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Sample records for striated muscle sarcomere

  1. Dynamic Regulation of Sarcomeric Actin Filaments in Striated Muscle

    OpenAIRE

    Ono, Shoichiro

    2010-01-01

    In striated muscle, the actin cytoskeleton is differentiated into myofibrils. Actin and myosin filaments are organized in sarcomeres and specialized for producing contractile forces. Regular arrangement of actin filaments with uniform length and polarity is critical for the contractile function. However, the mechanisms of assembly and maintenance of sarcomeric actin filaments in striated muscle are not completely understood. Live imaging of actin in striated muscle has revealed that actin sub...

  2. Expression of various sarcomeric tropomyosin isoforms in equine striated muscles

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    Syamalima Dube

    2017-06-01

    Full Text Available In order to better understand the training and athletic activity of horses, we must have complete understanding of the isoform diversity of various myofibrillar protein genes like tropomyosin. Tropomyosin (TPM, a coiled-coil dimeric protein, is a component of thin filament in striated muscles. In mammals, four TPM genes (TPM1, TPM2, TPM3, and TPM4 generate a multitude of TPM isoforms via alternate splicing and/or using different promoters. Unfortunately, our knowledge of TPM isoform diversity in the horse is very limited. Hence, we undertook a comprehensive exploratory study of various TPM isoforms from horse heart and skeletal muscle. We have cloned and sequenced two sarcomeric isoforms of the TPM1 gene called TPM1α and TPM1κ, one sarcomeric isoform of the TPM2 and one of the TPM3 gene, TPM2α and TPM3α respectively. By qRT-PCR using both relative expression and copy number, we have shown that TPM1α expression compared to TPM1κ is very high in heart. On the other hand, the expression of TPM1α is higher in skeletal muscle compared to heart. Further, the expression of TPM2α and TPM3α are higher in skeletal muscle compared to heart. Using western blot analyses with CH1 monoclonal antibody we have shown the high expression levels of sarcomeric TPM proteins in cardiac and skeletal muscle. Due to the paucity of isoform specific antibodies we cannot specifically detect the expression of TPM1κ in horse striated muscle. To the best of our knowledge this is the very first report on the characterization of sarcmeric TPMs in horse striated muscle.

  3. Expression of various sarcomeric tropomyosin isoforms in equine striated muscles

    OpenAIRE

    Dube, Syamalima; Chionuma, Henry; Matoq, Amr; Alshiekh-Nasany, Ruham; Abbott, Lynn; Poiesz, Bernard J.; Dube, Dipak K.

    2017-01-01

    In order to better understand the training and athletic activity of horses, we must have complete understanding of the isoform diversity of various myofibrillar protein genes like tropomyosin. Tropomyosin (TPM), a coiled-coil dimeric protein, is a component of thin filament in striated muscles. In mammals, four TPM genes (TPM1, TPM2, TPM3, and TPM4) generate a multitude of TPM isoforms via alternate splicing and/or using different promoters. Unfortunately, our knowledge of TPM isoform diversi...

  4. Isoform composition, gene expression and sarcomeric protein phosphorylation in striated muscle of mice after space flight

    Science.gov (United States)

    Vikhlyantsev, Ivan; Ulanova, Anna; Salmov, Nikolay; Gritsyna, Yulia; Bobylev, Alexandr; Rogachevsky, Vadim; Shenkman, Boris; Podlubnaya, Zoya

    Using RT-PCR and SDS-PAGE, changes in isoform composition, gene expression, titin and nebulin phosphorylation, as well as changes in isoform composition of myosin heavy chains in striated muscles of mice were studied after 30-day-long space flight onboard the Russian spacecraft “BION-M” No. 1. The muscle fibre-type shift from slow-to-fast was observed in m. gastrocnemius and m. tibialis anterior of animals from “Flight” group. A decrease in the content of the NT and N2A titin isoforms and nebulin in the skeletal muscles of animals from “Flight” group was found. Meanwhile, significant differences in gene expression of these proteins in skeletal muscles of mice from “Flight” and “Control” groups were not observed. Using Pro-Q Diamond stain, an increase in titin phosphorylation in m. gastrocnemius of mice from “Flight” group was detected. The content of the NT, N2BA and N2B titin isoforms in cardiac muscle of mice from “Flight” and “Control” groups did not differ, nevertheless an increase in titin gene expression in the myocardium of the “Flight” group animals was found. The observed changes will be discussed in the context of theirs role in contractile activity of striated muscles of mice in conditions of weightlessness. This work was supported by the Russian Foundation for Basic Research (grants No. 14-04-32240, 14-04-00112). Acknowledgement. We express our gratitude to the teams of Institute of Biomedical Problems RAS and “PROGRESS” Corporation involved in the preparation of the “BION-M” mission.

  5. S100A1: A Regulator of Striated Muscle Sarcoplasmic Reticulum Ca2+ Handling, Sarcomeric, and Mitochondrial Function

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    Mirko Völkers

    2010-01-01

    S100A1 has further been detected at different sites within the cardiac sarcomere indicating potential roles in myofilament function. More recently, a study reported a mitochondrial location of S100A1 in cardiomyocytes. Additionally, normalizing the level of S100A1 protein by means of viral cardiac gene transfer in animal heart failure models resulted in a disrupted progression towards cardiac failure and enhanced survival. This brief review is confined to the physiological and pathophysiological relevance of S100A1 in cardiac and skeletal muscle Ca2+ handling with a particular focus on its potential as a molecular target for future therapeutic interventions.

  6. Striated Muscle Function, Regeneration, and Repair

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    Shadrin, I.Y.; Khodabukus, A.; Bursac, N.

    2016-01-01

    As the only striated muscle tissues in the body, skeletal and cardiac muscle share numerous structural and functional characteristics, while exhibiting vastly different size and regenerative potential. Healthy skeletal muscle harbors a robust regenerative response that becomes inadequate after large muscle loss or in degenerative pathologies and aging. In contrast, the mammalian heart loses its regenerative capacity shortly after birth, leaving it susceptible to permanent damage by acute injury or chronic disease. In this review, we compare and contrast the physiology and regenerative potential of native skeletal and cardiac muscles, mechanisms underlying striated muscle dysfunction, and bioengineering strategies to treat muscle disorders. We focus on different sources for cellular therapy, biomaterials to augment the endogenous regenerative response, and progress in engineering and application of mature striated muscle tissues in vitro and in vivo. Finally, we discuss the challenges and perspectives in translating muscle bioengineering strategies to clinical practice. PMID:27271751

  7. Polarization gating enables sarcomere length measurements by laser diffraction in fibrotic muscle

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    Young, Kevin W.; Dayanidhi, Sudarshan; Lieber, Richard L.

    2014-11-01

    Sarcomere length is a key parameter commonly measured in muscle physiology since it dictates striated muscle active force. Laser diffraction (LD)-based measurements of sarcomere length are time-efficient and sample a greater number of sarcomeres compared with traditional microscopy-based techniques. However, a limitation to LD techniques is that signal quality is severely degraded by scattering events as photons propagate through tissue. Consequently, sarcomere length measurements are unattainable when the number of scattering events is sufficiently large in muscle tissue with a high scattering probability. This occurs in fibrotic skeletal muscle seen in muscular dystrophies and secondary to tissue trauma, thus eliminating the use of LD to study these skeletal muscle ailments. Here, we utilize polarization gating to extract diffracted signals that are buried in noise created by scattering. Importantly, we demonstrate that polarization-gated laser diffraction (PGLD) enables sarcomere length measurements in muscles from chronically immobilized mice hind limbs; these muscles have a substantial increase of intramuscular connective tissue that scatter light and disable sarcomere length measurements by traditional LD. Further, we compare PGLD sarcomere lengths to those measured by bright field (BF) and confocal microscopy as positive controls and reveal a significant bias of BF but not of confocal microscopy.

  8. Muscle lim protein isoform negatively regulates striated muscle actin dynamics and differentiation.

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    Vafiadaki, Elizabeth; Arvanitis, Demetrios A; Papalouka, Vasiliki; Terzis, Gerasimos; Roumeliotis, Theodoros I; Spengos, Konstantinos; Garbis, Spiros D; Manta, Panagiota; Kranias, Evangelia G; Sanoudou, Despina

    2014-07-01

    Muscle lim protein (MLP) has emerged as a critical regulator of striated muscle physiology and pathophysiology. Mutations in cysteine and glycine-rich protein 3 (CSRP3), the gene encoding MLP, have been directly associated with human cardiomyopathies, whereas aberrant expression patterns are reported in human cardiac and skeletal muscle diseases. Increasing evidence suggests that MLP has an important role in both myogenic differentiation and myocyte cytoarchitecture, although the full spectrum of its intracellular roles has not been delineated. We report the discovery of an alternative splice variant of MLP, designated as MLP-b, showing distinct expression in neuromuscular disease and direct roles in actin dynamics and muscle differentiation. This novel isoform originates by alternative splicing of exons 3 and 4. At the protein level, it contains the N-terminus first half LIM domain of MLP and a unique sequence of 22 amino acids. Physiologically, it is expressed during early differentiation, whereas its overexpression reduces C2C12 differentiation and myotube formation. This may be mediated through its inhibition of MLP/cofilin-2-mediated F-actin dynamics. In differentiated striated muscles, MLP-b localizes to the sarcomeres and binds directly to Z-disc components, including α-actinin, T-cap and MLP. The findings of the present study unveil a novel player in muscle physiology and pathophysiology that is implicated in myogenesis as a negative regulator of myotube formation, as well as in differentiated striated muscles as a contributor to sarcomeric integrity. © 2014 FEBS.

  9. In vivo sarcomere lengths and sarcomere elongations are not uniform across an intact muscle

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    Eng Kuan eMoo

    2016-05-01

    Full Text Available Sarcomere lengths have been a crucial outcome measure for understanding and explaining basic muscle properties and muscle function. Sarcomere lengths for a given muscle are typically measured at a single spot, often in the mid-belly of the muscle, and at a given muscle length. It is then assumed implicitly that the sarcomere length measured at this single spot represents the sarcomere lengths at other locations within the muscle, and force-length, force-velocity and power-velocity properties of muscles are often implied based on these single sarcomere length measurements. Although intuitively appealing, this assumption is yet to be supported by systematic evidence. The objective of this study was to measure sarcomere lengths at defined locations along and across an intact muscle, at different muscle lengths. Using second harmonic generation imaging technique, sarcomere patterns in passive mouse tibialis anterior (TA were imaged in a non-contact manner at five selected locations (‘proximal’, ‘distal’, ‘middle’, ‘medial’ and ‘lateral’ TA sites and at three different lengths encompassing the anatomical range of motion of the TA. We showed that sarcomere lengths varied substantially within small regions of the muscle and also for different sites across the entire TA. Also, sarcomere elongations with muscle lengthening were non-uniform across the muscle, with the highest sarcomere stretches occurring near the myotendinous junction. We conclude that muscle mechanics derived from sarcomere length measured from a small region of a muscle may not well represent the sarcomere length and associated functional properties of the entire muscle.

  10. Chaperones and the Proteasome System: Regulating the Construction and Demolition of Striated Muscle

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    Casey Carlisle

    2017-12-01

    Full Text Available Protein folding factors (chaperones are required for many diverse cellular functions. In striated muscle, chaperones are required for contractile protein function, as well as the larger scale assembly of the basic unit of muscle, the sarcomere. The sarcomere is complex and composed of hundreds of proteins and the number of proteins and processes recognized to be regulated by chaperones has increased dramatically over the past decade. Research in the past ten years has begun to discover and characterize the chaperones involved in the assembly of the sarcomere at a rapid rate. Because of the dynamic nature of muscle, wear and tear damage is inevitable. Several systems, including chaperones and the ubiquitin proteasome system (UPS, have evolved to regulate protein turnover. Much of our knowledge of muscle development focuses on the formation of the sarcomere but recent work has begun to elucidate the requirement and role of chaperones and the UPS in sarcomere maintenance and disease. This review will cover the roles of chaperones in sarcomere assembly, the importance of chaperone homeostasis and the cooperation of chaperones and the UPS in sarcomere integrity and disease.

  11. Systems Biology Approaches to Discerning Striated Muscle Pathologies

    OpenAIRE

    Mukund, Kavitha

    2016-01-01

    The human muscular system represents nearly 75% of the body mass and encompasses two major muscle forms- striated and smooth. Striated muscle, composed broadly of myofibers, accompanying membrane systems, cytoskeletal networks together with the metabolic and regulatory machinery, have revealed complexities in composition, structure and function. A disruption to any component within this complex system of interactions lead to disorders of the muscle, typically characterized by muscle fiber los...

  12. Poorly Understood Aspects of Striated Muscle Contraction

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    Alf Månsson

    2015-01-01

    Full Text Available Muscle contraction results from cyclic interactions between the contractile proteins myosin and actin, driven by the turnover of adenosine triphosphate (ATP. Despite intense studies, several molecular events in the contraction process are poorly understood, including the relationship between force-generation and phosphate-release in the ATP-turnover. Different aspects of the force-generating transition are reflected in the changes in tension development by muscle cells, myofibrils and single molecules upon changes in temperature, altered phosphate concentration, or length perturbations. It has been notoriously difficult to explain all these events within a given theoretical framework and to unequivocally correlate observed events with the atomic structures of the myosin motor. Other incompletely understood issues include the role of the two heads of myosin II and structural changes in the actin filaments as well as the importance of the three-dimensional order. We here review these issues in relation to controversies regarding basic physiological properties of striated muscle. We also briefly consider actomyosin mutation effects in cardiac and skeletal muscle function and the possibility to treat these defects by drugs.

  13. Compositional studies of myofibrils from rabbit striated muscle

    Energy Technology Data Exchange (ETDEWEB)

    Etlinger, J.D.; Zak, R.; Fischman, D.A.

    1976-01-01

    The localization of high-molecular-weight (80,000-200,000-daltons) proteins in the sarcomere of striated muscle has been studied by coordinated electron-microscopic and sodium dodecyl sulfate (SDS) gel electrophoretic analysis of native myofilaments and extracted and digested myofibrils. Methods were developed for the isolation of thick and thin filaments and of uncontracted myofibrils which are devoid of endoproteases and membrane fragments. Treatment of crude myofibrils with 0.5% Triton X-100 results in the release of a 110,000-dalton component without affecting the myofibrillar structure. Extraction of uncontracted myofibrils with a relaxing solution of high ionic strength results in the complete disappearance of the A band and M line. In this extract, five other protein bands in addition to myosin are resolved on SDS gels: bands M 1 (190,000 daltons) and M 2 (170,000 daltons), which are suggested to be components of the M line; M 3 (150,000 daltons), a degradation product; and a doublet M 4, M 5 (140,000 daltons), thick-filament protein having the same mobility as C protein.

  14. Sarcomere lattice geometry influences cooperative myosin binding in muscle.

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    Bertrand C W Tanner

    2007-07-01

    Full Text Available In muscle, force emerges from myosin binding with actin (forming a cross-bridge. This actomyosin binding depends upon myofilament geometry, kinetics of thin-filament Ca(2+ activation, and kinetics of cross-bridge cycling. Binding occurs within a compliant network of protein filaments where there is mechanical coupling between myosins along the thick-filament backbone and between actin monomers along the thin filament. Such mechanical coupling precludes using ordinary differential equation models when examining the effects of lattice geometry, kinetics, or compliance on force production. This study uses two stochastically driven, spatially explicit models to predict levels of cross-bridge binding, force, thin-filament Ca(2+ activation, and ATP utilization. One model incorporates the 2-to-1 ratio of thin to thick filaments of vertebrate striated muscle (multi-filament model, while the other comprises only one thick and one thin filament (two-filament model. Simulations comparing these models show that the multi-filament predictions of force, fractional cross-bridge binding, and cross-bridge turnover are more consistent with published experimental values. Furthermore, the values predicted by the multi-filament model are greater than those values predicted by the two-filament model. These increases are larger than the relative increase of potential inter-filament interactions in the multi-filament model versus the two-filament model. This amplification of coordinated cross-bridge binding and cycling indicates a mechanism of cooperativity that depends on sarcomere lattice geometry, specifically the ratio and arrangement of myofilaments.

  15. Accuracy of measurement by laser diffraction method of length of contracting muscle sarcomeres

    Energy Technology Data Exchange (ETDEWEB)

    Klimov, A.A.; Andreyev, O.A.

    The method of laser beam diffraction was used on striate muscle fibers to study changes in sarcomere length during contraction, as determined by the angular distance between (+1)-(left) and (-1)-(right) diffraction maximums. The course of force development was recorded simultaneously. The absolute margin of error in determining the mean length of sarcomeres did not exceed 0.05 ..mu..m in the range of 1.8 to 2.8 ..mu..m lengths, and 0.1 ..mu..m in the range of 2.8 to 3.3. Changes in mean length of sarcomeres were recorded with accuracy to 0.003 ..mu..m with concurrent monitoring of positions (+1) and (-1) maximums. It was demonstrated that during fiber contraction there is shifting not only of (+1) and (-1) maximums, but (0) maximum also, which is attributable to the effects of light refraction with change in shape of illuminated segment of fiber. This change can be caused by redistribution of sarcomere lengths along the axis of the fiber during contraction and, accordingly, local changes in fiber diameter.

  16. Neurohypophyseal hormones: novel actors of striated muscle development and homeostasis

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    Alessandra Costa

    2014-09-01

    Full Text Available Since the 1980's, novel functional roles of the neurohypophyseal hormones vasopressin and oxytocin have emerged. Several studies have investigated the effects of these two neurohormones on striated muscle tissues, both in vitro and in vivo. The effects of vasopressin on skeletal myogenic cells, developing muscle and muscle homeostasis have been documented. Oxytocin appears to have a greater influence on cardiomyocite differentiation and heart homeostasis. This review summarizes the studies on these novel roles of the two neurohypophyseal hormones, and open the possibility of new therapeutic approaches for diseases affecting striated muscle.

  17. Mean sarcomere length-force relationship of rat muscle fibre bundles

    NARCIS (Netherlands)

    Zuurbier, C. J.; Heslinga, J. W.; Lee-de Groot, M. B.; van der Laarse, W. J.

    1995-01-01

    To study how sarcomere length inhomogeneities and the duration of activation affect sarcomere length-force characteristics of muscle, the mean sarcomere length-force relationship was determined for twitches and at 100 and 300 ms during tetanic activation for rat extensor digitorum longus and

  18. Preparation of developing Xenopus muscle for sarcomeric protein localization by high-resolution imaging.

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    Nworu, Chinedu U; Krieg, Paul A; Gregorio, Carol C

    2014-04-01

    Mutations in several sarcomeric proteins have been linked to various human myopathies. Therefore, having an in vivo developmental model available that develops quickly and efficiently is key for investigators to elucidate the critical steps, components and signaling pathways involved in building a myofibril; this is the pivotal foundation for deciphering disease mechanisms as well as the development of myopathy-related therapeutics. Although striated muscle cell culture studies have been extremely informative in providing clues to both the distribution and functions of sarcomeric proteins, myocytes in vivo develop in an irreproducible 3D environment. Xenopus laevis (frog) embryos are cost effective, compliant to protein level manipulations and develop relatively quickly (⩽ a week) in a petri dish, thus providing a powerful system for de novo myofibrillogenesis studies. Although fluorophore-conjugated phalloidin labeling is the gold standard approach for investigating actin-thin filament architecture, it is well documented that phalloidin-labeling can be challenging and inconsistent within Xenopus embryos. Therefore we highlight several techniques that can be utilized to preserve both antibody and fluorophore-conjugated phalloidin labeling within Xenopus embryos for high-resolution fluorescence microscopy. Copyright © 2013. Published by Elsevier Inc.

  19. Autoradiographic analysis of protein regeneration in striated skeleton muscle

    International Nuclear Information System (INIS)

    Dadoune, J.P.

    1977-01-01

    An autoradiographic study was conducted of protein regeneration in striated muscles aimed at clarifying the contradictions in the literature: while some authors hold that the regeneration rate is identical for all types of myofibril proteins and the myofibril is thus regenerated as a whole, others claim that the regeneration rate differs depending on the type of the myofibril protein. Tritium-labelled leucine incorporation experiments showed the existence of at least 2 pools of newly formed proteins in striated muscles in both adult and young animals. One pool is regenerated in 1 to 2 weeks, the other roughly in a month. The regeneration of proteins is initially more significant in red fibres; thus the rate of myofibril protein regeneration is not uniform. In adult animals regeneration seems to be slower in filaments than in the sarcoplasm and in the mitochondria. (A.K.)

  20. In Vivo Sarcomere Length Measurement in Whole Muscles during Passive Stretch and Twitch Contractions

    OpenAIRE

    Young, Kevin W.; Kuo, Bill P.-P.; O’Connor, Shawn M.; Radic, Stojan; Lieber, Richard L.

    2017-01-01

    Muscle force is dictated by micrometer-scale contractile machines called sarcomeres. Whole-muscle force drops from peak force production to zero with just a few micrometers of sarcomere length change. No current technology is able to capture adequate dynamic sarcomere data in vivo, and thus we lack fundamental data needed to understand human movement and movement disorders. Methods such as diffraction, endoscopy, and optical coherence tomography have been applied to muscle but are prohibitive...

  1. Contracture of Slow Striated Muscle during Calcium Deprivation

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    Irwin, Richard L.; Hein, Manfred M.

    1963-01-01

    When deprived of calcium the slow striated muscle fibers of the frog develop reversible contractures in either hypertonic or isotonic solutions. While calcium deprivation continues because of a flowing calcium-free solution the muscles relax slowly and completely. Restoration of calcium during contracture relaxes the muscle promptly to initial tension. When relaxed during calcium lack the return of calcium does not change tension and the muscle stays relaxed. When contractures are induced by solutions containing small amounts of calcium relaxation does not occur or requires several hours. The rate of tension development depends upon the rate at which calcium moves outward since the contractures develop slower in low concentrations of calcium and are absent or greatly slowed in a stagnant calcium-free solution. Withdrawal of calcium prevents the contractile responses to ACh, KCl, or electrical stimulation through the nerve. Muscles return to their original excitability after calcium is restored. Origin of the contractures is unrelated to nerve activity since they are maximal during transmission failure from calcium lack, occur in denervated muscles, and are not blocked by high concentrations of d-tubocurarine, procaine, or atropine. The experiments also indicate that the contractures do not originate from repetitive activity of muscle membranes. The findings are most simply explained by relating the outward movement of calcium as a link for initiating contraction in slow type striated muscle. PMID:14065284

  2. Force produced after stretch in sarcomeres and half-sarcomeres isolated from skeletal muscles

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    Minozzo, Fábio C.; Baroni, Bruno M.; Correa, José A.; Vaz, Marco A.; Rassier, Dilson E.

    2013-07-01

    The goal of this study was to evaluate if isolated sarcomeres and half-sarcomeres produce a long-lasting increase in force after a stretch is imposed during activation. Single and half-sarcomeres were isolated from myofibrils using micro-needles, which were also used for force measurements. After full force development, both preparations were stretched by different magnitudes. The sarcomere length (SL) or half-sarcomere length variations (HSL) were extracted by measuring the initial and final distances from the Z-line to the adjacent Z-line or to a region externally adjacent to the M-line of the sarcomere, respectively. Half-sarcomeres generated approximately the same amount of isometric force (29.0 +/- SD 15.5 nN.μm-2) as single sarcomeres (32.1 +/- SD 15.3 nN.μm-2) when activated. In both cases, the steady-state forces after stretch were higher than the forces during isometric contractions at similar conditions. The results suggest that stretch-induced force enhancement is partly caused by proteins within the half-sarcomere.

  3. Effects of epimuscular myofascial force transmission on sarcomere length of passive muscles in the rat hindlimb

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    Tijs, C.; van Dieen, J.H.; Maas, H.

    2015-01-01

    Results from imaging studies and finite element models suggest epimuscular myofascial effects on sarcomere lengths in series within muscle fibers. However, experimental evidence is lacking. We evaluated epimuscular myofascial effects on (1) muscle belly, fiber, and mean sarcomere length and (2)

  4. Large-scale Models Reveal the Two-component Mechanics of Striated Muscle

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    Robert Jarosch

    2008-12-01

    Full Text Available This paper provides a comprehensive explanation of striated muscle mechanics and contraction on the basis of filament rotations. Helical proteins, particularly the coiled-coils of tropomyosin, myosin and α-actinin, shorten their H-bonds cooperatively and produce torque and filament rotations when the Coulombic net-charge repulsion of their highly charged side-chains is diminished by interaction with ions. The classical “two-component model” of active muscle differentiated a “contractile component” which stretches the “series elastic component” during force production. The contractile components are the helically shaped thin filaments of muscle that shorten the sarcomeres by clockwise drilling into the myosin cross-bridges with torque decrease (= force-deficit. Muscle stretch means drawing out the thin filament helices off the cross-bridges under passive counterclockwise rotation with torque increase (= stretch activation. Since each thin filament is anchored by four elastic α-actinin Z-filaments (provided with forceregulating sites for Ca2+ binding, the thin filament rotations change the torsional twist of the four Z-filaments as the “series elastic components”. Large scale models simulate the changes of structure and force in the Z-band by the different Z-filament twisting stages A, B, C, D, E, F and G. Stage D corresponds to the isometric state. The basic phenomena of muscle physiology, i. e. latency relaxation, Fenn-effect, the force-velocity relation, the length-tension relation, unexplained energy, shortening heat, the Huxley-Simmons phases, etc. are explained and interpreted with the help of the model experiments.

  5. In Vivo Sarcomere Length Measurement in Whole Muscles during Passive Stretch and Twitch Contractions.

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    Young, Kevin W; Kuo, Bill P-P; O'Connor, Shawn M; Radic, Stojan; Lieber, Richard L

    2017-02-28

    Muscle force is dictated by micrometer-scale contractile machines called sarcomeres. Whole-muscle force drops from peak force production to zero with just a few micrometers of sarcomere length change. No current technology is able to capture adequate dynamic sarcomere data in vivo, and thus we lack fundamental data needed to understand human movement and movement disorders. Methods such as diffraction, endoscopy, and optical coherence tomography have been applied to muscle but are prohibitively invasive, sensitive to motion artifact, and/or imprecise. Here, we report dynamic sarcomere length measurement in vivo using a combination of our recently validated resonant reflection spectroscopy method combined with optical frequency domain interferometry. Using a 250-μm-wide fiber optic probe, we captured nanometer sarcomere length changes from thousands of sarcomeres on the sub-millisecond timescale during whole-muscle stretch and twitch contraction. We believe that this demonstrates the first large-scale sensing of sarcomere dynamics in vivo, which is a necessary first step to understand movement disorders and to create patient-specific surgical interventions and rehabilitation. Published by Elsevier Inc.

  6. Sarcomere Imaging by Quantum Dots for the Study of Cardiac Muscle Physiology

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    Fuyu Kobirumaki-Shimozawa

    2012-01-01

    Full Text Available We here review the use of quantum dots (QDs for the imaging of sarcomeric movements in cardiac muscle. QDs are fluorescence substances (CdSe that absorb photons and reemit photons at a different wavelength (depending on the size of the particle; they are efficient in generating long-lasting, narrow symmetric emission profiles, and hence useful in various types of imaging studies. Recently, we developed a novel system in which the length of a particular, single sarcomere in cardiomyocytes can be measured at ~30 nm precision. Moreover, our system enables accurate measurement of sarcomere length in the isolated heart. We propose that QDs are the ideal tool for the study of sarcomere dynamics during excitation-contraction coupling in healthy and diseased cardiac muscle.

  7. Muscle contraction phenotypic analysis enabled by optogenetics reveals functional relationships of sarcomere components in Caenorhabditis elegans

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    Hwang, Hyundoo; Barnes, Dawn E.; Matsunaga, Yohei; Benian, Guy M.; Ono, Shoichiro; Lu, Hang

    2016-01-01

    The sarcomere, the fundamental unit of muscle contraction, is a highly-ordered complex of hundreds of proteins. Despite decades of genetics work, the functional relationships and the roles of those sarcomeric proteins in animal behaviors remain unclear. In this paper, we demonstrate that optogenetic activation of the motor neurons that induce muscle contraction can facilitate quantitative studies of muscle kinetics in C. elegans. To increase the throughput of the study, we trapped multiple worms in parallel in a microfluidic device and illuminated for photoactivation of channelrhodopsin-2 to induce contractions in body wall muscles. Using image processing, the change in body size was quantified over time. A total of five parameters including rate constants for contraction and relaxation were extracted from the optogenetic assay as descriptors of sarcomere functions. To potentially relate the genes encoding the sarcomeric proteins functionally, a hierarchical clustering analysis was conducted on the basis of those parameters. Because it assesses physiological output different from conventional assays, this method provides a complement to the phenotypic analysis of C. elegans muscle mutants currently performed in many labs; the clusters may provide new insights and drive new hypotheses for functional relationships among the many sarcomere components.

  8. Overexpression of TEAD-1 in transgenic mouse striated muscles produces a slower skeletal muscle contractile phenotype.

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    Tsika, Richard W; Schramm, Christine; Simmer, Gretchen; Fitzsimons, Daniel P; Moss, Richard L; Ji, Juan

    2008-12-26

    TEA domain (TEAD) transcription factors serve important functional roles during embryonic development and in striated muscle gene expression. Our previous work has implicated a role for TEAD-1 in the fast-to-slow fiber-type transition in response to mechanical overload. To investigate whether TEAD-1 is a modulator of slow muscle gene expression in vivo, we developed transgenic mice expressing hemagglutinin (HA)-tagged TEAD-1 under the control of the muscle creatine kinase promoter. We show that striated muscle-restricted HA-TEAD-1 expression induced a transition toward a slow muscle contractile protein phenotype, slower shortening velocity (Vmax), and longer contraction and relaxation times in adult fast twitch extensor digitalis longus muscle. Notably, HA-TEAD-1 overexpression resulted in an unexpected activation of GSK-3alpha/beta and decreased nuclear beta-catenin and NFATc1/c3 protein. These effects could be reversed in vivo by mechanical overload, which decreased muscle creatine kinase-driven TEAD-1 transgene expression, and in cultured satellite cells by TEAD-1-specific small interfering RNA. These novel in vivo data support a role for TEAD-1 in modulating slow muscle gene expression.

  9. Interactions between connected half-sarcomeres produce emergent mechanical behavior in a mathematical model of muscle.

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    Kenneth S Campbell

    2009-11-01

    Full Text Available Most reductionist theories of muscle attribute a fiber's mechanical properties to the scaled behavior of a single half-sarcomere. Mathematical models of this type can explain many of the known mechanical properties of muscle but have to incorporate a passive mechanical component that becomes approximately 300% stiffer in activating conditions to reproduce the force response elicited by stretching a fast mammalian muscle fiber. The available experimental data suggests that titin filaments, which are the mostly likely source of the passive component, become at most approximately 30% stiffer in saturating Ca2+ solutions. The work described in this manuscript used computer modeling to test an alternative systems theory that attributes the stretch response of a mammalian fiber to the composite behavior of a collection of half-sarcomeres. The principal finding was that the stretch response of a chemically permeabilized rabbit psoas fiber could be reproduced with a framework consisting of 300 half-sarcomeres arranged in 6 parallel myofibrils without requiring titin filaments to stiffen in activating solutions. Ablation of inter-myofibrillar links in the computer simulations lowered isometric force values and lowered energy absorption during a stretch. This computed behavior mimics effects previously observed in experiments using muscles from desmin-deficient mice in which the connections between Z-disks in adjacent myofibrils are presumably compromised. The current simulations suggest that muscle fibers exhibit emergent properties that reflect interactions between half-sarcomeres and are not properties of a single half-sarcomere in isolation. It is therefore likely that full quantitative understanding of a fiber's mechanical properties requires detailed analysis of a complete fiber system and cannot be achieved by focusing solely on the properties of a single half-sarcomere.

  10. In vivo functional and morphological characterization of bone and striated muscle microcirculation in NSG mice.

    Directory of Open Access Journals (Sweden)

    Haider Mussawy

    Full Text Available Organ-specific microcirculation plays a central role in tumor growth, tumor cell homing, tissue engineering, and wound healing. Mouse models are widely used to study these processes; however, these mouse strains often possess unique microhemodynamic parameters, making it difficult to directly compare experiments. The full functional characterization of bone and striated muscle microcirculatory parameters in non-obese diabetic-severe combined immunodeficiency/y-chain; NOD-Prkds IL2rg (NSG mice has not yet been reported. Here, we established either a dorsal skinfold chamber or femur window in NSG mice (n = 23, allowing direct analysis of microcirculatory parameters in vivo by intravital fluorescence microscopy at 7, 14, 21, and 28 days after chamber preparation. Organ-specific differences were observed. Bone had a significantly lower vessel density but a higher vessel diameter than striated muscle. Bone also showed higher effective vascular permeability than striated muscle. The centerline velocity values were similar in the femur window and dorsal skinfold chamber, with a higher volumetric blood flow in bone. Interestingly, bone and striated muscle showed similar tissue perfusion rates. Knowledge of physiological microhemodynamic values of bone and striated muscle in NSG mice makes it possible to analyze pathophysiological processes at these anatomic sites, such as tumor growth, tumor metastasis, and tumor microcirculation, as well as the response to therapeutic agents.

  11. Embracing change: striated-for-smooth muscle replacement in esophagus development.

    Science.gov (United States)

    Krauss, Robert S; Chihara, Daisuke; Romer, Anthony I

    2016-01-01

    The esophagus functions to transport food from the oropharyngeal region to the stomach via waves of peristalsis and transient relaxation of the lower esophageal sphincter. The gastrointestinal tract, including the esophagus, is ensheathed by the muscularis externa (ME). However, while the ME of the gastrointestinal tract distal to the esophagus is exclusively smooth muscle, the esophageal ME of many vertebrate species comprises a variable amount of striated muscle. The esophageal ME is initially composed only of smooth muscle, but its developmental maturation involves proximal-to-distal replacement of smooth muscle with striated muscle. This fascinating phenomenon raises two important questions: what is the developmental origin of the striated muscle precursor cells, and what are the cellular and morphogenetic mechanisms underlying the process? Studies addressing these questions have provided controversial answers. In this review, we discuss the development of ideas in this area and recent work that has shed light on these issues. A working model has emerged that should permit deeper understanding of the role of ME development and maturation in esophageal disorders and in the functional and evolutionary underpinnings of the variable degree of esophageal striated myogenesis in vertebrate species.

  12. Morphology of lesions in striated muscle fibres from the beige mouse

    DEFF Research Database (Denmark)

    Kirkeby, S

    1982-01-01

    Lesions in striated muscle fibres from the beige mouse are described at both the light- and electronmicroscopical levels. The muscles have two types of lesions, one is well defined cores in the fibres and the other is diffusely enlarged intermyofibrillar spaces (IMS). The cores can be filled...... with membrane-like structures or a fluffy unstructured material. In the areas with enlarged IMS comparatively few organelles are present and the muscle fibres seem to be fragmented....

  13. In Vivo Sarcomere Lengths Become More Non-uniform upon Activation in Intact Whole Muscle

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    Eng Kuan Moo

    2017-12-01

    Full Text Available The sarcomere force-length relationship has been extensively used to predict muscle force potential. The common practice is to measure the mean sarcomere length (SL in a relaxed muscle at a single location and at a given length, and this mean SL is assumed to represent the SLs at other locations across the muscle. However, in a previous study, we found that SLs are highly non-uniform across an intact passive muscle. Moreover, SL non-uniformity increases during activation in single myofibril experiments. Myofibrils lack some structural proteins that comprise an intact muscle, and therefore, the increased SL dispersion upon activation seen in myofibrils may not occur in intact whole muscle. The objectives of the current study were (i to measure the distribution of SLs in an activated intact muscle; and (ii to assess the feasibility of using the mean SL measured at a specific location of the muscle to predict muscle force. Using state-of-the-art multi-photon microscopy and a miniature tendon force transducer, in vivo sarcomeres in the mouse tibialis anterior were imaged simultaneously with muscle force during isometric tetanic contractions. We found that in vivo SL dispersion increased substantially during activation and reached average differences of ~1.0 μm. These differences in SL are associated with theoretical force differences of 70–100% of the maximal isometric force. Furthermore, SLs measured at a single location in the passive muscle were poor predictors of active force potential. Although mean SLs in the activated muscle were better predictors of force potential, predicted forces still differed by as much as 35% from the experimentally measured maximal isometric forces.

  14. Muscle sarcomere lesions and thrombosis after spaceflight and suspension unloading

    Energy Technology Data Exchange (ETDEWEB)

    Riley, D.A.; Ellis, S.; Giometti, C.S.; Hoh, J.F.Y.; Ilyina-Kakueva, E.I.; Oganov, V.S.; Slocum, G.R.; Bain, J.L.W.; Sedlak, F.R. (Argonne National Lab., IL (United States))

    1992-08-01

    Extended exposure of humans to spaceflight produces a progressive loss of skeletal muscle strength. This process must be understood to design effective countermeasures. The present investigation examined hindlimb muscles from flight rats killed as close to landing as possible. Spaceflight and tail suspension-hindlimb unloading (unloaded) produced significant decreases in fiber cross-sectional areas of the adductor longus (AL), a slow-twitch antigravity muscle. However, the mean wet weight of the flight AL muscles was near normal, whereas that of the suspension unloaded AL muscles was significantly reduced. Interstitial edema within the flight AL, but not in the unloaded AL, appeared to account for this apparent disagreement.In both conditions, the slow-twitch oxidative fibers atrophied more than the fast-twitch oxidative-glycolytic fibers. Microcirculation was also compromised by spaceflight, such that there was increased formation of thrombi in the postcapillary venules and capillaries.

  15. Embracing change: striated-for-smooth muscle replacement in esophagus development

    OpenAIRE

    Krauss, Robert S.; Chihara, Daisuke; Romer, Anthony I.

    2016-01-01

    The esophagus functions to transport food from the oropharyngeal region to the stomach via waves of peristalsis and transient relaxation of the lower esophageal sphincter. The gastrointestinal tract, including the esophagus, is ensheathed by the muscularis externa (ME). However, while the ME of the gastrointestinal tract distal to the esophagus is exclusively smooth muscle, the esophageal ME of many vertebrate species comprises a variable amount of striated muscle. The esophageal ME is initia...

  16. Cellular mechanism of eccentric-induced muscle injury and its relationship with sarcomere heterogeneity.

    Science.gov (United States)

    Choi, Seung Jun

    2014-08-01

    Activity-induced muscle injury and dysfunction have been identified as key components of musculoskeletal injuries. These injuries often occur following eccentric contractions, when the muscle is under tension and stretched by a force that is greater than the force generated by the muscle. Many daily activities require muscles to perform eccentric contractions, including walking (or running) downhill or down stairs, lowering heavy objects, and landing from a jump. Injuries often occur when these activities are performed at high intensity or for prolonged periods of time. General features of eccentric-induced muscle injury are well documented and include disruption of intracellular muscle structure, prolonged muscle weakness and dysfunction, a delayed-onset muscle soreness, and inflammation. Several weeks are required for the affected tissue to fully regenerate and recover from eccentric-induced muscle injury. Possible mechanisms responsible for eccentric-induced muscle injury are activation impairment and structural disruption of the sarcomere. These two factors seem to be the main sources of eccentric-induced muscle injury. Rather than being separate mechanisms they may be complimentary and interact with each other. Therefore, in this review we will focus on the two main cellular mechanism of muscle cell injury following accustomed eccentric contraction.

  17. Sarcomere length-dependence of activity-dependent twitch potentiation in mouse skeletal muscle

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    MacIntosh Brian R

    2002-12-01

    Full Text Available Abstract Background It has been reported that potentiation of a skeletal muscle twitch response is proportional to muscle length with a negative slope during staircase, and a positive slope during posttetanic potentiation. This study was done to directly compare staircase and posttetanic responses with measurement of sarcomere length to compare their length-dependence. Methods Mouse extensor digitorum longus (EDL muscles were dissected to small bundles of fibers, which permit measurement of sarcomere length (SL, by laser diffraction. In vitro fixed-end contractions of EDL fiber bundles were elicited at 22°C and 35°C at sarcomere lengths ranging from 2.35 μm to 3.85 μm. Twitch contractions were assessed before and after 1.5 s of 75 Hz stimulation at 22°C or during 10 s of 10 Hz stimulation at 22°C or 35°C. Results Staircase potentiation was greater at 35°C than 22°C, and the relative magnitude of the twitch contraction (Pt*/Pt was proportional to sarcomere length with a negative slope, over the range 2.3 μm – 3.7 μm. Linear regression yielded the following: Pt*/Pt = -0.59·SL+3.27 (r2 = 0.74; Pt*/Pt = -0.39·SL+2.34 (r2 = 0.48; and Pt*/Pt = -0.50·SL+2.45 (r2 = 0.80 for staircase at 35°C, and 22°C and posttetanic response respectively. Posttetanic depression rather than potentiation was present at long SL. This indicates that there may be two processes operating in these muscles to modulate the force: one that enhances and a second that depresses the force. Either or both of these processes may have a length-dependence of its mechanism. Conclusion There is no evidence that posttetanic potentiation is fundamentally different from staircase in these muscles.

  18. Still Heart Encodes a Structural HMT, SMYD1b, with Chaperone-Like Function during Fast Muscle Sarcomere Assembly.

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    Kendal Prill

    Full Text Available The vertebrate sarcomere is a complex and highly organized contractile structure whose assembly and function requires the coordination of hundreds of proteins. Proteins require proper folding and incorporation into the sarcomere by assembly factors, and they must also be maintained and replaced due to the constant physical stress of muscle contraction. Zebrafish mutants affecting muscle assembly and maintenance have proven to be an ideal tool for identification and analysis of factors necessary for these processes. The still heart mutant was identified due to motility defects and a nonfunctional heart. The cognate gene for the mutant was shown to be smyd1b and the still heart mutation results in an early nonsense codon. SMYD1 mutants show a lack of heart looping and chamber definition due to a lack of expression of heart morphogenesis factors gata4, gata5 and hand2. On a cellular level, fast muscle fibers in homozygous mutants do not form mature sarcomeres due to the lack of fast muscle myosin incorporation by SMYD1b when sarcomeres are first being assembled (19hpf, supporting SMYD1b as an assembly protein during sarcomere formation.

  19. Structural studies of the waves in striated muscle fibres shortened passively below their slack length.

    Science.gov (United States)

    Brown, L M; González-Serratos, H; Huxley, A F

    1984-06-01

    Isolated skeletal muscle fibres of Rana pipiens were shortened below their slack length by longitudinal compression in a gelatine block, and examined by light and electron microscopy. Waves appeared sharply when the striation spacing (S) reached a critical value (about 2 microns) and increased in height with further compression down to S = 1.6 microns while the resting band pattern was maintained. The waves were plane, helical or irregular, with wave lengths of 5-15 striations. The Z lines usually ran perpendicular to the direction of the myofibrils to form wedge-shaped sarcomeres. The bending occurred mainly in the I band. The thin filaments ran stiffly for about 30 nm from the Z line and then bent toward the A band. The thick filaments bent very slightly, particularly at their tips. The edges of the A band were indistinct, and there were no dense lines at the A-I junction. The appearance of the individual sarcomeres resembled those in relaxed myofibrils at slack length, with no Cm bands. The H zone was only seen occasionally in the slack and wavy fibres examined. In very thin sections the individual thin filaments were seen to end in the pseudo-H zone, and not to cross the M line. There was a single array of not more than six thin filaments round each thick one in transverse sections of the M-line region. These observations suggest that the narrowing of the bands observed in fresh wavy fibres is due mainly to the obliquity of the myofibrils, and that the sarcomere length measured parallel to their axis is longer than the striation spacing. The relationship between sarcomere length and the length of the thin-filament complex is discussed.

