocular vestibular evoked myogenic potentials in patients with episodic lateral tilt sensation.

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    Murofushi, Toshihisa; Nakahara, Haruka; Yoshimura, Eriko

    2012-05-02

    The otolith-ocular reflex in patients with episodic lateral tilt sensation without any other vestibular symptoms was assessed using ocular vestibular evoked myogenic potentials (oVEMP). Ten patients (6 men and 4 women, mean age=53.5) were enrolled. All patients had episodic lateral tilt sensation. Patients with a medical history of rotatory vertigo, loss of consciousness, head trauma, or symptoms or signs of central nervous dysfunction or proprioceptive dysfunction and those who had been definitely diagnosed with a disease that causes disequilibrium were excluded. All of the 10 patients had oVEMP tests and cervical VEMP (cVEMP) tests and underwent caloric tests. Eight of the 10 patients showed unilateral absence of oVEMP, one displayed a bilateral absence, and one displayed normal oVEMP. Concerning cVEMP, 4 patients showed a unilateral absence of cVEMP, one displayed unilaterally decreased responses and 5 displayed normal cVEMP. All patients showed normal bilateral caloric responses. The present study showed that patients with episodic lateral tilt sensation displayed abnormal otolith-ocular reflexes, as shown by their oVEMP, suggesting that these patients were suffering from utricular dysfunction. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  17. Correlation of Superior Canal Dehiscence Surface Area With Vestibular Evoked Myogenic Potentials, Audiometric Thresholds, and Dizziness Handicap.

    Science.gov (United States)

    Hunter, Jacob B; O'Connell, Brendan P; Wang, Jianing; Chakravorti, Srijata; Makowiec, Katie; Carlson, Matthew L; Dawant, Benoit; McCaslin, Devin L; Noble, Jack H; Wanna, George B

    2016-09-01

    To correlate objective measures of vestibular and audiometric function as well as subjective measures of dizziness handicap with the surface area of the superior canal dehiscence (SCD). Retrospective chart review and radiological analysis. Single tertiary academic referral center. Preoperative computed tomography imaging, patient survey, audiometric thresholds, and vestibular evoked myogenic potential (VEMP) testing in patients with confirmed SCD. Image analysis techniques were developed to measure the surface area of each SCD in computed tomography imaging. Preoperative ocular and cervical VEMPs, air and bone conduction thresholds, air-bone gap, dizziness handicap inventory scores, and surface area of the SCD. Fifty-three patients (mean age 52.7 yr) with 84 SCD were analyzed. The median surface area of dehiscence was 1.44 mm (0.068-8.23 mm). Ocular VEMP amplitudes (r = 0.61, p cervical VEMP amplitudes (r = 0.62, p cervical VEMP thresholds (r = -0.56, p ocular and cervical VEMP amplitudes, cervical VEMP thresholds, and air conduction thresholds at 250 Hz are significantly correlated with the surface area of the dehiscence.

  18. Brainstem lesion in benign paroxysmal vertigo children: Evaluated by a combined ocular and cervical vestibular-evoked myogenic potential test.

    Science.gov (United States)

    Lin, Kuei-You; Hsu, Ying-Shuo; Young, Yi-Ho

    2010-05-01

    This study utilized a combined ocular vestibular-evoked myogenic potential (oVEMP) and cervical VEMP (cVEMP) test in children with benign paroxysmal vertigo (BPV) to investigate whether the upper or lower brainstem is more frequently affected in BPV children. Fifteen BPV children aged 4-14 years, and 15 age- and sex-matched healthy children were enrolled. All subjects underwent pure tone audiometry, stabilometry, and a combined oVEMP and cVEMP test using acoustic stimulation. All BPV patients displayed normal hearing and clear oVEMPs. However, 11 (73%) of 15 BPV patients had delayed cVEMPs, showing significant difference when compared with 100% normal cVEMPs in healthy children. The sway path and sway area in stabilometry were significantly different between BPV and healthy children, regardless of whether their eyes were open or closed. However, neither the sway path nor sway area correlated significantly with cVEMP results. Normal oVEMPs in BPV children indicate an intact vestibulo-ocular reflex pathway, which travels through the upper brainstem. In contrast, delayed cVEMPs in BPV children reflect a retrolabyrinthine lesion along the sacculo-collic reflex pathway, which descends via the lower brainstem. Hence, the lower brainstem is more frequently affected than the upper brainstem in children with BPV. Copyright (c) 2010 Elsevier Ireland Ltd. All rights reserved.

  19. Degeneration of the vestibular nerve in unilateral Meniere's disease evaluated by galvanic vestibular-evoked myogenic potentials.

    Science.gov (United States)

    Chang, Chih-Ming; Young, Yi-Ho; Jaw, Fu-Shan; Wang, Chi-Te; Cheng, Po-Wen

    2017-09-01

    The staging system of Meniere's disease utilizes audiograms to probe cochlear dysfunction. We explored the addition of galvanic vestibular-evoked myogenic potentials (VEMP) to further explore vestibular function. Seventy patients with unilateral Meniere's disease were enrolled in this study. Within 2weeks of diagnosis, all subjects underwent pure tone audiometry, cervical and ocular VEMP, and caloric test. The prevalence of abnormal tests and the VEMP characteristic parameters such as latencies and amplitudes were analyzed. In affected ears, the abnormal rate of acoustic cVEMPs, galvanic cVEMPs, vibratory oVEMPs and galvanic oVEMPs was 37%, 17%, 20%, and 9%, respectively. No significant differences existed in VEMP latencies and amplitudes between affected ears and unaffected ears. The impairment of otolithic organs was found to be more than that of vestibular afferents. The deterioration of the saccule was more than that of the utricle, whereas retrolabyrinthine degeneration of sacculo-collic reflex and vestibulo-ocular reflex was similar. This study is the first to use an electrophysiological test to evaluate the retrolabyrinthine function of patients with unilateral Meniere's disease. Copyright © 2017 International Federation of Clinical Neurophysiology. Published by Elsevier B.V. All rights reserved.

  20. Feasibility of simultaneous recording of cervical and ocular vestibular-evoked myogenic potentials via galvanic vestibular stimulation.

    Science.gov (United States)

    Chang, Chih-Ming; Young, Yi-Ho; Cheng, Po-Wen

    2013-12-01

    Simultaneous galvanic vestibular stimulation (GVS)-cervical vestibular-evoked myogenic potential (cVEMP) and GVS-ocular (oVEMP) tests yielded similar information to that obtained in individual tests. This study compared the characteristic parameters of cVEMPs and oVEMPs via GVS between individual and simultaneous recording patterns in healthy and elderly subjects. Consequently, the effectiveness of simultaneous GVS-cVEMP and GVS-oVEMP tests was assessed. A total of 24 healthy and 16 elderly subjects were enrolled in this study. All participants underwent individual cVEMP, individual oVEMP, and simultaneous cVEMP and oVEMP testing via GVS mode in a random order. The response rates and characteristic parameters of cVEMPs and oVEMPs between individual and simultaneous tests, including latencies, intervals, and amplitudes, were measured. The VEMP parameters, including latencies, intervals, and amplitudes, all demonstrated no significant differences between individual and simultaneous tests (p > 0.05, paired t test), either in healthy or elderly subjects. Pearson's correlation analyses also revealed significant positive correlations in all parameters between these two tests (p < 0.05).

  1. Electrophysiologic vestibular evaluation in type 2 diabetic and prediabetic patients: Air conduction ocular and cervical vestibular evoked myogenic potentials.

    Science.gov (United States)

    Konukseven, Ozlem; Polat, Sefika Burcak; Karahan, Sevilay; Konukseven, Ertan; Ersoy, Reyhan; Cakir, Bekir; Kutluhan, Ahmet; Aksoy, Songul

    2015-08-01

    Chronically increased blood glucose levels may affect the vestibular system by damaging cells and neural structures in diabetes mellitus (DM). We aimed to search the effects of neurovascular degeneration on the vestibular system in type 2 DM and prediabetic patients by using air-conducted ocular (oVEMP) and cervical (cVEMP) vestibular evoked myogenic potentials. Prospective study. Thirty diabetic, 30 prediabetic patients, and 31 age- and sex-matched controls having no peripheral or central vestibular disease, were enrolled. All participants were evaluated by audiovestibular tests, oVEMP, and cVEMP. In the diabetic group, mean values of both oVEMP and cVEMP p1, n1 latencies were significantly longer compared to the prediabetic group and the control group, whereas latencies were similar in prediabetic and the control groups. Bilateral neural dysfunction was recognized in both tests and lateralization was not seen in VEMP asymmetric ratios. In the diabetic group, prevalence of pathological p1 and n1 latencies in oVEMP were 30.4% and 37.5%, whereas they were 53.7%, 59.3% in cVEMP, respectively. p1 latencies of cVEMP and oVEMP were positively correlated with HbA1c and fasting plasma glucose level in the diabetic group. Subclinical vestibular neuropathy can be a newly defined diabetes-related complication.

  2. Ocular vestibular evoked myogenic potentials induced by bone-conducted vibration in patients with unilateral inner ear disease.

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    Nagai, Noriko; Ogawa, Yasuo; Hagiwara, Akira; Otsuka, Koji; Inagaki, Taro; Shimizu, Shigetaka; Suzuki, Mamoru

    2014-02-01

    Patients with vestibular neuritis (VN) with complete canal paresis (CP) showed a higher rate of abnormal ocular vestibular evoked myogenic potential (oVEMP) than those with partial CP. From these results, it is speculated that the superior vestibular nerve function mainly affects oVEMP. Significant correlation was found between the grades of the hearing outcome and oVEMP in sudden sensorineural hearing loss (SSHL). We attempted to correlate the results of oVEMP with the results of cervical VEMP (cVEMP), results of subjective visual vertical (SVV), and clinical course in patients with various vestibular disorders. Twenty-two patients with VN, 65 with SSHL, and 22 with Meniere's disease (MD), were enrolled in this study. We compared the results of oVEMP with those of cVEMP, SVV, and the caloric test. Furthermore, the oVEMP results were compared with the initial hearing threshold, presence of vertigo, and hearing recovery in the patients with SSHL. The patients with VN with complete CP showed a higher rate of abnormal oVEMP than those with partial CP. In the patients with SSHL, the hearing recovery rate was lower in the patients with abnormal oVEMP than in those with normal oVEMP.

  3. Effect of personal music system use on sacculocollic reflex assessed by cervical vestibular-evoked myogenic potential: A preliminary investigation.

    Science.gov (United States)

    Singh, Niraj Kumar; Sasidharan, Chithra Sobha

    2016-01-01

    Listening to music through a portable personal music system (PMS) is a growing trend, especially among the youth. The preferred listening level in such kinds of PMS has been reported to cross the safe levels and its impact on the auditory system was demonstrated in several previous investigations. Owing to the commonality in several aspects between the auditory and the vestibular systems, it appears likely that the deleterious effects of PMS use could also be impinging on the vestibular system, which has never been investigated. The present study therefore, aimed at evaluating the effects of PMS use on the sacculocollic reflex assessed by the cervical vestibular-evoked myogenic potential (cVEMP) technique. Thirty-two regular PMS users and 32 nonregular PMS users underwent cVEMP testing using alternating polarity 500 Hz tone bursts. The results revealed no significant group difference in latencies and interaural asymmetry ratio. However, the cVEMP was significantly reduced in the group of individuals in whom the diffused field equivalent sound pressure levels (SPLs) were above the damage risk criteria (DRC) compared to those with diffused field equivalent SPLs below it (P< 0.01). Therefore, the use of PMS at high levels of volume controls could be deleterious to the vestibular well-being of an individual.

  4. Imaging and Outcomes for a New Entity: Low-Grade Sinonasal Sarcoma with Neural and Myogenic Features

    Science.gov (United States)

    Cannon, Richard B.; Wiggins, Richard H.; Witt, Benjamin L.; Dundar, Yusuf; Johnston, Tawni M.; Hunt, Jason P.

    2017-01-01

    Objectives Low-grade sinonasal sarcoma with neural and myogenic features (LGSSNMF) is a new, rare tumor. Our goal is to describe the imaging characteristics and surgical outcomes of this unique skull base malignancy. Design Retrospective case series. Setting Academic medical center. Participants There were three patients who met inclusion criteria with a confirmed LGSSNMF. Main Outcome Measures Imaging and histopathological characteristics, treatments, survival and recurrence outcomes, complications, morbidity, and mortality. Results Patients presented with diplopia, facial discomfort, a supraorbital mass, and nasal obstruction. Magnetic resonance imaging and computed tomography imaging in all cases showed an enhancing sinonasal mass with associated hyperostotic bone formation that involved the frontal sinus, invaded the lamina papyracea and anterior skull base, and had intracranial extension. One patient underwent a purely endoscopic surgical resection and the second underwent a craniofacial resection, while the last is pending treatment. All patients recovered well, without morbidity or long-term complications, and are currently without evidence of disease (mean follow-up of 2.1 years). One patient recurred after 17 months and underwent a repeat endoscopic skull base and dural resection. Conclusions The surgical outcomes and imaging of this unique, locally aggressive skull base tumor are characterized. PMID:28229035

  5. Effect of personal music system use on sacculocollic reflex assessed by cervical vestibular-evoked myogenic potential: A preliminary investigation

    Directory of Open Access Journals (Sweden)

    Niraj Kumar Singh

    2016-01-01

    Full Text Available Listening to music through a portable personal music system (PMS is a growing trend, especially among the youth. The preferred listening level in such kinds of PMS has been reported to cross the safe levels and its impact on the auditory system was demonstrated in several previous investigations. Owing to the commonality in several aspects between the auditory and the vestibular systems, it appears likely that the deleterious effects of PMS use could also be impinging on the vestibular system, which has never been investigated. The present study therefore, aimed at evaluating the effects of PMS use on the sacculocollic reflex assessed by the cervical vestibular-evoked myogenic potential (cVEMP technique. Thirty-two regular PMS users and 32 nonregular PMS users underwent cVEMP testing using alternating polarity 500 Hz tone bursts. The results revealed no significant group difference in latencies and interaural asymmetry ratio. However, the cVEMP was significantly reduced in the group of individuals in whom the diffused field equivalent sound pressure levels (SPLs were above the damage risk criteria (DRC compared to those with diffused field equivalent SPLs below it (P< 0.01. Therefore, the use of PMS at high levels of volume controls could be deleterious to the vestibular well-being of an individual.

  6. MITOGENIC SIGNALS CONTROL TRANSLATION OF THE EARLY GROWTH-RESPONSE GENE-1 IN MYOGENIC CELLS

    NARCIS (Netherlands)

    MAASS, A; GROHE, C; OBERDORF, S; SUKHATME, VP; VETTER, H; NEYSES, L

    1994-01-01

    Muscle is a major site of expression of the early growth reponse gene-1 (Egr-1). To investigate its role in muscle proliferation and/or differentiation we studied the effect of a variety of growth factors on cultured mouse muscle So18 cells. Three groups of responses could be distinguished: 1. AII,

  7. Maternal nutrient restriction in early gestation upregulates myogenic genes in cattle fetal muscle tissue

    Science.gov (United States)

    Prenatal myogenesis is a critical factor in determining the muscle growth potential of cattle. We hypothesized that maternal nutrient restriction during early gestation would alter the transcriptome of fetal primordial muscle tissue in cattle. A total of 14 Angus-cross heifers were estrus synchroniz...

  8. Vestibular evoked myogenic potentials are heavily dependent on type I hair cell activity of the saccular macula in guinea pigs.

    Science.gov (United States)

    Lue, June-Horng; Day, An-Shiou; Cheng, Po-Wen; Young, Yi-Ho

    2009-01-01

    This study applied the vestibular evoked myogenic potential (VEMP) test to guinea pigs coupled with electronic microscopic examination to determine whether VEMPs are dependent on type I or II hair cell activity of the saccular macula. An amount of 0.05 ml of gentamicin (40 mg/ml) was injected directly overlaying, but not through, the round window membrane of the left ear in guinea pigs.One week after surgery, auditory brainstem response test revealed normal responses in 12 animals (80%), and elevated thresholds in 3 animals (20%). The VEMP test using click stimulation showed absent responses in all 15 animals (100%). Another 6 gentamicin-treated animals underwent the VEMP test using galvanic stimulation and all 6 also displayed absent responses. Ultrathin sections of the saccular macula in the gentamicin-treated ears displayed morphologic alterations in type I or II hair cells, including shrinkage and/or vacuolization in the cytoplasm, increased electron density of the cytoplasm and nuclear chromatin, and cellular lucency. However, extrusion degeneration was rare and only present in type II hair cells. Quantitative analysis demonstrated that the histological density of intact type I hair cells was 1.1 +/- 1.2/4000 microm(2) in the gentamicin-treated ears, showing significantly less than that in control ears (4.5 +/- 1.8/4000 microm(2)). However, no significant difference was observed in the densities of intact type II hair cells and supporting cells between treated and control ears. Furthermore, the calyx terminals surrounding the damaged type I hair cells were swollen and disrupted, while the button afferents contacting the damaged type II hair cells were not obviously deformed. Based on the above results, we therefore conclude that VEMPs are heavily dependent on type I hair cell activity of the saccular macula in guinea pigs.

  9. The role of cervical and ocular vestibular-evoked myogenic potentials in the follow-up of vestibular neuritis.

    Science.gov (United States)

    Adamec, Ivan; Skorić, Magdalena Krbot; Handžić, Jadranka; Barušić, Anabella Karla; Bach, Ivo; Gabelić, Tereza; Habek, Mario

    2014-04-01

    This study evaluates the recovery of vestibular nerve function after vestibular neuritis (VN) by vestibular-evoked myogenic potentials (VEMPs). Twenty-six patients with the diagnosis of VN were included. All patients underwent ocular VEMP (oVEMP) and cervical VEMP (cVEMP) recordings, at 6 days and 6 months from the onset of the symptoms. Of the 26 patients, 14 showed improvement on oVEMP at month 6 (group 1), and 12 showed no change or worsening on oVEMP at 6 months (group 2). At the same time, there was no change in the amplitudes of the cVEMP on either healthy or affected sides in both groups. Inability to perform the Fukuda test, and chronic white matter supratentorial lesions present on brain magnetic resonance imaging (MRI) were more frequent in patients with worse outcome on oVEMP (P = 0.044 and 0.045, respectively). Although involvement of the inferior branch of the vestibular nerve was not associated with oVEMP outcome, oVEMP latencies (N10 and P13) were associated with improvement or worsening in oVEMP amplitudes, showing that prolonged latencies correlate with 6-month improvement in oVEMP amplitudes (Pearson correlation -0.472, P = 0.041 and -0.580, P = 0.009, respectively). This study identified clinical, MRI and neurophysiological predictors of recovery in patients with superior VN, and offers additional insight into, and better understanding of, the role of VEMP in diagnosis and prognosis of patients with VN. Further studies are needed to validate this diagnostic procedure and to assess its clinical usefulness in VN management.

  10. Test-retest reliability and age-related characteristics of the ocular and cervical vestibular evoked myogenic potential tests.