  20. Kinetic isoforms of intramembrane charge in intact amphibian striated muscle.

    Science.gov (United States)

    Huang, C L

    1996-04-01

    The effects of the ryanodine receptor (RyR) antagonists ryanodine and daunorubicin on the kinetic and steady-state properties of intramembrane charge were investigated in intact voltage-clamped frog skeletal muscle fibers under conditions that minimized time-dependent ionic currents. A hypothesis that RyR gating is allosterically coupled to configurational changes in dihydropyridine receptors (DHPRs) would predict that such interactions are reciprocal and that RyR modification should influence intramembrane charge. Both agents indeed modified the time course of charging transients at 100-200-microM concentrations. They independently abolished the delayed charging phases shown by q gamma currents, even in fibers held at fully polarized, -90-mV holding potentials; such waveforms are especially prominent in extracellular solutions containing gluconate. Charge movements consistently became exponential decays to stable baselines in the absence of intervening inward or other time-dependent currents. The steady-state charge transfers nevertheless remained equal through the ON and the OFF parts of test voltage steps. The charge-voltage function, Q(VT), shifted by approximately +10 mV, particularly through those test potentials at which delayed q gamma currents normally took place but retained steepness factors (k approximately 8.0 to 10.6 mV) that indicated persistent, steeply voltage-dependent q gamma contributions. Furthermore, both RyR antagonists preserved the total charge, and its variation with holding potential, Qmax (VH), which also retained similarly high voltage sensitivities (k approximately 7.0 to 9.0 mV). RyR antagonists also preserved the separate identities of q gamma and q beta species, whether defined by their steady-state voltage dependence or inactivation or pharmacological properties. Thus, tetracaine (2 mM) reduced the available steady-state charge movement and gave shallow Q(VT) (k approximately 14 to 16 mV) and Qmax (VH) (k approximately 14 to 17 m

  1. Striated muscle fiber size, composition and capillary density in diabetes in relation to neuropathy and muscle strength

    DEFF Research Database (Denmark)

    Andreassen, Christer Swan; Jensen, Jacob Malte; Jakobsen, Johannes

    2014-01-01

    OBJECTIVE: Diabetic polyneuropathy (DPN) leads to progressive loss of muscle strength in the lower extremities due to muscular atrophy. Changes in vascularization occur in diabetic striated muscle; however, the relationship between these changes and DPN is as yet unexplored. The aim of the present...... study was to evaluate histologic properties and capillarization of diabetic skeletal muscle in relation to DPN and muscle strength. METHODS: Twenty type 1 and 20 type 2 diabetic (T1D and T2D, respectively) patients underwent biopsy of the gastrocnemic muscle, isokinetic dynamometry at the ankle......, electrophysiological studies, clinical examination, and quantitative sensory examinations. Muscle biopsies were stained immunohistochemically and muscle fiber diameter, fiber type distribution, and capillary density determined. Twenty control subjects were also included in the study. RESULTS: No relationship was found...

  2. Muscleblind, BSF and TBPH are mislocalized in the muscle sarcomere of a Drosophila myotonic dystrophy model

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    Beatriz Llamusi

    2013-01-01

    Myotonic dystrophy type 1 (DM1 is a genetic disease caused by the pathological expansion of a CTG trinucleotide repeat in the 3′ UTR of the DMPK gene. In the DMPK transcripts, the CUG expansions sequester RNA-binding proteins into nuclear foci, including transcription factors and alternative splicing regulators such as MBNL1. MBNL1 sequestration has been associated with key features of DM1. However, the basis behind a number of molecular and histological alterations in DM1 remain unclear. To help identify new pathogenic components of the disease, we carried out a genetic screen using a Drosophila model of DM1 that expresses 480 interrupted CTG repeats, i(CTG480, and a collection of 1215 transgenic RNA interference (RNAi fly lines. Of the 34 modifiers identified, two RNA-binding proteins, TBPH (homolog of human TAR DNA-binding protein 43 or TDP-43 and BSF (Bicoid stability factor; homolog of human LRPPRC, were of particular interest. These factors modified i(CTG480 phenotypes in the fly eye and wing, and TBPH silencing also suppressed CTG-induced defects in the flight muscles. In Drosophila flight muscle, TBPH, BSF and the fly ortholog of MBNL1, Muscleblind (Mbl, were detected in sarcomeric bands. Expression of i(CTG480 resulted in changes in the sarcomeric patterns of these proteins, which could be restored by coexpression with human MBNL1. Epistasis studies showed that Mbl silencing was sufficient to induce a subcellular redistribution of TBPH and BSF proteins in the muscle, which mimicked the effect of i(CTG480 expression. These results provide the first description of TBPH and BSF as targets of Mbl-mediated CTG toxicity, and they suggest an important role of these proteins in DM1 muscle pathology.

  3. The Popeye Domain Containing Genes and Their Function in Striated Muscle

    Science.gov (United States)

    Schindler, Roland F. R.; Scotton, Chiara; French, Vanessa; Ferlini, Alessandra; Brand, Thomas

    2016-01-01

    The Popeye domain containing (POPDC) genes encode a novel class of cAMP effector proteins, which are abundantly expressed in heart and skeletal muscle. Here, we will review their role in striated muscle as deduced from work in cell and animal models and the recent analysis of patients carrying a missense mutation in POPDC1. Evidence suggests that POPDC proteins control membrane trafficking of interacting proteins. Furthermore, we will discuss the current catalogue of established protein-protein interactions. In recent years, the number of POPDC-interacting proteins has been rising and currently includes ion channels (TREK-1), sarcolemma-associated proteins serving functions in mechanical stability (dystrophin), compartmentalization (caveolin 3), scaffolding (ZO-1), trafficking (NDRG4, VAMP2/3) and repair (dysferlin) or acting as a guanine nucleotide exchange factor for Rho-family GTPases (GEFT). Recent evidence suggests that POPDC proteins might also control the cellular level of the nuclear proto-oncoprotein c-Myc. These data suggest that this family of cAMP-binding proteins probably serves multiple roles in striated muscle. PMID:27347491

  4. Intermittent stretch training of rabbit plantarflexor muscles increases soleus mass and serial sarcomere number.

    Science.gov (United States)

    De Jaeger, Dominique; Joumaa, Venus; Herzog, Walter

    2015-06-15

    In humans, enhanced joint range of motion is observed after static stretch training and results either from an increased stretch tolerance or from a change in the biomechanical properties of the muscle-tendon unit. We investigated the effects of an intermittent stretch training on muscle biomechanical and structural variables. The left plantarflexors muscles of seven anesthetized New Zealand (NZ) White rabbits were passively and statically stretched three times a week for 4 wk, while the corresponding right muscles were used as nonstretched contralateral controls. Before and after the stretching protocol, passive torque produced by the left plantarflexor muscles as a function of the ankle angle was measured. The left and right plantarflexor muscles were harvested from dead rabbits and used to quantify possible changes in muscle structure. Significant mass and serial sarcomere number increases were observed in the stretched soleus but not in the plantaris or medial gastrocnemius. This difference in adaptation between the plantarflexors is thought to be the result of their different fiber type composition and pennation angles. Neither titin isoform nor collagen amount was modified in the stretched compared with the control soleus muscle. Passive torque developed during ankle dorsiflexion was not modified after the stretch training on average, but was decreased in five of the seven experimental rabbits. Thus, an intermittent stretching program similar to those used in humans can produce a change in the muscle structure of NZ White rabbits, which was associated in some rabbits with a change in the biomechanical properties of the muscle-tendon unit. Copyright © 2015 the American Physiological Society.

  5. Immunocytochemical electron microscopic study and western blot analysis of myosin, paramyosin and miniparamyosin in the striated muscle of the fruit fly Drosophila melanogaster and in obliquely striated and smooth muscles of the earthworm Eisenia foetida.

    Science.gov (United States)

    Royuela, M; Fraile, B; Cervera, M; Paniagua, R

    1997-04-01

    Miniparamyosin is a paramyosin isoform (55-60 kDa) that has been isolated in insects (Drosophila) and immunolocalized in several species of arthropods, molluscs, annelids and nematodes. In this study, the presence and distribution of this protein, in comparison with that of paramyosin and myosin, has been examined in the striated muscle (tergal depressor of trochanter) of Drosophila melanogaster, and the obliquely striated muscle (body wall) and the smooth muscle (outer layer of the pseudoheart) of the earthworm Eisenia foetida by means of immunocytochemical electron microscopic study and Western blot analysis miniparamyosin paramyosin and myosin antibodies from Drosophila. In the striated muscle of D. melanogaster, the three proteins were immunolocalized along the length of the thick filaments (A-bands). The distribution of immunogold particles along these filaments was uniform. The relative proportions miniparamyosin/paramyosin/myosin (calculated by counting the number of immunogold particles) were: 1/10/68. In the obliquely striated muscle of E. foetida, immunoreactions to the three proteins were also found in the thick filaments, and the relative proportions miniparamyosin/paramyosin/myosin were 1/2.4/6.9. However, whereas the distribution of both myosin and miniparamyosin along the thick filament length was uniform, paramyosin immunolabelling was more abundant in the extremes of thick filaments (the outer zones of A-bands in the obliquely striated muscle), where the thick filaments become thinner than in the centre (the central zone of A-bands), where these filaments are thicker. The relative proportions of paramyosin in the outer and of paramyosin in the central zones of A-bands were 4/1. This irregular distribution of paramyosin along the thick filament length might be actual but it may also be explained by the fusiform shape of thick filaments in the earthworm: assuming that paramyosin is covered by myosin, paramyosin antigens would be more exposed in the

  6. Cannabinoid CB1 Receptors Are Localized in Striated Muscle Mitochondria and Regulate Mitochondrial Respiration

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    Juan Mendizabal-Zubiaga

    2016-10-01

    Full Text Available The cannabinoid type 1 (CB1 receptor is widely distributed in the brain and peripheral organs where it regulates cellular functions and metabolism. In the brain, CB1 is mainly localized on presynaptic axon terminals but is also found on mitochondria (mtCB1, where it regulates cellular respiration and energy production. Likewise, CB1 is localized on muscle mitochondria, but very little is known about it. The aim of this study was to further investigate in detail the distribution and functional role of mtCB1 in three different striated muscles. Immunoelectron microscopy for CB1 was used in skeletal muscles (gastrocnemius and rectus abdominis and myocardium from wild-type and CB1-KO mice. Functional assessments were performed in mitochondria purified from the heart of the mice and the mitochondrial oxygen consumption upon application of different acute delta-9-tetrahidrocannabinol (Δ9-THC concentrations (100 nM or 200 nM was monitored. About 26% of the mitochondrial profiles in gastrocnemius, 22% in the rectus abdominis and 17% in the myocardium expressed CB1. Furthermore, the proportion of mtCB1 versus total CB1 immunoparticles was about 60% in the gastrocnemius, 55% in the rectus abdominis and 78% in the myocardium. Importantly, the CB1 immunolabeling pattern disappeared in muscles of CB1-KO mice. Functionally, acute 100 nM or 200 nM THC treatment specifically decreased mitochondria coupled respiration between 12% and 15% in wild-type isolated mitochondria of myocardial muscles but no significant difference was noticed between THC treated and vehicle in mitochondria isolated from CB1-KO heart. Furthermore, gene expression of key enzymes involved in pyruvate synthesis, tricarboxylic acid (TCA cycle and mitochondrial respiratory chain was evaluated in the striated muscle of CB1-WT and CB1-KO. CB1-KO showed an increase in the gene expression of Eno3, Pkm2, and Pdha1, suggesting an increased production of pyruvate. In contrast, no significant

  7. Overexpression of Striated Muscle Activator of Rho Signaling (STARS) Increases C2C12 Skeletal Muscle Cell Differentiation.

    Science.gov (United States)

    Wallace, Marita A; Della Gatta, Paul A; Ahmad Mir, Bilal; Kowalski, Greg M; Kloehn, Joachim; McConville, Malcom J; Russell, Aaron P; Lamon, Séverine

    2016-01-01

    Skeletal muscle growth and regeneration depend on the activation of satellite cells, which leads to myocyte proliferation, differentiation and fusion with existing muscle fibers. Skeletal muscle cell proliferation and differentiation are tightly coordinated by a continuum of molecular signaling pathways. The striated muscle activator of Rho signaling (STARS) is an actin binding protein that regulates the transcription of genes involved in muscle cell growth, structure and function via the stimulation of actin polymerization and activation of serum-response factor (SRF) signaling. STARS mediates cell proliferation in smooth and cardiac muscle models; however, whether STARS overexpression enhances cell proliferation and differentiation has not been investigated in skeletal muscle cells. We demonstrate for the first time that STARS overexpression enhances differentiation but not proliferation in C2C12 mouse skeletal muscle cells. Increased differentiation was associated with an increase in the gene levels of the myogenic differentiation markers Ckm, Ckmt2 and Myh4, the differentiation factor Igf2 and the myogenic regulatory factors (MRFs) Myf5 and Myf6. Exposing C2C12 cells to CCG-1423, a pharmacological inhibitor of SRF preventing the nuclear translocation of its co-factor MRTF-A, had no effect on myotube differentiation rate, suggesting that STARS regulates differentiation via a MRTF-A independent mechanism. These findings position STARS as an important regulator of skeletal muscle growth and regeneration.

  8. Revealing t-tubules in striated muscle with new optical super-resolution microscopy techniques

    Directory of Open Access Journals (Sweden)

    Isuru D. Jayasinghe

    2014-12-01

    Full Text Available The t-tubular system plays a central role in the synchronisation of calcium signalling and excitation-contraction coupling in most striated muscle cells. Light microscopy has been used for imaging t-tubules for well over 100 years and together with electron microscopy (EM, has revealed the three-dimensional complexities of the t-system topology within cardiomyocytes and skeletal muscle fibres from a range of species. The emerging super-resolution single molecule localisation microscopy (SMLM techniques are offering a near 10-fold improvement over the resolution of conventional fluorescence light microscopy methods, with the ability to spectrally resolve nanometre scale distributions of multiple molecular targets. In conjunction with the next generation of electron microscopy, SMLM has allowed the visualisation and quantification of intricate t-tubule morphologies within large areas of muscle cells at an unprecedented level of detail. In this paper, we review recent advancements in the t-tubule structural biology with the utility of various microscopy techniques. We outline the technical considerations in adapting SMLM to study t-tubules and its potential to further our understanding of the molecular processes that underlie the sub-micron scale structural alterations observed in a range of muscle pathologies.

  9. Detection of a troponin I-like protein in non-striated muscle of the tardigrades (water bears).

    Science.gov (United States)

    Obinata, Takashi; Ono, Kanako; Ono, Shoichiro

    2011-03-01

    Tardigrades, also known as water bears, have somatic muscle fibers that are responsible for movement of their body and legs. These muscle fibers contain thin and thick filaments in a non-striated pattern. However, the regulatory mechanism of muscle contraction in tardigrades is unknown. In the absence of extensive molecular and genomic information, we detected a protein of 31 kDa in whole lysates of tardigrades that cross-reacted with the antibody raised against nematode troponin I (TnI). TnI is a component of the troponin complex that regulates actin-myosin interaction in a Ca(2+)-dependent and actin-linked manner. This TnI-like protein was co-extracted with actin in a buffer containing ATP and EGTA, which is known to induce relaxation of a troponin-regulated contractile system. The TnI-like protein was specifically expressed in the somatic muscle fibers in adult animals and partially co-localized with actin filaments in a non-striated manner. Interestingly, the pharyngeal muscle did not express this protein. These observations suggest that the non-striated somatic muscle of tardigrades has an actin-linked and troponin-regulated system for muscle contraction.

  10. Attenuation of cardiac remodeling after myocardial infarction by muscle LIM protein-calcineurin signaling at the sarcomeric Z-disc

    OpenAIRE

    Heineke, Joerg; Ruetten, Hartmut; Willenbockel, Christian; Gross, Sandra C.; Naguib, Marian; Schaefer, Arnd; Kempf, Tibor; Hilfiker-Kleiner, Denise; Caroni, Pico; Kraft, Theresia; Kaiser, Robert A.; Molkentin, Jeffery D.; Drexler, Helmut; Wollert, Kai C.

    2005-01-01

    Adverse left ventricular (LV) remodeling after myocardial infarction (MI) is a major cause for heart failure. Molecular modifiers of the remodeling process remain poorly defined. Patients with heart failure after MI have reduced LV expression levels of muscle LIM protein (MLP), a component of the sarcomeric Z-disk that is involved in the integration of stress signals in cardiomyocytes. By using heterozygous MLP mutant (MLP+/—) mice, we explored the role of MLP in post-MI remodeling. LV dimens...

  11. Effects of strain on contractile force and number of sarcomeres in series of Xenopus laevis single muscle fibres during long-term culture

    NARCIS (Netherlands)

    Jaspers, R.T.; Feenstra, Hiske; Verheyen, A.K.; van der Laarse, W.J.; Huijing, P.A.J.B.M.

    2004-01-01

    The aim of the present study is to test whether mechanical strain uniquely regulates muscle fibre atrophy/hypertrophy and adaptation of the number of sarcomeres in series within mature muscle fibres in vitro. Mature single muscle fibres from Xenopus laevis illiofibularis muscle were cultured (4--97

  12. Effects of strain on contractile force and number of sarcomeres in series of Xenopus laevis single muscle fibres during long-term culture

    NARCIS (Netherlands)

    Jaspers, R.T.; Feenstra, H.M.; Verheyen, A.K.; van der Laarse, W.J.; Huijing, P.A.J.B.M.

    2004-01-01

    The aim of the present study is to test whether mechanical strain uniquely regulates muscle fibre atrophy/hypertrophy and adaptation of the number of sarcomeres in series within mature muscle fibres in vitro. Mature single muscle fibres from Xenopus laevis illiofibularis muscle were cultured (4 - 97

  13. Splicing transitions of the anchoring protein ENH during striated muscle development.

    Science.gov (United States)

    Ito, Jumpei; Hashimoto, Taiki; Nakamura, Sho; Aita, Yusuke; Yamazaki, Tomoko; Schlegel, Werner; Takimoto, Koichi; Maturana, Andrés D

    2012-05-04

    The ENH (PDLIM5) protein acts as a scaffold to tether various functional proteins at subcellular sites via PDZ and three LIM domains. Splicing of the ENH primary transcript generates various products with different repertories of protein interaction modules. Three LIM-containing ENH predominates in neonatal cardiac tissue, whereas LIM-less ENHs are abundant in adult hearts, as well as skeletal muscles. Here we examine the timing of splicing transitions of ENH gene products during postnatal heart development and C2C12 myoblast differentiation. Real-time PCR analysis shows that LIM-containing ENH1 mRNA is gradually decreased during postnatal heart development, whereas transcripts with the short exon 5 appear in the late postnatal period and continues to increase until at least one month after birth. The splicing transition from LIM-containing ENH1 to LIM-less ENHs is also observed during the early period of C2C12 differentiation. This transition correlates with the emergence of ENH transcripts with the short exon 5, as well as the expression of myogenin mRNA. In contrast, the shift from the short exon 5 to the exon 7 occurs in the late differentiation period. The timing of this late event corresponds to the appearance of mRNA for the skeletal myosin heavy chain MYH4. Thus, coordinated and stepwise splicing transitions result in the production of specific ENH transcripts in mature striated muscles. Copyright © 2012 Elsevier Inc. All rights reserved.

  14. The Intriguing Dual Lattices of the Myosin Filaments in Vertebrate Striated Muscles: Evolution and Advantage

    Directory of Open Access Journals (Sweden)

    Pradeep K. Luther

    2014-12-01

    Full Text Available Myosin filaments in vertebrate striated muscle have a long roughly cylindrical backbone with cross-bridge projections on the surfaces of both halves except for a short central bare zone. In the middle of this central region the filaments are cross-linked by the M-band which holds them in a well-defined hexagonal lattice in the muscle A-band. During muscular contraction the M-band-defined rotation of the myosin filaments around their long axes influences the interactions that the cross-bridges can make with the neighbouring actin filaments. We can visualise this filament rotation by electron microscopy of thin cross-sections in the bare-region immediately adjacent to the M-band where the filament profiles are distinctly triangular. In the muscles of teleost fishes, the thick filament triangular profiles have a single orientation giving what we call the simple lattice. In other vertebrates, for example all the tetrapods, the thick filaments have one of two orientations where the triangles point in opposite directions (they are rotated by 60° or 180° according to set rules. Such a distribution cannot be developed in an ordered fashion across a large 2D lattice, but there are small domains of superlattice such that the next-nearest neighbouring thick filaments often have the same orientation. We believe that this difference in the lattice forms can lead to different contractile behaviours. Here we provide a historical review, and when appropriate cite recent work related to the emergence of the simple and superlattice forms by examining the muscles of several species ranging back to primitive vertebrates and we discuss the functional differences that the two lattice forms may have.

  15. Role of dystroglycan in limiting contraction-induced injury to the sarcomeric cytoskeleton of mature skeletal muscle.

    Science.gov (United States)

    Rader, Erik P; Turk, Rolf; Willer, Tobias; Beltrán, Daniel; Inamori, Kei-Ichiro; Peterson, Taylor A; Engle, Jeffrey; Prouty, Sally; Matsumura, Kiichiro; Saito, Fumiaki; Anderson, Mary E; Campbell, Kevin P

    2016-09-27

    Dystroglycan (DG) is a highly expressed extracellular matrix receptor that is linked to the cytoskeleton in skeletal muscle. DG is critical for the function of skeletal muscle, and muscle with primary defects in the expression and/or function of DG throughout development has many pathological features and a severe muscular dystrophy phenotype. In addition, reduction in DG at the sarcolemma is a common feature in muscle biopsies from patients with various types of muscular dystrophy. However, the consequence of disrupting DG in mature muscle is not known. Here, we investigated muscles of transgenic mice several months after genetic knockdown of DG at maturity. In our study, an increase in susceptibility to contraction-induced injury was the first pathological feature observed after the levels of DG at the sarcolemma were reduced. The contraction-induced injury was not accompanied by increased necrosis, excitation-contraction uncoupling, or fragility of the sarcolemma. Rather, disruption of the sarcomeric cytoskeleton was evident as reduced passive tension and decreased titin immunostaining. These results reveal a role for DG in maintaining the stability of the sarcomeric cytoskeleton during contraction and provide mechanistic insight into the cause of the reduction in strength that occurs in muscular dystrophy after lengthening contractions.

  16. VAPB/ALS8 MSP ligands regulate striated muscle energy metabolism critical for adult survival in caenorhabditis elegans.

    Directory of Open Access Journals (Sweden)

    Sung Min Han

    Full Text Available Mutations in VAPB/ALS8 are associated with amyotrophic lateral sclerosis (ALS and spinal muscular atrophy (SMA, two motor neuron diseases that often include alterations in energy metabolism. We have shown that C. elegans and Drosophila neurons secrete a cleavage product of VAPB, the N-terminal major sperm protein domain (vMSP. Secreted vMSPs signal through Roundabout and Lar-like receptors expressed on striated muscle. The muscle signaling pathway localizes mitochondria to myofilaments, alters their fission/fusion balance, and promotes energy production. Here, we show that neuronal loss of the C. elegans VAPB homolog triggers metabolic alterations that appear to compensate for muscle mitochondrial dysfunction. When vMSP levels drop, cytoskeletal or mitochondrial abnormalities in muscle induce elevated DAF-16, the Forkhead Box O (FoxO homolog, transcription factor activity. DAF-16 promotes muscle triacylglycerol accumulation, increases ATP levels in adults, and extends lifespan, despite reduced muscle mitochondria electron transport chain activity. Finally, Vapb knock-out mice exhibit abnormal muscular triacylglycerol levels and FoxO target gene transcriptional responses to fasting and refeeding. Our data indicate that impaired vMSP signaling to striated muscle alters FoxO activity, which affects energy metabolism. Abnormalities in energy metabolism of ALS patients may thus constitute a compensatory mechanism counterbalancing skeletal muscle mitochondrial dysfunction.

  17. Distribution of Myosin Attachment Times Predicted from Viscoelastic Mechanics of Striated Muscle

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    Bradley M. Palmer

    2011-01-01

    Full Text Available We demonstrate that viscoelastic mechanics of striated muscle, measured as elastic and viscous moduli, emerge directly from the myosin crossbridge attachment time, tatt, also called time-on. The distribution of tatt was modeled using a gamma distribution with shape parameter, p, and scale parameter, β. At 5 mM MgATP, β was similar between mouse α-MyHC (16.0±3.7 ms and β-MyHC (17.9±2.0 ms, and p was higher (P<0.05 for β-MyHC (5.6±0.4 no units compared to α-MyHC (3.2±0.9. At 1 mM MgATP, p approached a value of 10 in both isoforms, but β rose only in the β-MyHC (34.8±5.8 ms. The estimated mean tatt (i.e., pβ product was longer in the β-MyHC compared to α-MyHC, and became prolonged in both isoforms as MgATP was reduced as expected. The application of our viscoelastic model to these isoforms and varying MgATP conditions suggest that tatt is better modeled as a gamma distribution due to its representing multiple temporal events occurring within tatt compared to a single exponential distribution which assumes only one temporal event within tatt.

  18. Increased hypoxia-inducible factor-1α in striated muscle of tumor-bearing mice.

    Science.gov (United States)

    Devine, Raymond D; Bicer, Sabahattin; Reiser, Peter J; Wold, Loren E

    2017-06-01

    that occur during cancer cachexia. NEW & NOTEWORTHY We used proteomics and metadata analysis software to identify contributors to metabolic changes in striated muscle during cancer cachexia. We found increased expression of hypoxia-inducible factor-1α in the heart and skeletal muscle, suggesting a potential target for the therapeutic treatment of cancer cachexia. Copyright © 2017 the American Physiological Society.

  19. Testosterone enhances C-14 2-deoxyglucose uptake by striated muscle. [sex hormones and muscle

    Science.gov (United States)

    Toop, J.; Max, S. R.

    1982-01-01

    The effect of testosterone propionate (TP) on C-14 2-deoxyglucose (C-14 2DG) uptake was studied in the rat levator ani muscle in vivo using the autoradiographic technique. Following a delay of 1 to 3 h after injecting TP, the rate of C-14 2DG uptake in experimental animals began to increase and continued to increase for at least 20 h. The label, which corresponds to C-14 2-deoxyglucose 6-phosphate, as demonstrated by chromatographic analysis of muscle extracts, was uniformly distributed over the entire muscle and was predominantly in muscle fibers, although nonmuscular elements were also labeled. The 1 to 3 h time lag suggests that the TP effect may be genomic, acting via androgen receptors, rather than directly on muscle membranes. Acceleration of glucose uptake may be an important early event in the anabolic response of the rat levator ani muscle to androgens.

  20. Expression of TPM1κ, a Novel Sarcomeric Isoform of the TPM1 Gene, in Mouse Heart and Skeletal Muscle

    OpenAIRE

    Dube, Syamalima; Panebianco, Lauren; Matoq, Amr A.; Chionuma, Henry N.; Denz, Christopher R.; Poiesz, Bernard J.; Dube, Dipak K.

    2014-01-01

    We have investigated the expression of TPM1 α and TPM1 κ in mouse striated muscles. TPM1 α and TMP1 κ were amplified from the cDNA of mouse heart by using conventional RT-PCR. We have cloned the PCR amplified DNA and determined the nucleotide sequences. Deduced amino acid sequences show that there are three amino acid changes in mouse exon 2a when compared with the human TPM1 κ . However, the deduced amino acid sequences of human TPM1 α and mouse TPM1 α are identical. Conventional RT-PCR data...

  1. Immunocytochemical electron microscopic study and Western blot analysis of caldesmon and calponin in striated muscle of the fruit fly Drosophila melanogaster and in several muscle cell types of the earthworm Eisenia foetida.

    Science.gov (United States)

    Royuela, M; Fraile, B; Picazo, M L; Paniagua, R

    1997-01-01

    Caldesmon and calponin are two proteins that are characteristic of vertebrate smooth muscle. In invertebrates, caldesmon has only been studied in some molluscan muscles, and no previous references to calponin have been found. The aim of this paper was to investigate the presence and distribution of caldesmon and calponin in several invertebrate muscle cell types, classified according to their ultrastructural pattern: transversely striated muscle (flight muscle from Drosophila melanogaster), obliquely striated muscle (muscular body wall and inner muscular layer of the pseudoheart from the earthworm Eisenia foetida), and a muscle of doubtful classification which seems to be intermediate between smooth muscle and obliquely striated muscle (outer muscular layer of the pseudoheart, from E. foetida), using electron microscopy immunocytochemistry and Western blot analysis. Immunoreactions to both caldesmon and calponin were observed in the outer muscular layer cells from the earthworm pseudoheart but neither in the transversely striated muscle of D. melanogaster nor in the obliquely striated muscle from the earthworm. Present findings suggest that caldesmon- and calponin-like proteins are also present in invertebrate muscle cells, but only in those that are ultrastructurally similar to the vertebrate smooth muscle cells. Since discrepancies in the classification of some invertebrate muscles are common in the literature, the use of distinctive markers, such as troponin, caldesmon and calponin may improve our understanding of the nature and properties of many invertebrate muscles showing an ultrastructural pattern that does not resemble any of the classic muscle types.

  2. Contracture Coupling of Slow Striated Muscle in Non-Ionic Solutions and Replacement of Calcium, Sodium, and Potassium

    Science.gov (United States)

    Irwin, Richard L.; Hein, Manfred M.

    1964-01-01

    The development of contracture related to changes of ionic environment (ionic contracture coupling) has been studied in the slowly responding fibers of frog skeletal muscle. When deprived of external ions for 30 minutes by use of solutions of sucrose, mannitol, or glucose, the slow skeletal muscle fibers, but not the fast, develop pronounced and easily reversible contractures. Partial replacement of the non-ionic substance with calcium or sodium reduces the development of the contractures but replacement by potassium does not. The concentration of calcium necessary to prevent contracture induced by a non-ionic solution is greater than that needed to maintain relaxation in ionic solutions. To suppress the non-ionic-induced contractures to the same extent as does calcium requires several fold higher concentrations of sodium. Two types of ionic contracture coupling occur in slow type striated muscle fibers: (a) a calcium deprivation type which develops maximally at full physiological concentration of external sodium, shows a flow rate dependency for the calcium-depriving fluid, and is lessened when the sodium concentration is decreased by replacement with sucrose; (b) a sodium deprivation type which occurs maximally without external sodium, is lessened by increasing the sodium concentration, and has no flow rate dependency for ion deprivation. Both types of contracture are largely prevented by the presence of sufficient calcium. There thus seem to be calcium- and sodium-linked processes at work in the ionic contracture coupling of slow striated muscle. PMID:14127603

  3. Multi-tasking role of the mechanosensing protein Ankrd2 in the signaling network of striated muscle.

    Directory of Open Access Journals (Sweden)

    Anna Belgrano

    Full Text Available Ankrd2 (also known as Arpp together with Ankrd1/CARP and DARP are members of the MARP mechanosensing proteins that form a complex with titin (N2A/calpain 3 protease/myopalladin. In muscle, Ankrd2 is located in the I-band of the sarcomere and moves to the nucleus of adjacent myofibers on muscle injury. In myoblasts it is predominantly in the nucleus and on differentiation shifts from the nucleus to the cytoplasm. In agreement with its role as a sensor it interacts both with sarcomeric proteins and transcription factors.Expression profiling of endogenous Ankrd2 silenced in human myotubes was undertaken to elucidate its role as an intermediary in cell signaling pathways. Silencing Ankrd2 expression altered the expression of genes involved in both intercellular communication (cytokine-cytokine receptor interaction, endocytosis, focal adhesion, tight junction, gap junction and regulation of the actin cytoskeleton and intracellular communication (calcium, insulin, MAPK, p53, TGF-β and Wnt signaling. The significance of Ankrd2 in cell signaling was strengthened by the fact that we were able to show for the first time that Nkx2.5 and p53 are upstream effectors of the Ankrd2 gene and that Ankrd1/CARP, another MARP member, can modulate the transcriptional ability of MyoD on the Ankrd2 promoter. Another novel finding was the interaction between Ankrd2 and proteins with PDZ and SH3 domains, further supporting its role in signaling. It is noteworthy that we demonstrated that transcription factors PAX6, LHX2, NFIL3 and MECP2, were able to bind both the Ankrd2 protein and its promoter indicating the presence of a regulatory feedback loop mechanism.In conclusion we demonstrate that Ankrd2 is a potent regulator in muscle cells affecting a multitude of pathways and processes.

  4. A mechanism accounting for independence on starting length of tension increase in ramp stretches of active skeletal muscle at short half-sarcomere lengths.

    Science.gov (United States)

    Till, Olaf; Siebert, Tobias; Blickhan, Reinhard

    2010-09-07

    Based on previous experimental results of independence on starting length of the tension gradient in constant-velocity stretches of active skeletal muscle at muscle lengths including the ascending limb and the plateau of the tension-length relation, a possible physiological mechanism determining the tension increase in lengthening active muscle is discussed. Considering the sliding filament theory, it is suggested that the tension-length relation of a half-sarcomere in lengthening contractions is different from that in isometric contractions. The assumed mechanism predicts, among others, that the thick filament retains its shortened length in lengthening contractions starting from a half-sarcomere length where this filament is compressed. An example model is implemented and checked with simulations. Copyright 2010 Elsevier Ltd. All rights reserved.

  5. Acute effects of intramuscular aponeurotomy on rat GM: Force transmission, muscle force and sarcomere length

    NARCIS (Netherlands)

    Jaspers, R.T.; Brunner, R.; Pel, J.J.M.; Huijing, P.A.J.B.M.

    1999-01-01

    Acute effects of intramuscular aponeurotomy on muscle force and geometry as a function to muscle length were studied in rat m. gastrocnemius medialis (GM). Acutely after aponeurotomy, activation of the muscle at increasing lengths (acute trajectory) showed a spontaneous and progressive but partial

  6. Role of active contraction and tropomodulins in regulating actin filament length and sarcomere structure in developing zebrafish skeletal muscle

    Directory of Open Access Journals (Sweden)

    Lise eMazelet

    2016-03-01

    Full Text Available Whilst it is recognised that contraction plays an important part in maintaining the structure and function of mature skeletal muscle, its role during development remains undefined. In this study the role of movement in skeletal muscle maturation was investigated in intact zebrafish embryos using a combination of genetic and pharmacological approaches. An immotile mutant line (cacnb1ts25 which lacks functional voltage-gated calcium channels (dihydropyridine receptors in the muscle and pharmacological immobilisation of embryos with a reversible anaesthetic (Tricaine, allowed the study of paralysis (in mutants and anaesthetised fish and recovery of movement (reversal of anaesthetic treatment. The effect of paralysis in early embryos (aged between 17-24 hours post fertilisation, hpf on skeletal muscle structure at both myofibrillar and myofilament level was determined using both immunostaining with confocal microscopy and small angle X-ray diffraction. The consequences of paralysis and subsequent recovery on the localisation of the actin capping proteins Tropomodulin 1 &4 (Tmod in fish aged from 17hpf until 42hpf was also assessed. The functional consequences of early paralysis were investigated by examining the mechanical properties of the larval muscle. The length-force relationship, active and passive tension, was measured in immotile, recovered and control skeletal muscle at 5 and 7 day post fertilisation (dpf. Recovery of muscle function was also assessed by examining swimming patterns in recovered and control fish. Inhibition of the initial embryonic movements (up to 24 hpf resulted in an increase in myofibril length and a decrease in width followed by almost complete recovery in both moving and paralysed fish by 42hpf. In conclusion, myofibril organisation is regulated by a dual mechanism involving movement-dependent and movement-independent processes. The initial contractile event itself drives the localisation of Tmod1 to its sarcomeric

  7. (−)-EPICATECHIN IMPROVES MITOCHONDRIAL RELATED PROTEIN LEVELS AND AMELIORATES OXIDATIVE STRESS IN DYSTROPHIC DELTA SARCOGLYCAN NULL MOUSE STRIATED MUSCLE

    Science.gov (United States)

    Ramirez-Sanchez, Israel; De los Santos, Sergio; Gonzalez-Basurto, Silvia; Canto, Patricia; Mendoza-Lorenzo, Patricia; Palma-Flores, Carlos; Ceballos-Reyes, Guillermo; Villarreal, Francisco; Zentella-Dehesa, Alejandro; Coral-Vazquez, Ramon

    2014-01-01

    Muscular dystrophies (MD) are a group of heterogeneous genetic disorders characterized by progressive striated muscle wasting and degeneration. Although the genetic basis for many of these disorders has been identified, the exact mechanism for disease pathogenesis remains unclear. The presence of oxidative stress (OS) is known to contribute to the pathophysiology and severity of the MD. Mitochondrial dysfunction is observed in MD and likely represents an important determinant of increased OS. Experimental antioxidant therapies have been implemented with the aim of protecting against disease progression, but results from clinical trials have been disappointing. In this study, we explored the capacity of the cacao flavonoid (−)-epicatechin (Epi) to mitigate OS by acting as a positive regulator of mitochondrial structure/function endpoints and redox balance control systems in skeletal and cardiac muscles of dystrophic, δ-sarcoglycan (δ-SG) null mice. Wild type or δ-SG null 2.5 month old male mice were treated via oral gavage with either water (control animals) or Epi (1 mg/kg, twice/day) for 2 weeks. Results evidence a significant normalization of total protein carbonylation, recovery of reduced/oxidized glutathione (GSH/GSSG ratio) and enhanced superoxide dismutase 2, catalase and citrate synthase activities with Epi treatment. These effects were accompanied by increases in protein levels for thiolredoxin, glutathione peroxidase, superoxide dismutase 2, catalase and mitochondrial endpoints. Furthermore, we evidence decreases in heart and skeletal muscle fibrosis, accompanied with an improvement in skeletal muscle function with treatment. These results warrant the further investigation of Epi as a potential therapeutic agent to mitigate MD associated muscle degeneration. PMID:25284161

  8. The ‘Goldilocks Zone’ from a redox perspective - Adaptive versus deleterious responses to oxidative stress in striated muscle

    Directory of Open Access Journals (Sweden)

    Rick J Alleman

    2014-09-01

    Full Text Available Consequences of oxidative stress may be beneficial or detrimental in physiological systems. An organ system’s position on the ‘hormetic curve’ is governed by the source and temporality of reactive oxygen species (ROS production, proximity of ROS to moieties most susceptible to damage, and the capacity of the endogenous cellular ROS scavenging mechanisms. Most importantly, the resilience of the tissue (the capacity to recover from damage is a decisive factor, and this is reflected in the disparate response to ROS in cardiac and skeletal muscle. In myocytes, a high oxidative capacity invariably results in a significant ROS burden which in homeostasis, is rapidly neutralized by the robust antioxidant network. The up-regulation of key pathways in the antioxidant network is a central component of the hormetic response to ROS. Despite such adaptations, persistent oxidative stress over an extended time-frame (e.g. months to years inevitably leads to cumulative damages, maladaptation and ultimately the pathogenesis of chronic diseases. Indeed, persistent oxidative stress in heart and skeletal muscle has been repeatedly demonstrated to have causal roles in the etiology of heart disease and insulin resistance, respectively. Deciphering the mechanisms that underlie the divergence between adaptive and maladaptive responses to oxidative stress remains an active area of research for basic scientists and clinicians alike, as this would undoubtedly lead to novel therapeutic approaches. Here, we provide an overview of major types of ROS in striated muscle and the divergent adaptations that occur in response to them. Emphasis is placed on highlighting newly uncovered areas of research on this topic, with particular focus on the mitochondria, and the diverging roles that ROS play in muscle health (e.g., exercise or preconditioning and disease (e.g., cardiomyopathy, ischemia, metabolic syndrome.

  9. Creatine kinase deficiency in striated mouse muscle : biochemical and physiological studies

    NARCIS (Netherlands)

    Veld, Frank ter

    2003-01-01

    The balance between ATP energy demand and supply is essential in muscle cells. The creatine kinase system fulfils both a transporting and buffering role in muscle cells, whereby fluctuations in ATP free-energy demand can be counterbalanced. Removal of the creatine kinase proteins with the aid of

  10. Histochemical studies on striated muscle fibres of the beige mutant mouse

    DEFF Research Database (Denmark)

    Kirkeby, S

    1982-01-01

    A histological study of cylindric structures in skeletal muscle fibres from beige mice with the Chediak-Higashi syndrome was carried out. The muscle tissue was investigated morphologically with a differential interference contrast microscope and stained for glycogen, lipid, and basophile elements...

  11. Esterases in striated muscle from mice with the Chediak-Higashi syndrome

    DEFF Research Database (Denmark)

    Kirkeby, S; Moe, D

    1981-01-01

    In this paper a localized strong reaction for non-specific esterase forming cylindric structures is described within skeletal muscle fibres from the beige mouse. It seems from zymograms and protein electrophoresis that this esterase is membrane bound, highly reactive and present in rather small...... amounts within the muscle fibres....

  12. Development of Trichosomoides nasalis (Nematoda: Trichinelloidea) in the murid host: evidence for larval growth in striated muscle fibres

    Science.gov (United States)

    Fall, E.H.; Diagne, M.; Junker, K.; Duplantier, J.M.; Ba, K.; Vallée, I.; Bain, O.

    2012-01-01

    Trichosomoides nasalis (Trichinelloidea) is a parasite of Arvicanthis niloticus (Muridae) in Senegal. Female worms that harbour dwarf males in their uteri, occur in the epithelium of the nasal mucosa. Young laboratory-bred A. niloticus were either fed females containing larvated eggs or intraperitoneally injected with motile first-stage larvae recovered from female uteri. Both resulted in successful infection. Organs examined during rodent necropsy were blood and lymphatic circulatory systems (heart, large vessels, lymphnodes), lungs, liver, kidneys, thoracic and abdominal cavities, thoracic and abdominal muscular walls, diaphragm, tongue, and nasal mucosa. Development to adult nasal stages took three weeks. Recovery of newly hatched larvae from the peritoneal fluid at four-eight hours after oral infection suggests a direct passage from the stomach or intestinal wall to the musculature. However, dissemination through the blood, as observed with Trichinella spiralis, cannot be excluded even though newly hatched larvae of T. nasalis are twice as thick (15 μm). Developing larvae were found in histological sections of the striated muscle of the abdominal and thoracic walls, and larvae in fourth moult were dissected from these sites. Adult females were found in the deep nasal mucosa where mating occurred prior to worms settling in the nasal epithelium. The present study shows a remarkable similarity between T. nasalis and Trichinella species regarding muscle tropism, but the development of T. nasalis is not arrested at the late first-larval stage and does not induce transformation of infected fibres into nurse cells. T. nasalis seems a potential model to study molecular relations between trichinelloid larvae and infected muscle fibres. PMID:22314237

  13. Wheelchair marathon racing causes striated muscle distress in individuals with spinal cord injury.

    Science.gov (United States)

    Ide, M; Tajima, F; Furusawa, K; Mizushima, T; Ogata, H

    1999-03-01

    To assess the effects of wheelchair marathon racing in individuals with spinal cord injury (SCI) on circulating muscle enzymes and myoglobin. Thirty-one men with SCI, including 25 wheelchair marathon athletes and 6 sedentary men. Serum myoglobin (Mb), creatine kinase (CK) activity, and lactate dehydrogenase (LDH) were measured in participants of the 1995 Oita International Wheelchair Marathon Race (42.195 km). Blood samples were obtained 24 hours before, immediately after, 24 hours after, and 7 days after the race. Marathon racing resulted in significant increases in serum Mb, total CK activity, and LDH (pathletes with SCI. Completion of the marathon race did not cause cardiac muscle damage, however. Elevated muscle enzyme levels likely resulted from muscle distress rather than from dehydration.

  14. Impaired contractility of the circular striated urethral sphincter muscle may contribute to stress urinary incontinence in female zucker fatty rats.

    Science.gov (United States)

    Lee, Yung-Chin; Lin, Guiting; Wang, Guifang; Reed-Maldonado, Amanda; Lu, Zhihua; Wang, Lin; Banie, Lia; Lue, Tom F

    2017-08-01

    Obesity has been an independent risk factor for female stress urinary incontinence (SUI), the mechanism of this association remains unknown. The aim of this study is to validate the hypothesis that urethral dysfunction is a possible contributor to SUI in obese women. Ten Zucker Fatty (ZF) (ZUC-Lepr fa 185) and 10 Zucker Lean (ZL) (ZUC-Lepr fa 186) female rats at 12-week-old were used in this experiment. The urethral sphincter rings were harvested from the bladder neck through to the most proximal 2/3 regions. In the organ bath study, single pulses of electrical field stimulation (EFS) were applied. For the fatiguing stimulation, repeated multi-pulse EFS with 70 mA were applied at frequency of 5 Hz for 5 min. Caffeine-containing Krebs' solution was administrated to contract the urethra until the contraction began to reach a plateau for 10 min. We performed immunofluorescence staining of the urethra after the experiment was finished. Compared to ZL controls, ZF rats had significantly impaired muscle contractile activity (MCA) (P female rats had significantly impaired contractile properties of striated urethral sphincter, suggesting urethral dysfunction could be an important contributor to SUI in obesity. © 2016 Wiley Periodicals, Inc.

  15. Entropic elasticity in the generation of muscle Force - A theoretical model

    DEFF Research Database (Denmark)

    Nielsen, Bjørn Gilbert

    2002-01-01

    A novel simplified structural model of sarcomeric force production in striate muscle is presented. Using some simple assumptions regarding the distribution of myosin spring lengths at different sliding velocities it is possible to derive a very simple expression showing the main components...

  16. Psoas muscle architectural design, in vivo sarcomere length range, and passive tensile properties support its role as a lumbar spine stabilizer.

    Science.gov (United States)

    Regev, Gilad J; Kim, Choll W; Tomiya, Akihito; Lee, Yu Po; Ghofrani, Hossein; Garfin, Steven R; Lieber, Richard L; Ward, Samuel R

    2011-12-15

    Controlled laboratory and cross-sectional study designs. To determine psoas major (PM) muscle architectural properties, in vivo sarcomere-length operating range, and passive mechanical properties. PM is an important hip flexor but its role in lumbar spine function is not fully understood. Several investigators have detailed the gross anatomy of PM, but comprehensive architectural data and in vivo length-tension and passive mechanical behaviors have not been documented. PM was isolated in 13 cadaver specimens, permitting architectural measurements of physiological cross-sectional area (PCSA), normalized fiber length (Lf), and Lf:muscle length (Lm) ratio. Sarcomere lengths were measured in vivo from intraoperative biopsies taken with the hip joint in flexed and extended positions. Single-fiber and fiber bundle tensile properties and titin molecular weight were then measured from separate biopsies. Architecturally, average PCSA was 18.45 ± 1.32 cm2, average Lf was 12.70 ± 2 cm, and average Lf: Lm was 0.48 ± 0.06. Intraoperative sarcomere length measurements revealed that the muscle operates from 3.18 ± 0.20 μm with hip flexed at 10.7° ± 13.9° to 3.03 ± 0.22 μm with hip flexed at 55.9° ± 21.4°. Passive mechanical data demonstrated that the elastic modulus of the PM muscle fibers was 37.44 ± 9.11 kPa and of fiber bundles was 55.3 ± 11.8 kPa. Analysis of PM architecture demonstrates that its average Lf and passive biomechanical properties resemble those of the lumbar erector spinae muscles. In addition, PM sarcomere lengths were confined to the descending portion of the length-tension curve allowing the muscle to become stronger as the hip is flexed and the spine assumes a forward leaning posture. These findings suggest that the human PM has architectural and physiologic features that support its role as both a flexor of the hip and a dynamic stabilizer of the lumbar spine.

  17. Impact of a nickel-reduced stainless steel implant on striated muscle microcirculation: a comparative in vivo study.

    Science.gov (United States)

    Kraft, C N; Burian, B; Perlick, L; Wimmer, M A; Wallny, T; Schmitt, O; Diedrich, O

    2001-12-05

    The impairment of skeletal muscle microcirculation by a biomaterial may have profound consequences. With moderately good physical and corrosion characteristics, implant-quality stainless steel is particularly popular in orthopedic surgery. However, due to the presence of a considerable amount of nickel in the alloy, concern has been voiced in respect to local tissue responses. More recently a stainless steel alloy with a significant reduction of nickel has become commercially available. We, therefore, studied in vivo nutritive perfusion and leukocytic response of striated muscle to this nickel-reduced alloy, and compared these results with those of the materials conventional stainless steel and titanium. Using the hamster dorsal skinfold chamber preparation and intravital microscopy, we could demonstrate that reduction of the nickel quantity in a stainless steel implant has a positive effect on local microvascular parameters. Although the implantation of a conventional stainless steel sample led to a distinct and persistent activation of leukocytes combined with disruption of the microvascular endothelial integrity, marked leukocyte extravasation, and considerable venular dilation, animals with a nickel-reduced stainless steel implant showed only a moderate increase of these parameters, with a clear tendency of recuperation. Titanium implants merely caused a transient increase of leukocyte-endothelial cell interaction within the first 120 min, and no significant change in macromolecular leakage, leukocyte extravasation, or venular diameter. Pending biomechanical and corrosion testing, nickel-reduced stainless steel may be a viable alternative to conventional implant-quality stainless steel for biomedical applications. Concerning tolerance by the local vascular system, titanium currently remains unsurpassed. Copyright 2001 John Wiley & Sons, Inc. J Biomed Mater Res 57: 404-412, 2001

  18. Striated muscle activator of Rho signalling (STARS) is reduced in ageing human skeletal muscle and targeted by miR-628-5p.

    Science.gov (United States)

    Russell, A P; Wallace, M A; Kalanon, M; Zacharewicz, E; Della Gatta, P A; Garnham, A; Lamon, S

    2017-06-01

    The striated muscle activator of Rho signalling (STARS) is a muscle-specific actin-binding protein. The STARS signalling pathway is activated by resistance exercise and is anticipated to play a role in signal mechanotransduction. Animal studies have reported a negative regulation of STARS signalling with age, but such regulation has not been investigated in humans. Ten young (18-30 years) and 10 older (60-75 years) subjects completed an acute bout of resistance exercise. Gene and protein expression of members of the STARS signalling pathway and miRNA expression of a subset of miRNAs, predicted or known to target members of STARS signalling pathway, were measured in muscle biopsies collected pre-exercise and 2 h post-exercise. For the first time, we report a significant downregulation of the STARS protein in older subjects. However, there was no effect of age on the magnitude of STARS activation in response to an acute bout of exercise. Finally, we established that miR-628-5p, a miRNA regulated by age and exercise, binds to the STARS 3'UTR to directly downregulate its transcription. This study describes for the first time the resistance exercise-induced regulation of STARS signalling in skeletal muscle from older humans and identifies a new miRNA involved in the transcriptional control of STARS. © 2016 The Authors. Acta Physiologica published by John Wiley & Sons Ltd on behalf of Scandinavian Physiological Society.

  19. Role of mitochondria-cytoskeleton interactions in respiration regulation and mitochondrial organization in striated muscles.

    Science.gov (United States)

    Varikmaa, Minna; Bagur, Rafaela; Kaambre, Tuuli; Grichine, Alexei; Timohhina, Natalja; Tepp, Kersti; Shevchuk, Igor; Chekulayev, Vladimir; Metsis, Madis; Boucher, François; Saks, Valdur; Kuznetsov, Andrey V; Guzun, Rita

    2014-02-01

    The aim of this work was to study the regulation of respiration and energy fluxes in permeabilized oxidative and glycolytic skeletal muscle fibers, focusing also on the role of cytoskeletal protein tubulin βII isotype in mitochondrial metabolism and organization. By analyzing accessibility of mitochondrial ADP, using respirometry and pyruvate kinase-phosphoenolpyruvate trapping system for ADP, we show that the apparent affinity of respiration for ADP can be directly linked to the permeability of the mitochondrial outer membrane (MOM). Previous studies have shown that MOM permeability in cardiomyocytes can be regulated by VDAC interaction with cytoskeletal protein, βII tubulin. We found that in oxidative soleus skeletal muscle the high apparent Km for ADP is associated with low MOM permeability and high expression of non-polymerized βII tubulin. Very low expression of non-polymerized form of βII tubulin in glycolytic muscles is associated with high MOM permeability for adenine nucleotides (low apparent Km for ADP). © 2013.

  20. Expression and identification of 10 sarcomeric MyHC isoforms in human skeletal muscles of different embryological origin. Diversity and similarity in mammalian species.

    Science.gov (United States)

    Mascarello, Francesco; Toniolo, Luana; Cancellara, Pasqua; Reggiani, Carlo; Maccatrozzo, Lisa

    2016-09-01

    In the mammalian genome, among myosin heavy chain (MyHC) isoforms a family can be identified as sarcomeric based on their molecular structure which allows thick filament formation. In this study we aimed to assess the expression of the 10 sarcomeric isoforms in human skeletal muscles, adopting this species as a reference for comparison with all other mammalian species. To this aim, we set up the condition for quantitative Real Time PCR assay to detect and quantify MyHC mRNA expression in a wide variety of human muscles from somitic, presomitic and preotic origin. Specific patterns of expression of the following genes MYH1, MYH2, MYH3, MYH4, MYH6, MYH7, MYH8, MYH13, MYH14/7b and MYH15 were demonstrated in various muscle samples. On the same muscle samples which were analysed for mRNA expression, the corresponding MyHC proteins were studied with SDS PAGE and Western blot. The mRNA-protein comparison allowed the identification of 10 distinct proteins based on the electrophoretic migration rate. Three groups were formed based on the migration rate: fast migrating comprising beta/slow/1, alpha cardiac and fast 2B, slow migrating comprising fast 2X, fast 2A and two developmental isoforms (NEO and EMB), intermediate migrating comprising EO MyHC, slow B (product of MYH15), slow tonic (product of MYH14/7b). Of special interest was the demonstration of a protein band corresponding to 2B-MyHC in laryngeal muscles and the finding that all 10 isoforms are expressed in extraocular muscles. These latter muscles are the unique localization for extraocular, slow B (product of MYH15) and slow tonic (product of MYH14/7b). Copyright © 2016 Elsevier GmbH. All rights reserved.

  1. Protein Kinase CK2 Regulates Leukocyte-Endothelial Cell Interactions during Ischemia and Reperfusion in Striated Skin Muscle.

    Science.gov (United States)

    Ampofo, Emmanuel; Widmaier, Daniela; Montenarh, Mathias; Menger, Michael D; Laschke, Matthias W

    2016-01-01

    Ischemia and reperfusion (I/R) causes tissue injury by inflammatory processes. This involves the upregulation of endothelial surface proteins by phospho-regulated signaling pathways, resulting in enhanced interactions of leukocytes with endothelial cells. Recently, we found that protein kinase CK2 is a crucial regulator of leukocyte-mediated inflammation. Therefore, in this study we investigated the involvement of CK2 in leukocyte-endothelial cell interactions during I/R injury. We first analyzed the inhibitory action of (E)-3-(2,3,4,5-tetrabromophenyl)acrylic acid (TBCA) and CX-4945 on CK2 kinase activity and the viability of human dermal microvascular endothelial cells (HDMEC). To mimic I/R conditions in vitro, HDMEC were exposed to hypoxia and reoxygenation and the expression of adhesion molecules was analyzed by flow cytometry. Moreover, we analyzed in vivo the effect of CK2 inhibition on leukocyte-endothelial cell interactions in the dorsal skinfold chamber model of I/R injury by means of repetitive intravital fluorescence microscopy and immunohistochemistry. We found that TBCA and CX-4945 suppressed the activity of CK2 in HDMEC without affecting cell viability. This was associated with a significant downregulation of E-selectin and intercellular adhesion molecule (ICAM)-1 after in vitro hypoxia and reoxygenation. In vivo, CX-4945 treatment significantly decreased the numbers of adherent and transmigrated leukocytes in striated muscle tissue exposed to I/R. Our findings indicate that CK2 is involved in the regulation of leukocyte-endothelial cell interactions during I/R by mediating the expression of E-selectin and ICAM-1. © 2016 S. Karger AG, Basel.

  2. The length-tension curve in muscle depends on lattice spacing

    Energy Technology Data Exchange (ETDEWEB)

    Williams, C. D.; Salcedo, M. K.; Irving, T. C.; Regnier, M.; Daniel, T. L.

    2013-07-10

    Classic interpretations of the striated muscle length–tension curve focus on how force varies with overlap of thin (actin) and thick (myosin) filaments. New models of sarcomere geometry and experiments with skinned synchronous insect flight muscle suggest that changes in the radial distance between the actin and myosin filaments, the filament lattice spacing, are responsible for between 20% and 50% of the change in force seen between sarcomere lengths of 1.4 and 3.4 µm. Thus, lattice spacing is a significant force regulator, increasing the slope of muscle's force–length dependence.

  3. Isoform Composition and Gene Expression of Thick and Thin Filament Proteins in Striated Muscles of Mice after 30-Day Space Flight

    Directory of Open Access Journals (Sweden)

    Anna Ulanova

    2015-01-01

    Full Text Available Changes in isoform composition, gene expression of titin and nebulin, and isoform composition of myosin heavy chains as well as changes in titin phosphorylation level in skeletal (m. gastrocnemius, m. tibialis anterior, and m. psoas and cardiac muscles of mice were studied after a 30-day-long space flight onboard the Russian spacecraft “BION-M” number 1. A muscle fibre-type shift from slow-to-fast and a decrease in the content of titin and nebulin in the skeletal muscles of animals from “Flight” group was found. Using Pro-Q Diamond staining, an ~3-fold increase in the phosphorylation level of titin in m. gastrocnemius of mice from the “Flight” group was detected. The content of titin and its phosphorylation level in the cardiac muscle of mice from “Flight” and “Control” groups did not differ; nevertheless an increase (2.2 times in titin gene expression in the myocardium of flight animals was found. The observed changes are discussed in the context of their role in the contractile activity of striated muscles of mice under conditions of weightlessness.

  4. Tachykinins are involved in local reflex modulation of vagally mediated striated muscle contractions in the rat esophagus via tachykinin NK1 receptors.

    Science.gov (United States)

    Shiina, T; Shimizu, Y; Boudaka, A; Wörl, J; Takewaki, T

    2006-05-12

    The objective of the present study was to investigate the hypothesis of the presence of a local neural reflex modulating the vagally mediated contractions of striated muscle in the rat esophagus and to determine the possible involvement of tachykinins in such a local neural reflex. Electrical stimulation of the vagus nerve evoked twitch contractile responses that were abolished by d-tubocurarine (5 microM). Capsaicin (1-100 microM) inhibited the vagally mediated twitch contractions o f the normal rat esophageal preparations concentration-dependently but not those of the neonatally capsaicin-treated ones. NG-nitro-L-arginine methyl ester (100 microM), a nitric oxide synthase inhibitor, blocked the inhibitory effect of capsaicin and exogenous application of a nitric oxide donor (1 mM) inhibited the vagally mediated twitch contractions. Capsaicin suppressed acetylcholine release from the normal rat esophageal segments evoked by vagus nerve stimulation but not that from the neonatally capsaicin-treated ones. A selective tachykinin NK1 receptor antagonist (0.1 or 1 microM) attenuated the inhibitory effect of capsaicin. However, antagonists of tachykinin NK2, tachykinin NK3 and calcitonin gene-related peptide receptors (1 microM) did not have any effect. A tachykinin NK1 receptor agonist (1 or 5 microM) inhibited the vagally mediated twitch contractions, which was prevented by NG-nitro-L-arginine methyl ester (100 microM). These data suggest that the rat esophagus might have a local neural reflex inhibiting the vagally mediated striated muscle motility, which consists of capsaicin-sensitive sensory neurons and myenteric nitrergic neurons, and that tachykinins might be involved in the neural reflex through tachykinin NK1 receptors.

  5. Analysis of the ACTN3 heterozygous genotype suggests that α-actinin-3 controls sarcomeric composition and muscle function in a dose-dependent fashion.

    Science.gov (United States)

    Hogarth, Marshall W; Garton, Fleur C; Houweling, Peter J; Tukiainen, Taru; Lek, Monkol; Macarthur, Daniel G; Seto, Jane T; Quinlan, Kate G R; Yang, Nan; Head, Stewart I; North, Kathryn N

    2016-03-01

    A common null polymorphism (R577X) in ACTN3 causes α-actinin-3 deficiency in ∼ 18% of the global population. There is no associated disease phenotype, but α-actinin-3 deficiency is detrimental to sprint and power performance in both elite athletes and the general population. However, despite considerable investigation to date, the functional consequences of heterozygosity for ACTN3 are unclear. A subset of studies have shown an intermediate phenotype in 577RX individuals, suggesting dose-dependency of α-actinin-3, while others have shown no difference between 577RR and RX genotypes. Here, we investigate the effects of α-actinin-3 expression level by comparing the muscle phenotypes of Actn3(+/-) (HET) mice to Actn3(+/+) [wild-type (WT)] and Actn3(-/-) [knockout (KO)] littermates. We show reduction in α-actinin-3 mRNA and protein in HET muscle compared with WT, which is associated with dose-dependent up-regulation of α-actinin-2, z-band alternatively spliced PDZ-motif and myotilin at the Z-line, and an incremental shift towards oxidative metabolism. While there is no difference in force generation, HET mice have an intermediate endurance capacity compared with WT and KO. The R577X polymorphism is associated with changes in ACTN3 expression consistent with an additive model in the human genotype-tissue expression cohort, but does not influence any other muscle transcripts, including ACTN2. Overall, ACTN3 influences sarcomeric composition in a dose-dependent fashion in mouse skeletal muscle, which translates directly to function. Variance in fibre type between biopsies likely masks this phenomenon in human skeletal muscle, but we suggest that an additive model is the most appropriate for use in testing ACTN3 genotype associations. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  6. Seasonal changes in isoform composition of giant proteins of thick and thin filaments and titin (connectin) phosphorylation level in striated muscles of bears (Ursidae, Mammalia).

    Science.gov (United States)

    Salmov, N N; Vikhlyantsev, I M; Ulanova, A D; Gritsyna, Yu V; Bobylev, A G; Saveljev, A P; Makariushchenko, V V; Maksudov, G Yu; Podlubnaya, Z A

    2015-03-01

    Seasonal changes in the isoform composition of thick and thin filament proteins (titin, myosin heavy chains (MyHCs), nebulin), as well as in the phosphorylation level of titin in striated muscles of brown bear (Ursus arctos) and hibernating Himalayan black bear (Ursus thibetanus ussuricus) were studied. We found that the changes that lead to skeletal muscle atrophy in bears during hibernation are not accompanied by a decrease in the content of nebulin and intact titin-1 (T1) isoforms. However, a decrease (2.1-3.4-fold) in the content of T2 fragments of titin was observed in bear skeletal muscles (m. gastrocnemius, m. longissimus dorsi, m. biceps) during hibernation. The content of the stiffer N2B titin isoform was observed to increase relative to the content of its more compliant N2BA isoform in the left ventricles of hibernating bears. At the same time, in spite of the absence of decrease in the total content of T1 in the myocardium of hibernating brown bear, the content of T2 fragments decreased ~1.6-fold. The level of titin phosphorylation only slightly increased in the cardiac muscle of hibernating brown bear. In the skeletal muscles of brown bear, the level of titin phosphorylation did not vary between seasons. However, changes in the composition of MyHCs aimed at increasing the content of slow (I) and decreasing the content of fast (IIa) isoforms of this protein during hibernation of brown bear were detected. Content of MyHCs I and IIa in the skeletal muscles of hibernating Himalayan black bear corresponded to that in the skeletal muscles of hibernating brown bear.

  7. Characterization of the C-protein from posterior latissimus dorsi muscle of the adult chicken: heterogeneity within a single sarcomere

    OpenAIRE

    1983-01-01

    Specific isoforms of myofibrillar proteins are expressed in different muscles and in various fiber types within a single muscle. We have isolated and characterized monoclonal antibodies against C-proteins from slow tonic (anterior latissimus dorsi, ALD) and fast twitch (pectoralis major) muscles of the chicken. Although the antibody against "fast" C-protein (MF-1) did not bind to the "slow" isoform and the antibody to the "slow" C-protein (ALD-66) did not bind to the "fast" isoform, we observ...

  8. Spatially explicit, nano-mechanical models of the muscle half-sarcomere: Implications for biomechanical tuning in atrophy and fatigue

    Science.gov (United States)

    Kataoka, Aya; Tanner, Bertrand C. W.; Macpherson, J. Michael; Xu, Xiangrong; Wang, Qi; Regnier, Michael; Daniel, Thomas L.; Chase, P. Bryant

    2007-01-01

    Astronaut biomechanical performance depends on a wide variety of factors. Results from computational modelling suggest that muscle function—a key component of performance—could be modulated by compliance of the contractile filaments in muscle, especially when force is low such as transient Ca activation in a twitch, reduced activation in muscle fatigue encountered during EVA, or perhaps atrophy during prolonged space flight. We used Monte-Carlo models to investigate the hypotheses that myofilament compliance influences muscle function during a twitch, and also modulates the effects of cooperative interactions between contractile proteins on force generation. Peak twitch force and the kinetics of force decay were both decreased, while tension cost was increased, when myofilament compliance was increased relative to physiological values. Both the apparent Ca sensitivity and cooperativity of activation of steady-state isometric force were altered by myofilament compliance even when there were no explicit interactions included between binding sites. The effects of cooperative interactions between adjacent regulatory units were found to be greater than either the effect of myofilament compliance on apparent cooperativity of activation or that due to myosin cross-bridge-induced cooperativity. These results indicate that muscle function may be "tuned" at the molecular level, particularly under conditions of reduced Ca activation.

  9. Response of the JAK-STAT pathway to mammalian hibernation in 13-lined ground squirrel striated muscle.

    Science.gov (United States)

    Logan, Samantha M; Tessier, Shannon N; Tye, Joann; Storey, Kenneth B

    2016-03-01

    Over the course of the torpor-arousal cycle, hibernators must make behavioral, physiological, and molecular rearrangements in order to keep a very low metabolic rate and retain organ viability. 13-lined ground squirrels (Ictidomys tridecemlineatus) remain immobile during hibernation, and although the mechanisms of skeletal muscle survival are largely unknown, studies have shown minimal muscle loss in hibernating organisms. Additionally, the ground squirrel heart undergoes cold-stress, reversible cardiac hypertrophy, and ischemia-reperfusion without experiencing fatal impairment. This study examines the role of the Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway in the regulation of cell stress in cardiac and skeletal muscles, comparing euthermic and hibernating ground squirrels. Immunoblots showed a fivefold decrease in JAK3 expression during torpor in skeletal muscle, along with increases in STAT3 and 5 phosphorylation and suppressors of cytokine signaling-1 (SOCS1) protein levels. Immunoblots also showed coordinated increases in STAT1, 3 and 5 phosphorylation and STAT1 inhibitor protein expression in cardiac muscle during torpor. PCR analysis revealed that the activation of these pro-survival signaling cascades did not result in coordinate changes in downstream genes such as anti-apoptotic B-cell lymphoma-2 (Bcl-2) family gene expression. Overall, these results indicate activation of the JAK-STAT pathway in both cardiac and skeletal muscles, suggesting a response to cellular stress during hibernation.

  10. Characterization of muscle contraction with second harmonic generation microscopy

    Science.gov (United States)

    Prent, Nicole

    Muscle cells have the ability to change length and generate force due to orchestrated action of myosin nanomotors that cause sliding of actin filaments along myosin filaments in the sarcomeres, the fundamental contractile units, of myocytes. The correlated action of hundreds of sarcomeres is needed to produce the myocyte contractions. This study probes the molecular structure of the myofilaments and investigates the movement correlations between sarcomeres during contraction. In this study, second harmonic generation (SHG) microscopy is employed for imaging striated myocytes. Myosin filaments in striated myocytes inherently have a nonzero second-order susceptibility, [special characters omitted] and therefore generate efficient SHG. Employing polarization-in polarization-out (PIPO) SHG microscopy allows for the accurate determination of the characteristic ratio, [special characters omitted] in birefringent myocytes, which describes the structure of the myosin filament. Analysis shows that the b value at the centre of the myosin filament, where the nonlinear dipoles are better aligned, is slightly lower than the value at the edges of the filament, where there is more disorder in orientation of the nonlinear dipoles from the myosin heads. Forced stretching of myocytes resulted in an SHG intensity increase with the elongation of the sarcomere. SHG microscopy captured individual sarcomeres during contraction, allowing for the measurement of sarcomere length (SL) and SHG intensity (SI) fluctuations. The fluctuations also revealed higher SHG intensity in elongated sarcomeres. The sarcomere synchronization model (SSM) for contracting and quiescent myocytes was developed, and experimentally verified for three cases (isolated cardiomyocyte, embryonic chicken cardiomyocyte, and larva myocyte). During contraction, the action of SLs and SIs between neighbouring sarcomeres partially correlated, whereas in quiescent myocytes the SLs show an anti-correlation and the SIs have no

  11. Standardization of metachromatic staining method of myofibrillar ATPase activity of myosin to skeletal striated muscle of mules and donkeys

    Directory of Open Access Journals (Sweden)

    Flora H.F. D'Angelis

    2014-09-01

    Full Text Available This study aims at standardizing the pre-incubation and incubation pH and temperature used in the metachromatic staining method of myofibrillar ATPase activity of myosin (mATPase used for asses and mules. Twenty four donkeys and 10 mules, seven females and three males, were used in the study. From each animal, fragments from the Gluteus medius muscle were collected and percutaneous muscle biopsy was performed using a 6.0-mm Bergström-type needle. In addition to the metachromatic staining method of mATPase, the technique of nicotinamide adenine dinucleotide tetrazolium reductase (NADH-TR was also performed to confirm the histochemical data. The histochemical result of mATPase for acidic pre-incubation (pH=4.50 and alkaline incubation (pH=10.50, at a temperature of 37ºC, yielded the best differentiation of fibers stained with toluidine blue. Muscle fibers were identified according to the following colors: type I (oxidative, light blue, type IIA (oxidative-glycolytic, intermediate blue and type IIX (glycolytic, dark blue. There are no reports in the literature regarding the characterization and distribution of different types of muscle fibers used by donkeys and mules when performing traction work, cargo transportation, endurance sports (horseback riding and marching competitions. Therefore, this study is the first report on the standardization of the mATPase technique for donkeys and mules.

  12. Morphoquantitative effects on striated skeletal muscle of Wistar rats (Rattus norvegicus subjected to a diet utilized in young children from rural Mozambique

    Directory of Open Access Journals (Sweden)

    Catarina Tivane Nhamposse

    2016-12-01

    Full Text Available Mozambique is a country of sub-Saharan Africa where about 55% of the population lives below the absolute poverty line with less than one meal a day hardly surviving based on by donations. Food insecurity and precarious nutrition, especially in children, are factors that induce to levels of 44% of chronic malnutrition (CD in infants. The CD is responsible for one third of deaths in children under five years. The aim of this study was to evaluate the morphoquantitative effects in gastrocnemius muscle of Wistar rats fed with a diet utilized by people from rural areas of Mozambique. We used 75 Wistar rats weighing approximately 300 g divided in three groups: nourished or control (N, malnourished (D, and Mozambique or experimental group (M, measured at birth and at weaning. The animals were kept under the same housing conditions, temperature, humidity and light, but with different diets depending on the group: Group N with normal protein diet (20% casein, Group D with hypo-proteic diet (5% casein, and Group M with Mozambique diet. In all groups we evaluated the body mass at birth and weaning, and collected the right gastrocnemius muscle of male pups at weaning for analysis. Serial sections of 10 μm were performed in a cryostat prior to histology techniques of hematoxylin and eosin, picrosirius, NADH-tr and analysis in transmission electron microscope. Statistical evaluation was determined by analysis of variance (ANOVA and Tukey tests. Significant differences were found between groups N, D and M. In group M were observed a great variation of body mass that was approximately similar to group D; Group M also showed the same changes in muscle fiber which exhibited round-shaped contours, and predominance of type III collagen, similarly to malnourished group (D. Ultra-structurally, animals from Mozambique displayed a disorganization of the Z lines of sarcomeres, myofibrils disruption, decreased cross-sectional area and a smaller proportion of

  13. Microvascular response of striated muscle to metal debris. A comparative in vivo study with titanium and stainless steel.

    Science.gov (United States)

    Kraft, C N; Diedrich, O; Burian, B; Schmitt, O; Wimmer, M A

    2003-01-01

    Wear products of metal implants are known to induce biological events which may have profound consequences for the microcirculation of skeletal muscle. Using the skinfold chamber model and intravital microscopy we assessed microcirculatory parameters in skeletal muscle after confrontation with titanium and stainless-steel wear debris, comparing the results with those of bulk materials. Implantation of stainless-steel bulk and debris led to a distinct activation of leukocytes combined with a disruption of the microvascular endothelial integrity and massive leukocyte extravasation. While animals with bulk stainless steel showed a tendency to recuperation, stainless-steel wear debris induced such severe inflammation and massive oedema that the microcirculation broke down within 24 hours after implantation. Titanium bulk caused only a transient increase in leukocyte-endothelial cell interaction within the first 120 minutes and no significant change in macromolecular leakage, leukocyte extravasation or venular diameter. Titanium wear debris produced a markedly lower inflammatory reaction than stainless-steel bulk, indicating that a general benefit of bulk versus debris could not be claimed. Depending on its constituents, wear debris is capable of eliciting acute inflammation which may result in endothelial damage and subsequent failure of microperfusion. Our results indicate that not only the bulk properties of orthopaedic implants but also the microcirculatory implications of inevitable wear debris play a pivotal role in determining the biocompatibility of an implant.

  14. Determining the role of sarcomeric proteins in facioscapulohumeral muscular dystrophy: a study protocol

    NARCIS (Netherlands)

    Lassche, S.; Ottenheijm, C.A.C.; Voermans, N.C.; Westeneng, H.J.; Janssen, B.H.; Maarel, S.M. van der; Hopman, M.T.E.; Padberg, G.W.A.M.; Stienen, G.J.; Engelen, B.G.M. van

    2013-01-01

    BACKGROUND: Although muscle weakness is a hallmark of facioscapulohumeral muscular dystrophy (FSHD), the molecular mechanisms that lead to weakness in FSHD remain largely unknown. Recent studies suggest aberrant expression of genes involved in skeletal muscle development and sarcomere contractility,

  15. Dynamic Alterations to α-Actinin Accompanying Sarcomere Disassembly and Reassembly during Cardiomyocyte Mitosis.

    Directory of Open Access Journals (Sweden)

    Xiaohu Fan

    Full Text Available Although mammals are thought to lose their capacity to regenerate heart muscle shortly after birth, embryonic and neonatal cardiomyocytes in mammals are hyperplastic. During proliferation these cells need to selectively disassemble their myofibrils for successful cytokinesis. The mechanism of sarcomere disassembly is, however, not understood. To study this, we performed a series of immunofluorescence studies of multiple sarcomeric proteins in proliferating neonatal rat ventricular myocytes and correlated these observations with biochemical changes at different cell cycle stages. During myocyte mitosis, α-actinin and titin were disassembled as early as prometaphase. α-actinin (representing the sarcomeric Z-disk disassembly precedes that of titin (M-line, suggesting that titin disassembly occurs secondary to the collapse of the Z-disk. Sarcomere disassembly was concurrent with the dissolution of the nuclear envelope. Inhibitors of several intracellular proteases could not block the disassembly of α-actinin or titin. There was a dramatic increase in both cytosolic (soluble and sarcomeric α-actinin during mitosis, and cytosolic α-actinin exhibited decreased phosphorylation compared to sarcomeric α-actinin. Inhibition of cyclin-dependent kinase 1 (CDK1 induced the quick reassembly of the sarcomere. Sarcomere dis- and re-assembly in cardiomyocyte mitosis is CDK1-dependent and features dynamic differential post-translational modifications of sarcomeric and cytosolic α-actinin.

  16. The Effect of the Wooden Breast Myopathy on Sarcomere Structure and Organization.

    Science.gov (United States)

    Velleman, Sandra G; Clark, Daniel L; Tonniges, Jeffrey R

    2018-03-01

    The wooden breast (WB) has been classically identified by the phenotypic presence of a wood-like pectoralis major (p. major) muscle. The WB-affected p. major muscle is characterized by necrotic muscle fibers and the replacement of muscle with connective tissue, water, and fat. The objective of the current study was to determine the effect of the WB myopathy on sarcomere organization by transmission electron microscopy. Sarcomere structure and organization were examined in two broiler lines with a high incidence of WB (Lines A and B) and another broiler line without WB (Line C). Affected muscle had an increase in smaller myofibers with diameters of 20 μm or less. Sarcomere organization decreased with fiber diameter in both Lines A and B. The structure and organization of sarcomeres in Line C were similar to WB-unaffected muscle in Lines A and B. Taken together, these data demonstrate that the WB myopathy detrimentally affects sarcomere organization in a broiler line-specific manner. Disorganization of sarcomere structure will affect the function of the p. major muscle as well as meat quality.

  17. Fiber types in the striated urethral and anal sphincters

    DEFF Research Database (Denmark)

    Schrøder, H D; Reske-Nielsen, E

    1983-01-01

    Seven normal human striated urethral and anal sphincters obtained by autopsy were examined using histochemical techniques. In both the urethral sphincter and the subcutaneous (s.c.) and superficial part of the anal sphincter a characteristic pattern with two populations of muscle fibers, abundant...

  18. Effects of BTS (N-benzyl-p-toluene sulphonamide), an inhibitor for myosin-actin interaction, on myofibrillogenesis in skeletal muscle cells in culture.

    Science.gov (United States)

    Kagawa, Maiko; Sato, Naruki; Obinata, Takashi

    2006-11-01

    Actin filaments align around myosin filaments in the correct polarity and in a hexagonal arrangement to form cross-striated structures. It has been postulated that this myosin-actin interaction is important in the initial phase of myofibrillogenesis. It was previously demonstrated that an inhibitor of actin-myosin interaction, BDM (2,3-butanedione monoxime), suppresses myofibril formation in muscle cells in culture. However, further study showed that BDM also exerts several additional effects on living cells. In this study, we further examined the role of actin-myosin interaction in myofibril assembly in primary cultures of chick embryonic skeletal muscle by applying a more specific inhibitor, BTS (N-benzyl-p-toluene sulphonamide), of myosin ATPase and actin-myosin interaction. The assembly of sarcomeric structures from myofibrillar proteins was examined by immunocytochemical methods with the application of BTS to myotubes just after fusion. Addition of BTS (10-50 microM) significantly suppressed the organization of actin and myosin into cross-striated structures. BTS also interfered in the organization of alpha-actinin, C-protein (or MyBP-C), and connectin (or titin) into ordered striated structures, though the sensitivity was less. Moreover, when myotubes cultured in the presence of BTS were transferred to a control medium, sarcomeric structures were formed in 2-3 days, indicating that the inhibitory effect of BTS on myotubes is reversible. These results show that actin-myosin interaction plays a critical role in the process of myofibrillogenesis.

  19. Mechanical properties of non-sarcomeric cytoskeleton of mice cardiomyocytes and skeletal muscle fibers after 30-day spaceflight biosatellite BION-M1

    Science.gov (United States)

    Ogneva, Irina; Maximova, Maria; Larina, Irina

    The aim of this study was to determine transversal stiffness of the cortical cytoskeleton and cytoskeletal protein desmin content of the left ventricle cardiomyocytes, soleus and tibialis anterior muscle fibers of the mice after 30-days space flight biosatellite «BION-M1» (Russia, 2013). The dissection was made after 13-16.5 hours after landing. Transversal stiffness was measured in relaxed and calcium activated state by atomic force microscope. Desmin content was estimated by using western-blot, expression level of the gene, coding desmin, - by real time PCR. The transversal stiffness of the cortical cytoskeleton of the cardiomyocytes and fibers of the skeletal muscles (as measured using the atomic force microscopy) did not differ significantly within the study groups in comparison to the vivarium control group, except for its slight increase in tibialis anterior fibers muscle in the post-flight group of animals. The content of beta- and gamma-actin in the membranous fraction of proteins in the left ventricular cardiomyocytes (as detected using the western blot technique) did not differ significantly within all study groups and correlated with the transversal stiffness. Similar situation was revealed in fibers of the soleus muscle and tibialis anterior muscle, as well as correlation with the transversal stiffness of their cortical cytoskeleton was noted. At the same time, the content of beta-actin in the cytoplasmic fraction of proteins was found to be decreased in all types of studied tissues in comparison to the control levels in the post-flight group, as well as lowered beta-actin gene expression rates in the post-flight group of animals (as detected using the RT-PCR technique). After completion of the space flight, content of alpha-actinin-4 was found to be reduced in the membranous fraction of proteins of mouse cardiomyocytes, while its content in the cytoplasmic fraction of proteins did not change significantly. Furthermore, gene expression rates of this

  20. Dose-dependent separation of the hypertrophic and myotoxic effects of the β2-adrenergic receptor agonist clenbuterol in rat striated muscles.

    Science.gov (United States)

    Burniston, Jatin G; WA, Clark; Tan, Lip-Bun; Goldspink, David F

    2007-01-01

    Muscle growth in response to large doses (i.e., mg.kg-1) of β2-adrenergic receptor agonists has been consistently reported. However, such doses may also induce myocyte death in the heart and skeletal muscles and hence may not be applicable safe doses for humans. Here, we report the hypertrophic and myotoxic effects of different doses of clenbuterol. Rats were infused with clenbuterol (range, 1 μg to 1 mg.kg-1) for 14 days. Muscle protein content, myofiber cross-sectional area and myocyte death were then investigated. Infusions of ≥10 μg.kg-1.d-1 of clenbuterol significantly (Pclenbuterol in the absence of myocyte death. PMID:16411205

  1. The evolutionary origin of bilaterian smooth and striated myocytes

    Science.gov (United States)

    Brunet, Thibaut; Fischer, Antje HL; Steinmetz, Patrick RH; Lauri, Antonella; Bertucci, Paola; Arendt, Detlev

    2016-01-01

    The dichotomy between smooth and striated myocytes is fundamental for bilaterian musculature, but its evolutionary origin is unsolved. In particular, interrelationships of visceral smooth muscles remain unclear. Absent in fly and nematode, they have not yet been characterized molecularly outside vertebrates. Here, we characterize expression profile, ultrastructure, contractility and innervation of the musculature in the marine annelid Platynereis dumerilii and identify smooth muscles around the midgut, hindgut and heart that resemble their vertebrate counterparts in molecular fingerprint, contraction speed and nervous control. Our data suggest that both visceral smooth and somatic striated myocytes were present in the protostome-deuterostome ancestor and that smooth myocytes later co-opted the striated contractile module repeatedly – for example, in vertebrate heart evolution. During these smooth-to-striated myocyte conversions, the core regulatory complex of transcription factors conveying myocyte identity remained unchanged, reflecting a general principle in cell type evolution. DOI: http://dx.doi.org/10.7554/eLife.19607.001 PMID:27906129

  2. Three-dimensional stochastic model of actin–myosin binding in the sarcomere lattice

    Energy Technology Data Exchange (ETDEWEB)

    Mijailovich, Srboljub M.; Kayser-Herold, Oliver; Stojanovic, Boban; Nedic, Djordje; Irving, Thomas C.; Geeves, MA (Harvard); (IIT); (U. Kent); (Kragujevac)

    2016-11-18

    The effect of molecule tethering in three-dimensional (3-D) space on bimolecular binding kinetics is rarely addressed and only occasionally incorporated into models of cell motility. The simplest system that can quantitatively determine this effect is the 3-D sarcomere lattice of the striated muscle, where tethered myosin in thick filaments can only bind to a relatively small number of available sites on the actin filament, positioned within a limited range of thermal movement of the myosin head. Here we implement spatially explicit actomyosin interactions into the multiscale Monte Carlo platform MUSICO, specifically defining how geometrical constraints on tethered myosins can modulate state transition rates in the actomyosin cycle. The simulations provide the distribution of myosin bound to sites on actin, ensure conservation of the number of interacting myosins and actin monomers, and most importantly, the departure in behavior of tethered myosin molecules from unconstrained myosin interactions with actin. In addition, MUSICO determines the number of cross-bridges in each actomyosin cycle state, the force and number of attached cross-bridges per myosin filament, the range of cross-bridge forces and accounts for energy consumption. At the macroscopic scale, MUSICO simulations show large differences in predicted force-velocity curves and in the response during early force recovery phase after a step change in length comparing to the two simplest mass action kinetic models. The origin of these differences is rooted in the different fluxes of myosin binding and corresponding instantaneous cross-bridge distributions and quantitatively reflects a major flaw of the mathematical description in all mass action kinetic models. Consequently, this new approach shows that accurate recapitulation of experimental data requires significantly different binding rates, number of actomyosin states, and cross-bridge elasticity than typically used in mass action kinetic models to

  3. The Time Course of the Loss and Recovery of Contracture Ability in Frog Striated Muscle Following Exposure to Ca-Free Solutions

    Science.gov (United States)

    Milligan, J. V.

    1965-01-01

    Using area under the contracture curve to quantitate contractures, the diffusion coefficient of calcium ions within the frog toe muscle during washout in a calcium-free solution and subsequent recovery after reintroduction of calcium to the bathing solution was calculated to be about 2 x 10-6 cm2/sec. The diffusion coefficient measured during washout was found to be independent of temperature or initial calcium ion concentration. During recovery it was found to decrease if the temperature was lowered. This was likely due to the repolarization occurring after the depolarizing effect of the calcium-free solution. The relation between contracture area and [Ca]o was found to be useful over a wider range than that between maximum tension and [Ca]o. The normalized contracture areas were larger at lower calcium concentrations if the contractures were produced with cold potassium solutions or if NO3 replaced Cl in the bathing solutions. Decreasing the potassium concentration of the contracture solution to 50 mM from 115 mM did not change the relation between [Ca]o and the normalized area. If the K concentration of the bathing solution was increased, the areas were decreased at lower concentrations of Ca. PMID:14324991

  4. Interstitial cells of Cajal in the striated musculature of the mouse esophagus

    DEFF Research Database (Denmark)

    Rumessen, J J; de Kerchove d'Exaerde, A; Mignon, S

    2001-01-01

    Interstitial cells of Cajal (ICC) are important regulatory cells in the smooth muscle coats of the digestive tract. Expression of the Kit receptor tyrosine kinase was used in this study as a marker to study their distribution and development in the striated musculature of the mouse esophagus...... scarce in both muscle layers of the thoracic esophagus, while their number increased steeply toward the cardia in the striated portion of the intraabdominal esophagus. They did not form networks and had no relationship with intrinsic myenteric ganglia and motor end-plates. They were often close to nerve...... but absent in adult ICC-deficient KitW-lacZ/KitWv mice. Interstitial cells of Cajal were identified by electron microscopy by their ultrastructure in the striated muscle of the esophagus and exhibited Xgal labeling, while fibroblasts and muscle cells were unlabeled. Interstitial cells of Cajal are scattered...

  5. Disposition of the striated urethral sphincter and its relation to the prostate in human fetuses

    Directory of Open Access Journals (Sweden)

    Luciano A. Favorito

    2007-06-01

    Full Text Available OBJECTIVE: To describe the arrangement of the muscle fibers of the striated urethral sphincter and its relationship with the prostate during the fetal period in humans. MATERIALS AND METHODS: We analyzed 17 prostates from well preserved fresh human fetuses ranging in age from 10 to 31 weeks postconception (WPC. Transversal sections were obtained and stained with Gomori's trichrome and immunolabeled with anti alpha-actin antibody. RESULTS: We found that the urethral striated sphincter (rabdosphincter is located on the periphery of the smooth muscle and there was no merge between striated and smooth muscle fibers in any fetal period. In the prostate apex, the striated sphincter shows a circular arrangement and covers completely the urethra externally, whereas adjacent to verumontanum, it looks like a "horseshoe" and covers only the anterior and lateral surfaces of the urethra. Near the bladder neck, in fetuses younger than 20 WPC, we have found striated muscle fibers only at the anterior surface of the prostate, while in fetuses older than 20 WPC, the striated muscle covers the anterior and lateral surfaces of the prostate. CONCLUSIONS: The urethral sphincter muscle covers the anterior and lateral surfaces of the urethra in all fetuses older than 20 WPC, close to the bladder neck and at the distal prostate. In the region of the prostate apex, the urethral sphincter covers completely the urethra circularly. The knowledge of the normal anatomy of the urethral sphincter in fetuses could be important to understand its alterations in congenital anomalies involving the base of the bladder, the bladder neck and the proximal urethra.