    Science.gov (United States)

    Nguyen, Kimanh D; Welgampola, Miriam S; Carey, John P

    2010-07-01

    To determine the test-retest reliability and age-related trends of the cervical and ocular vestibular evoked myogenic potential (cVEMP and oVEMP, respectively) responses to air-conducted sound and bone-conducted vibration stimulation. Prospective study. Tertiary referral center. Fifty-three healthy adults with no hearing or vestibular deficits. All subjects underwent cVEMP and oVEMP testing in response to sounds (0.1-ms clicks and 500-Hz tone bursts) and vibration (midline forehead taps at the hairline, Fz, with a reflex hammer and a Brüel & Kjaer Mini-Shaker Type 4810). Twelve subjects underwent an additional testing session that was conducted at a mean of 10 weeks after the first one. Test-retest reliability for VEMP response parameters (latency, peak-to-peak amplitude, and asymmetry ratio) were assessed using the intraclass correlation coefficient (ICC). : oVEMP amplitudes had excellent test-retest reliability (ICC > 0.75) for all 4 stimuli; cVEMP amplitudes had excellent reliability for hammer taps and fair-to-good reliability for other stimuli. oVEMP asymmetry ratios had excellent reliability for clicks and fair-to-good reliability (ICC = 0.4-0.75) for other stimuli; cVEMP asymmetry ratios had fair-to-good reliability for clicks and hammer taps. Older subjects (>50 years old) were found to have significantly decreased cVEMP amplitudes in response to clicks, tones, and taps with a Mini-Shaker and significantly decreased oVEMP amplitudes in response to clicks, tones, and taps with a reflex hammer. No age-related changes were found for latencies or asymmetry ratios. Overall, oVEMP response parameters demonstrated better test-retest reliability than cVEMP response parameters, but oVEMPs and cVEMPs had similar age-related changes.

  11. Frequency and phase effects on cervical vestibular evoked myogenic potentials (cVEMPs) to air-conducted sound.

    Science.gov (United States)

    Govender, Sendhil; Dennis, Danielle L; Colebatch, James G

    2016-09-01

    Few previous studies of tuning using air-conducted (AC) stimuli and the cervical vestibular evoked myogenic potential (cVEMP) have compensated for the effects of middle ear (ME) attenuation. Zhang et al. (Exp Brain Res 213:111-116, 2011a) who did allow for ME effects were able to show a secondary peak around 100 Hz for the ocular VEMP (oVEMP). Recently, it has become clear that the otolith afferents responsible for the cVEMP and oVEMP differ and thus the nature of tuning may be more related to the reflex studied determining which otolith receptors are activated rather than the properties of the stimulus. We wished to reinvestigate the tuning for the cVEMP using AC stimuli, to establish whether the low-frequency peak is specific for the oVEMP or a consequence of the stimulus modality itself. In response to recent evidence using a 500 Hz AC stimulus that there was no effect of stimulus phase, we also investigated whether phase (condensation or rarefaction) had an effect at any frequency. We measured corrected cVEMP amplitudes and latencies in response to stimuli between 50 and 1200 Hz in 10 normal volunteers using an AC stimulus adjusted for ME attenuation. We confirmed earlier reports of the similarity of the tuning for both the cVEMP and oVEMP reflexes but found no separate 100 Hz peak for the cVEMP. AC stimulus phase did not affect either amplitude or latency. Both the tuning pattern and the phase effects contrast with those previously reported for bone-conducted (BC) stimuli. Unlike BC stimulation, which shows tuning consistent with an action on the otolith membrane, AC stimuli are likely to act through a different mechanism, most likely directly at the hair cell level.

  12. Frequency-Amplitude Ratio of Ocular Vestibular-Evoked Myogenic Potentials for Detecting Meniere's Disease: A Preliminary Investigation.

    Science.gov (United States)

    Singh, Niraj Kumar; Barman, Animesh

    2016-01-01

    Several parameters of ocular vestibular-evoked myogenic potential (oVEMP) have been used to identify Meniere's disease. Nonetheless frequency-amplitude ratio (FAR), which is the ratio of amplitude between two frequencies, is one among the parameters that has failed to attract researchers' attention despite proving its worth in diagnosis of Meniere's disease when used in conjunction with cervical VEMP. Thus, the present study aimed at investigating the utility of FAR of oVEMP in identifying Meniere's disease and finding out an optimum frequency pair for its diagnosis. Using a case-control design, oVEMPs were recorded for tone bursts of 500, 750, 1000, and 1500 Hz from 36 individuals with unilateral definite Meniere's disease in the age range of 15 to 50 years. For comparison purposes, oVEMP at the above frequencies were also obtained from an equal number of age- and gender-matched healthy individuals. The amplitudes of 750, 1000, 1500 Hz and tuned frequency, which was the frequency with the largest peak to peak amplitude among the above-mentioned frequencies, were divided by the amplitude of 500 Hz to obtain FARs for 750/500, 1000/500, 1500/500, and tuned frequency/500 frequency pairs. The results revealed significantly higher FAR in the Meniere's disease group than the healthy controls for all the frequency pairs (p < 0.05). The sensitivity of almost 90% and the specificity 100% was obtained for 1000/500 and 750/500, whereas the other frequency pairs produced a sensitivity of about 56% while still showing a specificity of 100%. High sensitivity and specificity, coupled with considerably lowered test duration when using only two frequencies, makes the use of FAR a more attractive prerogative, with 1000/500 as the frequency pair of choice.

  13. Assessment of the Clinical Utility of Cervical and Ocular Vestibular Evoked Myogenic Potential Testing in Elderly Patients.

    Science.gov (United States)

    Piker, Erin G; Baloh, Robert W; Witsell, David L; Garrison, Doug B; Lee, Walter T

    2015-08-01

    To assess whether patient age or sex was predictive of a bilaterally absent cervical or ocular vestibular evoked myogenic potential (cVEMP or oVEMP). Retrospective case review. Tertiary center. Patients presenting with normal vestibular tests (i.e. normal caloric and rotational chair) who underwent cVEMP and/or oVEMP testing. Patients with conductive hearing loss were excluded as were those with unilaterally abnormal VEMP results because they presented with evidence of a possible unilateral vestibular impairment. A total of 895 patients met criteria for cVEMPs and 297 for oVEMPs. The presence or absence of cVEMP and oVEMP responses elicited with a 500-Hz 125-dB pSPL air conduction stimulus. A logistic regression was performed including odd ratios and confidence intervals. Compared with adults in their 20s, the odds of bilaterally absent cVEMP responses are 6 times greater for patients in their 50s and 60s and over 22 times greater for patients in their 70s and 80s. A bilaterally absent oVEMP response is 6 times more likely for patients in their 40s, 50s, and 60 and 13 times greater for patients in their 70s. VEMPs in response to air conduction stimuli are bilaterally absent in a large percentage of older patients complaining of dizziness who otherwise have normal vestibular and auditory testing for their age. In combination with other abnormal vestibular findings, an absence of VEMP responses may be of value. However, the functional consequence of an isolated bilaterally absent VEMP is not known and may provide minimal information to an older patient's diagnostic picture. In cases where the response is bilaterally absent, a more intense AC stimulus should be used or bone conducted vibration should be considered.

  14. Evaluation of guinea pig model for ocular and cervical vestibular-evoked myogenic potentials for vestibular function test.

    Science.gov (United States)

    Yang, Ting-Hua; Liu, Shing-Hwa; Young, Yi-Ho

    2010-09-01

    This study used air-conducted sound (ACS) and bone-conducted vibration (BCV) stimuli in eliciting ocular vestibular-evoked myogenic potential (oVEMP) and cervical VEMP (cVEMP) in guinea pigs. Prospective study. Ten guinea pigs were treated with gentamicin (4 mg) on the left ear, whereas the right ear served as a control. One week after treatment, each animal underwent oVEMP and cVEMP tests using ACS and BCV modes in a randomized order, and was sacrificed for morphological study. Using ACS mode, oVEMPs were absent in all 10 (100%) animals despite the stimulus intensity increased up to 120 dB pe SPL. Conversely, using BCV mode, oVEMPs were present on the left (lesion) eye, and absent on the right (control) eye in all (100%) animals. For the cVEMPs via ACS mode, all right (control) necks had clear cVEMPs, and all (100%) left (lesion) necks revealed absent cVEMPs. However, via BCV mode, all right (control) necks and six (60%) left (lesion) necks showed clear cVEMPs. Morphological study demonstrated substantial loss of hair cells in the utricular and saccular macula. The cVEMP test via ACS mode is specific for investigating the saccular disorder, whereas the oVEMP test via BCV mode is preferable for investigating the utricular disorders in humans. The guinea pig model is consistent with the findings of humans. Restated, appropriate animal models for cVEMP and oVEMP in guinea pigs are via ACS and BCV modes, respectively.

  15. Ocular vestibular evoked myogenic potentials in response to air-conducted sound in Ménière's disease.

    Science.gov (United States)

    Winters, Stephanie M; Campschroer, Thijs; Grolman, Wilko; Klis, Sjaak F L

    2011-10-01

    Currently, Ménière's disease is predominantly diagnosed through clinical criteria. Additional standard vestibular testing, such as nystagmography, can show variable responses. In the last decade, the cervical vestibular evoked myogenic potential (VEMP) has shown to be of additive value in diagnosing Ménière's disease. In this study, the results of the ocular VEMP (oVEMP) in response to air-conducted sound will be discussed. To evaluate possible changes of the oVEMP in a large group (n = 37) of patients with Ménière's disease. In 55 subjects without Ménière's disease and 37 patients with Ménière's disease, oVEMPs in response to air-conducted sound stimulation (tone-burst, 500 Hz; maximum stimulus level, 120 dB sound pressure level) were studied. Recording was performed in upgaze with surface electrodes underneath both eyes. The burden of the test was scored by all subjects on a visual analogue scale. In patients with Ménière's disease the response rates are lower, the oVEMP amplitudes are smaller, and thresholds are higher than in subjects without Ménière's disease. This effect is observed in both ears of patients with Ménière's disease. The affected ear is more altered than the clinically unaffected ear. The air-conducted oVEMP can be a relevant addition to the current diagnostic workup of patients with possible Ménière's disease. A lower response rate, smaller amplitude, and higher threshold of the oVEMP indicate the pathologic disease in this population.

  16. Properties of rectified averaging of an evoked-type signal: theory and application to the vestibular-evoked myogenic potential.

    Science.gov (United States)

    Colebatch, J G

    2009-11-01

    The properties of rectified averages were investigated using the VEMP (vestibular-evoked myogenic potential) as an example of an evoked-type response. Recordings were made of surface EMG from the sternocleidomastoid (SCM) muscles of six volunteers, unstimulated, at different levels of tonic activation and then in response to clicks of different intensities. The stochastic properties of the surface EMG recorded were shown to be well modelled using a zero mean normal distribution with a standard deviation equivalent to the mean RMS (root mean squared) value (mean residual error variance 0.87%). Assuming a normal distribution, equations were derived for the expected value of both the rectified and RMS average with the addition of constant waveforms of different sizes. A simulation using recorded EMG and added sine waves of different amplitudes demonstrated that the equations predicted the rectified averages accurately. It also confirmed the importance of the relative amplitude of the added signal in determining whether it was detected using rectified averages. The same equations were then applied to actual data consisting of VEMPs of different relative amplitudes recorded from the volunteers. Whilst the signal-to-noise ratio (measured by corrected amplitude) was a major determinant of the nature of the rectified average, consistent deviations were detected between the predicted and actual rectified averages. Deviations from predicted values indicated that the VEMP did not behave simply like a constant signal added to tonic background EMG. A more complicated model, which included temporal jitter as well as inhibition of background EMG during the VEMP, was required to fit the physiological recordings. Rectified averages are sensitive to physiological properties, which are not apparent when using unrectified averages alone. Awareness of the properties of rectified averages should improve their interpretation.

  17. High intensity training may reverse the fiber type specific decline in myogenic stem cells in multiple sclerosis patients

    Directory of Open Access Journals (Sweden)

    Jean eFarup

    2016-05-01

    Full Text Available Multiple sclerosis (MS is associated with loss of skeletal muscle mass and function. The myogenic stem cells (satellite cells – SCs are instrumental to accretion of myonuclei, but remain to be investigated in MS. The present study aimed to compare the SC and myonuclei content between MS patients (n=23 and age matched healthy controls (HC, n=18. Furthermore, the effects of 12 weeks of high intensity training on SC and myonuclei content were explored in MS. Muscle biopsies were obtained from m. Vastus Lateralis at baseline (MS+HC and following 12 weeks of training (MS only. Frozen biopsies were sectioned followed by immunohistochemical analysis for fiber type specific SCs (Pax7+, myonuclei (MN and central nuclei content and fiber cross-sectional area (fCSA using ATPase histochemistry. At baseline the SCs per fiber was lower in type II compared to type I fiber in both MS (119%, p<0.01 and HC (69%, p<0.05, whereas the SCs per fCSA was lower in type II fibers compared to type I only in MS (72%, p<0.05. No differences were observed in MN or central nuclei between MS and HC. Following training the type II fiber SCs per fiber and fCSA in MS patients increased by 165% (p<0.05 and 135% (p<0.05, respectively. Furthermore, the type II fiber MN content increased by 35% (p<0.05 following training. In conclusion, the SC content is lower in type II compared to type I fibers in both MS and HC. Furthermore, high intensity training was observed to selectively increase the SC and myonuclei content in type II fibers in MS patients.

  18. Rotator cuff muscle degeneration and tear severity related to myogenic, adipogenic, and atrophy genes in human muscle.

    Science.gov (United States)

    Shah, Shivam A; Kormpakis, Ioannis; Cavinatto, Leonardo; Killian, Megan L; Thomopoulos, Stavros; Galatz, Leesa M

    2017-05-04

    Large rotator cuff tear size and advanced muscle degeneration can affect reparability of tears and compromise tendon healing. Clinicians often rely on direct measures of rotator cuff tear size and muscle degeneration from magnetic resonance imaging (MRI) to determine whether the rotator cuff tear is repairable. The objective of this study was to identify the relationship between gene expression changes in rotator cuff muscle degeneration to standard data available to clinicians. Radiographic assessment of preoperative rotator cuff tear severity was completed for 25 patients with varying magnitudes of rotator cuff tears. Tear width and retraction were measured using MRI, and Goutallier grade, tangent (tan) sign, and Thomazeau grade were determined. Expression of myogenic-, adipogenic-, atrophy-, and metabolism-related genes in biopsied muscles were correlated with tear width, tear retraction, Goutallier grade, tan sign, and Thomazeau grade. Tear width positively correlated with Goutallier grade in both the supraspinatus (r = 0.73) and infraspinatus (r = 0.77), along with tan sign (r = 0.71) and Thomazeau grade (r = 0.68). Decreased myogenesis (Myf5), increased adipogenesis (CEBPα, Lep, Wnt10b), and decreased metabolism (PPARα) correlated with radiographic assessments. Gene expression changes suggest that rotator cuff tears lead to a dramatic molecular response in an attempt to maintain normal muscle tissue, increase adipogenesis, and decrease metabolism. Fat accumulation and muscle atrophy appear to stem from endogenous changes rather than from changes mediated by infiltrating cells. Results suggest that chronic unloading of muscle, induced by rotator cuff tear, disrupts muscle homeostasis. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

  19. Myogenic reprogramming of bone marrow derived cells in a W⁴¹Dmd(mdx deficient mouse model.

    Directory of Open Access Journals (Sweden)

    Stuart Walsh

    Full Text Available Lack of expression of dystrophin leads to degeneration of muscle fibers and infiltration of connective and adipose tissue. Cell transplantation therapy has been proposed as a treatment for intractable muscle degenerative disorders. Several reports have demonstrated the ability of bone-marrow derived cells (BMDC to contribute to non-haematopoietic tissues including epithelium, heart, liver, skeletal muscle and brain following transplantation by means of fusion and reprogramming. A key issue is the extent to which fusion and reprogramming can occur in vivo, particularly under conditions of myogenic deterioration.To investigate the therapeutic potential of bone marrow transplantation in monogenetic myopathy, green fluorescent protein-positive (GFP+ bone marrow cells were transplanted into non-irradiated c-kit receptor-deficient (W⁴¹ mdx mice. This model allows BMDC reconstitution in the absence of irradiation induced myeloablation. We provide the first report of BMDC fusion in a W⁴¹Dmd(mdx deficient mouse model.In the absence of irradiation induced injury, few GFP+ cardiomyocytes and muscle fibres were detected 24 weeks post BMT. It was expected that the frequency of fusion in the hearts of W⁴¹Dmd(mdx mice would be similar to frequencies observed in infarcted mice. Although, it is clear from this study that individual cardiomyocytes with monogenetic deficiencies can be rescued by fusion, it is as clear that in the absence of irradiation, the formation of stable and reprogrammed fusion hybrids occurs, with the current techniques, at very low levels in non-irradiated recipients.

  20. Myogenic Potential of Whole Bone Marrow Mesenchymal Stem Cells In Vitro and In Vivo for Usage in Urinary Incontinence

    Science.gov (United States)

    Giammò, Alessandro; Boido, Marina; Rustichelli, Deborah; Mareschi, Katia; Errichiello, Edoardo; Parola, Maurizio; Ferrero, Ivana; Fagioli, Franca; Vercelli, Alessandro; Carone, Roberto

    2012-01-01

    Urinary incontinence, defined as the complaint of any involuntary loss of urine, is a pathological condition, which affects 30% females and 15% males over 60, often following a progressive decrease of rhabdosphincter cells due to increasing age or secondary to damage to the pelvic floor musculature, connective tissue and/or nerves. Recently, stem cell therapy has been proposed as a source for cell replacement and for trophic support to the sphincter. To develop new therapeutic strategies for urinary incontinence, we studied the interaction between mesenchymal stem cells (MSCs) and muscle cells in vitro; thereafter, aiming at a clinical usage, we analyzed the supporting role of MSCs for muscle cells in vitro and in in vivo xenotransplantation. MSCs can express markers of the myogenic cell lineages and give rise, under specific cell culture conditions, to myotube-like structures. Nevertheless, we failed to obtain mixed myotubes both in vitro and in vivo. For in vivo transplantation, we tested a new protocol to collect human MSCs from whole bone marrow, to get larger numbers of cells. MSCs, when transplanted into the pelvic muscles close to the external urethral sphincter, survived for a long time in absence of immunosuppression, and migrated into the muscle among fibers, and towards neuromuscular endplates. Moreover, they showed low levels of cycling cells, and did not infiltrate blood vessels. We never observed formation of cell masses suggestive of tumorigenesis. Those which remained close to the injection site showed an immature phenotype, whereas those in the muscle had more elongated morphologies. Therefore, MSCs are safe and can be easily transplanted without risk of side effects in the pelvic muscles. Further studies are needed to elucidate their integration into muscle fibers, and to promote their muscular transdifferentiation either before or after transplantation. PMID:23029081

  1. Myogenic potential of whole bone marrow mesenchymal stem cells in vitro and in vivo for usage in urinary incontinence.

    Directory of Open Access Journals (Sweden)

    Monica Gunetti

    Full Text Available Urinary incontinence, defined as the complaint of any involuntary loss of urine, is a pathological condition, which affects 30% females and 15% males over 60, often following a progressive decrease of rhabdosphincter cells due to increasing age or secondary to damage to the pelvic floor musculature, connective tissue and/or nerves. Recently, stem cell therapy has been proposed as a source for cell replacement and for trophic support to the sphincter. To develop new therapeutic strategies for urinary incontinence, we studied the interaction between mesenchymal stem cells (MSCs and muscle cells in vitro; thereafter, aiming at a clinical usage, we analyzed the supporting role of MSCs for muscle cells in vitro and in in vivo xenotransplantation. MSCs can express markers of the myogenic cell lineages and give rise, under specific cell culture conditions, to myotube-like structures. Nevertheless, we failed to obtain mixed myotubes both in vitro and in vivo. For in vivo transplantation, we tested a new protocol to collect human MSCs from whole bone marrow, to get larger numbers of cells. MSCs, when transplanted into the pelvic muscles close to the external urethral sphincter, survived for a long time in absence of immunosuppression, and migrated into the muscle among fibers, and towards neuromuscular endplates. Moreover, they showed low levels of cycling cells, and did not infiltrate blood vessels. We never observed formation of cell masses suggestive of tumorigenesis. Those which remained close to the injection site showed an immature phenotype, whereas those in the muscle had more elongated morphologies. Therefore, MSCs are safe and can be easily transplanted without risk of side effects in the pelvic muscles. Further studies are needed to elucidate their integration into muscle fibers, and to promote their muscular transdifferentiation either before or after transplantation.

  2. Stand-up exercise training facilitates muscle recovery from disuse atrophy by stimulating myogenic satellite cell proliferation in mice.

    Science.gov (United States)

    Itoh, Yuta; Hayakawa, Kimihide; Mori, Tomohiro; Agata, Nobuhide; Inoue-Miyazu, Masumi; Murakami, Taro; Sokabe, Masahiro; Kawakami, Keisuke

    2014-11-01

    Determining the cellular and molecular recovery processes in inactivity - or unloading -induced atrophied muscles should improve rehabilitation strategies. We assessed the effects of stand-up exercise (SE) training on the recovery of atrophied skeletal muscles in male mice. Mice were trained to stand up and press an elevated lever in response to a light-tone cue preceding an electric foot shock and then subjected to tail suspension (TS) for 2 weeks to induce disuse atrophy in hind limb muscles. After release from TS, mice were divided into SE-trained (SE cues: 25 times per set, two sets per day) and non-SE-trained groups. Seven days after the training, average myofiber cross-sectional area (CSA) of the soleus muscle was significantly greater in the SE-trained group than in the non-SE-trained group (1843 ± 194 μm(2) vs. 1315 ± 153 μm(2)). Mean soleus muscle CSA in the SE trained group was not different from that in the CON group subjected to neither TS nor SE training (2005 ± 196 μm(2)), indicating that SE training caused nearly complete recovery from muscle atrophy. The number of myonuclei per myofiber was increased by ~60% in the SE-trained group compared with the non-SE-trained and CON groups (0.92 ± 0.03 vs. 0.57 ± 0.03 and 0.56 ± 0.11, respectively). The number of proliferating myonuclei, identified by 5-ethynyl-2'-deoxyuridine staining, increased within the first few days of SE training. Thus, it is highly likely that myogenic satellite cells proliferated rapidly in atrophied muscles in response to SE training and fused with existing myofibers to reestablish muscle mass.

  3. FGFR4 and its novel splice form in myogenic cells: interplay of glycosylation and tyrosine phosphorylation

    OpenAIRE

    Kwiatkowski, Boguslaw A.; Kirillova, Irina; Richard, Robert E.; Israeli, David; Yablonka-Reuveni, Zipora

    2008-01-01

    The family of fibroblast growth factor receptors (FGFRs) is encoded by four distinct genes. FGFR1 and FGFR4 are both expressed during myogenesis, but whereas the function of FGFR1 in myoblast proliferation has been documented, the role of FGFR4 remains unknown. Here we report on a new splice form of FGFR4 cloned from primary cultures of mouse satellite cells. This form, named FGFR4(−16), lacks the entire exon 16, resulting in a deletion within the FGFR kinase domain. Expression of FGFR4(−16) ...

  4. Zinc-Finger Nuclease Knockout of Dual-Specificity Protein Phosphatase-5 Enhances the Myogenic Response and Autoregulation of Cerebral Blood Flow in FHH.1BN Rats

    Science.gov (United States)

    Fan, Fan; Geurts, Aron M.; Pabbidi, Mallikarjuna R.; Smith, Stanley V.; Harder, David R.; Jacob, Howard; Roman, Richard J.

    2014-01-01

    We recently reported that the myogenic responses of the renal afferent arteriole (Af-Art) and middle cerebral artery (MCA) and autoregulation of renal and cerebral blood flow (RBF and CBF) were impaired in Fawn Hooded hypertensive (FHH) rats and were restored in a FHH.1BN congenic strain in which a small segment of chromosome 1 from the Brown Norway (BN) containing 15 genes including dual-specificity protein phosphatase-5 (Dusp5) were transferred into the FHH genetic background. We identified 4 single nucleotide polymorphisms in the Dusp5 gene in FHH as compared with BN rats, two of which altered CpG sites and another that caused a G155R mutation. To determine whether Dusp5 contributes to the impaired myogenic response in FHH rats, we created a Dusp5 knockout (KO) rat in the FHH.1BN genetic background using a zinc-finger nuclease that introduced an 11 bp frame-shift deletion and a premature stop codon at AA121. The expression of Dusp5 was decreased and the levels of its substrates, phosphorylated ERK1/2 (p-ERK1/2), were enhanced in the KO rats. The diameter of the MCA decreased to a greater extent in Dusp5 KO rats than in FHH.1BN and FHH rats when the perfusion pressure was increased from 40 to 140 mmHg. CBF increased markedly in FHH rats when MAP was increased from 100 to 160 mmHg, and CBF was better autoregulated in the Dusp5 KO and FHH.1BN rats. The expression of Dusp5 was higher at the mRNA level but not at the protein level and the levels of p-ERK1/2 and p-PKC were lower in cerebral microvessels and brain tissue isolated from FHH than in FHH.1BN rats. These results indicate that Dusp5 modulates myogenic reactivity in the cerebral circulation and support the view that a mutation in Dusp5 may enhance Dusp5 activity and contribute to the impaired myogenic response in FHH rats. PMID:25397684

  5. Zinc-finger nuclease knockout of dual-specificity protein phosphatase-5 enhances the myogenic response and autoregulation of cerebral blood flow in FHH.1BN rats.

    Directory of Open Access Journals (Sweden)

    Fan Fan

    Full Text Available We recently reported that the myogenic responses of the renal afferent arteriole (Af-Art and middle cerebral artery (MCA and autoregulation of renal and cerebral blood flow (RBF and CBF were impaired in Fawn Hooded hypertensive (FHH rats and were restored in a FHH.1BN congenic strain in which a small segment of chromosome 1 from the Brown Norway (BN containing 15 genes including dual-specificity protein phosphatase-5 (Dusp5 were transferred into the FHH genetic background. We identified 4 single nucleotide polymorphisms in the Dusp5 gene in FHH as compared with BN rats, two of which altered CpG sites and another that caused a G155R mutation. To determine whether Dusp5 contributes to the impaired myogenic response in FHH rats, we created a Dusp5 knockout (KO rat in the FHH.1BN genetic background using a zinc-finger nuclease that introduced an 11 bp frame-shift deletion and a premature stop codon at AA121. The expression of Dusp5 was decreased and the levels of its substrates, phosphorylated ERK1/2 (p-ERK1/2, were enhanced in the KO rats. The diameter of the MCA decreased to a greater extent in Dusp5 KO rats than in FHH.1BN and FHH rats when the perfusion pressure was increased from 40 to 140 mmHg. CBF increased markedly in FHH rats when MAP was increased from 100 to 160 mmHg, and CBF was better autoregulated in the Dusp5 KO and FHH.1BN rats. The expression of Dusp5 was higher at the mRNA level but not at the protein level and the levels of p-ERK1/2 and p-PKC were lower in cerebral microvessels and brain tissue isolated from FHH than in FHH.1BN rats. These results indicate that Dusp5 modulates myogenic reactivity in the cerebral circulation and support the view that a mutation in Dusp5 may enhance Dusp5 activity and contribute to the impaired myogenic response in FHH rats.

  6. Satellite cell response to erythropoietin treatment and endurance training in healthy young men

    DEFF Research Database (Denmark)

    Hoedt, Andrea; Christensen, Britt; Nellemann, Birgitte

    2016-01-01

    KEY POINT: Erythropoietin (Epo) treatment may induce myogenic differentiation factor (MyoD) expression and prevent apoptosis in satellite cells (SCs) in murine and in vitro models. Endurance training stimulates SC proliferation in vivo in murine and human skeletal muscle. In the present study, we...... show, in human skeletal muscle, that treatment with an Epo-stimulating agent (darbepoetin-α) in vivo increases the content of MyoD(+) SCs in healthy young men. Moreover, we report that Epo receptor mRNA is expressed in adult human SCs, suggesting that Epo may directly target SCs through ligand......-term Epo treatment during disease conditions involving anaemia may impact SCs and warrants further investigation. Satellite cell (SC) proliferation is observed following erythropoitin treatment in vitro in murine myoblasts and endurance training in vivo in human skeletal muscle. The present study aimed...

  7. Maged1, a new regulator of skeletal myogenic differentiation and muscle regeneration

    Directory of Open Access Journals (Sweden)

    Achouri Younes

    2010-07-01

    Full Text Available Abstract Background In normal adult skeletal muscle, cell turnover is very slow. However, after an acute lesion or in chronic pathological conditions, such as primary myopathies, muscle stem cells, called satellite cells, are induced to proliferate, then withdraw definitively from the cell cycle and fuse to reconstitute functional myofibers. Results We show that Maged1 is expressed at very low levels in normal adult muscle but is strongly induced after injury, during the early phase of myoblast differentiation. By comparing in vitro differentiation of myoblasts derived from wild-type or Maged1 knockout mice, we observed that Maged1 deficiency results in reduced levels of p21CIP1/WAF1, defective cell cycle exit and impaired myotube maturation. In vivo, this defect results in delayed regeneration of injured muscle. Conclusions These data demonstrate for the first time that Maged1 is an important factor required for proper skeletal myoblast differentiation and muscle healing.

  8. Smad3 signaling is required for satellite cell function and myogenic differentiation of myoblasts

    Institute of Scientific and Technical Information of China (English)

    Xiaojia Ge; Ravi Kambadur; Craig McFarlane; Anuradha Vajjala; Sudarsanareddy Lokireddy; Zhi Hui Ng; Chek Kun Tan; Nguan Soon Tan; Walter Wahli; Mridula Sharma

    2011-01-01

    TGF-β and myostatin are the two most important regulators of muscle growth.Both growth factors have been shown to signal through a Smad3-dependent pathway.However to date,the role of Smad3 in muscle growth and differentiation is not investigated.Here,we demonstrate that Smad3-null mice have decreased muscle mass and pronounced skeletal muscle atrophy.Consistent with this,we also find increased protein ubiquitination and elevated levels of the ubiquitin E3 ligase MuRF1 in muscle tissue isolated from Smad3-null mice.Loss of Smad3 also led to defective satellite cell (SC) functionality.Smad3-null SCs showed reduced propensity for self-renewal,which may lead to a progressive loss of SC number.Indeed,decreased SC number was observed in skeletal muscle from Smad3- null mice showing signs of severe muscle wasting.Further in vitro analysis of primary myoblast cultures identified that Smad3-nuil myoblasts exhibit impaired proliferation,differentiation and fusion,resulting in the formation of atrophied myotubes.A search for the molecular mechanism revealed that loss of Smad3 results in increased myostatin expression in Smad3-null muscle and myoblasts.Given that myostatin is a negative regulator,we hypothesize that increased myostatin levels are responsible for the atrophic phenotype in Smad3-null mice.Consistent with this theory,inactivation of myostatin in Smad3-null mice rescues the muscle atrophy phenotype.

  9. Effects of type IV collagen on myogenic characteristics of IGF-I gene-engineered myoblasts.

    Science.gov (United States)

    Ito, Akira; Yamamoto, Masahiro; Ikeda, Kazushi; Sato, Masanori; Kawabe, Yoshinori; Kamihira, Masamichi

    2015-05-01

    Skeletal muscle regeneration requires migration, proliferation and fusion of myoblasts to form multinucleated myotubes. In our previous study, we showed that insulin-like growth factor (IGF)-I gene delivery stimulates the proliferation and differentiation of mouse myoblast C2C12 cells and promotes the contractile force generated by tissue-engineered skeletal muscles. The aim of this study was to investigate the effects of the extracellular matrix on IGF-I gene-engineered C2C12 cells in vitro. Retroviral vectors for doxycycline (Dox)-inducible expression of the IGF-I gene were transduced into C2C12 cells. When cultured on a type IV collagen-coated surface, we observed significant increases in the migration speed and number of IGF-I gene-engineered C2C12 cells with Dox addition, designated as C2C12/IGF (+) cells. Co-culture of C2C12/IGF (+) cells and parental C2C12 cells, which had been cultured in differentiation medium for 3 days, greatly enhanced myotube formation. Moreover, type IV collagen supplementation promoted the fusion of C2C12/IGF (+) cells with differentiated C2C12 cells and increased the number of myotubes with striations. Myotubes formed by C2C12/IGF (+) cells cultured on type IV collagen showed a dynamic contractile activity in response to electrical pulse stimulation. These findings indicate that type IV collagen promotes skeletal muscle regeneration mediated by IGF-I-expressing myoblasts, which may have important clinical implications in the design of myoblast-based therapies.

  10. TNF alpha inhibits myogenic differentiation of C2C12 cells through NF-κB activation and impairment of IGF-1 signaling pathway.

    Science.gov (United States)

    Zhao, Q; Yang, S T; Wang, J J; Zhou, J; Xing, S S; Shen, C C; Wang, X X; Yue, Y X; Song, J; Chen, M; Wei, Y Y; Zhou, Q P; Dai, T; Song, Y H

    2015-03-20

    Cachexia or muscle wasting is a common condition that occurs in many chronic diseases. The wasting conditions are characterized by increased levels of TNF-α which was also known as cachectin in the past. But how TNF-α exerts its cachetic effects remains controversial. To clarify this issue, we investigated the impact of TNF-α on C2C12 cell myogenic differentiation. Our results demonstrate that myotube formation was completely inhibited by TNF-α when added to differentiating C2C12 myoblasts. The inhibitory effect of TNF-α on differentiation was accompanied by activation of NF-κB and down regulation of myogenin and Akt. Importantly, TNF-α's effect on differentiation was abolished when IGF-1 was added to the culture. IGF-1 treatment also inhibited NF-κB reporter activity and restored Akt levels. Our data suggest that TNF-α inhibits myogenic differentiation through NF-κB activation and impairment of IGF-1 signaling pathway. The reversal of TNF-α induced inhibition of myogenesis by IGF-1 may have significant therapeutic potential.

  11. Phosphotyrosine phosphatase inhibitor bisperoxovanadium endows myogenic cells with enhanced muscle stem cell functions via epigenetic modulation of Sca-1 and Pw1 promoters.

    Science.gov (United States)

    Smeriglio, Piera; Alonso-Martin, Sonia; Masciarelli, Silvia; Madaro, Luca; Iosue, Ilaria; Marrocco, Valeria; Relaix, Frédéric; Fazi, Francesco; Marazzi, Giovanna; Sassoon, David A; Bouché, Marina

    2016-04-01

    Understanding the regulation of the stem cell fate is fundamental for designing novel regenerative medicine strategies. Previous studies have suggested that pharmacological treatments with small molecules provide a robust and reversible regulation of the stem cell program. Previously, we showed that treatment with a vanadium compound influences muscle cell fatein vitro In this study, we demonstrate that treatment with the phosphotyrosine phosphatase inhibitor bisperoxovanadium (BpV) drives primary muscle cells to a poised stem cell stage, with enhanced function in muscle regenerationin vivofollowing transplantation into injured muscles. Importantly, BpV-treated cells displayed increased self-renewal potentialin vivoand replenished the niche in both satellite and interstitial cell compartments. Moreover, we found that BpV treatment induces specific activating chromatin modifications at the promoter regions of genes associated with stem cell fate, includingSca-1andPw1 Thus, our findings indicate that BpV resets the cell fate program by specific epigenetic regulations, such that the committed myogenic cell fate is redirected to an earlier progenitor cell fate stage, which leads to an enhanced regenerative stem cell potential.-Smeriglio, P., Alonso-Martin, S., Masciarelli, S., Madaro, L., Iosue, I., Marrocco, V., Relaix, F., Fazi, F., Marazzi, G., Sassoon, D. A., Bouché, M. Phosphotyrosine phosphatase inhibitor bisperoxovanadium endows myogenic cells with enhanced muscle stem cell functionsviaepigenetic modulation of Sca-1 and Pw1 promoters.

  12. Creatine Prevents the Structural and Functional Damage to Mitochondria in Myogenic, Oxidatively Stressed C2C12 Cells and Restores Their Differentiation Capacity

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    Elena Barbieri

    2016-01-01

    Full Text Available Creatine (Cr is a nutritional supplement promoting a number of health benefits. Indeed Cr has been shown to be beneficial in disease-induced muscle atrophy, improve rehabilitation, and afford mild antioxidant activity. The beneficial effects are likely to derive from pleiotropic interactions. In accord with this notion, we previously demonstrated that multiple pleiotropic effects, including preservation of mitochondrial damage, account for the capacity of Cr to prevent the differentiation arrest caused by oxidative stress in C2C12 myoblasts. Given the importance of mitochondria in supporting the myogenic process, here we further explored the protective effects of Cr on the structure, function, and networking of these organelles in C2C12 cells differentiating under oxidative stressing conditions; the effects on the energy sensor AMPK, on PGC-1α, which is involved in mitochondrial biogenesis and its downstream effector Tfam were also investigated. Our results indicate that damage to mitochondria is crucial in the differentiation imbalance caused by oxidative stress and that the Cr-prevention of these injuries is invariably associated with the recovery of the normal myogenic capacity. We also found that Cr activates AMPK and induces an upregulation of PGC-1α expression, two events which are likely to contribute to the protection of mitochondrial quality and function.

  13. Age-dependent impact of CaV3.2 T-type calcium channel deletion on myogenic tone and flow-mediated vasodilatation in small arteries

    DEFF Research Database (Denmark)

    Mikkelsen, Miriam F.; Björling, Karl; Jensen, Lars Jørn

    2016-01-01

    .2-dependent and -independent effects. No changes in mRNA expression of several important K(+) and Ca(2+) channel genes were induced by CaV3.2 knock-out. However, the expression of the other T-type channel isoform (CaV3.1) was reduced at the mRNA and protein level in mature adult compared to young WT arteries......The myogenic response and flow-mediated vasodilatation are important regulators of local blood perfusion and total peripheral resistance, and are known to entail a calcium influx into vascular smooth muscle cells (VSMCs) and endothelial cells (ECs), respectively. CaV3.2 T-type calcium channels...... are expressed in both VSMCs and ECs of small arteries. The T-type channels are important drug targets but due to the lack of specific antagonists our understanding of the role of CaV3.2 channels in vasomotor tone at various ages is scarce. We evaluated the myogenic response, flow-mediated vasodilatation...