  6. Quantitative comparison of striated toolmarks.

    Science.gov (United States)

    Baiker, Martin; Keereweer, Isaac; Pieterman, René; Vermeij, Erwin; van der Weerd, Jaap; Zoon, Peter

    2014-09-01

    A comparison of striated toolmarks by human examiners is dependent on the experience of the expert and includes a subjective judgment within the process. In this article an automated method is presented for objective comparison of striated marks of screwdrivers. The combination of multi-scale registration (alignment) of toolmarks, that accounts for shift and scaling, with global cross correlation as objective toolmark similarity metric renders the approach robust with respect to large differences in angle of attack and moderate toolmark compression. In addition, a strategy to distinguish between relevant and non-relevant spatial frequency ranges (geometric details) is presented. The performance of the method is evaluated using 3D topography scans of experimental toolmarks of 50 unused screwdrivers. Known match and known non-match similarity distributions are estimated including a large range of angles of attack (15, 30, 45, 60 and 75°) for the known matches. It is demonstrated that the system has high discriminatory power, even if the toolmarks are made at a difference in angle of attack of larger than 15°. The probability distributions are subsequently employed to determine likelihood ratios. A comparison of the results of the automated method with the outcome of a toolmark comparison experiment involving three experienced toolmark examiners reveals, that the automated system is more powerful in correctly supporting the hypothesis of common origin for toolmarks with a large difference in angle of attack (30°). In return, the rate of toolmark comparisons that yield incorrect support for the hypothesis of common origin is higher for the automated system. In addition, a comparison between estimating known match and known non-match distributions using 2D and 3D data is presented and it is shown that for toolmarks of unused screwdrivers, relying on 3D is slightly better than relying on 2D data. Finally, a comparison between estimating known match and known non

  7. Abba is an essential TRIM/RBCC protein to maintain the integrity of sarcomeric cytoarchitecture.

    Science.gov (United States)

    Domsch, Katrin; Ezzeddine, Nader; Nguyen, Hanh T

    2013-08-01

    Organized sarcomeric striations are an evolutionarily conserved hallmark of functional skeletal muscles. Here, we demonstrate that the Drosophila Abba protein, a member of the TRIM/RBCC superfamily, has a pivotal regulatory role in maintaining proper sarcomeric cytoarchitecture during development and muscle usage. abba mutant embryos initially form muscles, but F-actin and Myosin striations become progressively disrupted when the muscles undergo growth and endure increased contractile forces during larval development. Abnormal Myosin aggregates and myofiber atrophy are also notable in the abba mutants. The larval defects result in compromised muscle function, and hence important morphogenetic events do not occur properly during pupation, leading to lethality. Abba is localized at larval Z-discs, and genetic evidence indicates that abba interacts with α-actinin, kettin/D-titin and mlp84B, genes that encode important Z-disc proteins for stable myofibrillar organization and optimal muscle function. RNAi experiments and ultrastructural analysis reveal that Abba has an additional crucial role in sarcomere maintenance in adult muscles. Abba is required to ensure the integrity and function of Z-discs and M-lines. Rescue experiments further show that Abba function is dependent upon its B-box/coiled-coil domain, NHL repeats and RING finger domain. The importance of these presumed protein-protein interactions and ubiquitin ligase-associated domains supports our hypothesis that Abba is needed for specific protein complex formation and stabilization at Z-discs and M-lines.

  8. Finite element modeling of aponeurotomy: altered intramuscular myofascial force transmission yields complex sarcomere length distributions determining acute effects

    NARCIS (Netherlands)

    Yucesoy, C.A.; Koopman, B.H.; Grootenboer, H.J.; Huijing, P.A.J.B.M.

    2007-01-01

    Finite element modeling of aponeurotomized rat extensor digitorium longus muscle was performed to investigate the acute effects of proximal aponeurotomy. The specific goal was to assess the changes in lengths of sarcomeres within aponeurotomized muscle and to explain how the intervention leads to

  9. Elastic energy storage and radial forces in the myofilament lattice depend on sarcomere length.

    Directory of Open Access Journals (Sweden)

    C David Williams

    Full Text Available We most often consider muscle as a motor generating force in the direction of shortening, but less often consider its roles as a spring or a brake. Here we develop a fully three-dimensional spatially explicit model of muscle to isolate the locations of forces and energies that are difficult to separate experimentally. We show the strain energy in the thick and thin filaments is less than one third the strain energy in attached cross-bridges. This result suggests the cross-bridges act as springs, storing energy within muscle in addition to generating the force which powers muscle. Comparing model estimates of energy consumed to elastic energy stored, we show that the ratio of these two properties changes with sarcomere length. The model predicts storage of a greater fraction of energy at short sarcomere lengths, suggesting a mechanism by which muscle function shifts as force production declines, from motor to spring. Additionally, we investigate the force that muscle produces in the radial or transverse direction, orthogonal to the direction of shortening. We confirm prior experimental estimates that place radial forces on the same order of magnitude as axial forces, although we find that radial forces and axial forces vary differently with changes in sarcomere length.

  10. Interstitial cells of Cajal in the striated musculature of the mouse esophagus

    DEFF Research Database (Denmark)

    Rumessen, J J; de Kerchove d'Exaerde, A; Mignon, S

    2001-01-01

    Interstitial cells of Cajal (ICC) are important regulatory cells in the smooth muscle coats of the digestive tract. Expression of the Kit receptor tyrosine kinase was used in this study as a marker to study their distribution and development in the striated musculature of the mouse esophagus. Sec...

  11. Geometrical Conditions Indispensable for Muscle Contraction

    Directory of Open Access Journals (Sweden)

    Ludmila Skubiszak

    2011-03-01

    Full Text Available Computer simulation has uncovered the geometrical conditions under which the vertebrate striated muscle sarcomere can contract. First, all thick filaments should have identical structure, namely: three myosin cross-bridges, building a crown, should be aligned at angles of 0°, 120°, 180°, and the successive crowns and the two filament halves should be turned around 120°. Second, all thick filaments should act simultaneously. Third, coordination in action of the myosin cross-bridges should exist, namely: the three cross-bridges of a crown should act simultaneously and the cross-bridge crowns axially 43 and 14.333 nm apart should act, respectively, simultaneously and with a phase shift. Fifth, six thin filaments surrounding the thick filament should be turned around 180° to each other in each sarcomere half. Sixth, thin filaments should be oppositely oriented in relation to the sarcomere middle. Finally, the structure of each of the thin filaments should change in consequence of strong interaction with myosin heads, namely: the axial distance and the angular alignment between neighboring actin monomers should be, respectively, 2.867 nm and 168° instead of 2.75 nm and 166.15°. These conditions ensure the stereo-specific interaction between actin and myosin and good agreement with the data gathered by electron microscopy and X-ray diffraction methods. The results suggest that the force is generated not only by the myosin cross-bridges but also by the thin filaments; the former acts by cyclical unwrapping and wrapping the thick filament backbone, and the latter byelongation.

  12. Electrostatic forces in muscle and cylindrical gel systems

    Energy Technology Data Exchange (ETDEWEB)

    Millman, B.M.; Nickel, B.G.

    1980-10-01

    Repulsive pressure has been measured as a function of lattice spacing in gels of tobacco mosaic virus (TMV) and in the filament lattice of vertebrate striated muscle. External pressures up to ten atm have been applied to these lattices by an osmotic stress method. Numerical solutions to the Poisson-Boltzmann equation in hexagonal lattices have been obtained and compared to the TMV and muscle data. The theoretical curves using values for kappa calculated from the ionic strength give a good fit to experimental data from TMV gels, and an approximate fit to that from the muscle lattice, provided that a charge radius for the muscle thick filaments of approx. 16 nm is assumed. Variations in ionic strength, sarcomere length and state of the muscle give results which agree qualitatively with the theory, though a good fit between experiment and theory in the muscle case will clearly require consideration of other types of forces. We conclude that Poisson-Boltzmann theory can provide a good first approximation to the long-range electrostatic forces operating in such biological gel systems.

  13. Mechanical strength of sarcomere structures of skeletal myofibrils studied by submicromanipulation.

    Science.gov (United States)

    Kayamori, Taisuke; Miyake, Norihito; Akiyama, Nao; Aimi, Momoko; Wakayama, Jun'ichi; Kunioka, Yuki; Yamada, Takenori

    2006-01-01

    The mechanical strength of sarcomere structures of skeletal muscle was studied by rupturing single myofibrils of rabbit psoas muscle by submicromanipulation techniques. Microbeads coated with alpha-actinin were attached to the surface of myofibrils immobilized to coverslip. By use of either optical tweezers or atomic force microscope, the attached beads were captured and detached from the myofibrils. During the detachment of the beads, the actin filaments bound specifically to the beads were peeled off from the bulk structures of myofibrils, thus rupturing the peripheral components of the myofibrils bound to the actin filaments. By analyzing the ruptures thus produced in various myofibril preparations, it was found that the sarcomere structure of myofibrils is maintained by numerous molecular components having the mechanical strength sufficient to sustain the contractile force produced by the actomyosin system. The present techniques could be applied to study the mechanical strength of cellular organelles containing actin filaments as their component.

  14. Hamstring contractures in children with spastic cerebral palsy result from a stiffer extracellular matrix and increased in vivo sarcomere length.

    Science.gov (United States)

    Smith, Lucas R; Lee, Ki S; Ward, Samuel R; Chambers, Henry G; Lieber, Richard L

    2011-05-15

    Cerebral palsy (CP) results from an upper motoneuron (UMN)lesion in the developing brain. Secondary to the UMNl esion,which causes spasticity, is a pathological response by muscle - namely, contracture. However, the elements within muscle that increase passive mechanical stiffness, and therefore result in contracture, are unknown. Using hamstring muscle biopsies from pediatric patients with CP (n =33) and control (n =19) patients we investigated passive mechanical properties at the protein, cellular, tissue and architectural levels to identify the elements responsible for contracture. Titin isoform, the major load-bearing protein within muscle cells, was unaltered in CP. Correspondingly, the passive mechanics of individual muscle fibres were not altered. However, CP muscle bundles, which include fibres in their constituent ECM, were stiffer than control bundles. This corresponded to an increase in collagen content of CP muscles measured by hydroxyproline assay and observed using immunohistochemistry. In vivo sarcomere length of CP muscle measured during surgery was significantly longer than that predicted for control muscle. The combination of increased tissue stiffness and increased sarcomere length interact to increase stiffness greatly of the contracture tissue in vivo. These findings provide evidence that contracture formation is not the result of stiffening at the cellular level, but stiffening of the ECM with increased collagen and an increase of in vivo sarcomere length leading to higher passive stresses.

  15. Muscle RING‐finger 2 and 3 maintain striated‐muscle structure and function

    Science.gov (United States)

    Lodka, Dörte; Pahuja, Aanchal; Geers‐Knörr, Cornelia; Scheibe, Renate J.; Nowak, Marcel; Hamati, Jida; Köhncke, Clemens; Purfürst, Bettina; Kanashova, Tamara; Schmidt, Sibylle; Glass, David J.; Morano, Ingo; Heuser, Arnd; Kraft, Theresia; Bassel‐Duby, Rhonda; Olson, Eric N.; Dittmar, Gunnar; Sommer, Thomas

    2015-01-01

    Abstract Background The Muscle‐specific RING‐finger (MuRF) protein family of E3 ubiquitin ligases is important for maintenance of muscular structure and function. MuRF proteins mediate adaptation of striated muscles to stress. MuRF2 and MuRF3 bind to microtubules and are implicated in sarcomere formation with noticeable functional redundancy. However, if this redundancy is important for muscle function in vivo is unknown. Our objective was to investigate cooperative function of MuRF2 and MuRF3 in the skeletal muscle and the heart in vivo. Methods MuRF2 and MuRF3 double knockout mice (DKO) were generated and phenotypically characterized. Skeletal muscle and the heart were investigated by morphological measurements, histological analyses, electron microscopy, immunoblotting, and real‐time PCR. Isolated muscles were subjected to in vitro force measurements. Cardiac function was determined by echocardiography and working heart preparations. Function of cardiomyocytes was measured in vitro. Cell culture experiments and mass‐spectrometry were used for mechanistic analyses. Results DKO mice showed a protein aggregate myopathy in skeletal muscle. Maximal force development was reduced in DKO soleus and extensor digitorum longus. Additionally, a fibre type shift towards slow/type I fibres occurred in DKO soleus and extensor digitorum longus. MuRF2 and MuRF3‐deficient hearts showed decreased systolic and diastolic function. Further analyses revealed an increased expression of the myosin heavy chain isoform beta/slow and disturbed calcium handling as potential causes for the phenotype in DKO hearts. Conclusions The redundant function of MuRF2 and MuRF3 is important for maintenance of skeletal muscle and cardiac structure and function in vivo. PMID:27493870

  16. Sarcomere Control Mechanisms and the Dynamics of the Cardiac Cycle

    Directory of Open Access Journals (Sweden)

    R. John Solaro

    2010-01-01

    Full Text Available This review focuses on recent developments in the molecular mechanisms by which Ca activates cardiac sarcomeres and how these mechanisms play out in the cardiac cycle. I emphasize the role of mechanisms intrinsic to the sarcomeres as significant determinants of systolic elastance and ventricular stiffening during ejection. Data are presented supporting the idea that processes intrinsic to the thin filaments may promote cooperative activation of the sarcomeres and be an important factor in maintaining and modifying systolic elastance. Application of these ideas to translational medicine and rationale drug design forms an important rationale for detailed understanding of these processes.

  17. Bones, Muscles, and Joints: The Musculoskeletal System

    Science.gov (United States)

    ... Skeletal muscles are called striated (pronounced: STRY-ay-ted) because they are made up of fibers that ... blood through your body. When we smile and talk, muscles are helping us communicate, and when we ...

  18. Contractile Dysfunction in Sarcomeric Hypertrophic Cardiomyopathy.

    Science.gov (United States)

    MacIver, David H; Clark, Andrew L

    2016-09-01

    The pathophysiological mechanisms underlying the clinical phenotype of sarcomeric hypertrophic cardiomyopathy are controversial. The development of cardiac hypertrophy in hypertension and aortic stenosis is usually described as a compensatory mechanism that normalizes wall stress. We suggest that an important abnormality in hypertrophic cardiomyopathy is reduced contractile stress (the force per unit area) generated by myocardial tissue secondary to abnormalities such as cardiomyocyte disarray. In turn, a progressive deterioration in contractile stress provokes worsening hypertrophy and disarray. A maintained or even exaggerated ejection fraction is explained by the increased end-diastolic wall thickness producing augmented thickening. We propose that the nature of the hemodynamic load in an individual with hypertrophic cardiomyopathy could determine its phenotype. Hypertensive patients with hypertrophic cardiomyopathy are more likely to develop exaggerated concentric hypertrophy; athletic individuals an asymmetric pattern; and inactive individuals a more apical hypertrophy. The development of a left ventricular outflow tract gradient and mitral regurgitation may be explained by differential regional strain resulting in mitral annular rotation. Copyright © 2016. Published by Elsevier Inc.

  19. Distinctive serum miRNA profile in mouse models of striated muscular pathologies.

    Directory of Open Access Journals (Sweden)

    Nicolas Vignier

    Full Text Available Biomarkers are critically important for disease diagnosis and monitoring. In particular, close monitoring of disease evolution is eminently required for the evaluation of therapeutic treatments. Classical monitoring methods in muscular dystrophies are largely based on histological and molecular analyses of muscle biopsies. Such biopsies are invasive and therefore difficult to obtain. The serum protein creatine kinase is a useful biomarker, which is however not specific for a given pathology and correlates poorly with the severity or course of the muscular pathology. The aim of the present study was the systematic evaluation of serum microRNAs (miRNAs as biomarkers in striated muscle pathologies. Mouse models for five striated muscle pathologies were investigated: Duchenne muscular dystrophy (DMD, limb-girdle muscular dystrophy type 2D (LGMD2D, limb-girdle muscular dystrophy type 2C (LGMD2C, Emery-Dreifuss muscular dystrophy (EDMD and hypertrophic cardiomyopathy (HCM. Two-step RT-qPCR methodology was elaborated, using two different RT-qPCR miRNA quantification technologies. We identified miRNA modulation in the serum of all the five mouse models. The most highly dysregulated serum miRNAs were found to be commonly upregulated in DMD, LGMD2D and LGMD2C mouse models, which all exhibit massive destruction of striated muscle tissues. Some of these miRNAs were down rather than upregulated in the EDMD mice, a model without massive myofiber destruction. The dysregulated miRNAs identified in the HCM model were different, with the exception of one dysregulated miRNA common to all pathologies. Importantly, a specific and distinctive circulating miRNA profile was identified for each studied pathological mouse model. The differential expression of a few dysregulated miRNAs in the DMD mice was further evaluated in DMD patients, providing new candidates of circulating miRNA biomarkers for DMD.

  20. Sarcomeres pattern proprioceptive sensory dendritic endings through Perlecan/UNC-52 in C. elegans

    Science.gov (United States)

    Liang, Xing; Dong, Xintong; Moerman, Donald G.; Shen, Kang; Wang, Xiangming

    2015-01-01

    Sensory dendrites innervate peripheral tissues through cell-cell interactions that are poorly understood. The proprioceptive neuron PVD in C. elegans extends regular terminal dendritic branches between muscle and hypodermis. We found that the PVD branch pattern was instructed by adhesion molecule SAX-7/L1CAM, which formed regularly spaced stripes on the hypodermal cell. The regularity of the SAX-7 pattern originated from the repeated and regularly spaced dense body of the sarcomeres in the muscle. The extracellular proteoglycan, UNC-52/Perlecan, links the dense body to the hemidesmosome on the hypodermal cells, which in turn instructed the SAX-7 stripes and PVD dendrites. Both UNC-52 and hemidesmosome components exhibited highly regular stripes that interdigitated with the SAX-7 stripe and PVD dendrites, reflecting the striking precision of subcellular patterning between muscle, hypodermis and dendrites. Hence, the muscular contractile apparatus provides the instructive cues to pattern proprioceptive dendrites. PMID:25982673

  1. Computing Average Passive Forces in Sarcomeres in Length-Ramp Simulations.

    Directory of Open Access Journals (Sweden)

    Gudrun Schappacher-Tilp

    2016-06-01

    Full Text Available Passive forces in sarcomeres are mainly related to the giant protein titin. Titin's extensible region consists of spring-like elements acting in series. In skeletal muscles these elements are the PEVK segment, two distinct immunoglobulin (Ig domain regions (proximal and distal, and a N2A portion. While distal Ig domains are thought to form inextensible end filaments in intact sarcomeres, proximal Ig domains unfold in a force- and time-dependent manner. In length-ramp experiments of single titin strands, sequential unfolding of Ig domains leads to a typical saw-tooth pattern in force-elongation curves which can be simulated by Monte Carlo simulations. In sarcomeres, where more than a thousand titin strands are arranged in parallel, numerous Monte Carlo simulations are required to estimate the resultant force of all titin filaments based on the non-uniform titin elongations. To simplify calculations, the stochastic model of passive forces is often replaced by linear or non-linear deterministic and phenomenological functions. However, new theories of muscle contraction are based on the hypothesized binding of titin to the actin filament upon activation, and thereby on a prominent role of the structural properties of titin. Therefore, these theories necessitate a detailed analysis of titin forces in length-ramp experiments. In our study we present a simple and efficient alternative to Monte Carlo simulations. Based on a structural titin model, we calculate the exact probability distributions of unfolded Ig domains under length-ramp conditions needed for rigorous analysis of expected forces, distribution of unfolding forces, etc. Due to the generality of our model, the approach is applicable to a wide range of stochastic protein unfolding problems.

  2. Uniquely identifying cell orientation and sarcomere length in the intact rodent heart with oblique plane remote focussing microscopy

    Science.gov (United States)

    Corbett, A. D.; Burton, R. A. B.; Bub, G.; Wilson, T.

    2015-07-01

    In cardiac imaging, the spacing between sub-cellular sarcomere structures is of great importance to physiologists in understanding muscle design and performance. Making accurate measurements of the sarcomere length (SL) presents a significant imaging challenge owing to the size of the SL (~2μm) and its naturally low variability (pathological models of chronic hypertension. As well as improving measurement precision, the distribution of α across the field of view provides additional structural information which can be related to disease morphology. To validate this new imaging protocol, the value of α calculated from the oblique planes provided the input to a rigid model cell which was used to predict the appearance of the cell in the conventional focal plane. The comparison of the model to the image data provided a confidence metric for our measurements. Finally, by considering the optical transfer function, the range of cell orientations for which the method is valid could be calculated.

  3. Hypothesis and theory: Mechanical instabilities and non-uniformities in hereditary sarcomere myopathies

    Directory of Open Access Journals (Sweden)

    Alf eMansson

    2014-09-01

    Full Text Available Familial hypertrophic cardiomyopathy (HCM, due to point mutations in genes for sarcomere proteins such as myosin, occurs in 1/500 people and is the most common cause of sudden death in young individuals. Similar mutations in skeletal muscle, e.g. in the MYH7 gene for slow myosin found in both the cardiac ventricle and slow skeletal muscle, may also cause severe disease but the severity and the morphological changes are often different. In HCM, the modified protein function leads, over years to decades, to secondary remodeling with substantial morphological changes, such as hypertrophy, myofibrillar disarray and extensive fibrosis associated with severe functional deterioration. Despite intense studies, it is unclear how the moderate mutation-induced changes in protein function cause the long-term effects. In hypertrophy of the heart due to pressure overload (e.g. hypertension, mechanical stress in the myocyte is believed to be major initiating stimulus for activation of relevant cell signaling cascades. Here it is considered how expression of mutated proteins, such as myosin or regulatory proteins, could have similar consequences through one or both of the following mechanisms: 1. contractile instabilities within each sarcomere (with more than one stable velocity for a given load, 2. different tension generating capacities of cells in series. These mechanisms would have the potential to cause increased tension and/or stretch of certain cells during parts of the cardiac cycle. Modeling studies are used to illustrate these ideas and experimental tests are proposed. The applicability of similar ideas to skeletal muscle is also postulated, and differences between heart and skeletal muscle are discussed.

  4. Subtle abnormalities in contractile function are an early manifestation of sarcomere mutations in dilated cardiomyopathy

    DEFF Research Database (Denmark)

    Lakdawala, Neal K; Thune, Jens J; Colan, Steven D

    2012-01-01

    Sarcomere mutations cause both dilated cardiomyopathy (DCM) and hypertrophic cardiomyopathy (HCM); however, the steps leading from mutation to disease are not well described. By studying mutation carriers before a clinical diagnosis develops, we characterize the early manifestations of sarcomere...

  5. Novel interactions of ankyrins-G at the costameres: The muscle-specific Obscurin/Titin-Binding-related Domain (OTBD) binds plectin and filamin C

    Energy Technology Data Exchange (ETDEWEB)

    Maiweilidan, Yimingjiang; Klauza, Izabela; Kordeli, Ekaterini, E-mail: ekaterini.kordeli@inserm.fr

    2011-04-01

    Ankyrins, the adapters of the spectrin skeleton, are involved in local accumulation and stabilization of integral proteins to the appropriate membrane domains. In striated muscle, tissue-dependent alternative splicing generates unique Ank3 gene products (ankyrins-G); they share the Obscurin/Titin-Binding-related Domain (OTBD), a muscle-specific insert of the C-terminal domain which is highly conserved among ankyrin genes, and binds obscurin and titin to Ank1 gene products. We previously proposed that OTBD sequences constitute a novel domain of protein-protein interactions which confers ankyrins with specific cellular functions in muscle. Here we searched for muscle proteins binding to ankyrin-G OTBD by yeast two hybrid assay, and we found plectin and filamin C, two organizing elements of the cytoskeleton with essential roles in myogenesis, muscle cell cytoarchitecture, and muscle disease. The three proteins coimmunoprecipitate from skeletal muscle extracts and colocalize at costameres in adult muscle fibers. During in vitro myogenesis, muscle ankyrins-G are first expressed in postmitotic myocytes undergoing fusion to myotubes. In western blots of subcellular fractions from C2C12 cells, the majority of muscle ankyrins-G appear associated with membrane compartments. Occasional but not extensive co-localization at nascent costameres suggested that ankyrin-G interactions with plectin and filamin C are not involved in costamere assembly; they would rather reinforce stability and/or modulate molecular interactions in sarcolemma microdomains by establishing novel links between muscle-specific ankyrins-G and the two costameric dystrophin-associated glycoprotein and integrin-based protein complexes. These results report the first protein-protein interactions involving the ankyrin-G OTBD domain and support the hypothesis that OTBD sequences confer ankyrins with a gain of function in vertebrates, bringing further consolidation and resilience of the linkage between sarcomeres

  6. Glutamine Synthetase in Muscle Is Required for Glutamine Production during Fasting and Extrahepatic Ammonia Detoxification

    NARCIS (Netherlands)

    He, Youji; Hakvoort, Theodorus B. M.; Köhler, S. Eleonore; Vermeulen, Jacqueline L. M.; de Waart, D. Rudi; de Theije, Chiel; ten Have, Gabrie A. M.; van Eijk, Hans M. H.; Kunne, Cindy; Labruyere, Wilhelmina T.; Houten, Sander M.; Sokolovic, Milka; Ruijter, Jan M.; Deutz, Nicolaas E. P.; Lamers, Wouter H.

    2010-01-01

    The main endogenous source of glutamine is de novo synthesis in striated muscle via the enzyme glutamine synthetase (GS). The mice in which GS is selectively but completely eliminated from striated muscle with the Cre-loxP strategy (GS-KO/M mice) are, nevertheless, healthy and fertile. Compared with

  7. Prediction of Sarcomere Mutations in Subclinical Hypertrophic Cardiomyopathy

    Science.gov (United States)

    Captur, Gabriella; Lopes, Luis R.; Mohun, Timothy J.; Patel, Vimal; Li, Chunming; Bassett, Paul; Finocchiaro, Gherardo; Ferreira, Vanessa M.; Esteban, Maite Tome; Muthurangu, Vivek; Sherrid, Mark V.; Day, Sharlene M.; Canter, Charles E.; McKenna, William J.; Seidman, Christine E.; Bluemke, David A.; Elliott, Perry M.; Ho, Carolyn Y.; Moon, James C.

    2014-01-01

    Background Sarcomere protein mutations in hypertrophic cardiomyopathy (HCM) induce subtle cardiac structural changes prior to the development of left ventricular hypertrophy (LVH). We have proposed that myocardial crypts are part of this phenotype and independently associated with the presence of sarcomere gene mutations. We tested this hypothesis in genetic HCM pre-LVH (G+LVH−). Methods and Results A multi-centre case-control study investigated crypts and 22 other cardiovascular magnetic resonance (CMR) parameters in subclinical HCM to determine their strength of association with sarcomere gene mutation carriage. The G+LVH− sample (n=73) was 29±13 years old and 51% male. Crypts were related to the presence of sarcomere mutations (for ≥1 crypt, β=2.5, 95% confidence interval [CI] 0.5-4.4, p=0.014; for ≥2 crypts, β=3.0, 95%CI 0.8-7.9, p=0.004). In combination with 3 other parameters: anterior mitral valve leaflet (AMVL) elongation (β=2.1, 95%CI 1.7-3.1, p<0.001), abnormal LV apical trabeculae (β=1.6, 95%CI 0.8-2.5, p<0.001), and smaller LV end-systolic volumes (β=1.4, 95%CI 0.5-2.3, p=0.001), multiple crypts indicated the presence of sarcomere gene mutations with 80% accuracy and an area under the curve of 0.85 (95%CI 0.8-0.9). In this G+LVH− population cardiac myosin-binding protein C mutation carriers had twice the prevalence of crypts when compared to the other combined mutations (47 vs. 23%; odds ratio, 2.9; 95%CI 1.1–7.9; p=0.045). Conclusions The subclinical HCM phenotype measured by CMR in a multi-center environment and consisting of crypts (particularly multiple), AMVL elongation, abnormal trabeculae and smaller LV systolic cavity, is indicative of the presence of sarcomere gene mutations and highlights the need for further study. PMID:25228707

  8. Insights from diploblasts; the evolution of mesoderm and muscle.

    Science.gov (United States)

    Burton, Patrick Michael

    2008-01-15

    The origin of both mesoderm and muscle are central questions in metazoan evolution. The majority of metazoan phyla are triploblasts, possessing three discrete germ layers. Attention has therefore been focused on two outgroups to triploblasts, Cnidaria and Ctenophora. Modern texts describe these taxa as diploblasts, lacking a mesodermal germ layer. However, some members of Medusozoa, one of two subphyla within Cnidaria, possess tissue independent of either the ectoderm or endoderm referred to as the entocodon. Furthermore, members of both Cnidaria and Ctenophora have been described as possessing striated muscle, a mesodermal derivative. While it is widely accepted that the ancestor of Eumetazoa was diploblastic, homology of the entocodon and mesoderm as well as striated muscle within Eumetazoa has been suggested. This implies a potential triploblastic ancestor of Eumetazoa possessing striated muscle. In the following review, I examine the evidence for homology of both muscle and mesoderm. Current data support a diploblastic ancestor of cnidarians, ctenophores, and triploblasts lacking striated muscle.

  9. Genetic advances in sarcomeric cardiomyopathies: state of the art.

    Science.gov (United States)

    Ho, Carolyn Y; Charron, Philippe; Richard, Pascale; Girolami, Francesca; Van Spaendonck-Zwarts, Karin Y; Pinto, Yigal

    2015-04-01

    Genetic studies in the 1980s and 1990s led to landmark discoveries that sarcomere mutations cause both hypertrophic and dilated cardiomyopathies. Sarcomere mutations also likely play a role in more complex phenotypes and overlap cardiomyopathies with features of hypertrophy, dilation, diastolic abnormalities, and non-compaction. Identification of the genetic cause of these important conditions provides unique opportunities to interrogate and characterize disease pathogenesis and pathophysiology, starting from the molecular level and expanding from there. With such insights, there is potential for clinical translation that may transform management of patients and families with inherited cardiomyopathies. If key pathways for disease development can be identified, they could potentially serve as targets for novel disease-modifying or disease-preventing therapies. By utilizing gene-based diagnostic testing, we can identify at-risk individuals prior to the onset of clinical disease, allowing for disease-modifying therapy to be initiated early in life, at a time that such treatment may be most successful. In this section, we review the current application of genetics in clinical management, focusing on hypertrophic cardiomyopathy as a paradigm; discuss state-of-the-art genetic testing technology; review emerging knowledge of gene expression in sarcomeric cardiomyopathies; and discuss both the prospects, as well as the challenges, of bringing genetics to medicine. © The Author 2015. Published by Oxford University Press on behalf of the European Society of Cardiology.

  10. The striated MR nephrogram, not a reflection of pathology

    Energy Technology Data Exchange (ETDEWEB)

    Trout, Andrew T.; Care, Marguerite M.; Towbin, Alexander J. [Cincinnati Children' s Hospital Medical Center, Department of Radiology - MLC 5031, Cincinnati, OH (United States); Zhang, Bin [Cincinnati Children' s Hospital Medical Center, Division of Biostatistics and Epidemiology, Cincinnati, OH (United States)

    2015-10-15

    We have intermittently observed low signal striations in the kidneys on delayed post-contrast MR exams of the spine. While we suspected these striations were due to concentrated gadolinium, the clinical importance of this finding was uncertain. To describe the striated MR nephrogram (low signal striations in the kidney) and assess its clinical relevance. Retrospective review of delayed post-contrast MRIs of the spine (mean: 45 min after contrast administration). The presence of the striated MR nephrogram was correlated with imaging parameters (field strength, time since contrast), and findings (gadolinium in the bladder, inferior vena cava and aorta diameters) and with clinical factors (history of renal disease, laboratory values). Seven hundred seventy-three exams performed on 229 patients, 8.3 ± 5.3 years of age, were reviewed. The striated MR nephrogram was observed in 102/773 examinations (13.2%) and was present on at least one study in 54/229 patients (23.6%). The presence of striations was associated with the specific magnet on which the exam was performed (P < 0.01) but not with magnet field strength. Serum creatinine was minimally lower in patients with striations (0.43 ± 0.12 vs. 0.49 ± 0.18 mg/dL, P = 0.002), but no other clinical or historical data, including time from contrast administration (P = 0.54), fluid status (P = 0.17) and clinical history of renal disease (P = 0.14), were predictive of the presence of striations. The striated MR nephrogram was observed in 13% of delayed post-contrast MR exams of the spine. Precipitating factors are unclear, but the striated nephrogram does not appear to be a marker of clinically apparent renal dysfunction. (orig.)

  11. Cloning, Sequencing, and the Expression of the Elusive Sarcomeric TPM4α Isoform in Humans

    Directory of Open Access Journals (Sweden)

    Dipak K. Dube

    2016-01-01

    Full Text Available In mammals, tropomyosin is encoded by four known TPM genes (TPM1, TPM2, TPM3, and TPM4 each of which can generate a number of TPM isoforms via alternative splicing and/or using alternate promoters. In humans, the sarcomeric isoform(s of each of the TPM genes, except for the TPM4, have been known for a long time. Recently, on the basis of computational analyses of the human genome sequence, the predicted sequence of TPM4α has been posted in GenBank. We designed primer-pairs for RT-PCR and showed the expression of the transcripts of TPM4α and a novel isoform TPM4δ in human heart and skeletal muscle. qRT-PCR shows that the relative expression of TPM4α and TPM4δ is higher in human cardiac muscle. Western blot analyses using CH1 monoclonal antibodies show the absence of the expression of TPM4δ protein (~28 kDa in human heart muscle. 2D western blot analyses with the same antibody show the expression of at least nine distinct tropomyosin molecules with a mass ~32 kD and above in adult heart. By Mass spectrometry, we determined the amino acid sequences of the extracted proteins from these spots. Spot “G” reveals the putative expression of TPM4α along with TPM1α protein in human adult heart.

  12. Fully automated muscle quality assessment by Gabor filtering of second harmonic generation images

    Science.gov (United States)

    Paesen, Rik; Smolders, Sophie; Vega, José Manolo de Hoyos; Eijnde, Bert O.; Hansen, Dominique; Ameloot, Marcel

    2016-02-01

    Although structural changes on the sarcomere level of skeletal muscle are known to occur due to various pathologies, rigorous studies of the reduced sarcomere quality remain scarce. This can possibly be explained by the lack of an objective tool for analyzing and comparing sarcomere images across biological conditions. Recent developments in second harmonic generation (SHG) microscopy and increasing insight into the interpretation of sarcomere SHG intensity profiles have made SHG microscopy a valuable tool to study microstructural properties of sarcomeres. Typically, sarcomere integrity is analyzed by fitting a set of manually selected, one-dimensional SHG intensity profiles with a supramolecular SHG model. To circumvent this tedious manual selection step, we developed a fully automated image analysis procedure to map the sarcomere disorder for the entire image at once. The algorithm relies on a single-frequency wavelet-based Gabor approach and includes a newly developed normalization procedure allowing for unambiguous data interpretation. The method was validated by showing the correlation between the sarcomere disorder, quantified by the M-band size obtained from manually selected profiles, and the normalized Gabor value ranging from 0 to 1 for decreasing disorder. Finally, to elucidate the applicability of our newly developed protocol, Gabor analysis was used to study the effect of experimental autoimmune encephalomyelitis on the sarcomere regularity. We believe that the technique developed in this work holds great promise for high-throughput, unbiased, and automated image analysis to study sarcomere integrity by SHG microscopy.

  13. Immunocytochemical electron microscopic study and western blot analysis of paramyosin in different invertebrate muscle cell types of the fruit fly Drosophila melanogaster, the earthworm Eisenia foetida, and the snail Helix aspersa.

    Science.gov (United States)

    Royuela, M; García-Anchuelo, R; Arenas, M I; Cervera, M; Fraile, B; Paniagua, R

    1996-04-01

    The presence and distribution pattern of paramyosin have been examined in different invertebrate muscle cell types by means of Western blot analysis and electron microscopy immunogold labelling. The muscles studied were: transversely striated muscle with continuous Z lines (flight muscle from Drosophila melanogaster), transversely striated muscle with discontinuous Z lines (heart muscle from the snail Helix aspersa), obliquely striated body wall muscle from the earthworm Eisenia foetida, and smooth muscles (retractor muscle from the snail and pseudoheart outer muscular layer from the earthworm). Paramyosin-like immunoreactivity was localized in thick filaments of all muscles studied. Immunogold particle density was similar along the whole thick filament length in insect flight muscle but it predominated in filament tips of fusiform thick filaments in both snail heart and earthworm body wall musculature when these filaments were observed in longitudinal sections. In obliquely sectioned thick filaments, immunolabelling was more abundant at the sites where filaments disappeared from the section. These results agree with the notion that paramyosin extended along the whole filament length, but that it can only be immunolabelled when it is not covered by myosin. In all muscles examined, immunolabelling density was lower in cross-sectioned myofilaments than in longitudinally sectioned myofilaments. This suggests that paramyosin does not form a continuous filament. The results of a semiquantitative analysis of paramyosin-like immunoreactivity indicated that it was more abundant in striated than in smooth muscles, and that, within striated muscles, transversely striated muscles contain more paramyosin than obliquely striated muscles.

  14. Design and optimization of multi-class series-parallel linear electromagnetic array artificial muscle.

    Science.gov (United States)

    Li, Jing; Ji, Zhenyu; Shi, Xuetao; You, Fusheng; Fu, Feng; Liu, Ruigang; Xia, Junying; Wang, Nan; Bai, Jing; Wang, Zhanxi; Qin, Xiansheng; Dong, Xiuzhen

    2014-01-01

    Skeletal muscle exhibiting complex and excellent precision has evolved for millions of years. Skeletal muscle has better performance and simpler structure compared with existing driving modes. Artificial muscle may be designed by analyzing and imitating properties and structure of skeletal muscle based on bionics, which has been focused on by bionic researchers, and a structure mode of linear electromagnetic array artificial muscle has been designed in this paper. Half sarcomere is the minimum unit of artificial muscle and electromagnetic model has been built. The structural parameters of artificial half sarcomere actuator were optimized to achieve better movement performance. Experimental results show that artificial half sarcomere actuator possesses great motion performance such as high response speed, great acceleration, small weight and size, robustness, etc., which presents a promising application prospect of artificial half sarcomere actuator.

  15. Differentiation and sarcomere formation in skeletal myocytes directly prepared from human induced pluripotent stem cells using a sphere-based culture.

    Science.gov (United States)

    Jiwlawat, Saowanee; Lynch, Eileen; Glaser, Jennifer; Smit-Oistad, Ivy; Jeffrey, Jeremy; Van Dyke, Jonathan M; Suzuki, Masatoshi

    Human induced-pluripotent stem cells (iPSCs) are a promising resource for propagation of myogenic progenitors. Our group recently reported a unique protocol for the derivation of myogenic progenitors directly (without genetic modification) from human pluripotent cells using free-floating spherical culture. Here we expand our previous efforts and attempt to determine how differentiation duration, culture surface coatings, and nutrient supplements in the medium influence progenitor differentiation and formation of skeletal myotubes containing sarcomeric structures. A long differentiation period (over 6 weeks) promoted the differentiation of iPSC-derived myogenic progenitors and subsequent myotube formation. These iPSC-derived myotubes contained representative sarcomeric structures, consisting of organized myosin and actin filaments, and could spontaneously contract. We also found that a bioengineering approach using three-dimensional (3D) artificial muscle constructs could facilitate the formation of elongated myotubes. Lastly, we determined how culture surface coating matrices and different supplements would influence terminal differentiation. While both Matrigel and laminin coatings showed comparable effects on muscle differentiation, B27 serum-free supplement in the differentiation medium significantly enhanced myogenesis compared to horse serum. Our findings support the possibility to create an in vitro model of contractile sarcomeric myofibrils for disease modeling and drug screening to study neuromuscular diseases. Copyright © 2017 International Society of Differentiation. Published by Elsevier B.V. All rights reserved.

  16. Evidence of skeletal muscle damage following electrically stimulated isometric muscle contractions in humans

    DEFF Research Database (Denmark)

    Mackey, Abigail; Bojsen-Moller, Jens; Qvortrup, Klaus

    2008-01-01

    It is unknown whether muscle damage at the level of the sarcomere can be induced without lengthening contractions. To investigate this, we designed a study where seven young, healthy men underwent 30 min of repeated electrical stimulated contraction of m. gastrocnemius medialis, with the ankle...... changes at the level of the z-lines were investigated by transmission electron microscopy. Blood samples were collected for measurement of creatine kinase activity, and muscle soreness was assessed in the days following stimulation. The biopsies from the stimulated muscle revealed macrophage infiltration...... of damage at the myofiber and sarcomere levels. These data support the hypothesis that muscle damage at the level of the sarcomere can be induced without lengthening muscle contractions....

  17. M line-deficient titin causes cardiac lethality through impaired maturation of the sarcomere.

    NARCIS (Netherlands)

    Weinert, S.; Bergmann, N.; Luo, X.; Erdmann, B.; Gotthardt, M.

    2006-01-01

    Titin, the largest protein known to date, has been linked to sarcomere assembly and function through its elastic adaptor and signaling domains. Titin's M-line region contains a unique kinase domain that has been proposed to regulate sarcomere assembly via its substrate titin cap (T-cap). In this

  18. A muscle stem cell for every muscle: variability of satellite cell biology among different muscle groups

    Directory of Open Access Journals (Sweden)

    Matthew Emerson Randolph

    2015-10-01

    Full Text Available The human body contains approximately 640 individual skeletal muscles. Despite the fact that all of these muscles are composed of striated muscle tissue, the biology of these muscles and their associated muscle stem cell populations are quite diverse. Skeletal muscles are affected differentially by various muscular dystrophies, such that certain genetic mutations specifically alter muscle function in only a subset of muscles. Additionally, defective muscle stem cells have been implicated in the pathology of some muscular dystrophies. The biology of muscle stem cells varies depending on their embryologic origins and the muscles with which they are associated. Here we review the biology of skeletal muscle stem cell populations of eight different muscle groups. Understanding the biological variation of skeletal muscles and their resident stem cells could provide valuable insight into mechanisms underlying the susceptibility of certain muscles to myopathic disease.

  19. A muscle stem cell for every muscle: variability of satellite cell biology among different muscle groups

    Science.gov (United States)

    Randolph, Matthew E.; Pavlath, Grace K.

    2015-01-01

    The human body contains approximately 640 individual skeletal muscles. Despite the fact that all of these muscles are composed of striated muscle tissue, the biology of these muscles and their associated muscle stem cell populations are quite diverse. Skeletal muscles are affected differentially by various muscular dystrophies (MDs), such that certain genetic mutations specifically alter muscle function in only a subset of muscles. Additionally, defective muscle stem cells have been implicated in the pathology of some MDs. The biology of muscle stem cells varies depending on the muscles with which they are associated. Here we review the biology of skeletal muscle stem cell populations of eight different muscle groups. Understanding the biological variation of skeletal muscles and their resident stem cells could provide valuable insight into mechanisms underlying the susceptibility of certain muscles to myopathic disease. PMID:26500547

  20. Virtual and simulated striated toolmarks for forensic applications.

    Science.gov (United States)

    Baiker, Martin; Petraco, Nicholas D K; Gambino, Carol; Pieterman, René; Shenkin, Peter; Zoon, Peter

    2016-04-01

    Large numbers of experimental toolmarks of screwdrivers are often required in casework of toolmark examiners and in research environments alike, to be able to recover the angle of attack of a crime scene mark and to determine statistically meaningful properties of toolmarks respectively. However, in practice the number of marks is limited by the time needed to create them. In this article, we present an approach to predict how a striated mark of a particular tool would look like, using 3D surface datasets of screwdrivers. We compare these virtual toolmarks qualitatively and quantitatively with real experimental marks in wax and show that they are very similar. In addition we study toolmark similarity, dependent on the angle of attack, with a very high angular resolution of 1°. The results show that for the tested type of screwdriver, our toolmark comparison framework yields known match similarity scores that are above the mean known non-match similarity scores, even for known match differences in angle of attack of up to 40°. In addition we demonstrate an approach to automatically recover the angle of attack of an experimental toolmark and experiments yield high accuracy and precision of 0.618 ± 4.179°. Furthermore, we present a strategy to study the structural elements of striated toolmarks using wavelet analysis, and show how to use the results to simulate realistic toolmarks. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  1. Echocardiographic strain imaging to assess early and late consequences of sarcomere mutations in hypertrophic cardiomyopathy

    DEFF Research Database (Denmark)

    Ho, Carolyn Y; Carlsen, Christian; Thune, Jens Jakob

    2009-01-01

    BACKGROUND: Genetic testing identifies sarcomere mutation carriers (G+) before clinical diagnosis of hypertrophic cardiomyopathy (HCM), allowing characterization of initial disease manifestations. Previous studies demonstrated that impaired relaxation develops before left ventricular hypertrophy...

  2. Study of the striated nature of a glow discharge

    International Nuclear Information System (INIS)

    Hernandez A, M.

    1995-01-01

    In an investigation in progress here, plasma diagnostics and detection of standing and moving striations is being made in a discharge in Argon at pressures of 2 x 10 -1 to 9 x 10 -1 mb and currents of 2 to 9 m-amp inside an discharge tube. Measurement of the temperature of the electrons, the concentration of electrons and the plasma potential are obtained in different places of the discharge by the double probe method, together with the computation system reported in [1]. In similar way an experimental work of the striated column in a discharge plasma to find the regimen of appearance of the standing and moving striations show some properties of moving striations (frequency and velocity) and standing striations. Two different oscilations are observed in motion in contrary directions along the discharge tube with a photomultiplier. (Author)

  3. At the Start of the Sarcomere: A Previously Unrecognized Role for Myosin Chaperones and Associated Proteins during Early Myofibrillogenesis

    Directory of Open Access Journals (Sweden)

    J. Layne Myhre

    2012-01-01

    Full Text Available The development of striated muscle in vertebrates requires the assembly of contractile myofibrils, consisting of highly ordered bundles of protein filaments. Myofibril formation occurs by the stepwise addition of complex proteins, a process that is mediated by a variety of molecular chaperones and quality control factors. Most notably, myosin of the thick filament requires specialized chaperone activity during late myofibrillogenesis, including that of Hsp90 and its cofactor, Unc45b. Unc45b has been proposed to act exclusively as an adaptor molecule, stabilizing interactions between Hsp90 and myosin; however, recent discoveries in zebrafish and C. elegans suggest the possibility of an earlier role for Unc45b during myofibrillogenesis. This role may involve functional control of nonmuscle myosins during the earliest stages of myogenesis, when premyofibril scaffolds are first formed from dynamic cytoskeletal actin. This paper will outline several lines of evidence that converge to build a model for Unc45b activity during early myofibrillogenesis.

  4. Alpha-actinin in different invertebrate muscle cell types of Drosophila melanogaster, the earthworm Eisenia foetida, and the snail Helix aspersa.

    Science.gov (United States)

    Royuela, M; Astier, C; Fraile, B; Paniagua, R

    1999-01-01

    The presence and distribution of alpha-actinin has been studied in several invertebrate muscle cell types. These comprised transversely striated muscle (flight muscle) from the fruit fly Drosophila melanogaster, transversely striated muscle (heart muscle) from the snail Helix aspersa, obliquely striated muscle (body wall muscle) from the earthworm Eisenia foetida, smooth muscle (retractor muscle) from H. aspersa, and smooth muscle (outer muscular layer of the pseudoheart) from E. foetida. The study was carried by means of Western blot analysis, ELISA, and immunohistochemical electron microscopy, using anti alpha-actinin antibody. Immunoreaction for a protein with the same molecular weight as that of mammalian alpha-actinin was detected in all muscle types studied, although the amount and intensity of immunoreaction varied among them. In the insect muscle, immunolabelling was found along the whole Z-line. In both the transversely striated muscle from the snail and the obliquely striated muscle from the earthworm, immunolabelling did not occupy the whole Z-line but showed discontinuous, orderly arranged patches along the Z-line course. In the two smooth muscles studied (snail and earthworm), immunolabelling was limited to small patches which did not show an apparently ordered distribution. Since it is assumed that alpha-actinin is located at the anchorage sites for actin filaments, present observations suggest that, only in the Drosophila muscle, actin filaments are parallelly arranged in all their course, whereas in the other invertebrate muscles studied these filaments converge on discontinuously distributed anchorage sites.

  5. Stretching skeletal muscle: chronic muscle lengthening through sarcomerogenesis.

    Directory of Open Access Journals (Sweden)

    Alexander M Zöllner

    Full Text Available Skeletal muscle responds to passive overstretch through sarcomerogenesis, the creation and serial deposition of new sarcomere units. Sarcomerogenesis is critical to muscle function: It gradually re-positions the muscle back into its optimal operating regime. Animal models of immobilization, limb lengthening, and tendon transfer have provided significant insight into muscle adaptation in vivo. Yet, to date, there is no mathematical model that allows us to predict how skeletal muscle adapts to mechanical stretch in silico. Here we propose a novel mechanistic model for chronic longitudinal muscle growth in response to passive mechanical stretch. We characterize growth through a single scalar-valued internal variable, the serial sarcomere number. Sarcomerogenesis, the evolution of this variable, is driven by the elastic mechanical stretch. To analyze realistic three-dimensional muscle geometries, we embed our model into a nonlinear finite element framework. In a chronic limb lengthening study with a muscle stretch of 1.14, the model predicts an acute sarcomere lengthening from 3.09[Formula: see text]m to 3.51[Formula: see text]m, and a chronic gradual return to the initial sarcomere length within two weeks. Compared to the experiment, the acute model error was 0.00% by design of the model; the chronic model error was 2.13%, which lies within the rage of the experimental standard deviation. Our model explains, from a mechanistic point of view, why gradual multi-step muscle lengthening is less invasive than single-step lengthening. It also explains regional variations in sarcomere length, shorter close to and longer away from the muscle-tendon interface. Once calibrated with a richer data set, our model may help surgeons to prevent muscle overstretch and make informed decisions about optimal stretch increments, stretch timing, and stretch amplitudes. We anticipate our study to open new avenues in orthopedic and reconstructive surgery and enhance

  6. Mechanical principles of effects of botulinum toxin on muscle length-force characteristics: an assessment by finite element modeling.

    Science.gov (United States)

    Turkoglu, Ahu N; Huijing, Peter A; Yucesoy, Can A

    2014-05-07

    Recent experiments involving muscle force measurements over a range of muscle lengths show that effects of botulinum toxin (BTX) are complex e.g., force reduction varies as a function of muscle length. We hypothesized that altered conditions of sarcomeres within active parts of partially paralyzed muscle is responsible for this effect. Using finite element modeling, the aim was to test this hypothesis and to study principles of how partial activation as a consequence of BTX affects muscle mechanics. In order to model the paralyzing effect of BTX, only 50% of the fascicles (most proximal, or middle, or most distal) of the modeled muscle were activated. For all muscle lengths, a vast majority of sarcomeres of these BTX-cases were at higher lengths than identical sarcomeres of the BTX-free muscle. Due to such "longer sarcomere effect", activated muscle parts show an enhanced potential of active force exertion (up to 14.5%). Therefore, a muscle force reduction originating exclusively from the paralyzed muscle fiber populations, is compromised by the changes of active sarcomeres leading to a smaller net force reduction. Moreover, such "compromise to force reduction" varies as a function of muscle length and is a key determinant of muscle length dependence of force reduction caused by BTX. Due to longer sarcomere effect, muscle optimum length tends to shift to a lower muscle length. Muscle fiber-extracellular matrix interactions occurring via their mutual connections along full peripheral fiber lengths (i.e., myofascial force transmission) are central to these effects. Our results may help improving our understanding of mechanisms of how the toxin secondarily affects the muscle mechanically. Copyright © 2014 Elsevier Ltd. All rights reserved.

  7. The relationship between shear force, compression, collagen characteristics, desmin degradation and sarcomere length in lamb biceps femoris.

    Science.gov (United States)

    Starkey, Colin P; Geesink, Geert H; van de Ven, Remy; Hopkins, David L

    2017-04-01

    This study aimed to identity the relationships between known variants of tenderness (collagen content (total and soluble), desmin degradation and sarcomere length) and shear force and compression in the biceps femoris aged for 14days from 112 mixed sex lambs. Desmin degradation was related to compression (Pcompression decreased. Sarcomere length (SL) was related to shear force (Pcompression (Pcompression, sarcomere length and soluble collagen. The findings from this experiment indicate that the known variants (soluble collagen, sarcomere length and desmin degradation) are related to shear force and compression in ovine biceps femoris. Crown Copyright © 2016. Published by Elsevier Ltd. All rights reserved.

  8. The role of titin in eccentric muscle contraction.

    Science.gov (United States)

    Herzog, Walter

    2014-08-15

    Muscle contraction and force regulation in skeletal muscle have been thought to occur exclusively through the relative sliding of and the interaction between the contractile filaments actin and myosin. While this two-filament sarcomere model has worked well in explaining the properties of isometrically and concentrically contracting muscle, it has failed miserably in explaining experimental observations in eccentric contractions. Here, I suggest, and provide evidence, that a third filament, titin, is involved in force regulation of sarcomeres by adjusting its stiffness in an activation-dependent (calcium) and active force-dependent manner. Upon muscle activation, titin binds calcium at specific sites, thereby increasing its stiffness, and cross-bridge attachment to actin is thought to free up binding sites for titin on actin, thereby reducing titin's free-spring length, thus increasing its stiffness and force upon stretch of active muscle. This role of titin as a third force regulating myofilament in sarcomeres, although not fully proven, would account for many of the unexplained properties of eccentric muscle contraction, while simultaneously not affecting the properties predicted by the two-filament cross-bridge model in isometric and concentric muscle function. Here, I identify the problems of the two-filament sarcomere model and demonstrate the advantages of the three-filament model by providing evidence of titin's contribution to active force in eccentric muscle function. © 2014. Published by The Company of Biologists Ltd.

  9. A multiscale chemo-electro-mechanical skeletal muscle model to analyze muscle contraction and force generation for different muscle fiber arrangements

    Directory of Open Access Journals (Sweden)

    Thomas eHeidlauf

    2014-12-01

    Full Text Available The presented chemo-electro-mechanical skeletal muscle model relies on a continuum mechanical formulation describing the muscle’s deformation and force generation on the macroscopic muscle level. Unlike other three-dimensional models, the description of the activation-induced behavior of the mechanical model is entirely based on chemo-electro-mechanical principles on the microscopic sarcomere level. Yet, the multiscale model reproduces key characteristics of skeletal muscles such as experimental force-length and force-velocity data on the macroscopic whole muscle level. The paper presents the methodological approaches required to obtain such a multiscale model, and demonstrates the feasibility of using such a model to analyze differences in the mechanical behavior of parallel-fibered muscles, in which the muscle fibers either span the entire length of the fascicles or terminate intrafascicularly. The presented results reveal that muscles, in which the fibers span the entire length of the fascicles, show lower peak forces, more dispersed twitches and fusion of twitches at lower stimulation frequencies. In detail, the model predicted twitch rise times of 38.2ms and 17.2ms for a 12 cm long muscle, in which the fibers span the entire length of the fascicles and with twelve fiber compartments in series, respectively. Further, the twelve-compartment model predicted peak twitch forces that were 19% higher than in the single-compartment model. The analysis of sarcomere lengths during fixed-end single twitch contractions at optimal length predicts rather small sarcomere length changes. The observed lengths range from 75 to 111% of the optimal sarcomere length, which corresponds to a region with maximum filament overlap. This result suggests that stability issues resulting from activation-induced stretches of non-activated sarcomeres are unlikely in muscles with passive forces appearing at short muscle length.

  10. Contribution of Post-translational Phosphorylation to Sarcomere-linked Cardiomyopathy Phenotypes

    Directory of Open Access Journals (Sweden)

    Margaret V Westfall

    2016-09-01

    Full Text Available Secondary shifts develop in post-translational phosphorylation of sarcomeric proteins in multi¬ple animal models of inherited cardiomyopathy. These signaling alterations together with the primary mutation are predicted to contribute to the overall cardiac phenotype. As a result, identification and integration of post-translational myofilament signaling responses are identified as priorities for gaining insights into sarcomeric cardiomyopathies. However, significant questions remain about the nature and contribution of post-translational phosphorylation to structural remodeling and cardiac dysfunction in animal models and human patients. This perspective essay discusses specific goals for filling critical gaps about post-translational signaling in response to these inherited mutations, especially within sarcomeric proteins. The discussion focuses primarily on pre-clinical analysis of animal models and defines challenges and future directions in this field.

  11. Protein Structure-Function Relationship at Work: Learning from Myopathy Mutations of the Slow Skeletal Muscle Isoform of Troponin T.

    Science.gov (United States)

    Mondal, Anupom; Jin, J-P

    2016-01-01

    Troponin T (TnT) is the sarcomeric thin filament anchoring subunit of the troponin complex in striated muscles. A nonsense mutation in exon 11 of the slow skeletal muscle isoform of TnT (ssTnT) gene ( TNNT1 ) was found in the Amish populations in Pennsylvania and Ohio. This single nucleotide substitution causes a truncation of the ssTnT protein at Glu 180 and the loss of the C-terminal tropomyosin (Tm)-binding site 2. As a consequence, it abolishes the myofilament integration of ssTnT and the loss of function causes an autosomal recessive nemaline myopathy (NM). More TNNT1 mutations have recently been reported in non-Amish ethnic groups with similar recessive NM phenotypes. A nonsense mutation in exon 9 truncates ssTnT at Ser 108 , deleting Tm-binding site 2 and a part of the middle region Tm-binding site 1. Two splicing site mutations result in truncation of ssTnT at Leu 203 or deletion of the exon 14-encoded C-terminal end segment. Another splicing mutation causes an internal deletion of the 39 amino acids encoded by exon 8, partially damaging Tm-binding site 1. The three splicing mutations of TNNT1 all preserve the high affinity Tm-binding site 2 but still present recessive NM phenotypes. The molecular mechanisms for these mutations to cause myopathy provide interesting models to study and understand the structure-function relationship of TnT. This focused review summarizes the current knowledge of TnT isoform regulation, structure-function relationship of TnT and how various ssTnT mutations cause recessive NM, in order to promote in depth studies for further understanding the pathogenesis and pathophysiology of TNNT1 myopathies toward the development of effective treatments.

  12. STARS knockout attenuates hypoxia-induced pulmonary arterial hypertension by suppressing pulmonary arterial smooth muscle cell proliferation.

    Science.gov (United States)

    Shi, Zhaoling; Wu, Huajie; Luo, Jianfeng; Sun, Xin

    2017-03-01

    STARS (STriated muscle Activator of Rho Signaling) is a sarcomeric protein, which expressed early in cardiac development and involved in pathological remodeling. Abundant evidence indicated that STARS could regulate cell proliferation, but it's exact function remains unclear. In this study, we aimed to investigate the role of STARS in the proliferation of pulmonary arterial smooth muscle cells (PASMC) and the potential effect on the progression of pulmonary arterial hypertension (PAH). In this study, we established a PAH mouse model through chronic hypoxia exposure as reflected by the increased RVSP and RVHI. Western blot and RT-qPCR detected the increased STARS protein and mRNA levels in PAH mice. Next, we cultured the primary PASMC from PAH mice. After STARS overexpression in PASMC, STARS, SRF and Egr-1 were up-regulated significantly. The MTT assay revealed an increase in cell proliferation. Flow cytometry showed a marked inhibition of cell apoptosis. However, STARS silence in PASMC exerted opposite effects with STARS overexpression. SRF siRNA transfection blocked the effects of STARS overexpression in PASMC. In order to further confirm the role of STARS in PAH mice in vivo, we exposed STARS knockout mice to hypoxia and found lower RVSP and RVHI in knockout mice as compared with controls. Our results not only suggest that STARS plays a crucial role in the development of PAH by increasing the proliferation of PASMC through activation of the SRF/Egr-1 pathway, but also provides a new mechanism for hypoxia-induced PAH. In addition, STARS may represent a potential treatment target. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  13. Roles of the troponin isoforms during indirect flight muscle ...

    Indian Academy of Sciences (India)

    Troponin proteins in cooperative interaction with tropomyosin are responsible for controlling the contraction of the striated muscles in response to changes in the intracellular calcium concentration. Contractility of the muscle is determined by the constituent protein isoforms, and the isoforms can switch over from one form to ...

  14. Sarcomeric myosin heavy chain is coded by a highly conserved multigene family.

    OpenAIRE

    Nguyen, H T; Gubits, R M; Wydro, R M; Nadal-Ginard, B

    1982-01-01

    pMHC25, a recombinant plasmid containing myosin heavy chain (MHC) cDNA sequences from differentiated myotubes of the L6E9 rat cell line, has been shown to hybridize to all sarcomeric MHC mRNAs so far tested but not to nonsarcomeric MHC mRNAs. In addition, pMHC25 hybridizes to multiple restriction endonuclease-digested fragments of rat genomic DNA corresponding to different MHC genomic sequences. Thus, the MHC gene represented by pMHC25 is a member of a sarcomeric MHC multigene family that has...

  15. High-frequency sarcomeric auto-oscillations induced by heating in living neonatal cardiomyocytes of the rat

    Energy Technology Data Exchange (ETDEWEB)

    Shintani, Seine A.; Oyama, Kotaro [Department of Pure and Applied Physics, School of Advanced Science and Engineering, Waseda University, Tokyo (Japan); Fukuda, Norio, E-mail: noriof@jikei.ac.jp [Department of Cell Physiology, The Jikei University School of Medicine, Tokyo (Japan); Ishiwata, Shin’ichi, E-mail: ishiwata@waseda.jp [Department of Pure and Applied Physics, School of Advanced Science and Engineering, Waseda University, Tokyo (Japan); WASEDA Bioscience Research Institute in Singapore (WABIOS) (Singapore)

    2015-02-06

    Highlights: • We tested the effects of infra-red laser irradiation on cardiac sarcomere dynamics. • A rise in temperature (>∼38 °C) induced high-frequency sarcomeric auto-oscillations. • These oscillations occurred with and without blockade of intracellular Ca{sup 2+} stores. • Cardiac sarcomeres can play a role as a temperature-dependent rhythm generator. - Abstract: In the present study, we investigated the effects of infra-red laser irradiation on sarcomere dynamics in living neonatal cardiomyocytes of the rat. A rapid increase in temperature to >∼38 °C induced [Ca{sup 2+}]{sub i}-independent high-frequency (∼5–10 Hz) sarcomeric auto-oscillations (Hyperthermal Sarcomeric Oscillations; HSOs). In myocytes with the intact sarcoplasmic reticular functions, HSOs coexisted with [Ca{sup 2+}]{sub i}-dependent spontaneous beating in the same sarcomeres, with markedly varying frequencies (∼10 and ∼1 Hz for the former and latter, respectively). HSOs likewise occurred following blockade of the sarcoplasmic reticular functions, with the amplitude becoming larger and the frequency lower in a time-dependent manner. The present findings suggest that in the mammalian heart, sarcomeres spontaneously oscillate at higher frequencies than the sinus rhythm at temperatures slightly above the physiologically relevant levels.

  16. Pathophysiology of muscle contractures in cerebral palsy.

    Science.gov (United States)

    Mathewson, Margie A; Lieber, Richard L

    2015-02-01

    Patients with cerebral palsy present with a variety of adaptations to muscle structure and function. These pathophysiologic symptoms include functional deficits such as decreased force production and range of motion, in addition to changes in muscle structure such as decreased muscle belly size, increased sarcomere length, and altered extracellular matrix structure and composition. On a cellular level, patients with cerebral palsy have fewer muscle stem cells, termed satellite cells, and altered gene expression. Understanding the nature of these changes may present opportunities for the development of new muscle treatment therapies. Published by Elsevier Inc.

  17. Electrocardiographic features of sarcomere mutation carriers with and without clinically overt hypertrophic cardiomyopathy

    DEFF Research Database (Denmark)

    Lakdawala, Neal K; Thune, Jens Jakob; Maron, Barry J

    2011-01-01

    In hypertrophic cardiomyopathy (HC), electrocardiographic (ECG) changes have been postulated to be an early marker of disease, detectable in sarcomere mutation carriers when left ventricular (LV) wall thickness is still normal. However, the ECG features of mutation carriers have not been fully...

  18. Disruption of a GATA4/Ankrd1 signaling axis in cardiomyocytes leads to sarcomere disarray: implications for anthracycline cardiomyopathy.

    Directory of Open Access Journals (Sweden)

    Billy Chen

    Full Text Available Doxorubicin (Adriamycin is an effective anti-cancer drug, but its clinical usage is limited by a dose-dependent cardiotoxicity characterized by widespread sarcomere disarray and loss of myofilaments. Cardiac ankyrin repeat protein (CARP, ANKRD1 is a transcriptional regulatory protein that is extremely susceptible to doxorubicin; however, the mechanism(s of doxorubicin-induced CARP depletion and its specific role in cardiomyocytes have not been completely defined. We report that doxorubicin treatment in cardiomyocytes resulted in inhibition of CARP transcription, depletion of CARP protein levels, inhibition of myofilament gene transcription, and marked sarcomere disarray. Knockdown of CARP with small interfering RNA (siRNA similarly inhibited myofilament gene transcription and disrupted cardiomyocyte sarcomere structure. Adenoviral overexpression of CARP, however, was unable to rescue the doxorubicin-induced sarcomere disarray phenotype. Doxorubicin also induced depletion of the cardiac transcription factor GATA4 in cardiomyocytes. CARP expression is regulated in part by GATA4, prompting us to examine the relationship between GATA4 and CARP in cardiomyocytes. We show in co-transfection experiments that GATA4 operates upstream of CARP by activating the proximal CARP promoter. GATA4-siRNA knockdown in cardiomyocytes inhibited CARP expression and myofilament gene transcription, and induced extensive sarcomere disarray. Adenoviral overexpression of GATA4 (AdV-GATA4 in cardiomyocytes prior to doxorubicin exposure maintained GATA4 levels, modestly restored CARP levels, and attenuated sarcomere disarray. Interestingly, siRNA-mediated depletion of CARP completely abolished the Adv-GATA4 rescue of the doxorubicin-induced sarcomere phenotype. These data demonstrate co-dependent roles for GATA4 and CARP in regulating sarcomere gene expression and maintaining sarcomeric organization in cardiomyocytes in culture. The data further suggests that concurrent

  19. Comparative Statistical Mechanics of Muscle and Non-Muscle Contractile Systems: Stationary States of Near-Equilibrium Systems in A Linear Regime

    Directory of Open Access Journals (Sweden)

    Yves Lecarpentier

    2017-10-01

    Full Text Available A. Huxley’s equations were used to determine the mechanical properties of muscle myosin II (MII at the molecular level, as well as the probability of the occurrence of the different stages in the actin–myosin cycle. It was then possible to use the formalism of statistical mechanics with the grand canonical ensemble to calculate numerous thermodynamic parameters such as entropy, internal energy, affinity, thermodynamic flow, thermodynamic force, and entropy production rate. This allows us to compare the thermodynamic parameters of a non-muscle contractile system, such as the normal human placenta, with those of different striated skeletal muscles (soleus and extensor digitalis longus as well as the heart muscle and smooth muscles (trachea and uterus in the rat. In the human placental tissues, it was observed that the kinetics of the actin–myosin crossbridges were considerably slow compared with those of smooth and striated muscular systems. The entropy production rate was also particularly low in the human placental tissues, as compared with that observed in smooth and striated muscular systems. This is partly due to the low thermodynamic flow found in the human placental tissues. However, the unitary force of non-muscle myosin (NMII generated by each crossbridge cycle in the myofibroblasts of the human placental tissues was similar in magnitude to that of MII in the myocytes of both smooth and striated muscle cells. Statistical mechanics represents a powerful tool for studying the thermodynamics of all contractile muscle and non-muscle systems.

  20. Architectural analysis and intraoperative measurements demonstrate the unique design of the multifidus muscle for lumbar spine stability.

    Science.gov (United States)

    Ward, Samuel R; Kim, Choll W; Eng, Carolyn M; Gottschalk, Lionel J; Tomiya, Akihito; Garfin, Steven R; Lieber, Richard L

    2009-01-01

    Muscular instability is an important risk factor for lumbar spine injury and chronic low-back pain. Although the lumbar multifidus muscle is considered an important paraspinal muscle, its design features are not completely understood. The purpose of the present study was to determine the architectural properties, in vivo sarcomere length operating range, and passive mechanical properties of the human multifidus muscle. We hypothesized that its architecture would be characterized by short fibers and a large physiological cross-sectional area and that it would operate over a relatively wide range of sarcomere lengths but would have very stiff passive material properties. The lumbar spines of eight cadaver specimens were excised en bloc from T12 to the sacrum. Multifidus muscles were isolated from each vertebral level, permitting the architectural measurements of mass, sarcomere length, normalized fiber length, physiological cross-sectional area, and fiber length-to-muscle length ratio. To determine the sarcomere length operating range of the muscle, sarcomere lengths were measured from intraoperative biopsy specimens that were obtained with the spine in the flexed and extended positions. The material properties of single muscle fibers were obtained from passive stress-strain tests of excised biopsy specimens. The average muscle mass (and standard error) was 146 +/- 8.7 g, and the average sarcomere length was 2.27 +/- 0.06 microm, yielding an average normalized fiber length of 5.66 +/- 0.65 cm, an average physiological cross-sectional area of 23.9 +/- 3.0 cm(2), and an average fiber length-to-muscle length ratio of 0.21 +/- 0.03. Intraoperative sarcomere length measurements revealed that the muscle operates from 1.98 +/- 0.15 microm in extension to 2.70 +/- 0.11 microm in flexion. Passive mechanical data suggested that the material properties of the muscle are comparable with those of muscles of the arm or leg. The architectural design (a high cross-sectional area and

  1. Animal electricity, Ca2+ and muscle contraction. A brief history of muscle research.

    Science.gov (United States)

    Martonosi, A N

    2000-01-01

    This brief review attempts to summarize some of the major phases of muscle research from Leeuwenhoek's description of sarcomeres in 1674, through Galvani's observation of "animal electricity" in 1791, to the discovery of Ca2+ as the key messenger in the coupling of nerve excitation to muscle contraction. The emerging molecular mechanism of the contraction process is one of the great achievements of biology, reflecting the intimate links between physics, chemistry and the life Sciences in the solution of biological problems.

  2. Isolation and individual electrical stimulation of single smooth-muscle cells from the urinary bladder of the pig

    NARCIS (Netherlands)

    J.J. Glerum (Jacobus); R. van Mastrigt (Ron); J.C. Romijn (Johannes); D.J. Griffiths (Derek)

    1987-01-01

    textabstractIn contrast to striated muscle, measurements on strips of smooth muscle cannot be uniquely interpreted in terms of an array of contractile units. Therefore scaling down to the single-cell level is necessary to gain detailed understanding of the contractile process in this type of muscle.

  3. Ultra-small angle X-ray diffraction from muscle

    Energy Technology Data Exchange (ETDEWEB)

    Nave, C.; Diakun, G.P.; Bordas, J.

    1986-05-15

    An ultra-small angle X-ray scattering instrument is described. It uses two channel cut crystals, one to monochromatise and collimate the beam and the other to analyse the scattered radiation. It has been used to collect diffraction data from muscle, in which the physiological unit cell, the sarcomere, has a repeat of 2000 nm or more.

  4. Spontaneous waves in muscle fibres

    Energy Technology Data Exchange (ETDEWEB)

    Guenther, Stefan; Kruse, Karsten [Department of Theoretical Physics, Saarland University, 66041 Saarbruecken (Germany); Max Planck Institute for the Physics of Complex Systems, Noethnitzer Street 38, 01187 Dresden (Germany)

    2007-11-15

    Mechanical oscillations are important for many cellular processes, e.g. the beating of cilia and flagella or the sensation of sound by hair cells. These dynamic states originate from spontaneous oscillations of molecular motors. A particularly clear example of such oscillations has been observed in muscle fibers under non-physiological conditions. In that case, motor oscillations lead to contraction waves along the fiber. By a macroscopic analysis of muscle fiber dynamics we find that the spontaneous waves involve non-hydrodynamic modes. A simple microscopic model of sarcomere dynamics highlights mechanical aspects of the motor dynamics and fits with the experimental observations.

  5. The striated muscles in pulmonary arterial hypertension: adaptations beyond the right ventricle

    NARCIS (Netherlands)

    Manders, E.; Rain, S.; Bogaard, H.J.; Handoko, M.L.; Stienen, G.J.M.; Vonk Noordegraaf, A.; Ottenheijm, C.A.C.; de Man, F.S.

    2015-01-01

    Pulmonary arterial hypertension (PAH) is a fatal lung disease characterised by progressive remodelling of the small pulmonary vessels. The daily-life activities of patients with PAH are severely limited by exertional fatigue and dyspnoea. Typically, these symptoms have been explained by right heart

  6. A striated muscle on the hard palate of rodents and rabbits

    Czech Academy of Sciences Publication Activity Database

    Pavlíková, H.; Witter, Kirsti; Míšek, Ivan

    2004-01-01

    Roč. 33, - (2004), s. 96-99 ISSN 0340-2096 R&D Projects: GA ČR GP304/01/P021; GA ČR GA304/02/0448; GA AV ČR KSK6005114 Institutional research plan: CEZ:AV0Z5045916 Keywords : hard palate * rabbits * rodents Subject RIV: EA - Cell Biology Impact factor: 0.625, year: 2004

  7. Effect of one stretch a week applied to the immobilized soleus muscle on rat muscle fiber morphology

    Directory of Open Access Journals (Sweden)

    Gomes A.R.S.

    2004-01-01

    Full Text Available We determined the effect of stretching applied once a week to the soleus muscle immobilized in the shortened position on muscle fiber morphology. Twenty-six male Wistar rats weighing 269 ± 26 g were divided into three groups. Group I, the left soleus was immobilized in the shortened position for 3 weeks; group II, the soleus was immobilized in the shortened position and stretched once a week for 3 weeks; group III, the soleus was submitted only to stretching once a week for 3 weeks. The medial part of the soleus muscle was frozen for histology and muscle fiber area evaluation and the lateral part was used for the determination of number and length of serial sarcomeres. Soleus muscle submitted only to immobilization showed a reduction in weight (44 ± 6%, P = 0.002, in serial sarcomere number (23 ± 15% and in cross-sectional area of the fibers (37 ± 31%, P < 0.001 compared to the contralateral muscles. The muscle that was immobilized and stretched showed less muscle fiber atrophy than the muscles only immobilized (P < 0.05. Surprisingly, in the muscles submitted only to stretching, fiber area was decreased compared to the contralateral muscle (2548 ± 659 vs 2961 ± 806 µm², respectively, P < 0.05. In conclusion, stretching applied once a week for 40 min to the soleus muscle immobilized in the shortened position was not sufficient to prevent the reduction of muscle weight and of serial sarcomere number, but provided significant protection against muscle fiber atrophy. In contrast, stretching normal muscles once a week caused a reduction in muscle fiber area.

  8. Comparative Biomechanics of Thick Filaments and Thin Filaments with Functional Consequences for Muscle Contraction

    Directory of Open Access Journals (Sweden)

    Mark S. Miller

    2010-01-01

    Full Text Available The scaffold of striated muscle is predominantly comprised of myosin and actin polymers known as thick filaments and thin filaments, respectively. The roles these filaments play in muscle contraction are well known, but the extent to which variations in filament mechanical properties influence muscle function is not fully understood. Here we review information on the material properties of thick filaments, thin filaments, and their primary constituents; we also discuss ways in which mechanical properties of filaments impact muscle performance.

  9. A Small Molecule Inhibitor of Sarcomere Contractility Acutely Relieves Left Ventricular Outflow Tract Obstruction in Feline Hypertrophic Cardiomyopathy.

    Directory of Open Access Journals (Sweden)

    Joshua A Stern

    Full Text Available Hypertrophic cardiomyopathy (HCM is an inherited disease of the heart muscle characterized by otherwise unexplained thickening of the left ventricle. Left ventricular outflow tract (LVOT obstruction is present in approximately two-thirds of patients and substantially increases the risk of disease complications. Invasive treatment with septal myectomy or alcohol septal ablation can improve symptoms and functional status, but currently available drugs for reducing obstruction have pleiotropic effects and variable therapeutic responses. New medical treatments with more targeted pharmacology are needed, but the lack of preclinical animal models for HCM with LVOT obstruction has limited their development. HCM is a common cause of heart failure in cats, and a subset exhibit systolic anterior motion of the mitral valve leading to LVOT obstruction. MYK-461 is a recently-described, mechanistically novel small molecule that acts at the sarcomere to specifically inhibit contractility that has been proposed as a treatment for HCM. Here, we use MYK-461 to test whether direct reduction in contractility is sufficient to relieve LVOT obstruction in feline HCM. We evaluated mixed-breed cats in a research colony derived from a Maine Coon/mixed-breed founder with naturally-occurring HCM. By echocardiography, we identified five cats that developed systolic anterior motion of the mitral valve and LVOT obstruction both at rest and under anesthesia when provoked with an adrenergic agonist. An IV MYK-461 infusion and echocardiography protocol was developed to serially assess contractility and LVOT gradient at multiple MYK-461 concentrations. Treatment with MYK-461 reduced contractility, eliminated systolic anterior motion of the mitral valve and relieved LVOT pressure gradients in an exposure-dependent manner. Our findings provide proof of principle that acute reduction in contractility with MYK-461 is sufficient to relieve LVOT obstruction. Further, these studies

  10. The gaseous plasmonic response of a one-dimensional photonic crystal composed of striated plasma layers

    Science.gov (United States)

    Wang, B.; Righetti, F.; Cappelli, M. A.

    2018-03-01

    We present simulations of the response of a one-dimensional striated plasma slab to incident electromagnetic waves that span regions both above and below the plasma frequency, ωp. Photonic bandgap modes are present throughout these regions, and volume and surface plasmon modes facilitate the response below ωp, where the dielectric constant, ɛp application of these structures as ultra-narrow tunable microwave transmission filters.

  11. Neural correlates of visual motion processing without awareness in patients with striate cortex and pulvinar lesions.

    Science.gov (United States)

    Barleben, Maria; Stoppel, Christian M; Kaufmann, Jörn; Merkel, Christian; Wecke, Thoralf; Goertler, Michael; Heinze, Hans-Jochen; Hopf, Jens-Max; Schoenfeld, Mircea A

    2015-04-01

    Patients with striate cortex lesions experience visual perception loss in the contralateral visual field. In few patients, however, stimuli within the blind field can lead to unconscious (blindsight) or even conscious perception when the stimuli are moving (Riddoch syndrome). Using functional magnetic resonance imaging (fMRI), we investigated the neural responses elicited by motion stimulation in the sighted and blind visual fields of eight patients with lesions of the striate cortex. Importantly, repeated testing ensured that none of the patients exhibited blindsight or a Riddoch syndrome. Three patients had additional lesions in the ipsilesional pulvinar. For blind visual field stimulation, great care was given that the moving stimulus was precisely presented within the borders of the scotoma. In six of eight patients, the stimulation within the scotoma elicited hemodynamic activity in area human middle temporal (hMT) while no activity was observed within the ipsilateral lesioned area of the striate cortex. One of the two patients in whom no ipsilesional activity was observed had an extensive lesion including massive subcortical damage. The other patient had an additional focal lesion within the lateral inferior pulvinar. Fiber-tracking based on anatomical and functional markers (hMT and Pulvinar) on individual diffusion tensor imaging (DTI) data from each patient revealed the structural integrity of subcortical pathways in all but the patient with the extensive subcortical lesion. These results provide clear evidence for the robustness of direct subcortical pathways from the pulvinar to area hMT in patients with striate cortex lesions and demonstrate that ipsilesional activity in area hMT is completely independent of conscious perception. © 2014 Wiley Periodicals, Inc.

  12. Hypertrophic Cardiomyopathy: A Vicious Cycle Triggered by Sarcomere Mutations and Secondary Disease Hits.

    Science.gov (United States)

    Wijnker, Paul J M; Sequeira, Vasco; Kuster, Diederik W D; Velden, Jolanda van der

    2018-04-11

    Hypertrophic cardiomyopathy (HCM) is a cardiac genetic disease characterized by left ventricular hypertrophy, diastolic dysfunction, and myocardial disarray. Disease onset occurs between 20 and 50 years of age, thus affecting patients in the prime of their life. HCM is caused by mutations in sarcomere proteins, the contractile building blocks of the heart. Despite increased knowledge of causal mutations, the exact path from genetic defect leading to cardiomyopathy is complex and involves additional disease hits. Recent Advances: Laboratory-based studies indicate that HCM development not only depends on the primary sarcomere impairment caused by the mutation but also on secondary disease-related alterations in the heart. Here we propose a vicious mutation-induced disease cycle, in which a mutation-induced energy depletion alters cellular metabolism with increased mitochondrial work, which triggers secondary disease modifiers that will worsen disease and ultimately lead to end-stage HCM. Evidence shows excessive cellular reactive oxygen species (ROS) in HCM patients and HCM animal models. Oxidative stress markers are increased in the heart (oxidized proteins, DNA, and lipids) and serum of HCM patients. In addition, increased mitochondrial ROS production and changes in endogenous antioxidants are reported in HCM. Mutant sarcomeric protein may drive excessive levels of cardiac ROS via changes in cardiac efficiency and metabolism, mitochondrial activation and/or dysfunction, impaired protein quality control, and microvascular dysfunction. Interventions restoring metabolism, mitochondrial function, and improved ROS balance may be promising therapeutic approaches. We discuss the effects of current HCM pharmacological therapies and potential future therapies to prevent and reverse HCM. Antioxid. Redox Signal. 00, 000-000.

  13. The Novel Desmin Mutant p.A120D Impairs Filament Formation, Prevents Intercalated Disk Localization, and Causes Sudden Cardiac Death

    DEFF Research Database (Denmark)

    Brodehl, Andreas; Dieding, Mareike; Klauke, Bärbel

    2013-01-01

    The intermediate filament protein desmin is encoded by the gene DES and contributes to the mechanical stabilization of the striated muscle sarcomere and cell contacts within the cardiac intercalated disk. DES mutations cause severe skeletal and cardiac muscle diseases with heterogeneous phenotype...

  14. Structure, function and evolution of insect flight muscle

    OpenAIRE

    Iwamoto, Hiroyuki

    2011-01-01

    Insects, the largest group of animals on the earth, owe their prosperity to their ability of flight and small body sizes. The ability of flight provided means for rapid translocation. The small body size allowed access to unutilized niches. By acquiring both features, however, insects faced a new problem: They were forced to beat their wings at enormous frequencies. Insects have overcome this problem by inventing asynchronous flight muscle, a highly specialized form of striated muscle capable...

  15. Oxygen dependence of respiration in rat spinotrapezius muscle in situ

    OpenAIRE

    Golub, Aleksander S.; Pittman, Roland N.

    2012-01-01

    The oxygen dependence of respiration in striated muscle in situ was studied by measuring the rate of decrease of interstitial Po2 [oxygen disappearance curve (ODC)] following rapid arrest of blood flow by pneumatic tissue compression, which ejected red blood cells from the muscle vessels and made the ODC independent from oxygen bound to hemoglobin. After the contribution of photo-consumption of oxygen by the method was evaluated and accounted for, the corrected ODCs were converted into the Po...

  16. T1 measurements identify extracellular volume expansion in hypertrophic cardiomyopathy sarcomere mutation carriers with and without left ventricular hypertrophy.

    Science.gov (United States)

    Ho, Carolyn Y; Abbasi, Siddique A; Neilan, Tomas G; Shah, Ravi V; Chen, Yucheng; Heydari, Bobak; Cirino, Allison L; Lakdawala, Neal K; Orav, E John; González, Arantxa; López, Begoña; Díez, Javier; Jerosch-Herold, Michael; Kwong, Raymond Y

    2013-05-01

    Myocardial fibrosis is a hallmark of hypertrophic cardiomyopathy (HCM) and a potential substrate for arrhythmias and heart failure. Sarcomere mutations seem to induce profibrotic changes before left ventricular hypertrophy (LVH) develops. To further evaluate these processes, we used cardiac magnetic resonance with T1 measurements on a genotyped HCM population to quantify myocardial extracellular volume (ECV). Sarcomere mutation carriers with LVH (G+/LVH+, n=37) and without LVH (G+/LVH-, n=29), patients with HCM without mutations (sarcomere-negative HCM, n=11), and healthy controls (n=11) underwent contrast cardiac magnetic resonance, measuring T1 times pre- and postgadolinium infusion. Concurrent echocardiography and serum biomarkers of collagen synthesis, hemodynamic stress, and myocardial injury were also available in a subset. Compared with controls, ECV was increased in patients with overt HCM, as well as G+/LVH- mutation carriers (ECV=0.36±0.01, 0.33±0.01, 0.27±0.01 in G+/LVH+, G+/LVH-, controls, respectively; P≤0.001 for all comparisons). ECV correlated with N-terminal probrain natriuretic peptide levels (r=0.58; P60% of overt patients with HCM but absent from G+/LVH- subjects. Both ECV and late gadolinium enhancement were more extensive in sarcomeric HCM than sarcomere-negative HCM. Myocardial ECV is increased in HCM sarcomere mutation carriers even in the absence of LVH. These data provide additional support that fibrotic remodeling is triggered early in disease pathogenesis. Quantifying ECV may help characterize the development of myocardial fibrosis in HCM and ultimately assist in developing novel disease-modifying therapy, targeting interstitial fibrosis.

  17. Persistent fibrosis, hypertrophy and sarcomere disorganisation after endoscopy-guided heart resection in adult Xenopus.

    Directory of Open Access Journals (Sweden)

    Lindsey Marshall

    Full Text Available Models of cardiac repair are needed to understand mechanisms underlying failure to regenerate in human cardiac tissue. Such studies are currently dominated by the use of zebrafish and mice. Remarkably, it is between these two evolutionary separated species that the adult cardiac regenerative capacity is thought to be lost, but causes of this difference remain largely unknown. Amphibians, evolutionary positioned between these two models, are of particular interest to help fill this lack of knowledge. We thus developed an endoscopy-based resection method to explore the consequences of cardiac injury in adult Xenopus laevis. This method allowed in situ live heart observation, standardised tissue amputation size and reproducibility. During the first week following amputation, gene expression of cell proliferation markers remained unchanged, whereas those relating to sarcomere organisation decreased and markers of inflammation, fibrosis and hypertrophy increased. One-month post-amputation, fibrosis and hypertrophy were evident at the injury site, persisting through 11 months. Moreover, cardiomyocyte sarcomere organisation deteriorated early following amputation, and was not completely recovered as far as 11 months later. We conclude that the adult Xenopus heart is unable to regenerate, displaying cellular and molecular marks of scarring. Our work suggests that, contrary to urodeles and teleosts, with the exception of medaka, adult anurans share a cardiac injury outcome similar to adult mammals. This observation is at odds with current hypotheses that link loss of cardiac regenerative capacity with acquisition of homeothermy.