  14. An experimental study of the myogenic component of joint contracture%挛缩膝关节中肌源性成分的实验研究

    Institute of Scientific and Technical Information of China (English)

    倪国新; 苏力; 唐军凯

    2002-01-01

    Objective To differentiate the myogenic and arthrogenic components of knee contracture resulting from being immobilized in extension for 4 weeks.Method Ten rabbits were used as experimental animals. The right hindlimbs were immobilized with the knee in extension for four weeks, while the left served as controls. The range of motion (ROM) of both knees was measured before and after myotomy.Result In immobilized and control hindlimbs, knee mean ROM were 79.5° and 138.5° respectively. The gain of ROM after myotomy in immobilized and control hindlimbs were (8.7± 0.45)° , (11.6± 0.58)° respectively. Only 14.75% of the limitation of motion in ROM was accounted for by the myotomy component. Conclusions After immobilized for 4 weeks, the ROM of knee was limited.However,the myogenic part was small and not different from normals.Because this study reports on only one time point,further investigations are needed.

  15. Critical Role Played by Cyclin D3 in the MyoD-Mediated Arrest of Cell Cycle during Myoblast Differentiation

    OpenAIRE

    1999-01-01

    During the terminal differentiation of skeletal myoblasts, the activities of myogenic factors regulate not only tissue-specific gene expressions but also the exit from the cell cycle. The induction of cell cycle inhibitors such as p21 and pRb has been shown to play a prominent role in the growth arrest of differentiating myoblasts. Here we report that, at the onset of differentiation, activation by MyoD of the Rb, p21, and cyclin D3 genes occurs in the absence of new protein synthesis and wit...

  16. Targeted Expression and Secretion of Human Apoprotein AI,Apoprotein E and Lecithin-choles-terol Acyltransferase from Myogenic Cell

    Institute of Scientific and Technical Information of China (English)

    范乐明; 张慧; 于书真; 陈琪; 魏恩会; 王南; 蔡海江

    2001-01-01

    Objective To investigate the possibility of heterologous expression for apoAI, apoE and LCAT by skeletal muscle cells and secretion into blood and to develop a safe and convenient gene therapy method for atherosclerosis. Methods Viral and nonviral vectors containing apoAI, apoE or LCAT genes were constructed and transfected into myogenic cells in vitro or injected directed into mouse skeletal muscle. The expression efficiencies of these vectors were investigated by ELISA assay for human apoAI and apoE3 and by the proteoliposome method for human LCAT. Genomic DNA was extracted from stable transduced myoblasts and analyzed for the presence of vector sequence by PCR amplifications. Immunocytochemistry assay was also performed to make an intuitionistic detection for the expression of transgene in myoblasts. Results All viral or nonviral vectors constructed in present study expressed the transgenes efficiently in mice skeletal muscles in vivo or cultured myoblasts in vitro. The transgene expression level of cells transfected with AAV-based plasmid vectors were 2-4 times higher then that of cells transfected with conventional plasmid vectors. Additionally, cells transfected with AAV-based bicistronic vector or tricistronic retroviral vector expressed both human apoAI and LCAT simultaneously. The sequences of retroviral or AAV-based plasmid vectors were found to be retained in host cells after transfection when that of conventional plasmid vectors were lost. Furthermore, transduced myoblasts maintained the ability for heterologous expression of human apoAI and LCAT even after differentiation into myotubes. For cells transfected with retroviral vectors, stable transduced clones can be selected by G418 and continued to efficiently express human apoAI and LCAT for 3 months. Conclusion These finds indicated that mice skeletal muscles or cultured myoblasts transduced with viral or non-viral vectors could efficiently express and secret human apoAI, apoE and LCAT. It suggested

  17. Voltage-gated K(+) channels sensitive to stromatoxin-1 regulate myogenic and neurogenic contractions of rat urinary bladder smooth muscle.

    Science.gov (United States)

    Chen, Muyan; Kellett, Whitney F; Petkov, Georgi V

    2010-07-01

    Members of the voltage-gated K(+) (K(V)) channel family are suggested to control the resting membrane potential and the repolarization phase of the action potential in urinary bladder smooth muscle (UBSM). Recent studies report that stromatoxin-1, a peptide isolated from tarantulas, selectively inhibits K(V)2.1, K(V)2.2, K(V)4.2, and K(V)2.1/9.3 channels. The objective of this study was to investigate whether K(V) channels sensitive to stromatoxin-1 participate in the regulation of rat UBSM contractility and to identify their molecular fingerprints. Stromatoxin-1 (100 nM) increased the spontaneous phasic contraction amplitude, muscle force, and tone in isolated UBSM strips. However, stromatoxin-1 (100 nM) had no effect on the UBSM contractions induced by depolarizing agents such as KCl (20 mM) or carbachol (1 microM). This indicates that, under conditions of sustained membrane depolarization, the K(V) channels sensitive to stromatoxin-1 have no further contribution to the membrane excitability and contractility. Stromatoxin-1 (100 nM) increased the amplitude of the electrical field stimulation-induced contractions, suggesting also a role for these channels in neurogenic contractions. RT-PCR experiments on freshly isolated UBSM cells showed mRNA expression of K(V)2.1, K(V)2.2, and K(V)9.3, but not K(V)4.2 channel subunits. Protein expression of K(V)2.1 and K(V)2.2 channels was detected using Western blot and was further confirmed by immunocytochemical detection in freshly isolated UBSM cells. These novel findings indicate that K(V)2.1 and K(V)2.2, but not K(V)4.2, channel subunits are expressed in rat UBSM and play a key role in opposing both myogenic and neurogenic UBSM contractions.

  18. TEAD transcription factors are required for normal primary myoblast differentiation in vitro and muscle regeneration in vivo

    OpenAIRE

    Shilpy Joshi; Guillaume Davidson; Stéphanie Le Gras; Shuichi Watanabe; Thomas Braun; Gabrielle Mengus; Irwin Davidson

    2017-01-01

    The TEAD family of transcription factors (TEAD1-4) bind the MCAT element in the regulatory elements of both growth promoting and myogenic differentiation genes. Defining TEAD transcription factor function in myogenesis has proved elusive due to overlapping expression of family members and their functional redundancy. We show that silencing of either Tead1, Tead2 or Tead4 did not effect primary myoblast (PM) differentiation, but that their simultaneous knockdown strongly impaired differentiati...

  19. Muscle Atrophy Reversed by Growth Factor Activation of Satellite Cells in a Mouse Muscle Atrophy Model

    DEFF Research Database (Denmark)

    Hauerslev, Simon; Vissing, John; Krag, Thomas O

    2014-01-01

    Muscular dystrophies comprise a large group of inherited disorders that lead to progressive muscle wasting. We wanted to investigate if targeting satellite cells can enhance muscle regeneration and thus increase muscle mass. We treated mice with hepatocyte growth factor and leukemia inhibitory...... factor under three conditions: normoxia, hypoxia and during myostatin deficiency. We found that hepatocyte growth factor treatment led to activation of the Akt/mTOR/p70S6K protein synthesis pathway, up-regulation of the myognic transcription factors MyoD and myogenin, and subsequently the negative growth...... control factor, myostatin and atrophy markers MAFbx and MuRF1. Hypoxia-induced atrophy was partially restored by hepatocyte growth factor combined with leukemia inhibitory factor treatment. Dividing satellite cells were three-fold increased in the treatment group compared to control. Finally, we...

  20. MicroRNA-17-92 Regulates the Transcription Factor E2F3b during Myogenesis In Vitro and In Vivo

    Directory of Open Access Journals (Sweden)

    Zhixiong Tang

    2017-03-01

    Full Text Available Myogenic differentiation, which occurs during muscle development, is a highly ordered process that can be regulated by E2F transcription factors. Available data show that E2F3b, but not E2F3a, is upregulated and required for myogenic differentiation. However, the regulation of E2F3b expression in myogenic differentiation is not well understood. To investigate whether E2Fb expression is controlled by miRNAs, we used bioinformatics to combine the database of microRNAs downregulated during myogenesis and those predicted to target E2F3. This identified miR-17 and miR-20a as miRNAs potentially involved in E2F3 regulation. We found that miR-17-92 controls the expression of E2F3b in C2C12 cells during myogenic differentiation. Moreover, we confirmed that miR-20a regulates the expression of E2F3b proteins in vivo using a muscle regeneration model.

  1. Spatiotemporal Mapping Techniques Show Clozapine Impairs Neurogenic and Myogenic Patterns of Activity in the Colon of the Rabbit in a Dose-Dependent Manner

    Directory of Open Access Journals (Sweden)

    Roger G. Lentle

    2017-04-01

    Full Text Available Background: Clozapine, an antipsychotic used in treatment-resistant schizophrenia, has adverse gastrointestinal effects with significant associated morbidity and mortality. However, its effects on defined patterns of colonic contractile activity have not been assessed.Method: We used novel radial and longitudinal spatiotemporal mapping techniques, combined with and monitoring of ambient lumen pressure, in ex vivo preparations of triply and of singly haustrated portions of rabbit colon. We identified the contractile patterns of mass peristalses, fast phasic, and ripple contractions and directly qualified the effects of clozapine, at concentrations of 10 μmol/L, 20 μmol/L, and 30 μmol/L, and of norclozapine, the main metabolite of clozapine, on contractile patterns. The effects of carbachol, serotonin and naloxone on clozapine-exposed preparations were also determined. Tetradotoxin was used to distinguish neurogenic from myogenic contractions.Results: At 10 μmol/L, clozapine temporarily abolished the longitudinal contractile components of mass peristalsis, which on return were significantly reduced in number and amplitude, as was maximal mass peristaltic pressure. These effects were reversed by carbachol (1 μmol/L and to some extent by serotonin (15 μmol/L. At 10 μmol/L, myogenic ripple contractions were not affected. At 20 μmol/L, clozapine had a similar but more marked effect on mass peristalses with both longitudinal and radial components and corresponding maximal pressure greatly reduced. At 30 μmol/L, clozapine suppressed the radial and longitudinal components of mass peristalses for over 30 min, as well as ripple contractions. Similar dose-related effects were observed on addition of clozapine to the mid colon. At 20 μmol/L, norclozapine had opposite effects to those of clozapine, causing an increase in the frequency of mass peristalsis with slight increases in basal tone. These slightly augmented contractions were abolished on

  2. Notch Signaling Rescues Loss of Satellite Cells Lacking Pax7 and Promotes Brown Adipogenic Differentiation.

    Science.gov (United States)

    Pasut, Alessandra; Chang, Natasha C; Rodriguez, Uxia Gurriaran; Faulkes, Sharlene; Yin, Hang; Lacaria, Melanie; Ming, Hong; Rudnicki, Michael A

    2016-07-12

    Pax7 is a nodal transcription factor that is essential for regulating the maintenance, expansion, and myogenic identity of satellite cells during both neonatal and adult myogenesis. Deletion of Pax7 results in loss of satellite cells and impaired muscle regeneration. Here, we show that ectopic expression of the constitutively active intracellular domain of Notch1 (NICD1) rescues the loss of Pax7-deficient satellite cells and restores their proliferative potential. Strikingly NICD1-expressing satellite cells do not undergo myogenic differentiation and instead acquire a brown adipogenic fate both in vivo and in vitro. NICD-expressing Pax7(-/-) satellite cells fail to upregulate MyoD and instead express the brown adipogenic marker PRDM16. Overall, these results show that Notch1 activation compensates for the loss of Pax7 in the quiescent state and acts as a molecular switch to promote brown adipogenesis in adult skeletal muscle.

  3. Evaluation of vestibular evoked myogenic potentials (VEMP) and electrocochleography for the diagnosis of Ménière's disease.

    Science.gov (United States)

    Lamounier, Pauliana; de Souza, Thiago Silva Almeida; Gobbo, Debora Aparecida; Bahmad, Fayez

    Ménière's disease (MD) is an inner ear disorder characterized by episodic vertigo, tinnitus, ear fullness, and fluctuating hearing. Its diagnosis can be especially difficult in cases where vestibular symptoms are present in isolation (vestibular MD). The definitive diagnosis is made histologically and can only be performed post-mortem, after analysis of the temporal bone. Endolymphatic hydrops is a histopathological finding of the disease and occurs more often in the cochlea and saccule, followed by the utricle and semicircular canals. Vestibular evoked myogenic potentials (VEMP) emerged as the method of assessment of vestibular function in 1994. Until then, there was no unique way of assessing saccular function and the inferior vestibular nerve. Given that the saccule is responsible for most cases of severe hydrops, VEMP appears as a new tool to assist in the diagnosis of MD. To evaluate the sensitivity and specificity of VEMP and electrocochleography (EcochG) in the diagnosis of definite MD compared with clinical diagnosis. The study includes 12 patients (24 ears) diagnosed with definite MD defined according to the clinical criteria proposed by the American Academy of Otolaryngology - Head and Neck Surgery (AAO-HNS) in 1995, as well as 12 healthy volunteers allocated to the control group (24 ears). A clinical diagnosis by the AAO-HNS criteria was considered as the gold standard. All patients underwent an otoneurological examination, including pure tone and speech audiometry, VEMP, and extratympanic EcochG. The sensitivity and specificity to detect the presence or absence of disease were calculated, as well as their 95% confidence intervals. The reliability of VEMP and EcochG in both ears was assessed using the kappa index. In both tests and in both ears, the ability to diagnose healthy cases was high, with specificity ranging from 84.6% to 100%. Moreover, the ability of the tests to diagnose the disease varied from low to moderate sensitivity, with values

  4. On the impact of examiners on latencies and amplitudes in cervical and ocular vestibular-evoked myogenic potentials evaluated over a large sample (N = 1,038).

    Science.gov (United States)

    Ertl, Matthias; Boegle, R; Kirsch, V; Dieterich, M

    2016-02-01

    Vestibular-evoked myogenic potentials (VEMPs) are frequently used in the clinical diagnosis and research of vertigo syndromes. Altered latencies or amplitudes are typically interpreted as an indication of disturbance in the processing of vestibular stimuli along the otolithic pathways. Correct interpretation, however, can be difficult as VEMP amplitudes can vary greatly across subjects and across laboratories, likely because they are very sensitive to measurement conditions. Here, we attempted to quantify the impact of examiner differences on VEMP data. We collected data from 1,038 people using eight different experimental examiners, and investigated the effect of examiner on VEMP latencies and amplitudes. We found that the examiner collecting the data had a strong effect on outcome measures with significant differences (p VEMPs are very sensitive to variables that may be influenced by the examiner. The field should therefore work on a better standard for VEMP recordings.

  5. Identification of myogenic regulatory genes in the muscle transcriptome of beltfish (Trichiurus lepturus: A major commercial marine fish species with robust swimming ability

    Directory of Open Access Journals (Sweden)

    Hui Zhang

    2016-06-01

    Full Text Available The beltfish (Trichiurus lepturus is considered as one of the most economically important marine fish in East Asia. It is a top predator with a robust swimming ability that is a good model to study muscle physiology in fish. In the present study, we used Illumina sequencing technology (NextSeq500 to sequence, assemble and annotate the muscle transcriptome of juvenile beltfish. A total of 57,509,280 clean reads (deposited in NCBI SRA database with accession number of SRX1674471 were obtained from RNA sequencing and 26,811 unigenes (with N50 of 1033 bp were obtained after de novo assembling with Trinity software. BLASTX against NR, GO, KEGG and eggNOG databases show 100%, 49%, 31% and 96% annotation rate, respectively. By mining beltfish muscle transcriptome, several key genes which play essential role on regulating myogenesis, including pax3, pax7, myf5, myoD, mrf4/myf6, myogenin and myostatin were identified with a low expression level. The muscle transcriptome of beltfish can provide some insight into the understanding of genome-wide transcriptome profile of teleost muscle tissue and give useful information to study myogenesis in juvenile/adult fish.

  6. Type I insulin-like growth factor receptor signaling in skeletal muscle regeneration and hypertrophy.

    Science.gov (United States)

    Philippou, A; Halapas, A; Maridaki, M; Koutsilieris, M

    2007-01-01

    Skeletal muscle is able not only to increase its mass as an adaptation to mechanical loading generated by and imposed upon muscle but also to regenerate after damage, via its intrinsic regulation of gene transcription. Both cellular processes, muscle regeneration and hypertrophy, are mediated by the activation, proliferation and differentiation of muscle satellite cells and appear to be modulated by the mitotic and myogenic activity of locally produced insulin-like growth factor 1 (IGF-1), which functions in an autocrine/paracrine mode. Differentiation of satellite cells into myoblasts involves the regulation of skeletal muscle-specific proteins belonging to the family of myogenic regulatory factors (MRFs). The endocrine, autocrine and paracrine functions of IGF-1 are mediated through binding to the type I IGF receptor (IGF-1.R), which is a ligand-activated receptor tyrosine kinase. The binding of IGF-1 to IGF-1.R induces its autophosphorylation, which recruits specific cytoplasmic molecules containing the Insulin Receptor Substrate Proteins (IRS). The recruitment of IRS proteins by IGF-1/IGF-1.R binding is a critical level at which the proliferative and differentiative actions of IGF-1 diverge. Specific signaling pathways downstream of IGF-1, potentially involved in the mitogenic and myogenic responses and mediating skeletal muscle protein synthesis and hypertrophy following exercise-induced muscle overloading and damage, are discussed. A potential alternative activation of different signaling pathway(s) via a different receptor remains to be demonstrated.

  7. Fetal skeletal muscle progenitors have regenerative capacity after intramuscular engraftment in dystrophin deficient mice.

    Directory of Open Access Journals (Sweden)

    Hiroshi Sakai

    Full Text Available Muscle satellite cells (SCs are stem cells that reside in skeletal muscles and contribute to regeneration upon muscle injury. SCs arise from skeletal muscle progenitors expressing transcription factors Pax3 and/or Pax7 during embryogenesis in mice. However, it is unclear whether these fetal progenitors possess regenerative ability when transplanted in adult muscle. Here we address this question by investigating whether fetal skeletal muscle progenitors (FMPs isolated from Pax3(GFP/+ embryos have the capacity to regenerate muscle after engraftment into Dystrophin-deficient mice, a model of Duchenne muscular dystrophy. The capacity of FMPs to engraft and enter the myogenic program in regenerating muscle was compared with that of SCs derived from adult Pax3(GFP/+ mice. Transplanted FMPs contributed to the reconstitution of damaged myofibers in Dystrophin-deficient mice. However, despite FMPs and SCs having similar myogenic ability in culture, the regenerative ability of FMPs was less than that of SCs in vivo. FMPs that had activated MyoD engrafted more efficiently to regenerate myofibers than MyoD-negative FMPs. Transcriptome and surface marker analyses of these cells suggest the importance of myogenic priming for the efficient myogenic engraftment. Our findings suggest the regenerative capability of FMPs in the context of muscle repair and cell therapy for degenerative muscle disease.

  8. IL-6 Impairs Myogenic Differentiation by Downmodulation of p90RSK/eEF2 and mTOR/p70S6K Axes, without Affecting AKT Activity

    Directory of Open Access Journals (Sweden)

    Michele Pelosi

    2014-01-01

    Full Text Available IL-6 is a multifaceted pleiotropic cytokine, which is produced by a variety of cell types and targets different cells and tissues. In physiological conditions, IL-6 can be locally and transiently produced by skeletal muscle and plays an important role in muscle homeostasis. Circulating IL-6 levels are normally very low or undetectable but are dramatically increased in several pathologic conditions. In this study, we aimed to define the potential molecular mechanisms underlying the effects of IL-6 on myogenic program. We explored the molecular mechanisms through which exogenous IL-6, or the conditioned medium from the murine C-26 adenocarcinoma cells (a cellular model that secretes high levels of IL-6 and induces cancer cachexia in mice, interferes with the myogenic program. Our study revealed that IL-6 induces the activation of the Stat3 signaling and promotes the downmodulation of the p90RSK/eEF2 and mTOR/p70S6K axes, while it does not affect the activation of AKT. We thus identified potential molecular mediators of the inhibitory effects of IL-6 on myogenic program.

  9. Myostatin acts as an autocrine/paracrine negative regulator in myoblast differentiation from human induced pluripotent stem cells

    Energy Technology Data Exchange (ETDEWEB)

    Gao, Fei; Kishida, Tsunao; Ejima, Akika [Department of Immunology, Kyoto Prefectural University of Medicine, Kyoto (Japan); Gojo, Satoshi [Department of Cardiac Support, Kyoto Prefectural University of Medicine, Kyoto (Japan); Mazda, Osam, E-mail: mazda@koto.kpu-m.ac.jp [Department of Immunology, Kyoto Prefectural University of Medicine, Kyoto (Japan)

    2013-02-08

    Highlights: ► iPS-derived cells express myostatin and its receptor upon myoblast differentiation. ► Myostatin inhibits myoblast differentiation by inhibiting MyoD and Myo5a induction. ► Silencing of myostatin promotes differentiation of human iPS cells into myoblasts. -- Abstract: Myostatin, also known as growth differentiation factor (GDF-8), regulates proliferation of muscle satellite cells, and suppresses differentiation of myoblasts into myotubes via down-regulation of key myogenic differentiation factors including MyoD. Recent advances in stem cell biology have enabled generation of myoblasts from pluripotent stem cells, but it remains to be clarified whether myostatin is also involved in regulation of artificial differentiation of myoblasts from pluripotent stem cells. Here we show that the human induced pluripotent stem (iPS) cell-derived cells that were induced to differentiate into myoblasts expressed myostatin and its receptor during the differentiation. An addition of recombinant human myostatin (rhMyostatin) suppressed induction of MyoD and Myo5a, resulting in significant suppression of myoblast differentiation. The rhMyostatin treatment also inhibited proliferation of the cells at a later phase of differentiation. RNAi-mediated silencing of myostatin promoted differentiation of human iPS-derived embryoid body (EB) cells into myoblasts. These results strongly suggest that myostatin plays an important role in regulation of myoblast differentiation from iPS cells of human origin. The present findings also have significant implications for potential regenerative medicine for muscular diseases.

  10. Ocular and Cervical Vestibular Evoked Myogenic Potentials (VEMPs) in healthy volunteers: the intra-, interobserver, and the test re-test reliability.

    Science.gov (United States)

    Venhovens, J; Meulstee, J; Verhagen, W I M

    2015-01-01

    The aims of the study are to determine the intra-, interobserver, and the test re-test reliability of the cervical and ocular vestibular evoked myogenic potentials (VEMPs). Twenty healthy subjects underwent acoustically and forehead tap elicited cervical and ocular VEMPs. The measurements were repeated one week later. The intra- and interobserver reliability of both ocular and cervical VEMPs is excellent. The test re-test reliability of the raw p13n23 peak-to-peak amplitudes of the cervical VEMPs is excellent (ICC: 0.76, 0.87) and the p13 latencies show a good reliability (ICC: 0.56, 0.73). The raw n1p1 peak-to-peak amplitudes of the ocular VEMPs show a fair-to-good test re-test reliability (ICC: 0.51, 0.64) and the n1 and p1 latencies show a poor reliability (ICC: -0.17 ≤ x ≤ 0.44). The intra- and interobserver reliability of the cervical and ocular VEMPs is excellent. The raw ocular and cervical VEMP peak-to-peak amplitudes are the most reliable parameters, followed by the cervical VEMP latencies. The ocular VEMP latencies shows a poor test re-test reliability. The individual VEMP results, however, remained within normal limits despite the test re-test variability.

  11. Effect of short-duration sleep deprivation on the vestibulo-ocular reflex system evaluated by ocular vestibular-evoked myogenic potential test.

    Science.gov (United States)

    Lin, Bing-Yi; Young, Yi-Ho

    2014-07-01

    The ocular vestibular-evoked myogenic potential (oVEMP) test yields the same information as the rotational test in sleep-deprived subjects as evidenced by increased vestibulo-ocular reflex (VOR) asymmetry. However, the duration of sleep deprivation (12 h) and testing time required for the oVEMP test are shorter than for the rotational test. This study utilized the oVEMP test in sleep-deprived subjects to investigate the effect of short-duration sleep deprivation on the VOR system. Twenty healthy resident physicians underwent oVEMP and cervical VEMP (cVEMP) tests twice in a randomized order; one test battery was performed after a normal sleep, and the other was performed after a night duty at the emergency service, which meant a state of 12 h sleep deprivation. All 20 subjects had clear oVEMPs regardless of whether testing was performed after a normal sleep or 12 h sleep deprivation condition. Significant differences were not identified between the two sleep conditions in terms of characteristic parameters, i.e. latencies and amplitude of oVEMPs. However, the mean asymmetry ratio after sleep deprivation (20 ± 13%) was significantly larger than that after normal sleep (8 ± 12%). Conversely, the cVEMP parameters did not differ significantly between the two sleep conditions.

  12. [The importance of vestibular evoked myogenic potentials for the assessment of the otolith function in the patients presenting with benign paroxysmal positional vertigo].

    Science.gov (United States)

    Kunel'skaya, N L; Baybakova, E V; Guseva, A L; Chugunova, M A; Manaenkova, E A

    2017-01-01

    The objective of the present study was to evaluate the otolith function in the patients presenting with idiopathic benign paroxysmal positional vertigo (pBPPV) attributable to the occlusion of the posterior semicircular canal (PSCC) of the inner ear with the use of vestibular evoked myogenic potentials (VEMP). Cervical (cVEMP) and ocular VEMP (oVEMP) were measured in 34 patients with idiopathic pBPPV before and 7 days after the treatment by means of reposition maneuvers. The results of the repeated Dix-Hallpike test performed 7 days after the repositioning maneuver were negative in 27 patients and positive in 7 patients. There was no statistically significant difference in the amplitude of cervical VEMP between the healthy and affected ears either before or after the repositioning treatment. The measurement of oVEMP revealed a reduction of the response amplitude on the affected side. The average values of the plnl on the healthy side were 12.84±1.09 and those on the affected side 4.62±0.69 (pocular VEMP on the affected side with intact cervical VEMP on both sides. The successful treatment of pBPPV of PSCC with the use of the liberatory maneuver results in the increase of the oVEMP amplitude on the affected side increases while the response asymmetry between both sides significantly decreases which indicates the repair of the utriculus otolith function.

  13. Gene expression deregulation in postnatal skeletal muscle of TK2 deficient mice reveals a lower pool of proliferating myogenic progenitor cells.

    Directory of Open Access Journals (Sweden)

    João A Paredes

    Full Text Available Loss of thymidine kinase 2 (TK2 causes a heterogeneous myopathic form of mitochondrial DNA (mtDNA depletion syndrome (MDS in humans that predominantly affects skeletal muscle tissue. In mice, TK2 deficiency also affects several tissues in addition to skeletal muscle, including brain, heart, adipose tissue, kidneys and causes death about 3 weeks after birth. We analysed skeletal muscle and heart muscle tissues of Tk2 knockout mice at postnatal development phase and observed that TK2 deficient pups grew slower and their skeletal muscles appeared significantly underdeveloped, whereas heart was close to normal in size. Both tissues showed mtDNA depletion and mitochondria with altered ultrastructure, as revealed by transmission electron microscopy. Gene expression microarray analysis showed a strong down-regulation of genes involved in cell cycle and cell proliferation in both tissues, suggesting a lower pool of undifferentiated proliferating cells. Analysis of isolated primary myoblasts from Tk2 knockout mice showed slow proliferation, less ability to differentiate and signs of premature senescence, even in absence of mtDNA depletion. Our data demonstrate that TK2 deficiency disturbs myogenic progenitor cells function in postnatal skeletal muscle and we propose this as one of the causes of underdeveloped phenotype and myopathic characteristic of the TK2 deficient mice, in addition to the progressive mtDNA depletion, mitochondrial damage and respiratory chain deficiency in post-mitotic differentiated tissue.

  14. cDNA Microarrays Detect Activation of a Myogenic Transcription Program by the PAX3-FKHR Fusion Oncogene

    National Research Council Canada - National Science Library

    Javed Khan; Michael L. Bittner; Lao H. Saal; Ulrike Teichmann; David O. Azorsa; Gerald C. Gooden; William J. Pavan; Jeffrey M. Trent; Paul S. Meltzer

    1999-01-01

    .... To investigate the actions of these transcription factors, both Pax3 and PAX3-FKHR were introduced into NIH 3T3 cells, and the resultant gene expression changes were analyzed with a murine cDNA micro...

  15. Pericyte response to contraction mode-specific resistance exercise training in human skeletal muscle.

    Science.gov (United States)

    Farup, Jean; De Lisio, Michael; Rahbek, Stine Klejs; Bjerre, Jonas; Vendelbo, Mikkel Holm; Boppart, Marni D; Vissing, Kristian

    2015-11-15

    Skeletal muscle satellite cells (SCs) are important for muscle repair and hypertrophy in response mechanical stimuli. Neuron-glial antigen 2-positive (NG2(+)) and alkaline phosphatase-positive (ALP(+)) pericytes may provide an alternative source of myogenic progenitors and/or secrete paracrine factors to induce Pax7(+) SC proliferation and differentiation. The purpose of this study was to investigate NG2(+) and ALP(+) cell quantity, as well as SC content and activation, in human skeletal muscle following prolonged concentric (Conc) or eccentric (Ecc) resistance training. Male subjects engaged in unilateral resistance training utilizing isolated Ecc or Conc contractions. After 12 wk, muscle biopsies were analyzed for NG2(+) and ALP(+) pericytes, total Pax7(+) SCs, activated SCs (Pax7(+)MyoD(+)), and differentiating myogenic cells (Pax7(-) MyoD(+)). NG2(+) cells localized to CD31(+) vessels and the majority coexpressed ALP. NG2(+) pericyte quantity decreased following both Conc and Ecc training (P < 0.05). ALP(+) pericyte quantity declined following Conc (P < 0.05) but not Ecc training. Conversely, total Pax7(+) SC content was elevated following Conc only (P < 0.001), while Pax7(+)MyoD(+) SC content was increased following Conc and Ecc (P < 0.001). Follow up analyses demonstrated that CD90(+) and platelet-derived growth factor receptor-α (PDGFRα)(+) mononuclear cell proliferation was also increased in response to both Conc and Ecc training (P < 0.01). In summary, resistance training results in a decline in pericyte quantity and an increase in mesenchymal progenitor cell proliferation, and these events likely influence SC pool expansion and increased activation observed posttraining.

  16. Parallel differentiation of embryonic stem cells into different cell types by a single gene-based differentiation system.

    Science.gov (United States)

    Thoma, Eva C; Maurus, Katja; Wagner, Toni U; Schartl, Manfred

    2012-04-01

    The generation of defined somatic cell types from pluripotent stem cells represents a promising system for many applications for regenerative therapy or developmental studies. Certain key developmental genes have been shown to be able to influence the fate determination of differentiating stem cells suggesting an alternative differentiation strategy to conventional medium-based methods. Here, we present a system allowing controlled, directed differentiation of embryonic stem cells (ESCs) solely by ectopic expression of single genes. We demonstrate that the myogenic master regulator myoD1 is sufficient to induce formation of skeletal muscle. In contrast to previous studies, our data suggest that myoD1-induced differentiation is independent of additional differentiation-inducing or lineage-promoting signals and occurs even under pluripotency-promoting conditions. Moreover, we demonstrate that single gene-induced differentiation enables the controlled formation of two distinct cell types in parallel. By mixing ES cell lines expressing myoD1 or the neural transcription factor ngn2, respectively, we generated a mixed culture of myocytes and neurons. Our findings provide new insights in the role of key developmental genes during cell fate decisions. Furthermore, this study represents an interesting strategy to obtain mixed cultures of different cells from stem cells, suggesting a valuable tool for cellular development and cell-cell interaction studies.

  17. Stomagenesis versus myogenesis: Parallels in intrinsic and extrinsic regulation of transcription factor mediated specialized cell-type differentiation in plants and animals.

    Science.gov (United States)

    Putarjunan, Aarthi; Torii, Keiko U

    2016-05-01

    Although the last common unicellular ancestor of plants and animals diverged several billion years ago, and while having developed unique developmental programs that facilitate differentiation and proliferation specific to plant and animal systems, there still exists a high degree of conservation in the logic regulating these developmental processes within these two seemingly diverse kingdoms. Stomatal differentiation in plants involves a series of orchestrated cell division events mediated by a family of closely related bHLH transcription factors (TFs) to create a pair of mature guard cells. These TFs are in turn regulated by a number of upstream signaling components that ultimately function to achieve lineage specific differentiation and organized tissue patterning on the plant epidermis. The logic involved in the specification of the myogenic differentiation program in animals is intriguingly similar to stomatal differentiation in plants: Closely-related myogenic bHLHs, known as MRFs (Myogenic Regulatory Factors) provide lineage specificity essential for cell-fate determination. These MRFs, similar to the bHLHs in plants, are regulated by several upstream signaling cascades that succinctly regulate each differentiation step, leading to the production of mature muscle fibers. This review aims at providing a perspective on the emerging parallels in the logic employed by key bHLH transcription factors and their upstream signaling components that function to precisely regulate key cell-state transition events in the stomatal as well as myogenic cell lineages. © 2016 The Authors. Development, Growth & Differentiation published by John Wiley & Sons Australia, Ltd on behalf of Japanese Society of Developmental Biologists.

  18. Biomimetic three-dimensional anisotropic geometries by uniaxial stretch of poly(ε-caprolactone) films for mesenchymal stem cell proliferation, alignment, and myogenic differentiation.

    Science.gov (United States)

    Wang, Zu-yong; Teo, Erin Yiling; Chong, Mark Seow Khoon; Zhang, Qin-yuan; Lim, Jing; Zhang, Zhi-yong; Hong, Ming-hui; Thian, Eng-san; Chan, Jerry Kok Yen; Teoh, Swee-hin

    2013-07-01

    Anisotropic geometries are critical for eliciting cell alignment to dictate tissue microarchitectures and biological functions. Current fabrication techniques are complex and utilize toxic solvents, hampering their applications for translational research. Here, we present a novel simple, solvent-free, and reproducible method via uniaxial stretching for incorporating anisotropic topographies on bioresorbable films with ambitions to realize stem cell alignment control. Uniaxial stretching of poly(ε-caprolactone) (PCL) films resulted in a three-dimensional micro-ridge/groove topography (inter-ridge-distance: ~6 μm; ridge-length: ~90 μm; ridge-depth: 200-900 nm) with uniform distribution and controllable orientation by the direction of stretch on the whole film surface. When stretch temperature (Ts) and draw ratio (DR) were increased, the inter-ridge-distance was reduced and ridge-length increased. Through modification of hydrolysis, increased surface hydrophilicity was achieved, while maintaining the morphology of PCL ridge/grooves. Upon seeding human mesenchymal stem cells (hMSCs) on uniaxial-stretched PCL (UX-PCL) films, aligned hMSC organization was obtained. Compared to unstretched films, hMSCs on UX-PCL had larger increase in cellular alignment (>85%) and elongation, without indication of cytotoxicity or reduction in cellular proliferation. This aligned hMSC organization was homogenous and stably maintained with controlled orientation along the ridges on the whole UX-PCL surface for over 2 weeks. Moreover, the hMSCs on UX-PCL had a higher level of myogenic genes' expression than that on the unstretched films. We conclude that uniaxial stretching has potential in patterning film topography with anisotropic structures. The UX-PCL in conjunction with hMSCs could be used as "basic units" to create tissue constructs with microscale control of cellular alignment and elongation for tissue engineering applications.

  19. Enhanced large conductance K+ channel activity contributes to the impaired myogenic response in the cerebral vasculature of Fawn Hooded Hypertensive rats

    Science.gov (United States)

    Pabbidi, Mallikarjuna R.; Mazur, Olga; Fan, Fan; Farley, Jerry M.; Gebremedhin, Debebe; Harder, David R.

    2014-01-01

    Recent studies have indicated that the myogenic response (MR) in cerebral arteries is impaired in Fawn Hooded Hypertensive (FHH) rats and that transfer of a 2.4 megabase pair region of chromosome 1 (RNO1) containing 15 genes from the Brown Norway rat into the FHH genetic background restores MR in a FHH.1BN congenic strain. However, the mechanisms involved remain to be determined. The present study examined the role of the large conductance calcium-activated potassium (BK) channel in impairing the MR in FHH rats. Whole-cell patch-clamp studies of cerebral vascular smooth muscle cells (VSMCs) revealed that iberiotoxin (IBTX; BK inhibitor)-sensitive outward potassium (K+) channel current densities are four- to fivefold greater in FHH than in FHH.1BN congenic strain. Inside-out patches indicated that the BK channel open probability (NPo) is 10-fold higher and IBTX reduced NPo to a greater extent in VSMCs isolated from FHH than in FHH.1BN rats. Voltage sensitivity of the BK channel is enhanced in FHH as compared with FHH.1BN rats. The frequency and amplitude of spontaneous transient outward currents are significantly greater in VSMCs isolated from FHH than in FHH.1BN rats. However, the expression of the BK-α and -β-subunit proteins in cerebral vessels as determined by Western blot is similar between the two groups. Middle cerebral arteries (MCAs) isolated from FHH rats exhibited an impaired MR, and administration of IBTX restored this response. These results indicate that there is a gene on RNO1 that impairs MR in the MCAs of FHH rats by enhancing BK channel activity. PMID:24464756

  20. Augmentation of ocular vestibular-evoked myogenic potentials via bone-conducted vibration stimuli in Ménière disease.

    Science.gov (United States)

    Wen, Ming-Hsun; Cheng, Po-Wen; Young, Yi-Ho

    2012-05-01

    The asymmetry ratio of ocular vestibular-evoked myogenic potential (oVEMP) >40% is interpreted as augmented or reduced response depending on whether the nI-pI amplitude of the lesion ear is larger or smaller than that of the opposite ear, respectively. This study compared unilateral Ménière disease patients with augmented oVEMPs vs reduced oVEMPs to elucidate the mechanism of augmented oVEMP. Case series with chart review. University hospital. Forty patients with unilateral definite Ménière disease were enrolled in this study, including 20 patients with augmented oVEMPs and another 20 patients with reduced oVEMPs in the hydropic side. All patients underwent audiometry, caloric test, and oVEMP and cervical VEMP (cVEMP) tests via bone-conducted vibration stimuli. Then, the oVEMP and cVEMP test results were compared with the stage of Ménière disease, respectively. The augmented group had earlier nI and pI latencies and larger nI-pI amplitude of oVEMPs compared with the reduced group. Caloric test also revealed a significant difference in abnormal responses between the augmented and reduced groups. However, both groups did not differ significantly in the abnormal percentage of cVEMP test results. A significant trend to decline in the prevalence of augmented oVEMPs was noted from stages I to III-IV but not in that of abnormal cVEMPs. The augmented oVEMPs have earlier latencies and larger amplitudes compared with the reduced oVEMPs, indicating that a relatively larger population of intact utricular afferents is activated during the early stage of Ménière disease.