  18. Cardiac Phosphoproteomics during Remote Ischemic Preconditioning: A Role for the Sarcomeric Z-Disk Proteins

    Directory of Open Access Journals (Sweden)

    Safa Abdul-Ghani

    2014-01-01

    Full Text Available Remote ischemic preconditioning (RIPC induced by brief ischemia/reperfusion cycles of remote organ (e.g., limb is cardioprotective. The myocardial cellular changes during RIPC responsible for this phenomenon are not currently known. The aim of this work was to identify the activation by phosphorylation of cardiac proteins following RIPC. To achieve our aim we used isobaric tandem mass tagging (TMT and reverse phase nanoliquid chromatography tandem spectrometry using a Linear Trap Quadropole (LTQ Orbitrap Velos mass spectrometer. Male C57/Bl6 mice were anesthetized by an intraperitoneal injection of Tribromoethanol. A cuff was placed around the hind limb and inflated at 200 mmHg to prevent blood flow as confirmed by Laser Doppler Flowmetry. RIPC was induced by 4 cycles of 5 min of limb ischemia followed by 5 min of reperfusion. Hearts were extracted for phosphoproteomics. We identified approximately 30 phosphoproteins that were differentially expressed in response to RIPC protocol. The levels of several phosphoproteins in the Z-disk of the sarcomere including phospho-myozenin-2 were significantly higher than control. This study describes and validates a novel approach to monitor the changes in the cardiac phosphoproteome following the cardioprotective intervention of RIPC and prior to index ischemia. The increased level of phosphorylated sarcomeric proteins suggests they may have a role in cardiac signaling during RIPC.

  19. The multiple roles of titin in muscle contraction and force production.

    Science.gov (United States)

    Herzog, Walter

    2018-01-20

    Titin is a filamentous protein spanning the half-sarcomere, with spring-like properties in the I-band region. Various structural, signaling, and mechanical functions have been associated with titin, but not all of these are fully elucidated and accepted in the scientific community. Here, I discuss the primary mechanical functions of titin, including its accepted role in passive force production, stabilization of half-sarcomeres and sarcomeres, and its controversial contribution to residual force enhancement, passive force enhancement, energetics, and work production in shortening muscle. Finally, I provide evidence that titin is a molecular spring whose stiffness changes with muscle activation and actin-myosin-based force production, suggesting a novel model of force production that, aside from actin and myosin, includes titin as a "third contractile" filament. Using this three-filament model of sarcomeres, the stability of (half-) sarcomeres, passive force enhancement, residual force enhancement, and the decrease in metabolic energy during and following eccentric contractions can be explained readily.

  20. Deletion of Drosophila muscle LIM protein decreases flight muscle stiffness and power generation

    OpenAIRE

    Clark, Kathleen A.; Lesage-Horton, Heather; Zhao, Cuiping; Beckerle, Mary C.; Swank, Douglas M.

    2011-01-01

    Muscle LIM protein (MLP) can be found at the Z-disk of sarcomeres where it is hypothesized to be involved in sensing muscle stretch. Loss of murine MLP results in dilated cardiomyopathy, and mutations in human MLP lead to cardiac hypertrophy, indicating a critical role for MLP in maintaining normal cardiac function. Loss of MLP in Drosophila (mlp84B) also leads to muscle dysfunction, providing a model system to examine MLP's mechanism of action. Mlp84B-null flies that survive to adulthood are...

  1. The human cardiac and skeletal muscle proteomes defined by transcriptomics and antibody-based profiling

    DEFF Research Database (Denmark)

    Lindskog, Cecilia; Linne, Jerker; Fagerberg, Linn

    2015-01-01

    Background: To understand cardiac and skeletal muscle function, it is important to define and explore their molecular constituents and also to identify similarities and differences in the gene expression in these two different striated muscle tissues. Here, we have investigated the genes and prot......Background: To understand cardiac and skeletal muscle function, it is important to define and explore their molecular constituents and also to identify similarities and differences in the gene expression in these two different striated muscle tissues. Here, we have investigated the genes...... genes are well in line with the physiological functions of cardiac and skeletal muscle, such as contraction, ion transport, regulation of membrane potential and actomyosin structure organization. A large fraction of the transcripts in both cardiac and skeletal muscle correspond to mitochondrial proteins...

  2. Single Muscle Fiber Proteomics Reveals Fiber-Type-Specific Features of Human Muscle Aging

    Directory of Open Access Journals (Sweden)

    Marta Murgia

    2017-06-01

    Full Text Available Skeletal muscle is a key tissue in human aging, which affects different muscle fiber types unequally. We developed a highly sensitive single muscle fiber proteomics workflow to study human aging and show that the senescence of slow and fast muscle fibers is characterized by diverging metabolic and protein quality control adaptations. Whereas mitochondrial content declines with aging in both fiber types, glycolysis and glycogen metabolism are upregulated in slow but downregulated in fast muscle fibers. Aging mitochondria decrease expression of the redox enzyme monoamine oxidase A. Slow fibers upregulate a subset of actin and myosin chaperones, whereas an opposite change happens in fast fibers. These changes in metabolism and sarcomere quality control may be related to the ability of slow, but not fast, muscle fibers to maintain their mass during aging. We conclude that single muscle fiber analysis by proteomics can elucidate pathophysiology in a sub-type-specific manner.

  3. Evaluation of muscle function of the extensor digitorum longus muscle ex vivo and tibialis anterior muscle in situ in mice.

    Science.gov (United States)

    Hakim, Chady H; Wasala, Nalinda B; Duan, Dongsheng

    2013-02-09

    Body movements are mainly provided by mechanical function of skeletal muscle. Skeletal muscle is composed of numerous bundles of myofibers that are sheathed by intramuscular connective tissues. Each myofiber contains many myofibrils that run longitudinally along the length of the myofiber. Myofibrils are the contractile apparatus of muscle and they are composed of repeated contractile units known as sarcomeres. A sarcomere unit contains actin and myosin filaments that are spaced by the Z discs and titin protein. Mechanical function of skeletal muscle is defined by the contractile and passive properties of muscle. The contractile properties are used to characterize the amount of force generated during muscle contraction, time of force generation and time of muscle relaxation. Any factor that affects muscle contraction (such as interaction between actin and myosin filaments, homeostasis of calcium, ATP/ADP ratio, etc.) influences the contractile properties. The passive properties refer to the elastic and viscous properties (stiffness and viscosity) of the muscle in the absence of contraction. These properties are determined by the extracellular and the intracellular structural components (such as titin) and connective tissues (mainly collagen) (1-2). The contractile and passive properties are two inseparable aspects of muscle function. For example, elbow flexion is accomplished by contraction of muscles in the anterior compartment of the upper arm and passive stretch of muscles in the posterior compartment of the upper arm. To truly understand muscle function, both contractile and passive properties should be studied. The contractile and/or passive mechanical properties of muscle are often compromised in muscle diseases. A good example is Duchenne muscular dystrophy (DMD), a severe muscle wasting disease caused by dystrophin deficiency (3). Dystrophin is a cytoskeletal protein that stabilizes the muscle cell membrane (sarcolemma) during muscle contraction (4). In the

  4. The pelvic floor muscles: muscle thickness in healthy and urinary-incontinent women measured by perineal ultrasonography with reference to the effect of pelvic floor training. Estrogen receptor studies

    DEFF Research Database (Denmark)

    Bernstein, Inge Thomsen

    1997-01-01

    depends on the structural components in the urethral wall, the position of the bladder neck and proximal urethra, the periurethral striated muscles, and the pelvic floor muscles. By means of pudendal blockade and simultaneous recordings of pressure and cross-sectional area in the urethra, it has been...... demonstrated that the striated periurethral muscles and the pelvic floor muscles are of paramount importance for the closure function. This emphasizes the importance of well-functioning pelvic floor muscles to obtain continence, and probably explains the rationale for the effect of pelvic floor training...... in treating urinary incontinence. This study presents a review of the literature on female urinary incontinence, continence mechanisms, pelvic floor muscles, and pelvic floor training. Furthermore, a review of the literature on estrogen receptors in the pelvic floor muscles is given. Perineal ultrasonography...

  5. Troponin activator augments muscle force in nemaline myopathy patients with nebulin mutations

    NARCIS (Netherlands)

    de Winter, J.M.; Buck, D.; Hidalgo, C.; Jasper, J.R.; Malik, F.I.; Clarke, N.F.; Stienen, G.J.M.; Lawlor, M.W.; Beggs, A.H.; Ottenheijm, C.A.C.; Granzier, H.

    2013-01-01

    Background: Nemaline myopathy-the most common non-dystrophic congenital myopathy-is caused by mutations in thin filament genes, of which the nebulin gene is the most frequently affected one. The nebulin gene codes for the giant sarcomeric protein nebuli which plays a crucial role in skeletal muscle

  6. Diagnostic yield, interpretation, and clinical utility of mutation screening of sarcomere encoding genes in Danish hypertrophic cardiomyopathy patients and relatives

    DEFF Research Database (Denmark)

    Andersen, Paal Skytt; Havndrup, Ole; Hougs, Lotte

    2008-01-01

    The American Heart Association (AHA) recommends family screening for hypertrophic cardiomyopathy (HCM). We assessed the outcome of family screening combining clinical evaluation and screening for sarcomere gene mutations in a cohort of 90 Danish HCM patients and their close relatives, in all 451...

  7. Effects of adenosine triphosphate concentration on motor force regulation during skeletal muscle contraction

    Science.gov (United States)

    Wei, J.; Dong, C.; Chen, B.

    2017-04-01

    We employ a mechanical model of sarcomere to quantitatively investigate how adenosine triphosphate (ATP) concentration affects motor force regulation during skeletal muscle contraction. Our simulation indicates that there can be negative cross-bridges resisting contraction within the sarcomere and higher ATP concentration would decrease the resistance force from negative cross-bridges by promoting their timely detachment. It is revealed that the motor force is well regulated only when ATP concentration is above a certain level. These predictions may provide insights into the role of ATP in regulating coordination among multiple motors.

  8. Mitochondrial affinity for ADP is twofold lower in creatine kinase knock-out muscles - Possible role in rescuing cellular energy homeostasis

    NARCIS (Netherlands)

    ter Veld, F; Jeneson, JAL; Nicolay, K

    Adaptations of the kinetic properties of mitochondria in striated muscle lacking cytosolic (M) and/or mitochondrial (Mi) creatine kinase (CK) isoforms in comparison to wild-type (WT) were investigated in vitro. Intact mitochondria were isolated from heart and gastrocnemius muscle of WT and single-

  9. Effect of passive stretching on the immobilized soleus muscle fiber morphology

    Directory of Open Access Journals (Sweden)

    Coutinho E.L.

    2004-01-01

    Full Text Available The aim of the present study was to determine the effect of stretching applied every 3 days to the soleus muscle immobilized in the shortened position on muscle fiber morphology. Eighteen 16-week-old Wistar rats were used and divided into three groups of 6 animals each: a the left soleus muscle was immobilized in the shortened position for 3 weeks; b during immobilization, the soleus was stretched for 40 min every 3 days; c the non-immobilized soleus was only stretched. Left and right soleus muscles were examined. One portion of the soleus was frozen for histology and muscle fiber area evaluation, while the other portion was used to identify the number and length of serial sarcomeres. Immobilized muscles (group A showed a significant decrease in weight (44 ± 6%, length (19 ± 7%, serial sarcomere number (23 ± 15%, and fiber area (37 ± 31% compared to the contralateral muscles (P < 0.05, paired Student t-test. The immobilized and stretched soleus (group B showed a similar reduction but milder muscle fiber atrophy compared to the only immobilized group (22 ± 40 vs 37 ± 31%, respectively; P < 0.001, ANOVA test. Muscles submitted only to stretching (group C significantly increased the length (5 ± 2%, serial sarcomere number (4 ± 4%, and fiber area (16 ± 44% compared to the contralateral muscles (P < 0.05, paired Student t-test. In conclusion, stretching applied every 3 days to immobilized muscles did not prevent the muscle shortening, but reduced muscle atrophy. Stretching sessions induced hypertrophic effects in the control muscles. These results support the use of muscle stretching in sports and rehabilitation.

  10. Optical reflectance in fibrous tissues and skeletal muscles

    Science.gov (United States)

    Ranasinghesagara, Janaka C.

    We studied two biological tissues with optically anisotropic structures: high moisture soy protein extrudates and skeletal muscles. High moisture extrusion has been used to produce vegetable meat analogs that resemble real animal meat and have significant health benefits. Since visual and textural properties are key factors for consumer acceptance, assessing fiber formation in the extruded soy protein product is important for quality control purpose. A non-destructive method based on photon migration was developed to measure fiber formation in extruded soy proteins. The measured fiber formation index in intact samples showed good agreement with that obtained from image analysis on peeled samples. By implementing this new method in a fast laser scanning system, we have acquired two dimensional mappings of fiber formation and orientation in the entire sample in real time. In addition to fibrous structures, skeletal muscles have a unique periodic sarcomere structure which produces strong light diffractions. However, inconsistent experimental results have been reported in single fiber diffraction studies. By applying the three-dimensional coupled wave theory in a physical sarcomere model, we found that a variety of experimental observations can be explained if inhomogeneous muscle morphological profiles are considered. We also discovered that the sarcomere structure produced a unique optical reflectance pattern in whole muscle. None of the existing light propagation theories are able to describe this pattern. We developed a Monte Carlo model incorporating the sarcomere diffraction effect. The simulated results quantitatively resemble the unique patterns observed in experiments. We used a set of parameters to quantify the optical reflectance profiles produced by a point incident light in whole muscle. Two parameters, q and B, were obtained by numerically fitting the equi-intensity contours of the reflectance pattern. Two spatial gradients were calculated along the

  11. Sarcomeric Myosin Expression in the Tongue Body of Humans, Macaques and Rats

    OpenAIRE

    Rahnert, Jill A.; Sokoloff, Alan J.; Burkholder, Thomas J.

    2009-01-01

    Expression of developmental and unconventional myosin heavy chain (MHC) isoforms in some adult head and neck muscles is thought to reflect specific contractile demands of muscle fibers active during kinematically complex movements. Mammalian tongue muscles are active during oromotor behaviors that encompass a wide range of tongue movement speeds and tongue shape changes (e.g. respiration, oral transport, swallowing, rejection), but the extent to which tongue muscles express developmental and ...

  12. Effect of distribution of striated laser hardening tracks on dry sliding wear resistance of biomimetic surface

    Science.gov (United States)

    Su, Wei; Zhou, Ti; Zhang, Peng; Zhou, Hong; Li, Hui

    2018-01-01

    Some biological surfaces were proved to have excellent anti-wear performance. Being inspired, Nd:YAG pulsed laser was used to create striated biomimetic laser hardening tracks on medium carbon steel samples. Dry sliding wear tests biomimetic samples were performed to investigate specific influence of distribution of laser hardening tracks on sliding wear resistance of biomimetic samples. After comparing wear weight loss of biomimetic samples, quenched sample and untreated sample, it can be suggested that the sample covered with dense laser tracks (3.5 mm spacing) has lower wear weight loss than the one covered with sparse laser tracks (4.5 mm spacing); samples distributed with only dense laser tracks or sparse laser tracks (even distribution) were proved to have better wear resistance than samples distributed with both dense and sparse tracks (uneven distribution). Wear mechanisms indicate that laser track and exposed substrate of biomimetic sample can be regarded as hard zone and soft zone respectively. Inconsecutive striated hard regions, on the one hand, can disperse load into small branches, on the other hand, will hinder sliding abrasives during wear. Soft regions with small range are beneficial in consuming mechanical energy and storing lubricative oxides, however, soft zone with large width (>0.5 mm) will be harmful to abrasion resistance of biomimetic sample because damages and material loss are more obvious on surface of soft phase. As for the reason why samples with even distributed bionic laser tracks have better wear resistance, it can be explained by the fact that even distributed laser hardening tracks can inhibit severe worn of local regions, thus sliding process can be more stable and wear extent can be alleviated as well.

  13. Comparison of echocardiographic and cardiac magnetic resonance imaging in hypertrophic cardiomyopathy sarcomere mutation carriers without left ventricular hypertrophy.

    Science.gov (United States)

    Valente, Anne Marie; Lakdawala, Neal K; Powell, Andrew J; Evans, Sarah P; Cirino, Allison L; Orav, E John; MacRae, Calum A; Colan, Steven D; Ho, Carolyn Y

    2013-06-01

    Left ventricular hypertrophy (LVH) typically manifests during or after adolescence in sarcomere mutation carriers at risk for developing hypertrophic cardiomyopathy. Guidelines recommend serial imaging of mutation carriers without LVH (G+/LVH-) to monitor for phenotypic evolution, but the optimal strategy is undefined. Compared with echocardiography (echo), cardiac MRI (CMR) offers improved endocardial visualization and potential to assess scar. However, the incremental advantage offered by CMR for early diagnosis of hypertrophic cardiomyopathy is unclear. Therefore, we systematically compared echo and CMR in G+/LVH- subjects. A total of 40 sarcomere mutation carriers with normal echo wall thickness (hypertrophic cardiomyopathy, including 1 with mild LVH by CMR at baseline. Echo is unlikely to miss substantial LVH; however, CMR identified mild hypertrophy in ≈10% of mutation carriers with normal echo wall thickness. CMR may be a useful adjunct in hypertrophic cardiomyopathy family screening, particularly in higher risk situations, or if echocardiographic images are suboptimal or suggest borderline LVH.

  14. Drosophila small heat shock protein CryAB ensures structural integrity of developing muscles, and proper muscle and heart performance.

    Science.gov (United States)

    Wójtowicz, Inga; Jabłońska, Jadwiga; Zmojdzian, Monika; Taghli-Lamallem, Ouarda; Renaud, Yoan; Junion, Guillaume; Daczewska, Malgorzata; Huelsmann, Sven; Jagla, Krzysztof; Jagla, Teresa

    2015-03-01

    Molecular chaperones, such as the small heat shock proteins (sHsps), maintain normal cellular function by controlling protein homeostasis in stress conditions. However, sHsps are not only activated in response to environmental insults, but also exert developmental and tissue-specific functions that are much less known. Here, we show that during normal development the Drosophila sHsp CryAB [L(2)efl] is specifically expressed in larval body wall muscles and accumulates at the level of Z-bands and around myonuclei. CryAB features a conserved actin-binding domain and, when attenuated, leads to clustering of myonuclei and an altered pattern of sarcomeric actin and the Z-band-associated actin crosslinker Cheerio (filamin). Our data suggest that CryAB and Cheerio form a complex essential for muscle integrity: CryAB colocalizes with Cheerio and, as revealed by mass spectrometry and co-immunoprecipitation experiments, binds to Cheerio, and the muscle-specific attenuation of cheerio leads to CryAB-like sarcomeric phenotypes. Furthermore, muscle-targeted expression of CryAB(R120G), which carries a mutation associated with desmin-related myopathy (DRM), results in an altered sarcomeric actin pattern, in affected myofibrillar integrity and in Z-band breaks, leading to reduced muscle performance and to marked cardiac arrhythmia. Taken together, we demonstrate that CryAB ensures myofibrillar integrity in Drosophila muscles during development and propose that it does so by interacting with the actin crosslinker Cheerio. The evidence that a DRM-causing mutation affects CryAB muscle function and leads to DRM-like phenotypes in the fly reveals a conserved stress-independent role of CryAB in maintaining muscle cell cytoarchitecture. © 2015. Published by The Company of Biologists Ltd.

  15. A novel electrical model of nerve and muscle using Pspice

    CERN Document Server

    Peasgood, W; Lam, C K; Armstrong, A G; Wood, W

    2003-01-01

    In this work, a model is developed to simulate the biological processes involved in nerve fibre transmission and subsequent muscle contraction. The model has been based on approximating biological structure and function to electrical circuits and as such was implemented on an electronics simulation software package called Pspice. Models of nerve, the nerve-muscle interface and muscle fibre have been implemented. The time dependent ionic properties of the nerve and muscle membranes have been simulated using the Hodgkin-Huxley equations and for the muscle fibre, the implementation of the Huxley sliding filament theory for muscular contraction. The results show that nerve may be considered as a fractal transmission line and that the amplitude of the nerve membrane depolarization is dependent on the dimensions of the fibre. Additionally, simulation of the nerve-muscle interface allows the fractal nerve model to be connected to the muscle fibre model and it is shown that a two sarcomere molecular simulation can pr...

  16. The role of variable muscle adaptation to limb lengthening in the development of joint contractures: an experimental study in the goat.

    Science.gov (United States)

    Makarov, Marina; Birch, John; Samchukov, Mikhail

    2009-03-01

    Muscle stiffness and joint contractures are currently regarded as the most common complications of limb lengthening. To better understand the mechanisms of joint contractures, architectural changes of all involved muscles were analyzed in 9 goats after 20% tibial lengthening with standard distraction protocol.All 13 muscles of the goat's tibia were found to be organized into an anterior compartment with 2 longitudinal and 4 pennate muscles and a posterior compartment with 1 longitudinal and 6 pennate muscles. Longitudinal muscles showed better compliance to distraction than pinnate muscles. Although muscle-to-bone lengthening ratio ranged widely (0-1.2), most of the muscles and especially those located in the posterior compartment showed much less lengthening than the bone. Muscular portions of the muscles lengthened more substantially (average, 17%) than their associated tendons (average, 7%). Muscle fiber length changes varied greatly between muscles (range, 0%-88%). Normalization of muscle fiber length revealed considerable elongation of anterior muscles fibers (25%) that was associated with an addition of new sarcomeres in series. Fiber length increase of all posterior muscles but one occurred by stretching of existing sarcomeres, with little addition or even dissolution of sarcomeres in series. This correlated with muscle mass changes showing significant muscle atrophy in the posterior compartment and better mass preservation in the anterior compartment.The study revealed striking difference in response to limb lengthening between individual muscles and muscles from antagonistic compartments in particular. Poor sarcomerogenesis in the posterior muscles leading to their insufficient length increase seems to play major role in the development of joint contractures.

  17. Muscle trade-offs in a power-amplified prey capture system.

    Science.gov (United States)

    Blanco, M Mendoza; Patek, S N

    2014-05-01

    Should animals operating at great speeds and accelerations use fast or slow muscles? The answer hinges on a fundamental trade-off: muscles can be maximally fast or forceful, but not both. Direct lever systems offer a straightforward manifestation of this trade-off, yet the fastest organisms use power amplification, not direct lever action. Power-amplified systems typically use slow, forceful muscles to preload springs, which then rapidly release elastic potential energy to generate high speeds and accelerations. However, a fast response to a stimulus may necessitate fast spring-loading. Across 22 mantis shrimp species (Stomatopoda), this study examined how muscle anatomy correlates with spring mechanics and appendage type. We found that muscle force is maximized through physiological cross-sectional area, but not through sarcomere length. Sit-and-wait predators (spearers) had the shortest sarcomere lengths (fastest contractions) and the slowest strike speeds. The species that crush shells (smashers) had the fastest speeds, most forceful springs, and longest sarcomeres. The origin of the smasher clade yielded dazzlingly high accelerations, perhaps due to the release from fast spring-loading for evasive prey capture. This study offers a new window into the dynamics of force-speed trade-offs in muscles in the biomechanical, comparative evolutionary framework of power-amplified systems. © 2014 The Author(s). Evolution © 2014 The Society for the Study of Evolution.

  18. Determination of the source of SHG verniers in zebrafish skeletal muscle

    Science.gov (United States)

    Dempsey, William P.; Hodas, Nathan O.; Ponti, Aaron; Pantazis, Periklis

    2015-12-01

    SHG microscopy is an emerging microscopic technique for medically relevant imaging because certain endogenous proteins, such as muscle myosin lattices within muscle cells, are sufficiently spatially ordered to generate detectable SHG without the use of any fluorescent dye. Given that SHG signal is sensitive to the structural state of muscle sarcomeres, SHG functional imaging can give insight into the integrity of muscle cells in vivo. Here, we report a thorough theoretical and experimental characterization of myosin-derived SHG intensity profiles within intact zebrafish skeletal muscle. We determined that “SHG vernier” patterns, regions of bifurcated SHG intensity, are illusory when sarcomeres are staggered with respect to one another. These optical artifacts arise due to the phase coherence of SHG signal generation and the Guoy phase shift of the laser at the focus. In contrast, two-photon excited fluorescence images obtained from fluorescently labeled sarcomeric components do not contain such illusory structures, regardless of the orientation of adjacent myofibers. Based on our results, we assert that complex optical artifacts such as SHG verniers should be taken into account when applying functional SHG imaging as a diagnostic readout for pathological muscle conditions.

  19. Detection of ultrastructural changes in genetically altered and exercised skeletal muscle using PS-OCT

    Science.gov (United States)

    Pasquesi, James J.; Schlachter, Simon C.; Boppart, Marni D.; Chaney, Eric; Kaufman, Stephen J.; Boppart, Stephen A.

    2006-02-01

    Birefringence of skeletal muscle has been associated with the ultrastructure of individual sarcomeres, specifically the arrangement of A-bands corresponding to the thick myosin filaments. Murine skeletal muscle (gastrocnemius) was imaged with a fiber-based PS-OCT imaging system to determine the level of birefringence present in the tissue under various conditions. In addition to muscle controls from wild-type mice, muscle from abnormal mice included: genetically-modified (mdx) mice which model human muscular dystrophy, transgenic mice exhibiting an overexpression of integrin (α7β1), and transgenic integrin (α7β1)knockout mice. Comparisons were also made between rested and exercised muscles to determine the effects of exercise on muscle birefringence for each of these normal and abnormal conditions. The PS-OCT images revealed that the presence of birefringence was similar in the rested muscle with dystrophy-like features (i.e., lacking the structural protein dystrophin - mdx) and in the integrin (α7β1)knockout muscle when compared to the normal (wild-type) control. However, exercising these abnormal muscle tissues drastically reduced the presence of birefringence detected by the PS-OCT system. The muscle exhibiting an overexpression of integrin (α7β1) remained heavily birefringent before and after exercise, similar to the normal (wild-type) muscle. These results suggest that there is a distinct relationship between the degree of birefringence detected using PS-OCT and the sarcomeric ultrastructure present within skeletal muscle.

  20. Architectural differences between the hamstring muscles.

    Science.gov (United States)

    Kellis, Eleftherios; Galanis, Nikiforos; Kapetanos, George; Natsis, Konstantinos

    2012-08-01

    The purpose of this study was to understand the detailed architectural properties of the human hamstring muscles. The long (BFlh) and short (BFsh) head of biceps femoris, semimembranosus (SM) and semitendinosus (ST) muscles were dissected and removed from their origins in eight cadaveric specimens (age 67.8±4.3 years). Mean fiber length, sarcomere length, physiological cross-section area and pennation angle were measured. These data were then used to calculate a similarity index (δ) between pairs of muscles. The results indicated moderate similarity between BFlh and BFsh (δ=0.54) and between BFlh and SM (δ=0.35). In contrast, similarity was low between SM and ST (δ=0.98) and between BFlh and SM (δ=1.17). The fascicle length/muscle length ratio was higher for the ST (0.58) and BFsh (0.50) compared with the BFlh (0.27) and SM (0.22). There were, however, high inter-correlations between individual muscle architecture values, especially for muscle thickness and fascicle length data sets. Prediction of the whole hamstring architecture was achieved by combining data from all four muscles. These data show different designs of the hamstring muscles, especially between the SM and ST (medial) and BFlh and BFsh (lateral) muscles. Modeling the hamstrings as one muscle group by assuming uniform inter-muscular architecture yields less accurate representation of human hamstring muscle function. Copyright © 2012 Elsevier Ltd. All rights reserved.

  1. Endocrine alterations from concentric vs. eccentric muscle actions: a brief review.

    Science.gov (United States)

    Kraemer, Robert R; Castracane, V Daniel

    2015-02-01

    Resistance exercise has a positive effect on many tissues, including heart, bone, skeletal muscle, and nervous tissue. Eccentric muscle actions offer a unique and a potentially beneficial form of exercise for maintaining and improving health. During resistance exercise, the effects of gravity, and mechanical properties of the sarcomere and connective tissue in skeletal muscle allow a greater muscle load during an eccentric (lengthening) muscle contraction than a concentric (shortening) muscle contraction. Consequently, older patients, patients with muscle or limb movement limitations or injuries, as well as cancer patients may be able to benefit from isolated eccentric muscle actions. There are specific physiological responses to eccentric muscle contractions. This review will describe the effects of different eccentric muscle contraction protocols on endocrine responses that could have positive effects on different tissues and recommend direction for future research. Copyright © 2015 Elsevier Inc. All rights reserved.

  2. Intermittent stretching induces fibrosis in denervated rat muscle.

    Science.gov (United States)

    Faturi, Fernanda M; Franco, Rúbia C; Gigo-Benato, Davilene; Turi, Andriette C; Silva-Couto, Marcela A; Messa, Sabrina P; Russo, Thiago L

    2016-01-01

    Stretching (St) has been used for treating denervated muscles. However, its effectiveness and safety claims require further study. Rats were divided into: (1) those with denervated (D) muscles, evaluated 7 or 15 days after sciatic nerve crush injury; (2) those with D muscles submitted to St during 7 or 15 days; and (3) those with normal muscles. Muscle fiber cross-sectional area, serial sarcomere number, sarcomere length, and connective tissue density were measured. MMP-2, MMP-9, TIMP-1, TGF-β1, and myostatin mRNAs were determined by real-time polymerase chain reaction. MMP-2 and MMP-9 activity was evaluated by zymography. Collagen I was localized using immunofluorescence. St did not prevent muscle atrophy due to denervation, but it increased fibrosis and collagen I deposition at day 15. St also upregulated MMP-9 and TGF-β1 gene expressions at day 7, and myostatin at day 15. Stretching denervated muscle does not prevent atrophy, but it increases fibrosis via temporal modulation of TGF-β1/myostatin and MMP-9 cascades. © 2015 Wiley Periodicals, Inc.

  3. PLASTICITY OF SKELETAL MUSCLE STUDIED BY STEREOLOGY

    Directory of Open Access Journals (Sweden)

    Ida Eržen

    2011-05-01

    Full Text Available The present contribution provides an overview of stereological methods applied in the skeletal muscle research at the Institute of Anatomy of the Medical Faculty in Ljubljana. Interested in skeletal muscle plasticity we studied three different topics: (i expression of myosin heavy chain isoforms in slow and fast muscles under experimental conditions, (ii frequency of satellite cells in young and old human and rat muscles and (iii capillary supply of rat fast and slow muscles. We analysed the expression of myosin heavy chain isoforms within slow rat soleus and fast extensor digitorum longus muscles after (i homotopic and heterotopic transplantation of both muscles, (ii low frequency electrical stimulation of the fast muscle and (iii transposition of the fast nerve to the slow muscle. The models applied were able to turn the fast muscle into a completely slow muscle, but not vice versa. One of the indicators for the regenerative potential of skeletal muscles is its satellite cell pool. The estimated parameters, number of satellite cells per unit fibre length, corrected to the reference sarcomere length (Nsc/Lfib and number of satellite cells per number of nuclei (myonuclei and satellite cell nuclei (Nsc/Nnucl indicated that the frequency of M-cadherin stained satellite cells declines in healthy old human and rat muscles compared to young muscles. To access differences in capillary densities among slow and fast muscles and slow and fast muscle fibres, we have introduced Slicer and Fakir methods, and tested them on predominantly slow and fast rat muscles. Discussing three different topics that require different approach, the present paper reflects the three decades of the development of stereological methods: 2D analysis by simple point counting in the 70's, the disector in the 80's and virtual spatial probes in the 90's. In all methods the interactive computer assisted approach was utilised.

  4. YAP-mediated mechanotransduction in skeletal muscle

    Directory of Open Access Journals (Sweden)

    Martina eFischer

    2016-02-01

    Full Text Available Skeletal muscle is not only translating chemical energy into mechanical work, it is also a highly adaptive and regenerative tissue whose architecture and functionality is determined by its mechanical and physical environment. Processing intra- and extracellular mechanical signaling cues contributes to the regulation of cell growth, survival, migration and differentiation. Yes-associated Protein (YAP, a transcriptional coactivator downstream of the Hippo pathway and its paralogue, the transcriptional co-activator with PDZ-binding motif (TAZ, were recently found to play a key role in mechanotransduction in various tissues including skeletal muscle. Furthermore, YAP/TAZ modulate myogenesis and muscle regeneration and abnormal YAP activity has been reported in muscular dystrophy and rhabdomyosarcoma. Here, we summarize the current knowledge of mechanosensing and -signaling in striated muscle. We highlight the role of YAP signaling and discuss the different routes and hypotheses of its regulation in the context of mechanotransduction.

  5. Muscle microanatomy and its changes during contraction: the legacy of William Bowman (1816-1892).

    Science.gov (United States)

    Frixione, Eugenio

    2006-01-01

    Striated muscle fine structure began to be really understood following a comprehensive survey of the matter carried out by William Bowman in the late 1830s. The publications resulting from such a study, the first of which earned for the author a precocious election as Fellow of the Royal Society, are herewith examined in the context of contemporary views on the subject as well as of their subsequent repercussion and current knowledge today. It is shown that not only Bowman succeeded in establishing the true architecture of striated muscle fibres to the extent possible with the most advanced technology available in his day--explaining and eradicating alternative erroneous concepts in the process--but also in correctly describing the basic microstructural changes associated with contraction. In addition, although unrecognized by him or others at the time, his experiments with muscle provided direct evidence for the existence of a selectively permeable cell membrane--in the present meaning of the word--over half a century before its officially accepted discovery. Yet, in spite of these remarkable advances, Bowman arrived at the conclusion that the structure of striated muscle fibres is essentially irrelevant for the mechanism of contraction. Possible reasons behind Bowman's breakthrough accomplishments as a pioneer of modern muscle research, and his failure to understand their significance for muscle physiology, are discussed.

  6. Structural adaptations of rat lateral gastrocnemius muscle-tendon complex to a chronic stretching program and their quantification based on ultrasound biomicroscopy and optical microscopic images.

    Science.gov (United States)

    Peixinho, Carolina Carneiro; Martins, Natália Santos Fonseca; de Oliveira, Liliam Fernandes; Machado, João Carlos

    2014-01-01

    A chronic regimen of flexibility training can increase range of motion, with the increase mechanisms believed to be a change in the muscle material properties or in the neural components associated with this type of training. This study followed chronic structural adaptations of lateral gastrocnemius muscle of rats submitted to stretching training (3 times a week during 8weeks), based on muscle architecture measurements including pennation angle, muscle thickness and tendon length obtained from ultrasound biomicroscopic images, in vivo. Fiber length and sarcomere number per 100μm were determined in 3 fibers of each muscle (ex vivo and in vitro, respectively), using conventional optical microscopy. Stretching training resulted in a significant pennation angle reduction of the stretched leg after 12 sessions (25%, P=0.002 to 0.024). Muscle thickness and tendon length presented no significant changes. Fiber length presented a significant increase for the stretched leg (8.5%, P=0.00006), with the simultaneous increase in sarcomere length (5%, P=0.041) since the stretched muscles presented less sarcomeres per 100μm. A stretching protocol with characteristics similar to those applied in humans was sufficient to modify muscle architecture of rats with absence of a sarcomerogenesis process. The results indicate that structural adaptations take place in skeletal muscle tissue submitted to moderate-intensity stretching training. Copyright © 2013 Elsevier Ltd. All rights reserved.

  7. In vivo Phosphoproteome of Human Skeletal Muscle Revealed by Phosphopeptide Enrichment and HPLC-ESI-MS/MS

    DEFF Research Database (Denmark)

    Højlund, Kurt; Bowen, Benjamin P; Hwang, Hyonson

    2009-01-01

    Protein phosphorylation plays an essential role in signal transduction pathways that regulate substrate and energy metabolism, contractile function, and muscle mass in human skeletal muscle. Abnormal phosphorylation of signaling enzymes has been identified in insulin resistant muscle using phosph...... 240 phosphoserines, 53 phosphothreonines and 13 phosphotyrosines in at least 2 out of 3 subjects. In addition, 61 ambiguous phosphorylation sites were identified in at least 2 out of 3 subjects. The majority of phosphoproteins detected are involved in sarcomeric function, excitation......-contraction coupling (the Ca2+-cycle), glycolysis and glycogen metabolism. Of particular interest, we identified multiple novel phosphorylation sites on several sarcomeric Z-disc proteins known to be involved in signaling and muscle disorders. These results provide numerous new targets for the investigation of human...

  8. Genetic architecture of gene expression in ovine skeletal muscle

    DEFF Research Database (Denmark)

    Kogelman, Lisette Johanna Antonia; Byrne, Keren; Vuocolo, Tony

    2011-01-01

    weighted gene co-expression network analysis and a differential gene co-expression network analysis. The modules of genes revealed by these analyses were enriched for a number of functional terms summarised as muscle sarcomere organisation and development, protein catabolism (proteosome), RNA processing...... has potential, amongst other mechanisms, to alter gene expression via cis- or trans-acting mechanisms in a manner that impacts the functional activities of specific pathways that contribute to muscling traits. By integrating sire-based genetic merit information for a muscling trait with progeny......-based gene expression data we directly tested the hypothesis that there is genetic structure in the gene expression program in ovine skeletal muscle.Results: The genetic performance of six sires for a well defined muscling trait, longissimus lumborum muscle depth, was measured using extensive progeny testing...

  9. Effects of striated laser tracks on thermal fatigue resistance of cast iron samples with biomimetic non-smooth surface

    International Nuclear Information System (INIS)

    Tong, Xin; Zhou, Hong; Liu, Min; Dai, Ming-jiang

    2011-01-01

    In order to enhance the thermal fatigue resistance of cast iron materials, the samples with biomimetic non-smooth surface were processed by Neodymium:Yttrium Aluminum Garnet (Nd:YAG) laser. With self-controlled thermal fatigue test method, the thermal fatigue resistance of smooth and non-smooth samples was investigated. The effects of striated laser tracks on thermal fatigue resistance were also studied. The results indicated that biomimetic non-smooth surface was benefit for improving thermal fatigue resistance of cast iron sample. The striated non-smooth units formed by laser tracks which were vertical with thermal cracks had the best propagation resistance. The mechanisms behind these influences were discussed, and some schematic drawings were introduced to describe them.

  10. Deleting exon 55 from the nebulin gene induces severe muscle weakness in a mouse model for nemaline myopathy

    OpenAIRE

    Ottenheijm, Coen A. C.; Buck, Danielle; de Winter, Josine M.; Ferrara, Claudia; Piroddi, Nicoletta; Tesi, Chiara; Jasper, Jeffrey R.; Malik, Fady I.; Meng, Hui; Stienen, Ger J. M.; Beggs, Alan H.; Labeit, Siegfried; Poggesi, Corrado; Lawlor, Michael W.; Granzier, Henk

    2013-01-01

    Nebulin—a giant sarcomeric protein—plays a pivotal role in skeletal muscle contractility by specifying thin filament length and function. Although mutations in the gene encoding nebulin (NEB) are a frequent cause of nemaline myopathy, the most common non-dystrophic congenital myopathy, the mechanisms by which mutations in NEB cause muscle weakness remain largely unknown. To better understand these mechanisms, we have generated a mouse model in which Neb exon 55 is deleted (NebΔExon55) to repl...

  11. The regulation of catch in molluscan muscle.

    Science.gov (United States)

    Twarog, B M

    1967-07-01

    Molluscan catch muscles are smooth muscles. As with mammalian smooth muscles, there is no transverse ordering of filaments or dense bodies. In contrast to mammalian smooth muscles, two size ranges of filaments are present. The thick filaments are long as well as large in diameter and contain paramyosin. The thin filaments contain actin and appear to run into and join the dense bodies. Vesicles are present which may be part of a sarcoplasmic reticulum. Neural activation of contraction in Mytilus muscle is similar to that observed in mammalian smooth muscles, and in some respects to frog striated muscle. The relaxing nerves, which reduce catch, are unique to catch muscles. 5-Hydroxytryptamine, which appears to mediate relaxation, specifically blocks catch tension but increases the ability of the muscle to fire spikes. It is speculated that Mytilus muscle actomyosin is activated by a Ca(++)-releasing mechanism, and that 5-hydroxytryptamine may reduce catch and increase excitability by influencing the rate of removal of intracellular free Ca(++).

  12. Calcium-dependence of Donnan potentials in glycerinated rabbit psoas muscle in rigor, at and beyond filament overlap; a role for titin in the contractile process

    DEFF Research Database (Denmark)

    Coomber, S J; Bartels, E M; Elliott, G F

    2011-01-01

    contracts and breaks the microelectrode. Therefore the rigor state was studied. There is no reason to suppose a priori that a similar voltage switch does not occur during contraction, however. Calcium dependence is still apparent in muscles stretched beyond overlap (sarcomere length>3.8 μm) and is also seen...

  13. Complete genome sequence of maize yellow striate virus, a new cytorhabdovirus infecting maize and wheat crops in Argentina.

    Science.gov (United States)

    Maurino, Fernanda; Dumón, Analía D; Llauger, Gabriela; Alemandri, Vanina; de Haro, Luis A; Mattio, M Fernanda; Del Vas, Mariana; Laguna, Irma Graciela; Giménez Pecci, María de la Paz

    2018-01-01

    A rhabdovirus infecting maize and wheat crops in Argentina was molecularly characterized. Through next-generation sequencing (NGS) of symptomatic leaf samples, the complete genome was obtained of two isolates of maize yellow striate virus (MYSV), a putative new rhabdovirus, differing by only 0.4% at the nucleotide level. The MYSV genome consists of 12,654 nucleotides for maize and wheat virus isolates, and shares 71% nucleotide sequence identity with the complete genome of barley yellow striate mosaic virus (BYSMV, NC028244). Ten open reading frames (ORFs) were predicted in the MYSV genome from the antigenomic strand and were compared with their BYSMV counterparts. The highest amino acid sequence identity of the MYSV and BYSMV proteins was 80% between the L proteins, and the lowest was 37% between the proteins 4. Phylogenetic analysis suggested that the MYSV isolates are new members of the genus Cytorhabdovirus, family Rhabdoviridae. Yellow striate, affecting maize and wheat crops in Argentina, is an emergent disease that presents a potential economic risk for these widely distributed crops.

  14. A functional magnetic resonance imaging investigation of visual hallucinations in the human striate cortex.

    Science.gov (United States)

    Abid, Hina; Ahmad, Fayyaz; Lee, Soo Y; Park, Hyun W; Im, Dongmi; Ahmad, Iftikhar; Chaudhary, Safee U

    2016-11-29

    Human beings frequently experience fear, phobia, migraine and hallucinations, however, the cerebral mechanisms underpinning these conditions remain poorly understood. Towards this goal, in this work, we aim to correlate the human ocular perceptions with visual hallucinations, and map them to their cerebral origins. An fMRI study was performed to examine the visual cortical areas including the striate, parastriate and peristriate cortex in the occipital lobe of the human brain. 24 healthy subjects were enrolled and four visual patterns including hallucination circle (HCC), hallucination fan (HCF), retinotopy circle (RTC) and retinotopy cross (RTX) were used towards registering their impact in the aforementioned visual related areas. One-way analysis of variance was used to evaluate the significance of difference between induced activations. Multinomial regression and and K-means were used to cluster activation patterns in visual areas of the brain. Significant activations were observed in the visual cortex as a result of stimulus presentation. The responses induced by visual stimuli were resolved to Brodmann areas 17, 18 and 19. Activation data clustered into independent and mutually exclusive clusters with HCC registering higher activations as compared to HCF, RTC and RTX. We conclude that small circular objects, in rotation, tend to leave greater hallucinating impressions in the visual region. The similarity between observed activation patterns and those reported in conditions such as epilepsy and visual hallucinations can help elucidate the cortical mechanisms underlying these conditions. Trial Registration 1121_GWJUNG.