  1. Optimizing stimulus repetition rate for recording ocular vestibular evoked myogenic potential elicited by air-conduction tone bursts of 500 Hz

    Directory of Open Access Journals (Sweden)

    Niraj Kumar Singh

    2014-03-01

    Full Text Available Amidst several publications reporting the effects of stimulus-related parameters on ocular vestibular evoked myogenic potential (oVEMP, the effect of the repetition rate on oVEMP responses has largely gone unexplored. Studies have used a repetition rate of ~5.1 Hz mainly due to a presumption that oVEMP, like cervical VEMP, should produce best responses for ~5 Hz, although there is paucity of experimental evidence to support this hypothesis. 52 healthy individuals in the age range of 17-35 years underwent air-conduction oVEMP elicited by 500 Hz tone-bursts using seven different repetition rates (3.1, 5.1, 10.1, 15.1, 20.1, 25.1 and 30.1 Hz. The results revealed a tendency for prolongation of latencies and reduction in amplitude with increasing repetition rate. However, significantly longer latencies were observed only for 20.1 Hz and larger amplitudes for 3.1 and 5.1 Hz (P<0.05. There was no significant difference between the rates of 3.1 Hz and 5.1 Hz. However 3.1 Hz produced poorer signal-to-noise ratio and required considerably longer time and thereby had lesser efficiency than 5.1 Hz (P<0.05. This would also result in higher fatigue and irritation levels considering the physical act of maintaining a supero-medial gaze. Thus the use of 5.1 Hz is recommended for clinical recording of oVEMP.

  2. Optimizing Stimulus Repetition Rate for Recording Ocular Vestibular Evoked Myogenic Potential Elicited by Air-Conduction Tone Bursts of 500 Hz.

    Science.gov (United States)

    Singh, Niraj Kumar; Kadisonga, Peter; Ashitha, Palliyath

    2014-03-06

    Amidst several publications reporting the effects of stimulus-related parameters on ocular vestibular evoked myogenic potential (oVEMP), the effect of the repetition rate on oVEMP responses has largely gone unexplored. Studies have used a repetition rate of ~5.1 Hz mainly due to a presumption that oVEMP, like cervical VEMP, should produce best responses for ~5 Hz, although there is paucity of experimental evidence to support this hypothesis. 52 healthy individuals in the age range of 17-35 years underwent air-conduction oVEMP elicited by 500 Hz tone-bursts using seven different repetition rates (3.1, 5.1, 10.1, 15.1, 20.1, 25.1 and 30.1 Hz). The results revealed a tendency for prolongation of latencies and reduction in amplitude with increasing repetition rate. However, significantly longer latencies were observed only for 20.1 Hz and larger amplitudes for 3.1 and 5.1 Hz (P<0.05). There was no significant difference between the rates of 3.1 Hz and 5.1 Hz. However 3.1 Hz produced poorer signal-to-noise ratio and required considerably longer time and thereby had lesser efficiency than 5.1 Hz (P<0.05). This would also result in higher fatigue and irritation levels considering the physical act of maintaining a supero-medial gaze. Thus the use of 5.1 Hz is recommended for clinical recording of oVEMP.

  3. Frequency tuning of the cervical vestibular-evoked myogenic potential (cVEMP) recorded from multiple sites along the sternocleidomastoid muscle in normal human subjects.

    Science.gov (United States)

    Wei, Wei; Jeffcoat, Ben; Mustain, William; Zhu, Hong; Eby, Thomas; Zhou, Wu

    2013-02-01

    Frequency tuning of tone burst-evoked myogenic potentials recorded from the sternocleidomastoid muscle (cervical VEMP or cVEMP) is used clinically to assess vestibular function. Understanding the characteristics of cVEMP is important for improving the specificity of cVEMP testing in diagnosing vestibular deficits. In the present study, we analyzed the frequency tuning properties of the cVEMPs by constructing detailed tuning curves and examining their morphology and dependence on SCM tonic level, sound intensity, and recording site along the SCM. Here we report two main findings. First, by employing nine tone frequencies between 125 and 4,000 Hz, some tuning curves exhibited two distinct peaks, which cannot be modeled by a single mass spring system as previously suggested. Instead, the observed tuning is better modeled as linear summation of two mass spring systems, with resonance frequencies at ~300 and ~1,000 Hz. Peak frequency of cVEMP tuning curves was not affected by SCM tonic level, sound intensity, and location of recording site on the SCM. However, sharpness of cVEMP tuning was increased at lower sound intensities. Second, polarity of cVEMP responses recorded from the lower quarter of the SCM was reversed as compared to that at the two upper sites. While more studies are needed, these results suggest that cVEMP tuning is mediated through multiple generators with different resonance frequencies. Future studies are needed to explore implications of these results on development of selective VEMP tests and determine the nature of polarity inversion at the lower quarter of SCM.

  4. Runx1 Transcription Factor Is Required for Myoblasts Proliferation during Muscle Regeneration.

    Directory of Open Access Journals (Sweden)

    Kfir Baruch Umansky

    2015-08-01

    Full Text Available Following myonecrosis, muscle satellite cells proliferate, differentiate and fuse, creating new myofibers. The Runx1 transcription factor is not expressed in naïve developing muscle or in adult muscle tissue. However, it is highly expressed in muscles exposed to myopathic damage yet, the role of Runx1 in muscle regeneration is completely unknown. Our study of Runx1 function in the muscle's response to myonecrosis reveals that this transcription factor is activated and cooperates with the MyoD and AP-1/c-Jun transcription factors to drive the transcription program of muscle regeneration. Mice lacking dystrophin and muscle Runx1 (mdx-/Runx1f/f, exhibit impaired muscle regeneration leading to age-dependent muscle waste, gradual decrease in motor capabilities and a shortened lifespan. Runx1-deficient primary myoblasts are arrested at cell cycle G1 and consequently differentiate. Such premature differentiation disrupts the myoblasts' normal proliferation/differentiation balance, reduces the number and size of regenerating myofibers and impairs muscle regeneration. Our combined Runx1-dependent gene expression, ChIP-seq, ATAC-seq and histone H3K4me1/H3K27ac modification analyses revealed a subset of Runx1-regulated genes that are co-occupied by MyoD and c-Jun in mdx-/Runx1f/f muscle. The data provide unique insights into the transcriptional program driving muscle regeneration and implicate Runx1 as an important participant in the pathology of muscle wasting diseases.

  5. Myogenic expression of an injectable protease-resistant growth hormone-releasing hormone augments long-term growth in pigs

    Science.gov (United States)

    Draghia-Akli, R.; Fiorotto, M. L.; Hill, L. A.; Malone, P. B.; Deaver, D. R.; Schwartz, R. J.

    1999-01-01

    Ectopic expression of a new serum protease-resistant porcine growth hormone-releasing hormone, directed by an injectable muscle-specific synthetic promoter plasmid vector (pSP-HV-GHRH), elicits growth in pigs. A single 10 mg intramuscular injection of pSP-HV-GHRH DNA followed by electroporation in three-week-old piglets elevated serum GHRH levels by twofold to fourfold, enhanced growth hormone secretion, and increased serum insulin-like growth factor-I by threefold to sixfold over control pigs. After 65 days the average body weight of the pigs injected with pSP-HV-GHRH was approximately 37% greater than the placebo-injected controls and resulted in a significant reduction in serum urea concentration, indicating a decrease in amino acid catabolism. Evaluation of body composition indicated a uniform increase in mass, with no organomegaly or associated pathology.

  6. The critical role of myostatin in differentiation of sheep myoblasts

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Chenxi [College of Life Science and Technology, Xinjiang University, Urumqi (China); Xinjiang Laboratory of Animal Biotechnology, Urumqi (China); Li, Wenrong; Zhang, Xuemei; Zhang, Ning; He, Sangang; Huang, Juncheng [Xinjiang Laboratory of Animal Biotechnology, Urumqi (China); Laboratory of Grass-fed Animal Genetics, Breeding and Reproduction of Ministry of Agriculture, Urumqi (China); Animal Biotechnological Research Center, Xinjiang Academy of Animal Science, Urumqi (China); Ge, Yubin [The State Engineering Laboratory of AIDS Vaccine, College of Life Science, Jilin University, Changchun (China); Liu, Mingjun, E-mail: xjlmj2004@yahoo.com.cn [Xinjiang Laboratory of Animal Biotechnology, Urumqi (China); Laboratory of Grass-fed Animal Genetics, Breeding and Reproduction of Ministry of Agriculture, Urumqi (China); Animal Biotechnological Research Center, Xinjiang Academy of Animal Science, Urumqi (China)

    2012-06-08

    Highlights: Black-Right-Pointing-Pointer Identification of the effective and specific shRNA to knockdown MSTN. Black-Right-Pointing-Pointer Overexpression of MSTN reversibly suppressed myogenic differentiation. Black-Right-Pointing-Pointer shRNA knockdown of endogenous MSTN promoted ovine myoblast differentiation. Black-Right-Pointing-Pointer MSTN inhibits myogenic differentiation through down-regulation of MyoD and Myogenin and up-regulation of Smad3. Black-Right-Pointing-Pointer Provides a promise for the generation of transgenic sheep to improve meat productivity. -- Abstract: Myostatin [MSTN, also known as growth differentiation factor 8 (GDF8)], is an inhibitor of skeletal muscle growth. Blockade of MSTN function has been reported to result in increased muscle mass in mice. However, its role in myoblast differentiation in farm animals has not been determined. In the present study, we sought to determine the role of MSTN in the differentiation of primary sheep myoblasts. We found that ectopic overexpression of MSTN resulted in lower fusion index in sheep myoblasts, which indicated the repression of myoblast differentiation. This phenotypic change was reversed by shRNA knockdown of the ectopically expressed MSTN in the cells. In contrast, shRNA knockdown of the endogenous MSTN resulted in induction of myogenic differentiation. Additional studies revealed that the induction of differentiation by knocking down the ectopically or endogenously expressed MSTN was accompanied by up-regulation of MyoD and myogenin, and down-regulation of Smad3. Our results demonstrate that MSTN plays critical role in myoblast differentiation in sheep, analogous to that in mice. This study also suggests that shRNA knockdown of MSTN could be a potentially promising approach to improve sheep muscle growth, so as to increase meat productivity.

  7. Deep Proteomics of Mouse Skeletal Muscle Enables Quantitation of Protein Isoforms, Metabolic Pathways, and Transcription Factors*

    Science.gov (United States)

    Deshmukh, Atul S.; Murgia, Marta; Nagaraj, Nagarjuna; Treebak, Jonas T.; Cox, Jürgen; Mann, Matthias

    2015-01-01

    Skeletal muscle constitutes 40% of individual body mass and plays vital roles in locomotion and whole-body metabolism. Proteomics of skeletal muscle is challenging because of highly abundant contractile proteins that interfere with detection of regulatory proteins. Using a state-of-the art MS workflow and a strategy to map identifications from the C2C12 cell line model to tissues, we identified a total of 10,218 proteins, including skeletal muscle specific transcription factors like myod1 and myogenin and circadian clock proteins. We obtain absolute abundances for proteins expressed in a muscle cell line and skeletal muscle, which should serve as a valuable resource. Quantitation of protein isoforms of glucose uptake signaling pathways and in glucose and lipid metabolic pathways provides a detailed metabolic map of the cell line compared with tissue. This revealed unexpectedly complex regulation of AMP-activated protein kinase and insulin signaling in muscle tissue at the level of enzyme isoforms. PMID:25616865

  8. Common and Distinctive Functions of the Hippo Effectors Taz and Yap in Skeletal Muscle Stem Cell Function.

    Science.gov (United States)

    Sun, Congshan; De Mello, Vanessa; Mohamed, Abdalla; Ortuste Quiroga, Huascar P; Garcia-Munoz, Amaya; Al Bloshi, Abdullah; Tremblay, Annie M; von Kriegsheim, Alexander; Collie-Duguid, Elaina; Vargesson, Neil; Matallanas, David; Wackerhage, Henning; Zammit, Peter S

    2017-08-01

    Hippo pathway downstream effectors Yap and Taz play key roles in cell proliferation and regeneration, regulating gene expression especially via Tead transcription factors. To investigate their role in skeletal muscle stem cells, we analyzed Taz in vivo and ex vivo in comparison with Yap. Small interfering RNA knockdown or retroviral-mediated expression of wild-type human or constitutively active TAZ mutants in satellite cells showed that TAZ promoted proliferation, a function shared with YAP. However, at later stages of myogenesis, TAZ also enhanced myogenic differentiation of myoblasts, whereas YAP inhibits such differentiation. Functionally, while muscle growth was mildly affected in Taz (gene Wwtr1(-/-) ) knockout mice, there were no overt effects on regeneration. Conversely, conditional knockout of Yap in satellite cells of Pax7(Cre-ERT2/+) : Yap(fl) °(x/fl) °(x) :Rosa26(Lacz) mice produced a regeneration deficit. To identify potential mechanisms, microarray analysis showed many common TAZ/YAP target genes, but TAZ also regulates some genes independently of YAP, including myogenic genes such as Pax7, Myf5, and Myod1 (ArrayExpress-E-MTAB-5395). Proteomic analysis revealed many novel binding partners of TAZ/YAP in myogenic cells, but TAZ also interacts with proteins distinct from YAP that are often involved in myogenesis and aspects of cytoskeleton organization (ProteomeXchange-PXD005751). Neither TAZ nor YAP bind members of the Wnt destruction complex but both regulated expression of Wnt and Wnt-cross talking genes with known roles in myogenesis. Finally, TAZ operates through Tead4 to enhance myogenic differentiation. In summary, Taz and Yap have overlapping functions in promoting myoblast proliferation but Taz then switches to enhance myogenic differentiation. Stem Cells 2017;35:1958-1972. © 2017 The Authors. The Authors Stem Cells published by Wiley Periodicals, Inc. on behalf of AlphaMed Press.

  9. Efficacy of cervical and ocular vestibular-evoked myogenic potentials in evaluation of benign paroxysmal positional vertigo of posterior semicircular canal.

    Science.gov (United States)

    Singh, Niraj Kumar; Apeksha, Kumari

    2016-09-01

    Benign paroxysmal positional vertigo (BPPV) constitutes a major proportion of the population with peripheral vestibulopathies. Although the freely floating otoconia within the semicircular canals is responsible for the symptoms of BPPV, the source of the otoconia debris is mainly believed to be the otolith organs. Therefore, the pathology in either or both the otolith organs appears a logical proposition. Cervical and ocular vestibular-evoked myogenic potentials (cVEMP and oVEMP), being the tests for functional integrity of the otolith organs, appear promising for investigating otolith involvement in BPPV. While recent evidences are suggestive of equivocal findings for cVEMP, there are only a few studies on oVEMP. Additionally, both these potentials have never been explored in the same set of individuals with BPPV. Therefore, the present study aimed to evaluate the functional integrity of the otolith organs through cVEMP and oVEMP in individuals with posterior canal BPPV. Thirty-one individuals with unilateral posterior canal BPPV and 31 age- and gender-matched healthy controls underwent 500 Hz tone-burst-evoked cVEMP and oVEMP. The results demonstrated no significant group difference on any of the cVEMP parameters (p > 0.05). A similar trend was noticed for the latency-related parameters of oVEMP. However, the peak-to-peak amplitude was significantly smaller in the affected ears of individuals with BPPV than their unaffected ears and the ears of healthy controls (p < 0.05). The BPPV group showed significantly higher inter-aural amplitude difference ratio than the healthy controls (p < 0.05). Further, the sensitivity and specificity of oVEMP were also found to be far superior to those of cVEMP. Thus, the outcome of the present study revealed involvement of utricle rather than saccule in posterior canal BPPV, and therefore, oVEMP appears to be better suited to clinical investigation than cVEMP in individuals with posterior canal BPPV.

  10. JAZF1 promotes proliferation of C2C12 cells, but retards their myogenic differentiation through transcriptional repression of MEF2C and MRF4—Implications for the role of Jazf1 variants in oncogenesis and type 2 diabetes

    Energy Technology Data Exchange (ETDEWEB)

    Yuasa, Katsutoshi; Aoki, Natsumi; Hijikata, Takao, E-mail: hijikata@musashino-u.ac.jp

    2015-08-15

    Single-nucleotide polymorphisms associated with type 2 diabetes (T2D) have been identified in Jazf1, which is also involved in the oncogenesis of endometrial stromal tumors. To understand how Jazf1 variants confer a risk of tumorigenesis and T2D, we explored the functional roles of JAZF1 and searched for JAZF1 target genes in myogenic C2C12 cells. Consistent with an increase of Jazf1 transcripts during myoblast proliferation and their decrease during myogenic differentiation in regenerating skeletal muscle, JAZF1 overexpression promoted cell proliferation, whereas it retarded myogenic differentiation. Examination of myogenic genes revealed that JAZF1 overexpression transcriptionally repressed MEF2C and MRF4 and their downstream genes. AMP deaminase1 (AMPD1) was identified as a candidate for JAZF1 target by gene array analysis. However, promoter assays of Ampd1 demonstrated that mutation of the putative binding site for the TR4/JAZF1 complex did not alleviate the repressive effects of JAZF1 on promoter activity. Instead, JAZF1-mediated repression of Ampd1 occurred through the MEF2-binding site and E-box within the Ampd1 proximal regulatory elements. Consistently, MEF2C and MRF4 expression enhanced Ampd1 promoter activity. AMPD1 overexpression and JAZF1 downregulation impaired AMPK phosphorylation, while JAZF1 overexpression also reduced it. Collectively, these results suggest that aberrant JAZF1 expression contributes to the oncogenesis and T2D pathogenesis. - Highlights: • JAZF1 promotes cell cycle progression and proliferation of myoblasts. • JAZF1 retards myogenic differentiation and hypertrophy of myotubes. • JAZF1 transcriptionally represses Mef2C and Mrf4 expression. • JAZF1 has an impact on the phosphorylation of AMPK.

  11. 三黄膏与生肌膏联合治疗Ⅲ、Ⅳ期压疮的疗效分析%The effect of Sanhuang plaster and Myogenic ointment on stage m and Ⅳ bedsore

    Institute of Scientific and Technical Information of China (English)

    蔡淑珍; 梁华; 谭贵英

    2012-01-01

    Objective To discuss the effect of Sanhuang plaster and Myogenic ointment on stage Ⅲ and Ⅳ bedsore.Methods Sixty - eight patients with bedsore were divided into experimental group of 34 patients and control group of 34 patients.The patients of two groups were treated with sponge or alternate inflatable cushion for decompression,keeping skin clean and dry,helping rolling massage,nutrition intervention,debridement to remove necrotic tissue and clean the wound with normal saline and related whole - body treatment.The experimental group was also added with Sanhuang plaster and Myogenic ointment,while the control group was given external apphcation with antibiotics.The treatment analysis was made on the 21st day and the 35th day of the treatment.Results The treatment efficiency and the cure rate of the experimental group were obviously higher than those of the control group,the healing time was significantly shorter than that of control group ( both are p < 0.05 ).Conclusions Applying Sanhuang plaster and Myogenic ointment to the treatment of stage Ⅲ and Ⅳ bedsore can obviously shorten the healing time and improve the cure rate.%目的 探讨三黄膏与生肌膏联合治疗Ⅲ、Ⅳ期压疮的效果.方法 将68例压疮患者分为实验组和对照组各34例,两组患者均给予海绵垫或交替式充气垫减压、保持皮肤清洁干燥、协助翻身按摩、营养干预、清创去除坏死组织、生理盐水清洗创面等压疮护理措施及相应的全身治疗,实验组加用三黄膏与生肌膏外敷,对照组加用抗生素外敷.于治疗第21天、第35天进行治疗效果分析.结果 实验组患者治疗有效率及治愈率明显高于对照组,愈合时间明显短于对照组(均为P<0.05),差异有统计学意义.结论 应用三黄膏与生肌膏联合治疗Ⅲ、Ⅳ期压疮可明显缩短愈合时间,提高治愈率.