  15. A pilot study of muscle plasma protein changes after exercise

    DEFF Research Database (Denmark)

    Dahlqvist, Julia R; Voss, Line G; Lauridsen, Thomas

    2014-01-01

    profiles were measured before and after exercise in 3 groups: subjects affected by either Becker muscular dystrophy or McArdle disease, and healthy subjects. RESULTS: Mb and TnI appeared early in the blood, and the increase of TnI was only observed in patients with muscle disease. The CK increase was more...... delayed in plasma. Tcap was not detectable at any time. CONCLUSIONS: Our results suggest that TnI is a marker of more severe damage signifying sarcomeric damage, and it could therefore be an important supplement to CK and Mb in clinical practice. Tcap is not useful as a marker for skeletal muscle damage....

  16. Studies of Infection and Dissemination of Rift Valley Fever Virus in Mosquitoes.

    Science.gov (United States)

    1991-10-15

    sarcomere (A-band) microfilaments actin & myosin (Note: all insect muscles are striated; visceral muscles will be mentioned in association with the various...granule zonula pellucida 4 Male Reproductive (Ln. 9, 10; Em. 37 & 38) testes follicular epithelium spermatogonia spermatozoa vas efferens vas...rectum 53. salivary gland 54. seminal vesicle 55. spermatheca 56. spermatozoa 57.---------- 58. tergosternal flight muscles 59. testicle 60. thoracic

  17. Differential Sarcomere and Electrophysiological Maturation of Human iPSC-Derived Cardiac Myocytes in Monolayer vs. Aggregation-Based Differentiation Protocols

    Directory of Open Access Journals (Sweden)

    Dorota Jeziorowska

    2017-06-01

    Full Text Available Human induced pluripotent stem cells (iPSCs represent a powerful human model to study cardiac disease in vitro, notably channelopathies and sarcomeric cardiomyopathies. Different protocols for cardiac differentiation of iPSCs have been proposed either based on embroid body formation (3D or, more recently, on monolayer culture (2D. We performed a direct comparison of the characteristics of the derived cardiomyocytes (iPSC-CMs on day 27 ± 2 of differentiation between 3D and 2D differentiation protocols with two different Wnt-inhibitors were compared: IWR1 (inhibitor of Wnt response or IWP2 (inhibitor of Wnt production. We firstly found that the level of Troponin T (TNNT2 expression measured by FACS was significantly higher for both 2D protocols as compared to the 3D protocol. In the three methods, iPSC-CM show sarcomeric structures. However, iPSC-CM generated in 2D protocols constantly displayed larger sarcomere lengths as compared to the 3D protocol. In addition, mRNA and protein analyses reveal higher cTNi to ssTNi ratios in the 2D protocol using IWP2 as compared to both other protocols, indicating a higher sarcomeric maturation. Differentiation of cardiac myocytes with 2D monolayer-based protocols and the use of IWP2 allows the production of higher yield of cardiac myocytes that have more suitable characteristics to study sarcomeric cardiomyopathies.

  18. An Electron Microscopic Study of the Irradiation Effects on the Striated Duct Cells of the Submandibular Gland in Rats

    International Nuclear Information System (INIS)

    Lee, Kyu Chan; Lee, Sang Rae

    1990-01-01

    The purpose of this study was to investigate the effects of irradiation on the striated duct cells of the rat submandibular gland ductal tissues which control the characteristics of saliva. For this study, the experimental group was composed of 36 irradiated Sprague Dawley strain rats divided into 8 subgroups- 1 hour, 2 hours, 3 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours after irradiation. 4 non-irradiated rats were used as the control group. The experimental animals were singly irradiated with a dose of 18 Gy gamma ray to their head and neck region by the Co-6-teletherapy unit and sacrificed after each experimental duration. The specimens were examined with a light microscope with an H-E stain and with a transmission electron microscope. The results of this study were as follows. 1. In the light micrograph, a severe atrophic change occurred in the striated duct cells at 2 hours after irradiation and gradual recovery occurred from 6 hours after irradiation. 2. The nuclear chromosomes of the striated duct cells were changed granular at 2 hours after irradiation. Recovery was observed at 6 hours after irradiation. Nuclear bodies were also observed from 3 hours after irradiation. 3. The mitochondria of the striated duct cells had indistinct cristae at 2 hours after irradiation, and were degenerated or swollen at 3 hours after irradiation. They recovered, however, from 6 hours, with an increasing number at 48 hours a regular arrangement was observed at 72 hours after irradiation. 4. The microvilli showed atrophic changes at 2 hours after irradiation and were almost lost at 3 hours after irradiation. They were observed again from 48 hours after irradiation. 5. The rough endoplasmic reticulum and golgi body were not apparent at 1 hours after irradiation and were dilated with degeneration 2 hours after, but intact rough endoplasmic reticulum were observed from 3 hours after irradiation and developed well at 24 hours after irradiation. By the result of this

  19. DAAM is required for thin filament formation and Sarcomerogenesis during muscle development in Drosophila.

    Directory of Open Access Journals (Sweden)

    Imre Molnár

    2014-02-01

    Full Text Available During muscle development, myosin and actin containing filaments assemble into the highly organized sarcomeric structure critical for muscle function. Although sarcomerogenesis clearly involves the de novo formation of actin filaments, this process remained poorly understood. Here we show that mouse and Drosophila members of the DAAM formin family are sarcomere-associated actin assembly factors enriched at the Z-disc and M-band. Analysis of dDAAM mutants revealed a pivotal role in myofibrillogenesis of larval somatic muscles, indirect flight muscles and the heart. We found that loss of dDAAM function results in multiple defects in sarcomere development including thin and thick filament disorganization, Z-disc and M-band formation, and a near complete absence of the myofibrillar lattice. Collectively, our data suggest that dDAAM is required for the initial assembly of thin filaments, and subsequently it promotes filament elongation by assembling short actin polymers that anneal to the pointed end of the growing filaments, and by antagonizing the capping protein Tropomodulin.

  20. Striated muscle microvascular response to silver implants: A comparative in vivo study with titanium and stainless steel.

    Science.gov (United States)

    Kraft, C N; Hansis, M; Arens, S; Menger, M D; Vollmar, B

    2000-02-01

    Local microvascular perfusion is the primary line of defense of tissue against microorganisms and plays a considerable role in reparative processes. The impairment of the microcirculation by a biomaterial may therefore have profound consequences. Silver is known to have excellent antimicrobial activity and, although regional and systemic toxic effects have been described, silver is regularly discussed as an implant material in bone surgery. Because little is known about the influence of silver implants on the adjacent host tissue microvasculature, we studied in vivo nutritive perfusion and leukocytic response, and compared these results with those of the conventionally used materials titanium and stainless steel. Using the hamster dorsal skinfold chamber preparation and intravital microscopy, the implantation of a commercially pure silver sample led to a distinct and persistent activation of leukocytes combined with a marked disruption of the microvascular endothelial integrity, massive leukocyte extravasation, and considerable venular dilation. Whereas animals with stainless-steel implants showed a moderate increase in these parameters with a tendency to recuperate, titanium implants caused only a transient increase of leukocyte-endothelial cell interaction within the first 120 min and no significant change in macromolecular leakage, leukocyte extravasation and venular diameter. After 3 days, five of six preparations with silver samples showed severe inflammation and massive edema. Thus, the use of silver as an implant material should be critically judged despite its bactericidal properties. The implant material titanium seems to be well tolerated by the local vascular system and currently represents the golden standard. Copyright 2000 John Wiley & Sons, Inc.

  1. Action of lovastatin, simvastatin, and pravastatin on sterol synthesis and their antiproliferative effect in cultured myoblasts from human striated muscle

    NARCIS (Netherlands)

    Vliet, A.K. van; Nègre-Arrariou, P.; Thiel, G.C.F. van; Bolhuis, P.A.; Cohen, L.H.

    1996-01-01

    Lovastatin, simvastatin, and pravastatin are fairly strong inhibitors of sterol synthesis in human myoblasts in culture. Lovastatin and simvastatin have IC50 values of 19 ± 6 nM and 4.0 ± 2.3 nM, respectively. Pravastatin is a weaker inhibitor of sterol synthesis (IC50 value of 110 ± 38 nM). Through

  2. Identification and Characterization of Wheat Yellow Striate Virus, a Novel Leafhopper-Transmitted Nucleorhabdovirus Infecting Wheat

    Directory of Open Access Journals (Sweden)

    Yan Liu

    2018-03-01

    Full Text Available A new wheat viral disease was found in China. Bullet-shaped viral particles within the nucleus of the infected wheat leave cells, which possessed 180–210 nm length and 35–40 nm width, were observed under transmission electron microscopy. A putative wheat-infecting rhabdovirus vectored by the leafhopper Psammotettix alienus was identified and tentatively named wheat yellow striate virus (WYSV. The full-length nucleotide sequence of WYSV was determined using transcriptome sequencing and RACE analysis of both wheat samples and leafhoppers P. alienus. The negative-sense RNA genome of WYSV contains 14,486 nucleotides (nt and seven open reading frames (ORFs encode deduced proteins in the order N-P-P3-M-P6-G-L on the antisense strand. In addition, WYSV genome has a 76-nt 3′ leader RNA and a 258-nt 5′ trailer, and the ORFs are separated by conserved intergenic sequences. The entire genome sequence shares 58.1 and 57.7% nucleotide sequence identity with two strains of rice yellow stunt virus (RYSV-A and RYSV-B genomes, respectively. The highest amino acid sequence identity was 63.8% between the L proteins of the WYSV and RYSV-B, but the lowest was 29.5% between the P6 proteins of these viruses. Phylogenetic analysis firmly established WYSV as a new member of the genus Nucleorhabdovirus. Collectively, this study provided evidence that WYSV is likely the first nucleorhabdovirus described infecting wheat via leafhopper P. alienus transmission.

  3. Calpain 3 Is Activated through Autolysis within the Active Site and Lyses Sarcomeric and Sarcolemmal Components

    Science.gov (United States)

    Taveau, Mathieu; Bourg, Nathalie; Sillon, Guillaume; Roudaut, Carinne; Bartoli, Marc; Richard, Isabelle

    2003-01-01

    Calpain 3 (Capn3) is known as the skeletal muscle-specific member of the calpains, a family of intracellular nonlysosomal cysteine proteases. This enigmatic protease has many unique features among the calpain family and, importantly, mutations in Capn3 have been shown to be responsible for limb girdle muscular dystrophy type 2A. Here we demonstrate that the Capn3 activation mechanism is similar to the universal activation of caspases and corresponds to an autolysis within the active site of the protease. We undertook a search for substrates in immature muscle cells, as several lines of evidence suggest that Capn3 is mostly in an inactive state in muscle and needs a signal to be activated. In this model, Capn3 proteolytic activity leads to disruption of the actin cytoskeleton and disorganization of focal adhesions through cleavage of several endogenous proteins. In addition, we show that titin, a previously identified Capn3 partner, and filamin C are further substrates of Capn3. Finally, we report that Capn3 colocalizes in vivo with its substrates at various sites along cytoskeletal structures. We propose that Capn3-mediated cleavage produces an adaptive response of muscle cells to external and/or internal stimuli, establishing Capn3 as a muscle cytoskeleton regulator. PMID:14645524

  4. Small differences in Drosophila tropomyosin expression have significant effects on muscle function

    Energy Technology Data Exchange (ETDEWEB)

    Tansey, T.; Miller, R.C. (Univ. of Illinois, Chicago (United States)); Schultz, J.R.; Storti, R.V. (Georgetown Univ., Washington, DC (United States))

    1991-12-01

    The effects of promoter deletions on Drosophila tropomyosin I (TmI) gene expression have been determined by measuring TmI RNA levels in transformed flies. Decreases in RNA levels have been correlated with rescue of flightless and jumpless mutant phenotypes in Ifm(3)3 mutant transformed flies and changes in muscle ultrastructure. The results of this analysis have allowed us to identify a region responsible for 20% of maximal TmI expression, estimate threshold levels of TmI RNA required for indirect flight and jump muscle function, and obtain evidence suggesting that sarcomere length may be an important determinant of flight muscle functions.

  5. Mesodermal iPSC–derived progenitor cells functionally regenerate cardiac and skeletal muscle

    Science.gov (United States)

    Quattrocelli, Mattia; Swinnen, Melissa; Giacomazzi, Giorgia; Camps, Jordi; Barthélemy, Ines; Ceccarelli, Gabriele; Caluwé, Ellen; Grosemans, Hanne; Thorrez, Lieven; Pelizzo, Gloria; Muijtjens, Manja; Verfaillie, Catherine M.; Blot, Stephane; Janssens, Stefan; Sampaolesi, Maurilio

    2015-01-01

    Conditions such as muscular dystrophies (MDs) that affect both cardiac and skeletal muscles would benefit from therapeutic strategies that enable regeneration of both of these striated muscle types. Protocols have been developed to promote induced pluripotent stem cells (iPSCs) to differentiate toward cardiac or skeletal muscle; however, there are currently no strategies to simultaneously target both muscle types. Tissues exhibit specific epigenetic alterations; therefore, source-related lineage biases have the potential to improve iPSC-driven multilineage differentiation. Here, we determined that differential myogenic propensity influences the commitment of isogenic iPSCs and a specifically isolated pool of mesodermal iPSC-derived progenitors (MiPs) toward the striated muscle lineages. Differential myogenic propensity did not influence pluripotency, but did selectively enhance chimerism of MiP-derived tissue in both fetal and adult skeletal muscle. When injected into dystrophic mice, MiPs engrafted and repaired both skeletal and cardiac muscle, reducing functional defects. Similarly, engraftment into dystrophic mice of canine MiPs from dystrophic dogs that had undergone TALEN-mediated correction of the MD-associated mutation also resulted in functional striatal muscle regeneration. Moreover, human MiPs exhibited the same capacity for the dual differentiation observed in murine and canine MiPs. The findings of this study suggest that MiPs should be further explored for combined therapy of cardiac and skeletal muscles. PMID:26571398

  6. A role for muscle LIM protein (MLP) in vascular remodeling.

    Science.gov (United States)

    Wang, Xiaohong; Li, Qinglu; Adhikari, Neeta; Hall, Jennifer L

    2006-04-01

    Given the well-defined role of LIM-motif containing proteins in cytoskeletal organization, cell fate, and differentiation, we hypothesized that the regulation of LIM proteins played an integral role in vascular remodeling. We screened a compendium of cDNA microarray data from rat vascular smooth muscle cells (VSMC) for novel LIM-containing targets and identified muscle LIM protein (MLP), a gene previously thought to be only in striated muscle. Sequence analysis, RTQPCR and Western blotting reconfirmed expression of MLP in VSMC. MLP was elevated>10-fold 7 days following balloon injury in the rat carotid artery. Wire injury led to a significantly increased intima/media ratio in MLP -/- mice compared to wild-type controls (PMLP deficient VSMC (n=6, PMLP significantly restored apoptotic response (N=6, PMLP in vascular smooth muscle and demonstrate that it plays a critical role in vascular remodeling. This is consistent with earlier findings demonstrating a role for MLP in striated muscle remodeling in response to load and stretch.

  7. The Caenorhabditis elegans paxillin orthologue, PXL-1, is required for pharyngeal muscle contraction and for viability.

    Science.gov (United States)

    Warner, Adam; Qadota, Hiroshi; Benian, Guy M; Vogl, A Wayne; Moerman, Donald G

    2011-07-15

    We have identified the gene C28H8.6 (pxl-1) as the Caenorhabditis elegans orthologue of vertebrate paxillin. PXL-1 contains the four C-terminal LIM domains conserved in paxillin across all species and three of the five LD motifs found in the N-terminal half of most paxillins. In body wall muscle, PXL-1 antibodies and a full-length green fluorescent protein translational fusion localize to adhesion sites in the sarcomere, the functional repeat unit in muscle responsible for contraction. PXL-1 also localizes to ring-shaped structures near the sarcolemma in pharyngeal muscle corresponding to podosome-like sites of actin attachment. Our analysis of a loss-of-function allele of pxl-1, ok1483, shows that loss of paxillin leads to early larval arrested animals with paralyzed pharyngeal muscles and eventual lethality, presumably due to an inability to feed. We rescued the mutant phenotype by expressing paxillin solely in the pharynx and found that these animals survived and are essentially wild type in movement and body wall muscle structure. This indicates a differential requirement for paxillin in these two types of muscle. In pharyngeal muscle it is essential for contraction, whereas in body wall muscle it is dispensable for filament assembly, sarcomere stability, and ultimately movement.

  8. The Caenorhabditis elegans paxillin orthologue, PXL-1, is required for pharyngeal muscle contraction and for viability

    Science.gov (United States)

    Warner, Adam; Qadota, Hiroshi; Benian, Guy M.; Vogl, A. Wayne; Moerman, Donald G.

    2011-01-01

    We have identified the gene C28H8.6 (pxl-1) as the Caenorhabditis elegans orthologue of vertebrate paxillin. PXL-1 contains the four C-terminal LIM domains conserved in paxillin across all species and three of the five LD motifs found in the N-terminal half of most paxillins. In body wall muscle, PXL-1 antibodies and a full-length green fluorescent protein translational fusion localize to adhesion sites in the sarcomere, the functional repeat unit in muscle responsible for contraction. PXL-1 also localizes to ring-shaped structures near the sarcolemma in pharyngeal muscle corresponding to podosome-like sites of actin attachment. Our analysis of a loss-of-function allele of pxl-1, ok1483, shows that loss of paxillin leads to early larval arrested animals with paralyzed pharyngeal muscles and eventual lethality, presumably due to an inability to feed. We rescued the mutant phenotype by expressing paxillin solely in the pharynx and found that these animals survived and are essentially wild type in movement and body wall muscle structure. This indicates a differential requirement for paxillin in these two types of muscle. In pharyngeal muscle it is essential for contraction, whereas in body wall muscle it is dispensable for filament assembly, sarcomere stability, and ultimately movement. PMID:21633109

  9. A comparative study of various electrodes in electromyography of the striated urethral and anal sphincter in children

    DEFF Research Database (Denmark)

    Nielsen, K K; Kristensen, E S; Qvist, N

    1985-01-01

    The series comprised 41 children aged 6 to 14 years consecutively referred with recurrent urinary tract infection and/or enuresis. Carbon dioxide cystometry was carried out in the supine and the erect position and combined with simultaneous electromyography (EMG). The external urethral sphincter...... was examined with a ring electrode mounted on a urethral catheter, while recordings from the striated anal sphincter were based on an anal plug electrode and perianal electrocardiographic (ECG) skin electrodes: 211 EMG and cystometric examinations were performed and all three methods gave satisfactory results...

  10. The structure of Mytilus smooth muscle and the electrical constants of the resting muscle.

    Science.gov (United States)

    Twarog, B M; Dewey, M M; Hidaka, T

    1973-02-01

    The individual muscle fibers of the anterior byssus retractor muscle (ABRM) of Mytilus edulis L. are uninucleate, 1.2-1.8 mm in length, 5 microm in diameter, and organized into bundles 100-200 microm in diameter, surrounded by connective tissue. Some bundles run the length of the whole muscle. Adjacent muscle cell membranes are interconnected by nexuses at frequent intervals. Specialized attachments exist between muscle fibers and connective tissue. Electrical constants of the resting muscle membrane were measured with intracellular recording electrodes and both extracellular and intracellular current-passing electrodes. With an intracellular current-passing electrode, the time constant tau, was 4.3 +/- 1.5 ms. With current delivered via an extracellular electrode tau was 68.3 +/- 15 ms. The space constant, lambda, was 1.8 mm +/- 0.4. The membrane input resistance, R(eff), ranged from 23 to 51 MOmega. The observations that values of tau depend on the method of passing current, and that the value of lambda is large relative to fiber length and diameter are considered evidence that the individual muscle fibers are electrically interconnected within bundles in a three-dimensional network. Estimations are made of the membrane resistance, R(m), to compare the values to fast and slow striated muscle fibers and mammalian smooth muscles. The implications of this study in reinterpreting previous mechanical and electrical studies are discussed.

  11. Independent specialisation of myosin II paralogues in muscle vs. non-muscle functions during early animal evolution: a ctenophore perspective.

    Science.gov (United States)

    Dayraud, Cyrielle; Alié, Alexandre; Jager, Muriel; Chang, Patrick; Le Guyader, Hervé; Manuel, Michaël; Quéinnec, Eric

    2012-07-02

    Myosin II (or Myosin Heavy Chain II, MHCII) is a family of molecular motors involved in the contractile activity of animal muscle cells but also in various other cellular processes in non-muscle cells. Previous phylogenetic analyses of bilaterian MHCII genes identified two main clades associated respectively with smooth/non-muscle cells (MHCIIa) and striated muscle cells (MHCIIb). Muscle cells are generally thought to have originated only once in ancient animal history, and decisive insights about their early evolution are expected to come from expression studies of Myosin II genes in the two non-bilaterian phyla that possess muscles, the Cnidaria and Ctenophora. We have uncovered three MHCII paralogues in the ctenophore species Pleurobrachia pileus. Phylogenetic analyses indicate that the MHCIIa / MHCIIb duplication is more ancient than the divergence between extant metazoan lineages. The ctenophore MHCIIa gene (PpiMHCIIa) has an expression pattern akin to that of "stem cell markers" (Piwi, Vasa…) and is expressed in proliferating cells. We identified two MHCIIb genes that originated from a ctenophore-specific duplication. PpiMHCIIb1 represents the exclusively muscular form of myosin II in ctenophore, while PpiMHCIIb2 is expressed in non-muscle cells of various types. In parallel, our phalloidin staining and TEM observations highlight the structural complexity of ctenophore musculature and emphasize the experimental interest of the ctenophore tentacle root, in which myogenesis is spatially ordered and strikingly similar to striated muscle formation in vertebrates. MHCIIa expression in putative stem cells/proliferating cells probably represents an ancestral trait, while specific involvement of some MHCIIa genes in smooth muscle fibres is a uniquely derived feature of the vertebrates. That one ctenophore MHCIIb paralogue (PpiMHCIIb2) has retained MHCIIa-like expression features furthermore suggests that muscular expression of the other paralogue, PpiMHCIIb1, was

  12. Independent specialisation of myosin II paralogues in muscle vs. non-muscle functions during early animal evolution: a ctenophore perspective

    Science.gov (United States)

    2012-01-01

    Background Myosin II (or Myosin Heavy Chain II, MHCII) is a family of molecular motors involved in the contractile activity of animal muscle cells but also in various other cellular processes in non-muscle cells. Previous phylogenetic analyses of bilaterian MHCII genes identified two main clades associated respectively with smooth/non-muscle cells (MHCIIa) and striated muscle cells (MHCIIb). Muscle cells are generally thought to have originated only once in ancient animal history, and decisive insights about their early evolution are expected to come from expression studies of Myosin II genes in the two non-bilaterian phyla that possess muscles, the Cnidaria and Ctenophora. Results We have uncovered three MHCII paralogues in the ctenophore species Pleurobrachia pileus. Phylogenetic analyses indicate that the MHCIIa / MHCIIb duplication is more ancient than the divergence between extant metazoan lineages. The ctenophore MHCIIa gene (PpiMHCIIa) has an expression pattern akin to that of "stem cell markers" (Piwi, Vasa…) and is expressed in proliferating cells. We identified two MHCIIb genes that originated from a ctenophore-specific duplication. PpiMHCIIb1 represents the exclusively muscular form of myosin II in ctenophore, while PpiMHCIIb2 is expressed in non-muscle cells of various types. In parallel, our phalloidin staining and TEM observations highlight the structural complexity of ctenophore musculature and emphasize the experimental interest of the ctenophore tentacle root, in which myogenesis is spatially ordered and strikingly similar to striated muscle formation in vertebrates. Conclusion MHCIIa expression in putative stem cells/proliferating cells probably represents an ancestral trait, while specific involvement of some MHCIIa genes in smooth muscle fibres is a uniquely derived feature of the vertebrates. That one ctenophore MHCIIb paralogue (PpiMHCIIb2) has retained MHCIIa-like expression features furthermore suggests that muscular expression of the

  13. Independent specialisation of myosin II paralogues in muscle vs. non-muscle functions during early animal evolution: a ctenophore perspective

    Directory of Open Access Journals (Sweden)

    Dayraud Cyrielle

    2012-07-01

    Full Text Available Abstract Background Myosin II (or Myosin Heavy Chain II, MHCII is a family of molecular motors involved in the contractile activity of animal muscle cells but also in various other cellular processes in non-muscle cells. Previous phylogenetic analyses of bilaterian MHCII genes identified two main clades associated respectively with smooth/non-muscle cells (MHCIIa and striated muscle cells (MHCIIb. Muscle cells are generally thought to have originated only once in ancient animal history, and decisive insights about their early evolution are expected to come from expression studies of Myosin II genes in the two non-bilaterian phyla that possess muscles, the Cnidaria and Ctenophora. Results We have uncovered three MHCII paralogues in the ctenophore species Pleurobrachia pileus. Phylogenetic analyses indicate that the MHCIIa / MHCIIb duplication is more ancient than the divergence between extant metazoan lineages. The ctenophore MHCIIa gene (PpiMHCIIa has an expression pattern akin to that of "stem cell markers" (Piwi, Vasa… and is expressed in proliferating cells. We identified two MHCIIb genes that originated from a ctenophore-specific duplication. PpiMHCIIb1 represents the exclusively muscular form of myosin II in ctenophore, while PpiMHCIIb2 is expressed in non-muscle cells of various types. In parallel, our phalloidin staining and TEM observations highlight the structural complexity of ctenophore musculature and emphasize the experimental interest of the ctenophore tentacle root, in which myogenesis is spatially ordered and strikingly similar to striated muscle formation in vertebrates. Conclusion MHCIIa expression in putative stem cells/proliferating cells probably represents an ancestral trait, while specific involvement of some MHCIIa genes in smooth muscle fibres is a uniquely derived feature of the vertebrates. That one ctenophore MHCIIb paralogue (PpiMHCIIb2 has retained MHCIIa-like expression features furthermore suggests that muscular

  14. Role of calcium-dependent proteinase in molt-induced claw muscle atrophy

    Energy Technology Data Exchange (ETDEWEB)

    Mykles, D.L.; Skinner, D.M.

    1984-01-01

    The claw closer muscle of the Bermuda land crab Gecarcinus lateralis undergoes a sequential atrophy and restoration during each intermolt cycle. Muscle protein decreases 40% during proecdysis and is restored following ecdysis. Amino acid incorporation into protein of postecdysial muscle is five times greater than that in anecdysial muscle. Since the rates of protein synthesis in anecdysial and proecdysial muscle are the same it appears that proecdysial muscle atrophy is caused primarily by an increase in protein degradation. A calcium-dependent proteinase (CDP) active at neutral pH has been implicated in the nonlysosomal hydrolysis of myofibrillar proteins. We have examined the role of a CDP in atrophy of the claw closer muscle. The many similarities between crustacean and vertebrate CDPs have established this crustacean system as a simple and convenient model for the role of Ca/sup 2 +/-dependent proteolysis in myofibrillar protein turnover and its manifestation in the structure of the sarcomere. 16 references, 8 figures. (ACR)

  15. Striated nephrogram as an incidental finding in MRI examination of children; Streifiges Nephrogramm als Zufallsbefund nach Kontrastmittelgabe bei Kindern in der MRT

    Energy Technology Data Exchange (ETDEWEB)

    Strocka, S.; Sorge, I.; Ritter, L.; Hirsch, F.W. [Leipzig Univ. (Germany). Dept. of Pediatric Radiology

    2016-01-15

    A highly striated contrast pattern of the kidneys occasionally appears in abdominal MRI examinations of children following the administration of gadolinium. As this phenomenon is well known but has not yet been explicitly described in literature, we investigated how frequently and in which clinical context this occurred. 855 abdominal MRI examinations with contrast media of 362 children between 2006 and 2014 were analysed retrospectively. A striated renal parenchyma was found in a total of nine children and eleven examinations (1.3 % of examinations) and did only occur at a field strength of 3 Tesla. Of these children, seven had previously had tumors and chemotherapy. In two children there was no evidence of a previously serious condition with medications or a kidney disease. All of them had a normal renal function. A noticeably striated nephrogram in the later phase of an MRI examination following administration of gadolinium may appear as an incidental finding in examinations at 3 Tesla without pathological relevance.

  16. Making muscle elastic: the structural basis of myomesin stretching.

    Directory of Open Access Journals (Sweden)

    Larissa Tskhovrebova

    2012-02-01

    Full Text Available Skeletal and cardiac muscles are remarkable biological machines that support and move our bodies and power the rhythmic work of our lungs and hearts. As well as producing active contractile force, muscles are also passively elastic, which is essential to their performance. The origins of both active contractile and passive elastic forces can be traced to the individual proteins that make up the highly ordered structure of muscle. In this Primer, we describe the organization of sarcomeres--the structural units that produce contraction--and the nature of the proteins that make muscle elastic. In particular, we focus on an elastic protein called myomesin, whose novel modular architecture helps explain elasticity.

  17. Premature aging in skeletal muscle lacking serum response factor.

    Directory of Open Access Journals (Sweden)

    Charlotte Lahoute

    Full Text Available Aging is associated with a progressive loss of muscle mass, increased adiposity and fibrosis that leads to sarcopenia. At the molecular level, muscle aging is known to alter the expression of a variety of genes but very little is known about the molecular effectors involved. SRF (Serum Response Factor is a crucial transcription factor for muscle-specific gene expression and for post-natal skeletal muscle growth. To assess its role in adult skeletal muscle physiology, we developed a post-mitotic myofiber-specific and tamoxifen-inducible SRF knockout model. Five months after SRF loss, no obvious muscle phenotype was observed suggesting that SRF is not crucial for myofiber maintenance. However, mutant mice progressively developed IIB myofiber-specific atrophy accompanied by a metabolic switch towards a more oxidative phenotype, muscular lipid accumulation, sarcomere disorganization and fibrosis. After injury, mutant muscles exhibited an altered regeneration process, showing smaller regenerated fibers and persistent fibrosis. All of these features are strongly reminiscent of abnormalities encountered in aging skeletal muscle. Interestingly, we also observed an important age associated decrease in SRF expression in mice and human muscles. Altogether, these results suggest that a naturally occurring SRF down-regulation precedes and contributes to the muscle aging process. Indeed, triggering SRF loss in the muscles of mutant mice results in an accelerated aging process.

  18. Recent improvements in small angle x-ray diffraction for the study of muscle physiology

    Science.gov (United States)

    Reconditi, Massimo

    2006-10-01

    The molecular mechanism of muscle contraction is one of the most important unresolved problems in biology and biophysics. Notwithstanding the great advances of recent years, it is not yet known in detail how the molecular motor in muscle, the class II myosin, converts the free energy of ATP hydrolysis into work by interacting with its track, the actin filament; neither is it understood how the high efficiency in energy conversion depends on the cooperative action of myosin motors working in parallel along the actin filament. Research in muscle contraction involves the combination of mechanical, biochemical and structural methods in studies that span from tissue to single molecule. Therefore, more than for any other research field, progress in the comprehension of muscle contraction at the molecular level is related to, and in turn contributes to, the advancement of methods in biophysics. This review will focus on the progress achieved by time-resolved small angle x-ray scattering (SAXS) from muscle, an approach made possible by the highly ordered arrangement of both the contractile proteins myosin and actin in the ca 2 µm long structural unit, the sarcomere, that repeats along the whole length of the muscle cell. Among time-resolved structural techniques, SAXS has proved to be the most powerful method of investigation, as it allows the molecular motor to be studied in situ, in intact single muscle cells, where it is possible to combine the structural study with fast mechanical methods that synchronize the action of the molecular motors. The latest development of this technique allows Angstrom-scale measurements of the axial movement of the motors that pull the actin filament towards the centre of the sarcomere, by exploiting the x-ray interference between the two arrays of myosin motors in the two halves of the sarcomere.

  19. Recent improvements in small angle x-ray diffraction for the study of muscle physiology

    Energy Technology Data Exchange (ETDEWEB)

    Reconditi, Massimo [Universita di Firenze, Lab di Fisiologia - DBAG, c/o Dip. di Fisica, via Sansone 1, I-50019 Sesto Fiorentino (Italy)

    2006-10-15

    The molecular mechanism of muscle contraction is one of the most important unresolved problems in biology and biophysics. Notwithstanding the great advances of recent years, it is not yet known in detail how the molecular motor in muscle, the class II myosin, converts the free energy of ATP hydrolysis into work by interacting with its track, the actin filament; neither is it understood how the high efficiency in energy conversion depends on the cooperative action of myosin motors working in parallel along the actin filament. Research in muscle contraction involves the combination of mechanical, biochemical and structural methods in studies that span from tissue to single molecule. Therefore, more than for any other research field, progress in the comprehension of muscle contraction at the molecular level is related to, and in turn contributes to, the advancement of methods in biophysics. This review will focus on the progress achieved by time-resolved small angle x-ray scattering (SAXS) from muscle, an approach made possible by the highly ordered arrangement of both the contractile proteins myosin and actin in the ca 2 {mu}m long structural unit, the sarcomere, that repeats along the whole length of the muscle cell. Among time-resolved structural techniques, SAXS has proved to be the most powerful method of investigation, as it allows the molecular motor to be studied in situ, in intact single muscle cells, where it is possible to combine the structural study with fast mechanical methods that synchronize the action of the molecular motors. The latest development of this technique allows Angstrom-scale measurements of the axial movement of the motors that pull the actin filament towards the centre of the sarcomere, by exploiting the x-ray interference between the two arrays of myosin motors in the two halves of the sarcomere.

  20. Stiff muscle fibers in calf muscles of patients with cerebral palsy lead to high passive muscle stiffness.

    Science.gov (United States)

    Mathewson, Margie A; Chambers, Henry G; Girard, Paul J; Tenenhaus, Mayer; Schwartz, Alexandra K; Lieber, Richard L

    2014-12-01

    Cerebral palsy (CP), caused by an injury to the developing brain, can lead to alterations in muscle function. Subsequently, increased muscle stiffness and decreased joint range of motion are often seen in patients with CP. We examined mechanical and biochemical properties of the gastrocnemius and soleus muscles, which are involved in equinus muscle contracture. Passive mechanical testing of single muscle fibers from gastrocnemius and soleus muscle of patients with CP undergoing surgery for equinus deformity showed a significant increase in fiber stiffness (p<0.01). Bundles of fibers that included their surrounding connective tissues showed no stiffness difference (p=0.28).). When in vivo sarcomere lengths were measured and fiber and bundle stiffness compared at these lengths, both fibers and bundles of patients with CP were predicted to be much stiffer in vivo compared to typically developing (TD) individuals. Interestingly, differences in fiber and bundle stiffness were not explained by typical biochemical measures such as titin molecular weight (a giant protein thought to impact fiber stiffness) or collagen content (a proxy for extracellular matrix amount). We suggest that the passive mechanical properties of fibers and bundles are thus poorly understood. © 2014 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

  1. Titin force enhancement following active stretch of skinned skeletal muscle fibres.

    Science.gov (United States)

    Powers, Krysta; Joumaa, Venus; Jinha, Azim; Moo, Eng Kuan; Smith, Ian Curtis; Nishikawa, Kiisa; Herzog, Walter

    2017-09-01

    In actively stretched skeletal muscle sarcomeres, titin-based force is enhanced, increasing the stiffness of active sarcomeres. Titin force enhancement in sarcomeres is vastly reduced in mdm , a genetic mutation with a deletion in titin. Whether loss of titin force enhancement is associated with compensatory mechanisms at higher structural levels of organization, such as single fibres or entire muscles, is unclear. The aim of this study was to determine whether mechanical deficiencies in titin force enhancement are also observed at the fibre level, and whether mechanisms compensate for the loss of titin force enhancement. Single skinned fibres from control and mutant mice were stretched actively and passively beyond filament overlap to observe titin-based force. Mutant fibres generated lower contractile stress (force divided by cross-sectional area) than control fibres. Titin force enhancement was observed in control fibres stretched beyond filament overlap, but was overshadowed in mutant fibres by an abundance of collagen and high variability in mechanics. However, titin force enhancement could be measured in all control fibres and most mutant fibres following short stretches, accounting for ∼25% of the total stress following active stretch. Our results show that the partial loss of titin force enhancement in myofibrils is not preserved in all mutant fibres and this mutation likely affects fibres differentially within a muscle. An increase in collagen helps to reestablish total force at long sarcomere lengths with the loss in titin force enhancement in some mutant fibres, increasing the overall strength of mutant fibres. © 2017. Published by The Company of Biologists Ltd.

  2. Evaluation of carcass and muscle traits in Santa Ines female lambs finished with different agricultural products

    Directory of Open Access Journals (Sweden)

    A.M. Menezes

    Full Text Available ABSTRACT The aim of this study was to evaluate the effect of different agricultural products on quantitative aspects of carcass, body constituents, cooking loss, shear force and colorimetry of the Longissimus lumborum and Triceps brachii muscles in Santa Ines lambs. 24 Santa Ines female lambs received one of four diets which were isoproteic and isoenergetic with fixed levels of forage (60% and concentrate (40% of corn and soybean meal during 45 days. The forages per diet differed: coast-cross hay (HAY, cassava hay (CASS, dehydrated by-product of pea crop (PEA and sugarcane (SC. The average weight of the lambs at the beginning of the experiment was 26.35kg. Animals were slaughtered in a federally certified abattoir. Initial and final pH, cooking losses, color using the CIELAB system, shear force and the quantity of sarcomeres per 100μm were measured. Hot carcass, cold and half carcass weights were affected by treatments (P<0.05. The sarcomere length of Triceps brachii muscle 24 hours after slaughter differed between diets and coast-cross hay had the lowest value. The sarcomere length differed significantly between diets and the dehydrated by-product of pea crop had the lowest number of sarcomeres immediately after slaughter compared to other diets. There was no influence of diet on colorimetry, cooking loss and shear force. The decrease in pH followed the development of the process of rigor mortis in the Longissimus lumborum and Triceps brachii muscles in the first hour and up to 24 hours after slaughter. Diets did not alter the pH, water holding capacity, colorimetry or shear force. The pea by-product and sugarcane can replace traditional sources of fodder without depreciation of meat characteristics.

  3. Isolated abscess in superior rectus muscle in a child

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    Sushank Ashok Bhalerao

    2015-01-01

    Full Text Available Pyomyositis is a primary bacterial infection of striated muscles nearly always caused by Staphylococcus aureus. Development of the intramuscular abscess involving the extra-ocular muscles (EOMs remains an extremely rare process. We herein present a case of isolated EOM pyomyositis involving superior rectus muscle in a 2-year male child who was referred with complaints of swelling in left eye (LE and inability to open LE since last 1-month. Orbital computed tomography (CT scan showed a well-defined, hypo-dense, peripheral rim-enhancing lesion in relation to left superior rectus muscle suggestive of left superior rectus abscess. The abscess was drained through skin approach. We concluded that pyomyositis of EOM should be considered in any patient presenting with acute onset of orbital inflammation and characteristic CT or magnetic resonance imaging features. Management consists of incision and drainage coupled with antibiotic therapy.

  4. The effect of structural and biochemical changes of muscles during post-mortem process on meat tenderness

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    Ana KAIĆ

    2017-11-01

    Full Text Available The aim of this paper was to describe the main structural and biochemical changes of muscles during post-mortem process implicated with meat tenderness. Taking into account complexity of biochemical and structural changes in muscles, meat tenderness is associated with the amount and quality of connective tissue, intramuscular lipid content (marbling, sarcomere length, and myofibrillar protein degradation. In short, more tender meat is associated with lower connective tissue content, higher collagen solubility, greater intramuscular lipid content, longer sarcomeres and greater myofibrillar protein degradation. However, it must be considered that the interactions among the listed factors are complex and dependent on numerous processes during post-mortem such as chilling regime, electrical stimulation, carcass suspension, aging, mechanical tenderization, different marinades, and thermal processing conditions.

  5. Skeletal muscle atrophy and the E3 ubiquitin ligases MuRF1 and MAFbx/atrogin-1

    Science.gov (United States)

    Baehr, Leslie M.

    2014-01-01

    Muscle RING finger 1 (MuRF1) and muscle atrophy F-box (MAFbx)/atrogin-1 were identified more than 10 years ago as two muscle-specific E3 ubiquitin ligases that are increased transcriptionally in skeletal muscle under atrophy-inducing conditions, making them excellent markers of muscle atrophy. In the past 10 years much has been published about MuRF1 and MAFbx with respect to their mRNA expression patterns under atrophy-inducing conditions, their transcriptional regulation, and their putative substrates. However, much remains to be learned about the physiological role of both genes in the regulation of mass and other cellular functions in striated muscle. Although both MuRF1 and MAFbx are enriched in skeletal, cardiac, and smooth muscle, this review will focus on the current understanding of MuRF1 and MAFbx in skeletal muscle, highlighting the critical questions that remain to be answered. PMID:25096180

  6. Challenging muscle homeostasis uncovers novel chaperone interactions in Caenorhabditis elegans

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    Anna eFrumkin

    2014-11-01

    Full Text Available Proteome stability is central to cellular function and the lifespan of an organism. This is apparent in muscle cells, where incorrect folding and assembly of the sarcomere contributes to disease and aging. Apart from the myosin-assembly factor UNC-45, the complete network of chaperones involved in assembly and maintenance of muscle tissue is currently unknown. To identify additional factors required for sarcomere quality control, we performed genetic screens based on suppressed or synthetic motility defects in Caenorhabditis elegans. In addition to ethyl methyl sulfonate-based mutagenesis, we employed RNAi-mediated knockdown of candidate chaperones in unc-45 temperature-sensitive mutants and screened for impaired movement at permissive conditions. This approach confirmed the cooperation between UNC-45 and Hsp90. Moreover, the screens identified three novel co-chaperones, CeHop (STI-1, CeAha1 (C01G10.8 and Cep23 (ZC395.10, required for muscle integrity. The specific identification of Hsp90 and Hsp90 co-chaperones highlights the physiological role of Hsp90 in myosin folding. Our work thus provides a clear example of how a combination of mild perturbations to the proteostasis network can uncover specific quality control modules.

  7. Prenatal Co 60-irradiation effects on visual acuity, maturation of the fovea in the retina, and the striate cortex of squirrel monkey offspring

    International Nuclear Information System (INIS)

    Ordy, J.M.; Brizzee, K.R.; Young, R.