  12. Transposição da rima palpebral em ptose miogênica mitocondrial Tarsal switch levator for mitochondrial myogenic ptosis

    Directory of Open Access Journals (Sweden)

    Lucia Miriam Dumont Lucci

    2009-04-01

    Full Text Available OBJETIVO: Relatar o uso da ressecção do tarso da pálpebra superior e enxerto na pálpebra inferior para a elevação da rima palpebral e liberação do eixo visual, sem causar complicações corneanas em pacientes portadores de ptose miogênica mitocondrial. MÉTODOS: Estudo prospectivo. A técnica cirúrgica consiste na tarsectomia da pálpebra superior e autoenxerto do tarso na lamela posterior da pálpebra inferior. As cirurgias foram realizadas sob anestesia local. No caso de diplopia, a cirurgia foi realizada em apenas um olho. RESULTADOS: O procedimento foi realizado em 9 olhos de 6 pacientes com miopatia mitocondrial. Cinco pacientes eram do sexo feminino, a média de idade foi de 59,8 anos e o seguimento variou de 30 a 60 meses. A rima palpebral elevou em todos os pacientes, descobrindo o eixo visual na posição primária do olhar e melhorando a posição viciosa de cabeça. Não houve complicações decorrentes da exposição do globo ocular. CONCLUSÃO: A técnica de transposição da rima palpebral é útil na correção do mau posicionamento das pálpebras em pacientes sem mecanismos de proteção porque eleva a pálpebra superior e a inferior, diminuindo ou eliminando o risco de causar lagoftalmo com complicações corneanas.PURPOSE: To report the use of tarsal switch levator resection procedures that lift the palpebral fissure to visual axis without causing corneal complications in patients with mitochondrial myogenic ptosis. METHODS: Prospective study. The technique consists of an upper eyelid tarsectomy, with transfer of the autologous tarsoconjunctival graft to the posterior lamella of the lower eyelid. The surgeries were performed under local anesthesia. In case of diplopia, the surgery was performed in one eye. RESULTS: Tarsal switch procedure was performed in 9 eyes of 6 patients with mitochondrial myopathy. There were 5 women; the average age was 59.8 years and the follow-up ranged from 30 to 60 months. The palpebral

  13. The Secretory Leukocyte Protease Inhibitor mRNA expression is involved with inflammatory control after downhill exercise in the triceps brachii intermediary head in Wistar rats.

    Science.gov (United States)

    Minari, André Luis Araujo; Oyama, Lila Missae; Dos Santos, Ronaldo Vagner Thomatieli

    2017-06-30

    After severe skeletal muscle damage, communication between inflammatory macrophages, myogenic cells, and modulatory secretion factors is essential to induce re-establishment of skeletal muscle structure. To analyze when characteristic gene expression of macrophages, myogenic cells, and SLPI are modulated after an exercise-induced muscle damage (EIMD) downhill protocol. Twenty-six rats were exposed to an experimental protocol of exercise and euthanized before (CTRL), immediately after (G0), and 24 (G24) and 48 (G48) hours after the exercise. After euthanasia, the Triceps brachii were dissected and analyzed by enzyme-linked immunosorbent assay and real time polymerase chain reaction. The CD68 expression was higher in the G24 when compared with all groups (p value < 0.05), whereas the CD163 was inhibited compared with G0 (p value < 0.05). MyoD and Myogenin were higher in the G24 when compared with G0 and G48 (p value < 0.05). The mRNA Secretory Leukocyte Protease Inhibitor (SLPI) was higher in the G48 when compared with the CTRL and G0 (p value < 0.05). IL-6 and TNF-α cytokines did not significantly change, but IL-10 presented a trend to be lower in the G0 when compared with G24 (p value = 0.054). A significant negative correlation was observed between CD68/CD163 (C.C = -0.71) and positive correlations between CD68/Myogenin (C.C = 0.65); MyoD/Myogenin (C.C = 0.72); IL-10/MyoD (C.C = 0.46), IL-10/MYOGENIN (C.C = 0.59); and IL-6/IL-10 (C.C = 0.64). A higher expression of CD68, concomitant with an elevation in MyoD and Myogenin 24 h after exercise, along with some correlations, suggests macrophage communication with myogenic cells independent of CD163 elevation. Additionally, the reestablishment of IL-10 in 24 h with the SLPI increased until 48 h indicate that these molecules are involved with anti-inflammatory transition after downhill exercise in the TBIH of Wistar rats.

  14. Myostatin shows a specific expression pattern in pig skeletal and extraocular muscles during pre- and post-natal growth.

    Science.gov (United States)

    Patruno, Marco; Caliaro, Francesca; Maccatrozzo, Lisa; Sacchetto, Roberta; Martinello, Tiziana; Toniolo, Luana; Reggiani, Carlo; Mascarello, Francesco

    2008-02-01

    Myogenesis is driven by an extraordinary array of cellular signals that follow a common expression pattern among different animal phyla. Myostatin (mstn) is a secreted growth factor that plays a pivotal role in skeletal muscle mass regulation. The aim of the present study was to investigate mstn expression in a large mammal (the pig) in order to ascertain whether distinct expression changes of this factor might be linked to the fiber-type composition of the muscle examined and/or to specific developmental stages. To assess the expression pattern of mstn in relation to myogenic proliferative (Pax7 and MyoD) and differentiative (myogenin) markers, we evaluated muscles with different myosin heavy-chain compositions sampled during pre- and post-natal development and on myogenic cells isolated from the same muscles. Skeletal muscles showed higher levels of mRNA for mstn and all other genes examined during fetal development than after birth. The wide distribution of mstn was also confirmed by immunohistochemistry experiments supporting evidence for cytoplasmic staining in early fetal periods as well as the localization in type 1 fibers at the end of the gestation period. Extraocular muscles, in contrast, did not exhibit decreasing mRNA levels for mstn or other genes even in adult samples and expressed higher levels of both mstn mRNA and protein compared with skeletal muscles. Experiments carried out on myogenic cells showed that mstn mRNA levels decreased when myoblasts entered the differentiation program and that cells isolated at early post-natal stages maintained a high level of Pax7 expression. Our results showed that mstn had a specific expression pattern whose variations depended on the muscle type examined, thus supporting the hypothesis that at birth, porcine myogenic cells continue to be influenced by hyperplastic/proliferative mechanisms.

  15. Muscle atrophy reversed by growth factor activation of satellite cells in a mouse muscle atrophy model.

    Directory of Open Access Journals (Sweden)

    Simon Hauerslev

    Full Text Available Muscular dystrophies comprise a large group of inherited disorders that lead to progressive muscle wasting. We wanted to investigate if targeting satellite cells can enhance muscle regeneration and thus increase muscle mass. We treated mice with hepatocyte growth factor and leukemia inhibitory factor under three conditions: normoxia, hypoxia and during myostatin deficiency. We found that hepatocyte growth factor treatment led to activation of the Akt/mTOR/p70S6K protein synthesis pathway, up-regulation of the myognic transcription factors MyoD and myogenin, and subsequently the negative growth control factor, myostatin and atrophy markers MAFbx and MuRF1. Hypoxia-induced atrophy was partially restored by hepatocyte growth factor combined with leukemia inhibitory factor treatment. Dividing satellite cells were three-fold increased in the treatment group compared to control. Finally, we demonstrated that myostatin regulates satellite cell activation and myogenesis in vivo following treatment, consistent with previous findings in vitro. Our results suggest, not only a novel in vivo pharmacological treatment directed specifically at activating the satellite cells, but also a myostatin dependent mechanism that may contribute to the progressive muscle wasting seen in severely affected patients with muscular dystrophy and significant on-going regeneration. This treatment could potentially be applied to many conditions that feature muscle wasting to increase muscle bulk and strength.

  16. Experiment list: SRX029151 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available 60,93.0,55.4,1123 GSM611284: EV-10T1/2 source_name=C3H10T1/2 fibroblasts with empty vector, forced to underg...l (empty vector) || growth protocol=forced to undergo myogenic differentiation by expression of MyoD || samp

  17. Growth factor array fabrication using a color ink jet printer.

    Science.gov (United States)

    Watanabe, Kohei; Miyazaki, Takeshi; Matsuda, Ryoichi

    2003-04-01

    We have developed a novel method for growth factor analysis using a commercial color ink jet printer to fabricate substrata patterned with growth factors. We prepared substrata with insulin printed in a simple pattern or containing multiple areas of varying quantities of printed insulin. When we cultured the mouse myoblast cell line, C2C12, on the insulin-patterned substrata, the cells were grown in the same pattern with the insulin-printed pattern. Cell culture with the latter substrata demonstrated that quantity control of insulin deposition by a color ink jet printer is possible. For further applications, we developed substrata with insulin-like growth factor-I (IGF-I) and basic fibroblast growth factor (bFGF) spotted in 16 different areas in varying combinations and concentrations (growth factor array). With this growth factor array, C2C12 cells were cultured, and the onset of muscle cell differentiation was monitored for the expression of the myogenic regulator myogenin. The ratio of cells expressing myogenin varied with the doses of IGF-I and bFGF in the sections, demonstrating a feasibility of growth factor array fabrication by a color ink jet printer. Since a printer manipulates several colors, this method can be easily applied to multivariate analyses of growth factors and attachment factors affecting cell growth and differentiation. This method may provide a powerful tool for cell biology and tissue engineering, especially for stem cell research in investigating unknown conditions for differentiation.

  18. Massage therapy in the management of myogenic TMD: a pilot study Massoterapia para o tratamento da DTM miogênica: um estudo piloto

    Directory of Open Access Journals (Sweden)

    Verena Kise Capellini

    2006-01-01

    Full Text Available INTRODUCTION: The Temporomandibular disorder (TMD is greatly prevalent in the population and can be associated with bruxism. This disorder produces several signs and symptoms. Among them, pain is one of the most important because it reduces life quality and productivity of people who have such disorder. The aim of this research was to study if massage causes pain relief and/or electromyographic (EMG changes. MATERIALS AND METHODS: The subjects were chosen by a questionnaire and divided into 2 different groups. Their ages varied from 19 to 22 years. The experimental group consisted of 6 TMD patients, who were submitted to the massage treatment and 4 EMG-sessions (the 1st EMG-session occurred before the treatment and the others in the 1st, 15th and 30th days after the treatment. The control group consisted of 6 TMD patients, who were submitted to the same 4 EMG-sessions. While EMG activity was recorded, subjects were asked to keep mandibular rest position (MRP and to perform maximal voluntary clenching (MVC. The treatment consisted of 15 massage-sessions on face and neck and in application of Visual Analogue Scale (VAS for measuring pain level. The massage sessions had 30 minutes of duration and were performed daily. The EMG data were processed to obtain the Root Mean Square (RMS, which were normalized by MVC. RESULTS: It was demonstrated that (1 RMS-MRP of the right masseter in experimental group at the 1st EMG-session was higher than at the 2nd EMG-session and (2 statistically significant reduction was found for VAS values after massage session. CONCLUSION: Unfortunately the sample is insufficient to draw any conclusions, therefore, more studies regarding the use of massage in the management of myogenic TMD are necessary.INTRODUÇÃO: A disfunção temporomandibular (DTM tem grande prevalência na população e pode ser associada com bruxismo. Esta desordem produz vários sinais e sintomas. Entre eles, a dor é um do mais importantes porque

  19. Temperature effect on proliferation and differentiation of satellite cells from turkeys with different growth rates.

    Science.gov (United States)

    Clark, D L; Coy, C S; Strasburg, G M; Reed, K M; Velleman, S G

    2016-04-01

    Poultry selected for growth have an inefficient thermoregulatory system and are more sensitive to temperature extremes. Satellite cells are precursors to skeletal muscle and mediate all posthatch muscle growth. Their physiological functions are affected by temperature. The objective of the current study was to determine how temperature affects satellite cells isolated from the pectoralis major (p. major) muscle (breast muscle) of turkeys selected for increased 16 wk body weight (F line) in comparison to a randombred control line (RBC2) from which the F line originated. Pectoralis major muscle satellite cells were thermally challenged by culturing between 33°C and 43°C to analyze the effects of cold and heat on proliferation and differentiation as compared to control temperature of 38°C. Expression levels of myogenic regulatory factors: myogenic differentiation factor 1 (MYOD1) and myogenin (MYOG) were quantified by quantitative polymerase chain reaction (qPCR). At all sampling times, proliferation increased at a linear rate across temperature in both the RBC2 and F lines. Differentiation also increased at a linear rate across temperature from 33 to 41°C at all sampling times in both the F and RBC2 lines. Satellite cells isolated from F line turkeys were more sensitive to both hot and cold temperatures as proliferation and differentiation increased to a greater extent across temperature (33 to 43°C) when compared with the RBC2 line. Expression of MYOD1 and MYOG increased as temperatures increased from 33 to 41°C at all sampling times in both the F and RBC2 lines. These results demonstrate that satellite cell function is sensitive to both cold and hot temperatures and p. major muscle satellite cells from F line turkeys are more sensitive to temperature extremes than RBC2 satellite cells.

  20. Identification of Transcription Factors for Lineage-Specific ESC Differentiation

    Science.gov (United States)

    Yamamizu, Kohei; Piao, Yulan; Sharov, Alexei A.; Zsiros, Veronika; Yu, Hong; Nakazawa, Kazu; Schlessinger, David; Ko, Minoru S.H.

    2013-01-01

    Summary A network of transcription factors (TFs) determines cell identity, but identity can be altered by overexpressing a combination of TFs. However, choosing and verifying combinations of TFs for specific cell differentiation have been daunting due to the large number of possible combinations of ∼2,000 TFs. Here, we report the identification of individual TFs for lineage-specific cell differentiation based on the correlation matrix of global gene expression profiles. The overexpression of identified TFs—Myod1, Mef2c, Esx1, Foxa1, Hnf4a, Gata2, Gata3, Myc, Elf5, Irf2, Elf1, Sfpi1, Ets1, Smad7, Nr2f1, Sox11, Dmrt1, Sox9, Foxg1, Sox2, or Ascl1—can direct efficient, specific, and rapid differentiation into myocytes, hepatocytes, blood cells, and neurons. Furthermore, transfection of synthetic mRNAs of TFs generates their appropriate target cells. These results demonstrate both the utility of this approach to identify potent TFs for cell differentiation, and the unanticipated capacity of single TFs directly guides differentiation to specific lineage fates. PMID:24371809

  1. NOV/CCN3 impairs muscle cell commitment and differentiation.

    Science.gov (United States)

    Calhabeu, Frederico; Lafont, Jérome; Le Dreau, Gwenvael; Laurent, Maryvonne; Kazazian, Chantal; Schaeffer, Laurent; Martinerie, Cécile; Dubois, Catherine

    2006-06-10

    NOV (nephroblastoma overexpressed) is a member of a family of proteins which encodes secreted matrix-associated proteins. NOV is expressed during development in dermomyotome and limb buds, but its functions are still poorly defined. In order to understand the role of NOV in myogenic differentiation, C2C12 cells overexpressing NOV (C2-NOV) were generated. These cells failed to engage into myogenic differentiation, whereas they retained the ability to differentiate into osteoblasts. In differentiating conditions, C2-NOV cells remained proliferative, failed to express differentiation markers and lost their ability to form myotubes. Inhibition of differentiation by NOV was also observed with human primary muscle cells. Further examination of C2-NOV cells revealed a strong downregulation of the myogenic determination genes MyoD and Myf5 and of IGF-II expression. MyoD forced expression in C2-NOV was sufficient to restore differentiation and IGF-II induction whereas 10(-6) M insulin treatment had no effects. NOV therefore acts upstream of MyoD and does not affect IGF-II induction and signaling. HES1, a target of Notch, previously proposed to mediate NOV action, was not implicated in the inhibition of differentiation. We propose that NOV is a specific cell fate regulator in the myogenic lineage, acting negatively on key myogenic genes thus controlling the transition from progenitor cells to myoblasts.

  2. DNA damage-dependent regulation of MyoD function

    OpenAIRE

    Simonatto, Marta

    1980-01-01

    Durante la rigenerazione muscolare, le cellule progenitrici sono esposte ad una serie di stimoli extracellulari che coordinano la loro attivazione, proliferazione e differenziamento. Al tempo stesso, tuttavia, l’ambiente rigenerativo, stimolando la sintesi del DNA e aumentando lo stato di ossidazione intracellulare, costituisce una fonte di stress genotossico, come mostriamo in vivo in questo lavoro. Uno studio svolto in precedenza nel nostro laboratorio ha identificato e ca...

  3. Exercise training alters DNA methylation patterns in genes related to muscle growth and differentiation in mice.

    Science.gov (United States)

    Kanzleiter, Timo; Jähnert, Markus; Schulze, Gunnar; Selbig, Joachim; Hallahan, Nicole; Schwenk, Robert Wolfgang; Schürmann, Annette

    2015-05-15

    The adaptive response of skeletal muscle to exercise training is tightly controlled and therefore requires transcriptional regulation. DNA methylation is an epigenetic mechanism known to modulate gene expression, but its contribution to exercise-induced adaptations in skeletal muscle is not well studied. Here, we describe a genome-wide analysis of DNA methylation in muscle of trained mice (n = 3). Compared with sedentary controls, 2,762 genes exhibited differentially methylated CpGs (P 5%, coverage >10) in their putative promoter regions. Alignment with gene expression data (n = 6) revealed 200 genes with a negative correlation between methylation and expression changes in response to exercise training. The majority of these genes were related to muscle growth and differentiation, and a minor fraction involved in metabolic regulation. Among the candidates were genes that regulate the expression of myogenic regulatory factors (Plexin A2) as well as genes that participate in muscle hypertrophy (Igfbp4) and motor neuron innervation (Dok7). Interestingly, a transcription factor binding site enrichment study discovered significantly enriched occurrence of CpG methylation in the binding sites of the myogenic regulatory factors MyoD and myogenin. These findings suggest that DNA methylation is involved in the regulation of muscle adaptation to regular exercise training.

  4. Effect of eccentric contraction on satellite cell activation in human vastus lateralis muscle.

    Science.gov (United States)

    Imaoka, Yoko; Kawai, Minako; Mori, Futoshi; Miyata, Hirofumi

    2015-09-01

    We compared the time-course of satellite cell (SC) activation between eccentric and concentric contractions in the vastus lateralis (VL) muscle after step exercise. Young adults participated in a 30-min step up/down exercise which mainly involved concentric contractions with the right VL muscle and eccentric contractions with the left VL muscle. The concentric and eccentric contraction phases of the VL muscles were identified by changes in the electromyogram (EMG) and knee joint angle. Biopsy samples were taken from both VL muscles at three time periods: before the exercise and 2 and 5 days after the exercise. We found that the numbers of SCs were significantly increased in the type IIa fibers of the left VL at 2 and 5 days after the exercise. The expression of both hepatocyte growth factor (HGF) and myogenic differentiation 1 (MyoD) mRNA had significantly increased in the left VL at 2 and 5 days after the exercise and in the right VL at 5 days after the exercise. The expression of transient receptor potential canonical (TRPC) 1 mRNA also increased in the left VL at 2 days after exercise. These results indicate that eccentric contraction can effectively activate SC proliferation for up to 5 days after exercise. Similar changes in HGF, MyoD and TRPC1 mRNA expression suggest that HGF/c-Met signal activation through cation influx has a major impact on skeletal muscle SC activation in response to eccentric exercise.

  5. In vivo tendon engineering with skeletal muscle derived cells in a mouse model.

    Science.gov (United States)

    Chen, Bo; Wang, Bin; Zhang, Wen Jie; Zhou, Guangdong; Cao, Yilin; Liu, Wei

    2012-09-01

    Engineering a functional tendon with strong mechanical property remains an aim to be achieved for its eventual application. Both skeletal muscle and tendon are closely associated during their development and both can bear strong mechanical loading dynamically. This study explored the possibility of engineering stronger tendons with mouse skeletal muscle derived cells (MDCs) and with mouse tenocytes as a control. The results demonstrated that both MDCs and tenocytes shared the gene expression of growth differentiation factor-8 (GDF-8), collagens I, III, VI, scleraxis and tenomodulin, but with MyoD gene expression only in MDCs. Quantitatively, MDCs expressed higher levels of GDF-8, collagens III and VI (p Young's modulus (p < 0.05). Furthermore, with the increase of implantation time, MDCs gradually lost their expression of myogenic molecules of MyoD and desmin and gained the expression of tenomodulin, a marker for tenocytes. Collectively, these results indicate that MDCs may serve as a desirable alternative cell source for engineering functional tendon tissue. Copyright © 2012 Elsevier Ltd. All rights reserved.

  6. Mecanismos celulares e moleculares que controlam o desenvolvimento e o crescimento muscular Cellular and molecular mechanisms that control muscular development and growth

    Directory of Open Access Journals (Sweden)

    Maeli Dal Pai Silva

    2007-07-01

    crescimento muscular.Skeletal muscle consists predominantly of muscle fibers surrounded by a connective tissue layer. This tissue has a great plasticity, and the knowledge of the morphological characteristics, myogenesis, and growth dynamics is important to the understanding of its physiology and to the farm animal selection for meat production. Most skeletal muscles are derived from mesodermal precursor cells originated from the somites. During embryonic development, specification of mesodermal precursor cells to the myogenic lineage is regulated by positive and negative signals from surrounding tissues. Specification to the myogenic lineage requires the up-regulation of four muscle-specific transcriptional factors (MyoD, Myf5, myogenin, and MRF4, called myogenic regulatory factors (MRFs, which have a well-defined role in skeletal muscle development and differentiation. The MRFs belong to the basic helix-loop-helix (bHLH super family of transcription factors, that forms heterodimers and bind to the E-box (CANNTG DNA sequence motif found in the promoters of many muscle specific genes. Specification to the myogenic lineage requires the up-regulation of the MRFs MyoD and Myf5. Proliferative MyoD and/or Myf5 positive myogenic cells are termed myoblasts. Proliferating myoblasts withdraw from the cell cycle to become terminally differentiated myocytes that express the "late" MRFs, myogenin and MRF4. Specialized populations of myogenic stem cells, termed satellite cells, arise during the late stages of embryogenesis are highly active during the postnatal muscle growth, which may occur by muscle fiber hyperplasia or hypertrophy. Quiescent satellite cells do not express detectable levels of MRF; however, during muscle growth, in response to several growth factors, activated satellite cells proliferate and express the MRFs in a similar manner to muscle precursor cells during skeletal muscle development. Muscle growth mechanisms are controlled by the temporally expression of muscle

  7. Specific expression of myogenic,adipogenic and osteogenic gene in skeletal muscle of mdx mice%mdx小鼠骨骼肌组织成肌、成脂、成骨基因的特异性表达

    Institute of Scientific and Technical Information of China (English)

    冷雁; 张为西; 周琛; 郑振扬; 张成; 李秋玲

    2011-01-01

    BACKGROUND: Duchenne's muscular dystrophy (DMD) is a fatal recessive X-Iinked form of muscular dystrophy characterized by prog ressive muscular degeneration, and there is no cure for DMD currently. Stem cell transplantation may provide us with a creative perspective. But the myogenic differentiation rate of transplanted cells is quite low in skeletal muscle. OBJECTIVE: The differences of myogenic, adipogenic and osteogenic gene expression levels in skeletal muscle between mdx mice and C57BL/6J mice were compared, with the purpose of investigating the underlying mechanism of pathological changes in skeletal muscle from mdx mice.METHODS : Frozen sections of skeletal muscle from mdx and C57BL/6J mice were prepared, stained with hematoxylin-eosin staining and Vonkossa staining. The morphological changes of the muscles were observed under microscope. Additionally, total RNA of skeletal muscle from mdx and C57BL/6J mice was extracted, reversed, and the expression levels of myogenic,adipogenic and osteogenic characteristic genes were examined by real-time PCR.RESULTS AND CONCLUSION: Necrosis and regeneration were found in skeletal muscles of mdx mice, and mild adipose hyperplasia and fibrous connective tissue hyperplasia were also observed. Moreover, calcium deposition nodules were easily detected by Vonkossa staining. The form of skeletal muscle cells from C57 mice was clear, and the nuclei were located in the cell periphery. Osteogenic and adipogenic gene expressions of skeletal muscle from mdx mice were elevated to a certain degree by real-time PCR (P < 0.05), compared with C57 mice, whereas myogenic gene expression was decreased (P< 0.05). The reason why adipocyte and osteoblast in skeletal muscle of mdx mice overgrew may be due to degeneration and necrosis of skeletal muscle which caused by dystrophin gene deletion, and it differentiates into osteoblasts and adipocytes instead of myoblasts.%背景:干细胞移植是治疗肌营养不良症的有效方法之一,

  8. HES6 enhances the motility of alveolar rhabdomyosarcoma cells

    Energy Technology Data Exchange (ETDEWEB)

    Wickramasinghe, Caroline M [MRC Cancer Cell Unit, Hutchison-MRC Research centre, Addenbrooke' s Hospital Cambridge, CB2 0XZ (United Kingdom); MRC Laboratory of Molecular Biology, Addenbrooke' s Hospital Cambridge, CB2 0QH (United Kingdom); Domaschenz, Renae [MRC Cancer Cell Unit, Hutchison-MRC Research centre, Addenbrooke' s Hospital Cambridge, CB2 0XZ (United Kingdom); Gene Regulation and Chromatin Group, MRC Clinical Sciences Centre, Faculty of Medicine, Imperial College, Hammersmith Campus, Du Cane Road, London W12 ONN (United Kingdom); Amagase, Yoko [MRC Cancer Cell Unit, Hutchison-MRC Research centre, Addenbrooke' s Hospital Cambridge, CB2 0XZ (United Kingdom); Department of Pathophysiology, Faculty of Pharmaceutical Sciences, Doshisha Women' s College of Liberal Arts, Kodo, Kyotanabe, Kyoto 610-0395 (Japan); Williamson, Daniel [Molecular Cytogenetics, The Institute of Cancer Research, Sutton SM2 5NG (United Kingdom); Northern Institute for Cancer Research, Paul O' Gorman Building, Medical School, Newcastle University, Framlington Place, Newcastle upon Tyne, NE2 4HH (United Kingdom); Missiaglia, Edoardo; Shipley, Janet [Molecular Cytogenetics, The Institute of Cancer Research, Sutton SM2 5NG (United Kingdom); Murai, Kasumi [MRC Cancer Cell Unit, Hutchison-MRC Research centre, Addenbrooke' s Hospital Cambridge, CB2 0XZ (United Kingdom); Jones, Philip H, E-mail: phj20@cam.ac.uk [MRC Cancer Cell Unit, Hutchison-MRC Research centre, Addenbrooke' s Hospital Cambridge, CB2 0XZ (United Kingdom)

    2013-01-01

    Absract: HES6, a member of the hairy-enhancer-of-split family of transcription factors, plays multiple roles in myogenesis. It is a direct target of the myogenic transcription factor MyoD and has been shown to regulate the formation of the myotome in development, myoblast cell cycle exit and the organization of the actin cytoskeleton during terminal differentiation. Here we investigate the expression and function of HES6 in rhabdomyosarcoma, a soft tissue tumor which expresses myogenic genes but fails to differentiate into muscle. We show that HES6 is expressed at high levels in the subset of alveolar rhabdomyosarcomas expressing PAX/FOXO1 fusion genes (ARMSp). Knockdown of HES6 mRNA in the ARMSp cell line RH30 reduces proliferation and cell motility. This phenotype is rescued by expression of mouse Hes6 which is insensitive to HES6 siRNA. Furthermore, expression microarray analysis indicates that the HES6 knockdown is associated with a decrease in the levels of Transgelin, (TAGLN), a regulator of the actin cytoskeleton. Knockdown of TAGLN decreases cell motility, whilst TAGLN overexpression rescues the motility defect resulting from HES6 knockdown. These findings indicate HES6 contributes to the pathogenesis of ARMSp by enhancing both proliferation and cell motility.

  9. Hes6 is required for actin cytoskeletal organization in differentiating C2C12 myoblasts

    Energy Technology Data Exchange (ETDEWEB)

    Malone, Caroline M.P.; Domaschenz, Renae; Amagase, Yoko [MRC Cancer Cell Unit, Hutchison-MRC Research Centre, Addenbrooke' s Hospital, Cambridge CB2 0XZ (United Kingdom); Dunham, Ian [EMBL-European Bioinformatics Institute (EBI), Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1SD (United Kingdom); Murai, Kasumi [MRC Cancer Cell Unit, Hutchison-MRC Research Centre, Addenbrooke' s Hospital, Cambridge CB2 0XZ (United Kingdom); Jones, Philip H., E-mail: phj20@cam.ac.uk [MRC Cancer Cell Unit, Hutchison-MRC Research Centre, Addenbrooke' s Hospital, Cambridge CB2 0XZ (United Kingdom)

    2011-07-01

    Hes6 is a member of the hairy-enhancer-of-split family of transcription factors that regulate proliferating cell fate in development and is known to be expressed in developing muscle. Here we investigate its function in myogenesis in vitro. We show that Hes6 is a direct transcriptional target of the myogenic transcription factors MyoD and Myf5, indicating that it is integral to the myogenic transcriptional program. The localization of Hes6 protein changes during differentiation, becoming predominantly nuclear. Knockdown of Hes6 mRNA levels by siRNA has no effect on cell cycle exit or induction of myosin heavy chain expression in differentiating C2C12 myoblasts, but F-actin filament formation is disrupted and both cell motility and myoblast fusion are reduced. The knockdown phenotype is rescued by expression of Hes6 cDNA resistant to siRNA. These results define a novel role for Hes6 in actin cytoskeletal dynamics in post mitotic myoblasts.

  10. How to make rapid eye movements “rapid”: the role of growth factors for muscle contractile properties

    Science.gov (United States)

    Li, Tian; Feng, Cheng-Yuan

    2011-01-01

    Different muscle functions require different muscle contraction properties. Saccade-generating extraocular muscles (EOMs) are the fastest muscles in the human body, significantly faster than limb skeletal muscles. Muscle contraction speed is subjected to plasticity, i.e., contraction speed can be adjusted to serve different demands, but little is known about the molecular mechanisms that control contraction speed. Therefore, we examined whether myogenic growth factors modulate contractile properties, including twitch contraction time (onset of force to peak force) and half relaxation time (peak force to half relaxation). We examined effects of three muscle-derived growth factors: insulin-like growth factor 1 (IGF1), cardiotrophin-1 (CT1), and glial cell line-derived neurotrophic factor (GDNF). In gain-of-function experiments, CT1 or GDNF injected into the orbit shortened contraction time, and IGF1 or CT1 shortened half relaxation time. In loss-of-function experiments with binding proteins or neutralizing antibodies, elimination of endogenous IGFs prolonged both contraction time and half relaxation time, while eliminating endogenous GDNF prolonged contraction time, with no effect on half relaxation time. Elimination of endogenous IGFs or CT1, but not GDNF, significantly reduced contractile force. Thus, IGF1, CT1, and GDNF have partially overlapping but not identical effects on muscle contractile properties. Expression of these three growth factors was measured in chicken and/or rat EOMs by real-time PCR. The “fast” EOMs express significantly more message encoding these growth factors and their receptors than skeletal muscles with slower contractile properties. Taken together, these findings indicate that EOM contractile kinetics is regulated by the amount of myogenic growth factors available to the muscle. PMID:21279379

  11. Heart Failure-Induced Diaphragm Myopathy

    Directory of Open Access Journals (Sweden)

    Aline Regina Ruiz Lima

    2014-07-01

    Full Text Available Background: Intracellular signaling pathways involved in skeletal myosin heavy chain (MyHC isoform alterations during heart failure (HF are not completely understood. We tested the hypothesis that diaphragm expression of mitogen-activated protein kinases (MAPK and myogenic regulatory factors is changed in rats with myocardial infarction (MI induced HF. Methods: Six months after MI rats were subjected to transthoracic echocardiography. After euthanasia, infarcted rats were subdivided in MI/HF- group (with no HF evidence; n=10, and MI/HF+ (with right ventricular hypertrophy and lung congestion; n=10. Sham-operated rats were used as controls (n=10. MyHC isoforms were analyzed by electrophoresis. Statistical analysis: ANOVA and Pearson correlation. Results: MI/HF- had left cardiac chambers dilation with systolic and diastolic left ventricular dysfunction. Cardiac injury was more intense in MI/HF+ than MI/HF-. MyHC I isoform percentage was higher in MI/HF+ than MI/HF-, and IIb isoform lower in MI/HF+ than Sham. Left atrial diameter-to-body weight ratio positively correlated with MyHC I (p=0.005 and negatively correlated with MyHC IIb (p=0.02. TNF-a serum concentration positively correlated with MyHC I isoform. Total and phosphorylated ERK was lower in MI/HF- and MI/HF+ than Sham. Phosphorylated JNK was lower in MI/HF- than Sham. JNK and p38 did not differ between groups. Expression of NF-κB and the myogenic regulatory factors MyoD, myogenin, and MRF4 was similar between groups. Conclusion: Diaphragm MyHC fast-to-slow shift is related to cardiac dysfunction severity and TNF-a serum levels in infarcted rats. Reduced ERK expression seems to participate in MyHC isoform changes. Myogenic regulatory factors and NF-κB do not modulate diaphragm MyHC distribution during chronic HF.

  12. Insulin-like growth factor binding protein-5 modulates muscle differentiation through an insulin-like growth factor-dependent mechanism.

    Science.gov (United States)

    James, P L; Stewart, C E; Rotwein, P

    1996-05-01

    The insulin-like growth factor binding proteins (IGFBPs) are a family of six secreted proteins which bind to and modulate the actions of insulin-like growth factors-I and -II (IGF-I and -II). IGFBP-5 is more conserved than other IGFBPs characterized to date, and is expressed in adult rodent muscle and in the developing myotome. We have shown previously that C2 myoblasts secrete IGFBP-5 as their sole IGFBP. Here we use these cells to study the function of IGFBP-5 during myogenesis, a process stimulated by IGFs. We stably transfected C2 cells with IGFBP-5 cDNAs under control of a constitutively active promoter. Compared with vector-transfected control cells, C2 myoblasts expressing the IGFBP-5 transgene in the sense orientation exhibit increased IGFBP-5 levels in the extracellular matrix during proliferation, and subsequently fail to differentiate normally, as assessed by both morphological and biochemical criteria. Compared to controls, IGFBP-5 sense myoblasts show enhanced survival in low serum medium, remaining viable for at least four weeks in culture. By contrast, myoblasts expressing the IGFBP-5 antisense transcript differentiate prematurely and more extensively than control cells. The inhibition of myogenic differentiation by high level expression of IGFBP-5 could be overcome by exogenous IGFs, with des (1-3) IGF-I, an analogue with decreased affinity for IGFBP-5 but normal affinity for the IGF-I receptor, showing the highest potency. These results are consistent with a model in which IGFBP-5 blocks IGF-stimulated myogenesis, and indicate that sequestration of IGFs in the extracellular matrix could be a possible mechanism of action. Our observations also suggest that IGFBP-5 normally inhibits muscle differentiation, and imply a role for IGFBP-5 in regulating IGF action during myogenic development in vivo.

  13. Sex-Specific Muscular Maturation Responses Following Prenatal Exposure to Methylation-Related Micronutrients in Pigs

    Science.gov (United States)

    Oster, Michael; Trakooljul, Nares; Reyer, Henry; Zeyner, Annette; Muráni, Eduard; Ponsuksili, Siriluck; Wimmers, Klaus

    2017-01-01

    Supplementation of micronutrients involved in DNA methylation, particularly during pregnancy, is recommended because of its impacts on human health, but further evidence is needed regarding the effects of over-supplementation and differences between sexes. Here, a porcine model was used to assess effects of maternal supplementation with one-carbon-cycle compounds during prenatal and postnatal stages on offspring muscle development. Sows received either a standard diet (CON) or a standard diet supplemented with folate, B6, B12, methionine, choline, and zinc (MET) throughout gestation. Myogenesis-, growth-, and nutrient utilization-related transcript expression was assessed using quantitative PCR. Organismal phenotype and gene expression effects differed significantly between males and females. Male MET-offspring showed increased fetal weight during late pregnancy but decreased live weight postnatally, with compensatory transcriptional responses comprising myogenic key drivers (Pax7, MyoD1, myogenin). In contrast, female weights were unaffected by diet, and mRNA abundances corresponded to a phenotype of cellular reorganization via FABP3, FABP4, SPP1 and Insulin-like Growth Factor-signaling. These findings in an animal model suggest that supplementation during pregnancy with methylation-related micronutrients can promote sex-specific myogenic maturation processes related to organismal growth and muscle metabolism. The usage of maternal dietary supplements should be more carefully considered regarding its ability to promote fetal and postnatal health. PMID:28106759

  14. The effect of temperature on proliferation and differentiation of chicken skeletal muscle satellite cells isolated from different muscle types.

    Science.gov (United States)

    Harding, Rachel L; Halevy, Orna; Yahav, Shlomo; Velleman, Sandra G

    2016-04-01

    Skeletal muscle satellite cells are a muscle stem cell population that mediate posthatch muscle growth and repair. Satellite cells respond differentially to environmental stimuli based upon their fiber-type of origin. The objective of this study was to determine how temperatures below and above the in vitro control of 38°C affected the proliferation and differentiation of satellite cells isolated from the chicken anaerobic pectoralis major (p. major) or mixed fiber biceps femoris (b.femoris) muscles. The satellite cells isolated from the p. major muscle were more sensitive to both cold and hot temperatures compared to the b.femoris satellite cells during both proliferation and differentiation. The expressions of myogenic regulatory transcription factors were also different between satellite cells from different fiber types. MyoD expression, which partially regulates proliferation, was generally expressed at higher levels in p. major satellite cells compared to the b.femoris satellite cells from 33 to 43°C during proliferation and differentiation. Similarly, myogenin exp