    1982-01-01

    In the present study, foveal striate cortex depth increased significantly from 1400 μm to 1650 μm by 90 days, whereas prenatal 100 rad exposure resulted in a significant reduction of foveal striate cortex thickness at 90 days of age. From birth to 90 days, cell packing density decreased, whereas overall neuropil density increased in both control and 100 rad exposed offspring. Regarding the effects of prenatal radiation on Meynert cells, there was a significant difference in the time course of early postnatal spine frequency reduction on apical dendrites of Meynert cells, particularly in laminae V and IV. It seems possible that the significant differences in the time course of perinatal increases and subsequent decreases of spines and synapses on such pyramidal neurons as Meynert cells in the deep layers of the striate cortex may play an important role in the development of binocular acuity. Future follow-up studies will be essential from 90 days to 1 and 2 years to determine the extent of recovery from, and persistence of visual acuity impairments in relation to structural alterations in the foveal projection of the retino-geniculo-striate system of diurnal primates. (orig./MG)

  8. Expression of neurotrophic factors in diabetic muscle--relation to neuropathy and muscle strength.

    Science.gov (United States)

    Andreassen, C S; Jakobsen, J; Flyvbjerg, A; Andersen, H

    2009-10-01

    Diabetic polyneuropathy can lead to atrophy and weakness of distally located striated muscles due to denervation. Lack of neurotrophic support is believed to contribute to the development of diabetic neuropathy. In this study, we measured the expression of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin 3 (NT-3), neurotrophin 4 (NT-4) and ciliary neurotrophic factor (CNTF) in muscle biopsies taken from the gastrocnemic and deltoid muscles in 42 diabetic patients and 20 healthy control subjects. To express the distal neuropathic gradient and to reduce interindividual variation, a distal/proximal ratio between expression levels in the gastrocnemic and deltoid muscles was calculated for all neurotrophic factors. Neuropathic status was determined by clinical examination, electrophysiological studies and quantitative sensory examination in diabetic patients, and muscle strength at both the shoulder and ankle was assessed by isokinetic dynamometry. Distal/proximal ratios for NT-3 were lower in diabetic patients [median (range) 110.7 (39.8-546.8)] than in controls [157.6 (63.3-385.4); (P < 0.05)], and in neuropathic diabetic patients [107.1 (39.8-326.0)] versus patients without neuropathy [134.5 (46.6-546.8); (P < 0.005)]. Further, ratios for NT-3 were related to muscle strength (r(s) = 0.41, P < 0.01) and showed a tendency towards a negative relationship to the combined score of all measures of neuropathy [Neuropathy rank-sum score (NRSS)] (r(s) = -0.27, P = 0.09). Similar trends were observed for ratios for NT-4. Ratios for NGF (r(s) = -0.32, P < 0.05) and BDNF (r(s) = -0.32, P < 0.05) were related to NRSS, but not to muscle strength. Ratios for CNTF were higher in diabetic patients [64.6 (23.7-258.7)] compared with controls [50.2 (27.2-186.4); (P < 0.05)], but showed no relationship to neither NRSS nor muscle strength. Our results show that the expression of NT-3 is reduced in striated muscles in diabetic patients and is related to

  9. Effect of the bendiocarb on the ultrastructure of rabbit skeletal muscle

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    Katarína Holovská

    2017-01-01

    Full Text Available Bendiocarb belongs to the group of carbamate insecticides that inhibit acetylcholinesterase. In agriculture, it is used to control a variety of insects, therefore it is important to examine every potential aspect of its toxicology. The aim of this study was to observe the effect of bendiocarb on the ultrastructure of the skeletal muscle in rabbits. Rabbits in all experimental groups received capsules of bendiocarb (96% Bendiocarb, Bayer, Germany per os daily at a dose of 5 mg/kg body weight. Samples of skeletal muscles were collected on days 10 and 20. On day 10 of the experiment, muscle fibres were not affected consistently. The observed changes were moderate and focal. Electron microscopy revealed dilatation of sarcoplasmic reticulum, and myofilament disorganization. On day 20 of the experiment, the ultrastructural changes in muscle fibres were more intense and more frequent. The most important alteration was the disruption of the sarcomeres due to the lysis of both thick and thin myofilaments. However, in the unchanged regions of muscle fibres a prominent mitochondrial swelling was observed. Many mitochondria lacked cristae and thus appeared as large membrane-bound cytoplasmic vesicles. The results presented in this study indicate that bendiocarb affects the ultrastructure of skeletal muscles. The intensity of damage (dissolution of myofilaments and disruption of sarcomeres was related to the duration of administration of bendiocarb.

  10. In vivo detection of exercised-induced ultrastructural changes in genetically-altered murine skeletal muscle using polarization-sensitive optical coherence tomography

    Science.gov (United States)

    Boppart, Stephen

    2006-02-01

    Skeletal muscle fibers are a known source of form birefringence in biological tissue. The birefringence present in skeletal muscle is associated with the ultrastructure of individual sarcomeres, specifically the arrangement of A-bands corresponding to the thick myosin filaments. Certain structural proteins that prevent damage and maintain the structural and functional health of the muscle fiber preserve the organization of the Abands in skeletal muscle. Therefore, the level of birefringence detected can estimate the health of the muscle as well as the damage incurred during exercise. Murine skeletal muscle from both genetically-altered (mdx) and normal (wild-type) specimens were imaged in vivo with a fiber-based PSOCT imaging system to quantitatively determine the level of birefringence present in the tissue before and after exercise. The mdx muscle lacks dystrophin, a structural protein that is mutated in Duchenne muscular dystrophy in humans. Muscle from these mdx mice exhibited a marked decrease in birefringence after exercise, whereas the wild-type muscle was highly birefringent before and after exercise. The quantitative results from this tissue optics study suggest for the first time that there is a distinct relationship between the degree of birefringence detected using PS-OCT and the sarcomeric ultrastructure present within skeletal muscle.

  11. Muscle Cramps

    Science.gov (United States)

    Muscle cramps are sudden, involuntary contractions or spasms in one or more of your muscles. They often occur after ... It is a very common muscle problem. Muscle cramps can be caused by nerves that malfunction. Sometimes ...

  12. Muscle differentiation in a colonial ascidian: organisation, gene expression and evolutionary considerations

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    Burighel Paolo

    2009-09-01

    Full Text Available Abstract Background Ascidians are tunicates, the taxon recently proposed as sister group to the vertebrates. They possess a chordate-like swimming larva, which metamorphoses into a sessile adult. Several ascidian species form colonies of clonal individuals by asexual reproduction. During their life cycle, ascidians present three muscle types: striated in larval tail, striated in the heart, and unstriated in the adult body-wall. Results In the colonial ascidian Botryllus schlosseri, we investigated organisation, differentiation and gene expression of muscle beginning from early buds to adults and during zooid regression. We characterised transcripts for troponin T (BsTnT-c, adult muscle-type (BsMA2 and cytoplasmic-type (BsCA1 actins, followed by in situ hybridisation (ISH on sections to establish the spatio-temporal expression of BsTnT-c and BsMA2 during asexual reproduction and in the larva. Moreover, we characterised actin genomic sequences, which by comparison with other metazoans revealed conserved intron patterns. Conclusion Integration of data from ISH, phalloidin staining and TEM allowed us to follow the phases of differentiation of the three muscle kinds, which differ in expression pattern of the two transcripts. Moreover, phylogenetic analyses provided evidence for the close relationship between tunicate and vertebrate muscle genes. The characteristics and plasticity of muscles in tunicates are discussed.

  13. Explaining the variation in the shear force of lamb meat using sarcomere length, the rate of rigor onset and pH.

    Science.gov (United States)

    Hopkins, D L; Toohey, E S; Lamb, T A; Kerr, M J; van de Ven, R; Refshauge, G

    2011-08-01

    The temperature when the pH=6.0 (temp@pH6) impacts on the tenderness and eating quality of sheep meat. Due to the expense, sarcomere length is not routinely measured as a variable to explain variation in shear force, but whether measures such as temp@pH6 are as useful a parameter needs to be established. Measures of rigor onset in 261 carcases, including the temp@pH6, were evaluated in this study for their ability to explain some of the variation in shear force. The results show that for 1 day aged product combinations of the temp@pH6, the pH at 18 °C and the pH at 24 h provided a larger reduction (almost double) in total shear force variation than sarcomere length alone, with pH at 24 h being the single best measure. For 5 day aged product, pH at 18 °C was the single best measure. Inclusion of sarcomere length did represent some improvement, but the marginal increase would not be cost effective. Crown Copyright © 2011. Published by Elsevier Ltd. All rights reserved.

  14. In vivo cardiac nano-imaging: A new technology for high-precision analyses of sarcomere dynamics in the heart.

    Science.gov (United States)

    Shimozawa, Togo; Hirokawa, Erisa; Kobirumaki-Shimozawa, Fuyu; Oyama, Kotaro; Shintani, Seine A; Terui, Takako; Kushida, Yasuharu; Tsukamoto, Seiichi; Fujii, Teruyuki; Ishiwata, Shin'ichi; Fukuda, Norio

    2017-03-01

    The cardiac pump function is a result of a rise in intracellular Ca 2+ and the ensuing sarcomeric contractions [i.e., excitation-contraction (EC) coupling] in myocytes in various locations of the heart. In order to elucidate the heart's mechanical properties under various settings, cardiac imaging is widely performed in today's clinical as well as experimental cardiology by using echocardiogram, magnetic resonance imaging and computed tomography. However, because these common techniques detect local myocardial movements at a spatial resolution of ∼100 μm, our knowledge on the sub-cellular mechanisms of the physiology and pathophysiology of the heart in vivo is limited. This is because (1) EC coupling occurs in the μm partition in a myocyte and (2) cardiac sarcomeres generate active force upon a length change of ∼100 nm on a beat-to-beat basis. Recent advances in optical technologies have enabled measurements of intracellular Ca 2+ dynamics and sarcomere length displacements at high spatial and temporal resolution in the beating heart of living rodents. Future studies with these technologies are warranted to open a new era in cardiac research. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. Cardiac MRI assessed left ventricular hypertrophy in differentiating hypertensive heart disease from hypertrophic cardiomyopathy attributable to a sarcomeric gene mutation

    Energy Technology Data Exchange (ETDEWEB)

    Sipola, Petri [Kuopio University Hospital, Department of Clinical Radiology, Kuopio (Finland); University of Eastern Finland, Institute of Clinical Medicine, Faculty of Health Sciences, Kuopio (Finland); Magga, Jarkko; Peuhkurinen, Keijo [Kuopio University Hospital, Department of Medicine, Kuopio (Finland); Husso, Minna [Kuopio University Hospital, Department of Clinical Radiology, Kuopio (Finland); Jaeaeskelaeinen, Pertti; Kuusisto, Johanna [Kuopio University Hospital, Department of Medicine, Kuopio (Finland); Kuopio University Hospital, Heart Center, P.O. Box 1777, Kuopio (Finland)

    2011-07-15

    To evaluate the value of cardiac magnetic resonance imaging (CMRI)-assessed left ventricular hypertrophy (LVH) in differentiating between hypertensive heart disease and hypertrophic cardiomyopathy (HCM). 95 unselected subjects with mild-to-moderate hypertension, 24 patients with HCM attributable to the D175N mutation of the {alpha}-tropomyosin gene and 17 control subjects were studied by cine CMRI. Left ventricular (LV) quantitative and qualitative characteristics were evaluated. LV maximal end-diastolic wall thickness, wall thickness-to-LV volume ratio, end-diastolic septum thickness and septum-to-lateral wall thickness ratio were useful measures for differentiating between LVH due to hypertension and HCM. The most accurate measure for identifying patients with HCM was the LV maximal wall thickness {>=}17 mm, with a sensitivity, specificity, negative predictive value, positive predictive value, and accuracy of 90%, 93%, 86%, 95% and 91%, respectively. LV maximal wall thickness in the anterior wall, or regional bulging in left ventricular wall was found only in patients with HCM. LV mass index was not discriminant between patients with HCM and those with LVH due to hypertension. LV maximal thickness measured by CMRI is the best anatomical parameter in differentiating between LVH due to mild-to-moderate hypertension and HCM attributable to a sarcomeric mutation. CMRI assessment of location and quality of LVH is also of value in differential diagnosis. (orig.)

  16. Lack of Glycogenin Causes Glycogen Accumulation and Muscle Function Impairment.

    Science.gov (United States)

    Testoni, Giorgia; Duran, Jordi; García-Rocha, Mar; Vilaplana, Francisco; Serrano, Antonio L; Sebastián, David; López-Soldado, Iliana; Sullivan, Mitchell A; Slebe, Felipe; Vilaseca, Marta; Muñoz-Cánoves, Pura; Guinovart, Joan J

    2017-07-05

    Glycogenin is considered essential for glycogen synthesis, as it acts as a primer for the initiation of the polysaccharide chain. Against expectations, glycogenin-deficient mice (Gyg KO) accumulate high amounts of glycogen in striated muscle. Furthermore, this glycogen contains no covalently bound protein, thereby demonstrating that a protein primer is not strictly necessary for the synthesis of the polysaccharide in vivo. Strikingly, in spite of the higher glycogen content, Gyg KO mice showed lower resting energy expenditure and less resistance than control animals when subjected to endurance exercise. These observations can be attributed to a switch of oxidative myofibers toward glycolytic metabolism. Mice overexpressing glycogen synthase in the muscle showed similar alterations, thus indicating that this switch is caused by the excess of glycogen. These results may explain the muscular defects of GSD XV patients, who lack glycogenin-1 and show high glycogen accumulation in muscle. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. Fatigue resistance of rat extraocular muscles does not depend on creatine kinase activity

    Directory of Open Access Journals (Sweden)

    Hayeß Katrin

    2005-08-01

    Full Text Available Abstract Background Creatine kinase (CK links phosphocreatine, an energy storage system, to cellular ATPases. CK activity serves as a temporal and spatial buffer for ATP content, particularly in fast-twitch skeletal muscles. The extraocular muscles are notoriously fast and active, suggesting the need for efficient ATP buffering. This study tested the hypotheses that (1 CK isoform expression and activity in rat extraocular muscles would be higher, and (2 the resistance of these muscles to fatigue would depend on CK activity. Results We found that mRNA and protein levels for cytosolic and mitochondrial CK isoforms were lower in the extraocular muscles than in extensor digitorum longus (EDL. Total CK activity was correspondingly decreased in the extraocular muscles. Moreover, cytoskeletal components of the sarcomeric M line, where a fraction of CK activity is found, were downregulated in the extraocular muscles as was shown by immunocytochemistry and western blotting. CK inhibition significantly accelerated the development of fatigue in EDL muscle bundles, but had no major effect on the extraocular muscles. Searching for alternative ATP buffers that could compensate for the relative lack of CK in extraocular muscles, we determined that mRNAs for two adenylate kinase (AK isoforms were expressed at higher levels in these muscles. Total AK activity was similar in EDL and extraocular muscles. Conclusion These data indicate that the characteristic fatigue resistance of the extraocular muscles does not depend on CK activity.

  18. Comparative Skeletal Muscle Proteomics Using Two-Dimensional Gel Electrophoresis

    Science.gov (United States)

    Murphy, Sandra; Dowling, Paul; Ohlendieck, Kay

    2016-01-01

    The pioneering work by Patrick H. O’Farrell established two-dimensional gel electrophoresis as one of the most important high-resolution protein separation techniques of modern biochemistry (Journal of Biological Chemistry 1975, 250, 4007–4021). The application of two-dimensional gel electrophoresis has played a key role in the systematic identification and detailed characterization of the protein constituents of skeletal muscles. Protein changes during myogenesis, muscle maturation, fibre type specification, physiological muscle adaptations and natural muscle aging were studied in depth by the original O’Farrell method or slightly modified gel electrophoretic techniques. Over the last 40 years, the combined usage of isoelectric focusing in the first dimension and sodium dodecyl sulfate polyacrylamide slab gel electrophoresis in the second dimension has been successfully employed in several hundred published studies on gel-based skeletal muscle biochemistry. This review focuses on normal and physiologically challenged skeletal muscle tissues and outlines key findings from mass spectrometry-based muscle proteomics, which was instrumental in the identification of several thousand individual protein isoforms following gel electrophoretic separation. These muscle-associated protein species belong to the diverse group of regulatory and contractile proteins of the acto-myosin apparatus that forms the sarcomere, cytoskeletal proteins, metabolic enzymes and transporters, signaling proteins, ion-handling proteins, molecular chaperones and extracellular matrix proteins. PMID:28248237

  19. A novel electrical model of nerve and muscle using Pspice

    Energy Technology Data Exchange (ETDEWEB)

    Peasgood, W; Dissado, L A; Lam, C K; Armstrong, A; Wood, W [Department of Engineering, University of Leicester, Leicester (United Kingdom)

    2003-02-21

    In this work, a model is developed to simulate the biological processes involved in nerve fibre transmission and subsequent muscle contraction. The model has been based on approximating biological structure and function to electrical circuits and as such was implemented on an electronics simulation software package called Pspice. Models of nerve, the nerve-muscle interface and muscle fibre have been implemented. The time dependent ionic properties of the nerve and muscle membranes have been simulated using the Hodgkin-Huxley equations and for the muscle fibre, the implementation of the Huxley sliding filament theory for muscular contraction. The results show that nerve may be considered as a fractal transmission line and that the amplitude of the nerve membrane depolarization is dependent on the dimensions of the fibre. Additionally, simulation of the nerve-muscle interface allows the fractal nerve model to be connected to the muscle fibre model and it is shown that a two sarcomere molecular simulation can produce realistic macroscopic muscle force profiles.

  20. Application of microbeam technique for the small-angle diffraction studies of a living tissue

    Energy Technology Data Exchange (ETDEWEB)

    Drakopoulos, M. [ESRF, B.P. 220, 38043 Grenoble, Cedex (France); Sergienko, P.M. [Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, Pushchino 142290 (Russian Federation)]. E-mail: serg@iteb.ru; Snigireva, I. [ESRF, B.P. 220, 38043 Grenoble, Cedex (France); Snigirev, A. [ESRF, B.P. 220, 38043 Grenoble, Cedex (France); Vazina, A.A. [Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, Pushchino 142290 (Russian Federation)

    2005-05-01

    X-ray diffraction patterns from a single hair cross-section and from the functionally different submicron fragments of a frog striated skeletal muscle sarcomere were obtained for the first time by using a microbeam on the undulator beam line ID18 of the third-generation European Synchrotron Radiation Facility (ESRF, Grenoble, France). The microbeam technique proves to be a potential tool for studying the local microstructures of various biological objects.

  1. Fast skeletal muscle troponin activator tirasemtiv increases muscle function and performance in the B6SJL-SOD1G93A ALS mouse model.

    Directory of Open Access Journals (Sweden)

    Darren T Hwee

    Full Text Available Amyotrophic Lateral Sclerosis (ALS is a motor neuron disease characterized by progressive motor neuron loss resulting in muscle atrophy, declining muscle function, and eventual paralysis. Patients typically die from respiratory failure 3 to 5 years from the onset of symptoms. Tirasemtiv is a fast skeletal troponin activator that sensitizes the sarcomere to calcium; this mechanism of action amplifies the response of muscle to neuromuscular input producing greater force when nerve input is reduced. Here, we demonstrate that a single dose of tirasemtiv significantly increases submaximal isometric force, forelimb grip strength, grid hang time, and rotarod performance in a female transgenic mouse model (B6SJL-SOD1 G93A of ALS with functional deficits. Additionally, diaphragm force and tidal volume are significantly higher in tirasemtiv-treated female B6SJL-SOD1 G93A mice. These results support the potential of fast skeletal troponin activators to improve muscle function in neuromuscular diseases.

  2. Muscle Structure Influences Utrophin Expression in mdx Mice

    Science.gov (United States)

    Banks, Glen B.; Combs, Ariana C.; Odom, Guy L.; Bloch, Robert J.; Chamberlain, Jeffrey S.

    2014-01-01

    Duchenne muscular dystrophy (DMD) is a severe muscle wasting disorder caused by mutations in the dystrophin gene. To examine the influence of muscle structure on the pathogenesis of DMD we generated mdx4cv:desmin double knockout (dko) mice. The dko male mice died of apparent cardiorespiratory failure at a median age of 76 days compared to 609 days for the desmin−/− mice. An ∼2.5 fold increase in utrophin expression in the dko skeletal muscles prevented necrosis in ∼91% of 1a, 2a and 2d/x fiber-types. In contrast, utrophin expression was reduced in the extrasynaptic sarcolemma of the dko fast 2b fibers leading to increased membrane fragility and dystrophic pathology. Despite lacking extrasynaptic utrophin, the dko fast 2b fibers were less dystrophic than the mdx4cv fast 2b fibers suggesting utrophin-independent mechanisms were also contributing to the reduced dystrophic pathology. We found no overt change in the regenerative capacity of muscle stem cells when comparing the wild-type, desmin−/−, mdx4cv and dko gastrocnemius muscles injured with notexin. Utrophin could form costameric striations with α-sarcomeric actin in the dko to maintain the integrity of the membrane, but the lack of restoration of the NODS (nNOS, α-dystrobrevin 1 and 2, α1-syntrophin) complex and desmin coincided with profound changes to the sarcomere alignment in the diaphragm, deposition of collagen between the myofibers, and impaired diaphragm function. We conclude that the dko mice may provide new insights into the structural mechanisms that influence endogenous utrophin expression that are pertinent for developing a therapy for DMD. PMID:24922526

  3. Comparative decline of the protein profiles of nebulin in response to denervation in skeletal muscle

    Energy Technology Data Exchange (ETDEWEB)

    Wei, Jih-Hua [Department of Internal Medicine, Min-Sheng General Hospital, Taoyuan, Taiwan (China); Chang, Nen-Chung [Division of Cardiology, Department of Internal Medicine, College of Medicine, Taipei Medical University Hospital, Taipei, Taiwan (China); Chen, Sy-Ping [Department of Nursing, Chang Gung University of Science and Technology, Taoyuan, Taiwan (China); Geraldine, Pitchairaj [Department of Animal Science, School of Life Sciences, Bharathidasan University, Tiruchirappalli, Tamil Nadu (India); Jayakumar, Thanasekaran, E-mail: tjaya_2002@yahoo.co.in [Department of Pharmacology and Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan (China); Fong, Tsorng-Harn, E-mail: thfong@tmu.edu.tw [Department of Anatomy and Cell Biology, College of Medicine, Taipei Medical University, Taipei, Taiwan (China)

    2015-10-09

    The sliding filament model of the sarcomere was developed more than half a century ago. This model, consisting only of thin and thick filaments, has been efficacious in elucidating many, but not all, features of skeletal muscle. Work during the 1980s revealed the existence of two additional filaments: the giant filamentous proteins titin and nebulin. Nebulin, a giant myofibrillar protein, acts as a protein ruler to maintain the lattice arrays of thin filaments and plays a role in signal transduction and contractile regulation. However, the change of nebulin and its effect on thin filaments in denervation-induced atrophic muscle remains unclear. The purpose of this study is to examine the content and pattern of nebulin, myosin heavy chain (MHC), actin, and titin in innervated and denervated tibialis anterior (TA) muscles of rats using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), densitometry and electron microscopic (EM) analyses. The results revealed that denervation induced muscle atrophy is accompanied by decreased nebulin content in a time-dependent manner. For instant, the levels of nebulin in denervated muscles were markedly (P < 0.05) decreased, about 24.6% and 40.2% in comparison with innervated muscle after denervation of 28 and 56 days, respectively. The nebulin/MHC, nebulin/actin, and nebulin/titin ratios were decreased, suggesting a concomitant reduction of nebulin in denervated muscle. Moreover, a western blotting assay proved that nebulin declined faster than titin on 28 and 56 days of denervated muscle. In addition, EM study revealed that the disturbed arrangements of myofilaments and a disorganized contractile apparatus were also observed in denervated muscle. Overall, the present study provides evidence that nebulin is more sensitive to the effect of denervation than MHC, actin, and titin. Nebulin decline indeed resulted in disintegrate of thin filaments and shortening of sarcomeres. - Highlights: • We successfully

  4. Role of skeletal muscle in lung development.

    Science.gov (United States)

    Baguma-Nibasheka, Mark; Gugic, Dijana; Saraga-Babic, Mirna; Kablar, Boris

    2012-07-01

    Skeletal (striated) muscle is one of the four basic tissue types, together with the epithelium, connective and nervous tissues. Lungs, on the other hand, develop from the foregut and among various cell types contain smooth, but not skeletal muscle. Therefore, during earlier stages of development, it is unlikely that skeletal muscle and lung depend on each other. However, during the later stages of development, respiratory muscle, primarily the diaphragm and the intercostal muscles, execute so called fetal breathing-like movements (FBMs), that are essential for lung growth and cell differentiation. In fact, the absence of FBMs results in pulmonary hypoplasia, the most common cause of death in the first week of human neonatal life. Most knowledge on this topic arises from in vivo experiments on larger animals and from various in vitro experiments. In the current era of mouse mutagenesis and functional genomics, it was our goal to develop a mouse model for pulmonary hypoplasia. We employed various genetically engineered mice lacking different groups of respiratory muscles or lacking all the skeletal muscle and established the criteria for pulmonary hypoplasia in mice, and therefore established a mouse model for this disease. We followed up this discovery with systematic subtractive microarray analysis approach and revealed novel functions in lung development and disease for several molecules. We believe that our approach combines elements of both in vivo and in vitro approaches and allows us to study the function of a series of molecules in the context of lung development and disease and, simultaneously, in the context of lung's dependence on skeletal muscle-executed FBMs.

  5. Synergistic Activities of Abdominal Muscles Are Required for Efficient Micturition in Anesthetized Female Mice.

    Science.gov (United States)

    Zhang, Chuan; Zhang, Yingchun; Cruz, Yolanda; Boone, Timothy B; Munoz, Alvaro

    2018-03-01

    To characterize the electromyographic activity of abdominal striated muscles during micturition in urethane-anesthetized female mice, and to quantitatively evaluate the contribution of abdominal responses to efficient voiding. Cystometric and multichannel electromyographic recordings were integrated to enable a comprehensive evaluation during micturition in urethane-anesthetized female mice. Four major abdominal muscle domains were evaluated: the external oblique, internal oblique, and superior and inferior rectus abdominis. To further characterize the functionality of the abdominal muscles, pancuronium bromide (25 μg/mL or 50 μg/mL, abdominal surface) was applied as a blocking agent of neuromuscular junctions. We observed a robust activation of the abdominal muscles during voiding, with a consistent onset/offset concomitant with the bladder pressure threshold. Pancuronium was effective, in a dose-dependent fashion, for partial and complete blockage of abdominal activity. Electromyographic discharges during voiding were significantly inhibited by applying pancuronium. Decreased cystometric parameters were recorded, including the peak pressure, pressure threshold, intercontractile interval, and voiding duration, suggesting that the voiding efficiency was significantly compromised by abdominal muscle relaxation. The relevance of the abdominal striated musculature for micturition has remained a topic of debate in human physiology. Although the study was performed on anesthetized mice, these results support the existence of synergistic abdominal electromyographic activity facilitating voiding in anesthetized mice. Further, our study presents a rodent model that can be used for future investigations into micturition-related abdominal activity.

  6. Validity of Estimation of Pelvic Floor Muscle Activity from Transperineal Ultrasound Imaging in Men.

    Science.gov (United States)

    Stafford, Ryan E; Coughlin, Geoff; Lutton, Nicholas J; Hodges, Paul W

    2015-01-01

    To investigate the relationship between displacement of pelvic floor landmarks observed with transperineal ultrasound imaging and electromyography of the muscles hypothesised to cause the displacements. Three healthy men participated in this study, which included ultrasound imaging of the mid-urethra, urethra-vesical junction, ano-rectal junction and bulb of the penis. Fine-wire electromyography electrodes were inserted into the puborectalis and bulbocavernosus muscles and a transurethral catheter electrode recorded striated urethral sphincter electromyography. A nasogastric sensor recorded intra-abdominal pressure. Tasks included submaximal and maximal voluntary contractions, and Valsalva. The relationship between each of the parameters measured from ultrasound images and electromyography or intra-abdominal pressure amplitudes was described with nonlinear regression. Strong, non-linear relationships were calculated for each predicted landmark/muscle pair for submaximal contractions (R2-0.87-0.95). The relationships between mid-urethral displacement and striated urethral sphincter electromyography, and bulb of the penis displacement and bulbocavernosus electromyography were strong during maximal contractions (R2-0.74-0.88). Increased intra-abdominal pressure prevented shortening of puborectalis, which resulted in weak relationships between electromyography and anorectal and urethravesical junction displacement during all tasks. Displacement of landmarks in transperineal ultrasound imaging provides meaningful measures of activation of individual pelvic floor muscles in men during voluntary contractions. This method may aid assessment of muscle function or feedback for training.

  7. Validity of Estimation of Pelvic Floor Muscle Activity from Transperineal Ultrasound Imaging in Men.

    Directory of Open Access Journals (Sweden)

    Ryan E Stafford

    Full Text Available To investigate the relationship between displacement of pelvic floor landmarks observed with transperineal ultrasound imaging and electromyography of the muscles hypothesised to cause the displacements.Three healthy men participated in this study, which included ultrasound imaging of the mid-urethra, urethra-vesical junction, ano-rectal junction and bulb of the penis. Fine-wire electromyography electrodes were inserted into the puborectalis and bulbocavernosus muscles and a transurethral catheter electrode recorded striated urethral sphincter electromyography. A nasogastric sensor recorded intra-abdominal pressure. Tasks included submaximal and maximal voluntary contractions, and Valsalva. The relationship between each of the parameters measured from ultrasound images and electromyography or intra-abdominal pressure amplitudes was described with nonlinear regression.Strong, non-linear relationships were calculated for each predicted landmark/muscle pair for submaximal contractions (R2-0.87-0.95. The relationships between mid-urethral displacement and striated urethral sphincter electromyography, and bulb of the penis displacement and bulbocavernosus electromyography were strong during maximal contractions (R2-0.74-0.88. Increased intra-abdominal pressure prevented shortening of puborectalis, which resulted in weak relationships between electromyography and anorectal and urethravesical junction displacement during all tasks.Displacement of landmarks in transperineal ultrasound imaging provides meaningful measures of activation of individual pelvic floor muscles in men during voluntary contractions. This method may aid assessment of muscle function or feedback for training.

  8. Investigations into the Sarcomeric Protein and Ca2+-Regulation Abnormalities Underlying Hypertrophic Cardiomyopathy in Cats (Felix catus

    Directory of Open Access Journals (Sweden)

    Andrew E. Messer

    2017-06-01

    Full Text Available Hypertrophic cardiomyopathy (HCM is the most common single gene inherited cardiomyopathy. In cats (Felix catus HCM is even more prevalent and affects 16% of the outbred population and up to 26% in pedigree breeds such as Maine Coon and Ragdoll. Homozygous MYBPC3 mutations have been identified in these breeds but the mutations in other cats are unknown. At the clinical and physiological level feline HCM is closely analogous to human HCM but little is known about the primary causative mechanism. Most identified HCM causing mutations are in the genes coding for proteins of the sarcomere. We therefore investigated contractile and regulatory proteins in left ventricular tissue from 25 cats, 18 diagnosed with HCM, including a Ragdoll cat with a homozygous MYBPC3 R820W, and 7 non-HCM cats in comparison with human HCM (from septal myectomy and donor heart tissue. Myofibrillar protein expression was normal except that we observed 20–44% MyBP-C haploinsufficiency in 5 of the HCM cats. Troponin extracted from 8 HCM and 5 non-HCM cat hearts was incorporated into thin filaments and studied by in vitro motility assay. All HCM cat hearts had a higher (2.06 ± 0.13 fold Ca2+-sensitivity than non-HCM cats and, in all the HCM cats, Ca2+-sensitivity was not modulated by troponin I phosphorylation. We were able to restore modulation of Ca2+-sensitivity by replacing troponin T with wild-type protein or by adding 100 μM Epigallocatechin 3-gallate (EGCG. These fundamental regulatory characteristics closely mimic those seen in human HCM indicating a common molecular mechanism that is independent of the causative mutation. Thus, the HCM cat is a potentially useful large animal model.

  9. Investigations into the Sarcomeric Protein and Ca2+-Regulation Abnormalities Underlying Hypertrophic Cardiomyopathy in Cats (Felix catus)

    Science.gov (United States)

    Messer, Andrew E.; Chan, Jasmine; Daley, Alex; Copeland, O'Neal; Marston, Steven B.; Connolly, David J.

    2017-01-01

    Hypertrophic cardiomyopathy (HCM) is the most common single gene inherited cardiomyopathy. In cats (Felix catus) HCM is even more prevalent and affects 16% of the outbred population and up to 26% in pedigree breeds such as Maine Coon and Ragdoll. Homozygous MYBPC3 mutations have been identified in these breeds but the mutations in other cats are unknown. At the clinical and physiological level feline HCM is closely analogous to human HCM but little is known about the primary causative mechanism. Most identified HCM causing mutations are in the genes coding for proteins of the sarcomere. We therefore investigated contractile and regulatory proteins in left ventricular tissue from 25 cats, 18 diagnosed with HCM, including a Ragdoll cat with a homozygous MYBPC3 R820W, and 7 non-HCM cats in comparison with human HCM (from septal myectomy) and donor heart tissue. Myofibrillar protein expression was normal except that we observed 20–44% MyBP-C haploinsufficiency in 5 of the HCM cats. Troponin extracted from 8 HCM and 5 non-HCM cat hearts was incorporated into thin filaments and studied by in vitro motility assay. All HCM cat hearts had a higher (2.06 ± 0.13 fold) Ca2+-sensitivity than non-HCM cats and, in all the HCM cats, Ca2+-sensitivity was not modulated by troponin I phosphorylation. We were able to restore modulation of Ca2+-sensitivity by replacing troponin T with wild-type protein or by adding 100 μM Epigallocatechin 3-gallate (EGCG). These fundamental regulatory characteristics closely mimic those seen in human HCM indicating a common molecular mechanism that is independent of the causative mutation. Thus, the HCM cat is a potentially useful large animal model. PMID:28642712

  10. A striated, far travelled clast of rhyolitic tuff from Thames river deposits at Ardleigh, Essex, England : evidence for early Middle Pleistocene glaciation in the Thames catchment

    OpenAIRE

    Rose, J.; Carney, J.N.; Silva, B.N.; Booth, S.J.

    2010-01-01

    This paper reports the discovery of an in-situ striated, far-travelled, oversized clast in the Ardleigh Gravels of the Kesgrave Sands and Gravels of the River Thames at Ardleigh, east of Colchester in Essex, eastern England. The morphology, petrography and geochemistry of the clast, and the sedimentology of the host deposit are described. The striations are interpreted, on the basis of their sub-parallelism and the shape and subroundedness of the clast, as glacial and the clast is pr...

  11. Cytoskeleton, L-type Ca2+ and stretch activated channels in injured skeletal muscle

    Directory of Open Access Journals (Sweden)

    Fabio Francini

    2013-07-01

    Full Text Available The extra-sarcomeric cytoskeleton (actin microfilaments and anchoring proteins is involved in maintaining the sarco-membrane stiffness and integrity and in turn the mechanical stability and function of the intra- and sub-sarcoplasmic proteins. Accordingly, it regulates Ca2+ entry through the L-type Ca2+ channels and the mechano-sensitivity of the stretch activated channels (SACs. Moreover, being intra-sarcomeric cytoskeleton bound to costameric proteins and other proteins of the sarcoplasma by intermediate filaments, as desmin, it integrates the properties of the sarcolemma with the skeletal muscle fibres contraction. The aim of this research was to compare the cytoskeleton, SACs and the ECC alterations in two different types of injured skeletal muscle fibres: by muscle denervation and mechanical overload (eccentric contraction. Experiments on denervation were made in isolated Soleus muscle of male Wistar rats; forced eccentric-contraction (EC injury was achieved in Extensor Digitorum Longus muscles of Swiss mice. The method employed conventional intracellular recording with microelectrodes inserted in a single fibre of an isolated skeletal muscle bundle. The state of cytoskeleton was evaluated by recording SAC currents and by evaluating the resting membrane potential (RMP value determined in current-clamp mode. The results demonstrated that in both injured skeletal muscle conditions the functionality of L-type Ca2+ current, ICa, was affected. In parallel, muscle fibres showed an increase of the resting membrane permeability and of the SAC current. These issues, together with a more depolarized RMP are an index of altered cytoskeleton. In conclusion, we found a symilar alteration of ICa, SAC and cytoskeleton in both injured skeletal muscle conditions.

  12. Use of gene targeting for compromising energy homeostasis in neuromuscular tissues:The role of sarcomeric mitochondrial creatine kinase

    NARCIS (Netherlands)

    Steeghs, K.; Heerschap, A.; de Haan, A.; Ruitenbeek, W.; Oerlemans, F.; Deursen, J.; Perryman, B.; Pette, D.; Br|ckwilder, M.; Koudijs, J.; Jap, P.; Wieringa, B.

    1997-01-01

    We have introduced a single knock-out mutation in the mitochondrial creatine kinase gene (ScCKmit) in the mouse germ line via targeted mutagenesis in mouse embryonic stem (ES) cells. Surprisingly, ScCKmit -/- muscles, unlike muscles of mice with a deficiency of cytosolic M-type creatine kinase (M-CK

  13. Geologic continuous casting below continental and deep-sea detachment faults and at the striated extrusion of Sacsayhuaman, Peru

    Science.gov (United States)

    Spencer, J.E.

    1999-01-01

    In the common type of industrial continuous casting, partially molten metal is extruded from a vessel through a shaped orifice called a mold in which the metal assumes the cross-sectional form of the mold as it cools and solidifies. Continuous casting can be sustained as long as molten metal is supplied and thermal conditions are maintained. I propose that a similar process produced parallel sets of grooves in three geologic settings, as follows: (1) corrugated metamorphic core complexes where mylonized mid-crustal rocks were exhumed by movement along low-angle normal faults known as detachment faults; (2) corrugated submarine surfaces where ultramafic and mafic rocks were exhumed by normal faulting within oceanic spreading centers; and (3) striated magma extrusions exemplified by the famous grooved outcrops at the Inca fortress of Sacsayhuaman in Peru. In each case, rocks inferred to have overlain the corrugated surface during corrugation genesis molded and shaped a plastic to partially molten rock mass as it was extruded from a moderate- to high-temperature reservoir.

  14. Muscle Deoxygenation Causes Muscle Fatigue

    Science.gov (United States)

    Murthy, G.; Hargens, A. R.; Lehman, S.; Rempel, D.

    1999-01-01

    Muscle fatigue is a common musculoskeletal disorder in the work place, and may be a harbinger for more disabling cumulative trauma disorders. Although the cause of fatigue is multifactorial, reduced blood flow and muscle oxygenation may be the primary factor in causing muscle fatigue during low intensity muscle exertion. Muscle fatigue is defined as a reduction in muscle force production, and also occurs among astronauts who are subjected to postural constraints while performing lengthy, repetitive tasks. The objectives of this research are to: 1) develop an objective tool to study the role of decreased muscle oxygenation on muscle force production, and 2) to evaluate muscle fatigue during prolonged glovebox work.

  15. Titin Isoform Size is Not Correlated with Thin Filament Length in Rat Skeletal Muscle

    Directory of Open Access Journals (Sweden)

    Marion Lewis Greaser

    2014-02-01

    Full Text Available The mechanisms controlling thin filament length in muscle remain controversial. It was recently reported that thin filament length was related to titin size, and that the latter might be involved in thin filament length determination. Titin plays several crucial roles in the sarcomere, but its function as it pertains to the thin filament has not been explored. We tested this relationship using several muscles from wild type rats and from a mutant rat model which results in increased titin size. Myofibrils were isolated from skeletal muscles (extensor digitorum longus, external oblique, gastrocnemius, longissimus dorsi, psoas major, and tibialis anterior using both adult wild type (WT and homozygous mutant (HM rats. Phalloidin and antibodies against tropomodulin-4 and nebulin’s N-terminus were used to determine thin filament length. The WT rats studied express skeletal muscle titin sizes ranging from 3.2 to 3.7 MDa, while the HM rats express a giant titin isoform sized at 3.7 MDa. No differences in phalloidin-based thin filament length, nebulin N terminus distances from the Z line, or tropomodulin distances from the Z line were observed across genotypes. The data indicates that, although titin performs many sarcomeric functions, its correlation with thin filament length and structure could not be demonstrated in the rat. Current models of thin filament assembly are inadequate to explain the phalloidin, nebulin N terminus, and tropomodulin staining patterns in the myofibril.

  16. Muscle hypertrophy as the presenting sign in a patient with a complete FHL1 deletion.

    Science.gov (United States)

    Willis, T A; Wood, C L; Hudson, J; Polvikoski, T; Barresi, R; Lochmüller, H; Bushby, K; Straub, V

    2016-08-01

    Four and a half LIM protein 1 (FHL1/SLIM1) has recently been identified as the causative gene mutated in four distinct diseases affecting skeletal muscle that have overlapping features, including reducing body myopathy, X-linked myopathy, X-linked dominant scapuloperoneal myopathy and Emery-Dreifuss muscular dystrophy. FHL1 localises to the sarcomere and the sarcolemma and is believed to participate in muscle growth and differentiation as well as in sarcomere assembly. We describe in this case report a boy with a deletion of the entire FHL1 gene who is now 15 years of age and presented with muscle hypertrophy, reduced subcutaneous fat, rigid spine and short stature. This case is the first, to our knowledge, with a complete loss of the FHL1 protein and MAP7D3 in combination. It supports the theory that dominant negative effects (accumulation of cytotoxic-mutated FHL1 protein) worsen the pathogenesis. It extends the phenotype of FHL1-related myopathies and should prompt future testing in undiagnosed patients who present with unexplained muscle hypertrophy, contractures and rigid spine, particularly if male. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  17. Time-resolved x-ray diffraction from frog skeletal muscle during an isotonic twitch under a small load

    International Nuclear Information System (INIS)

    Sugi, Haruo; Amemiya, Yoshiyuki; Hashizume, Hiroo.

    1978-01-01

    A time-resolved x-ray diffraction technique was used to study the time course of change in the intensity ratio Isub(1,0)/Isub(1,1) during isotonic twitch (initial sarcomere, 2.4 μm) under a small load and to determine the kinetic properties of the crossbridges responsible for muscle contraction. Isotonic twitches in four other preparations with an initial sarcomere of 2.4 μm and in two with an initial sarcomere of 2.3 μm and 2.2 μm, respectively, were examined. In each case, the intensity ratio started to decrease at stimulation, reached a minimum value of 0.8 - 1.0 within the first 20 - 30% of the shortening phase, and maintained this value until the beginning of the relaxation phase. Gradual recovery of the intensity ratio to the resting value was seen during the relaxation phase. During the recovery phase, the intensity ratio appeared to exhibit oscillatory changes. Though the extent of shortening was reduced by about 30% at the end of each experiment, the duration of the shortening phase remained almost unchanged in all the preparations examined. The time course of change in the intensity ratio was also examined during an isometric twitch in four preparations (sarcomere, 2.4 μm) with the tibial end connected to a strain gauge. The extent of internal shortening of muscle fibres against the tendons and the recording system during an isometric twitch or a tetanus at low temperatures was estimated. The intensity ratio decreased to a minimum value of 0.5 - 0.6 during the rising phase of isometric tension and started to return to the resting value after the beginning of relaxation. In both isotonic and isometric twitches, a decrease in the intensity ratio resulted from both a decrease in the 1,0 intensity and an increase in the 1,1 intensity. (J.P.N.)

  18. Recent applications of multiwire proportional chambers for time resolved studies on muscle

    Energy Technology Data Exchange (ETDEWEB)

    Faruqi, A.R.; Huxley, H.E.; Kress, M.

    1986-12-01

    The use of multiwire proportional chambers has played an important role in recording time resolved data from vertebrate muscle under various physiological conditions from which we have selected two examples from recent work. The first example describes the measurement of the equatorial pattern along with measurements of the muscle sarcomere length showing that the delay in tension development relative to changes in the X-ray intensity are not due to ''internal'' shortening in the muscle. The second example describes time-resolved measurements on the thin filament activation process, studied with both a linear and an area detector. The results show clearly that changes in thin filament structure precede cross-bridge formation producing further strong evidence for the ''steric blocking model'' for muscle contraction.

  19. The Popeye domain containing 2 (popdc2) gene in zebrafish is required for heart and skeletal muscle development

    Science.gov (United States)

    Kirchmaier, Bettina C.; Poon, Kar Lai; Schwerte, Thorsten; Huisken, Jan; Winkler, Christoph; Jungblut, Benno; Stainier, Didier Y.; Brand, Thomas

    2013-01-01

    The Popeye domain containing (Popdc) genes encode a family of transmembrane proteins with an evolutionary conserved Popeye domain. These genes are abundantly expressed in striated muscle tissue, however their function is not well understood. In this study we have investigated the role of the popdc2 gene in zebrafish. Popdc2 transcripts were detected in the embryonic myocardium and transiently in the craniofacial and tail musculature. Morpholino oligonucleotide-mediated knockdown of popdc2 resulted in aberrant development of skeletal muscle and heart. Muscle segments in the trunk were irregularly shaped and craniofacial muscles were severely reduced or even missing. In the heart, pericardial edema was prevalent in the morphants and heart chambers were elongated and looping was abnormal. These pathologies in muscle and heart were alleviated after reducing the morpholino concentration. However the heart still was abnormal displaying cardiac arrhythmia at later stages of development. Optical recordings of cardiac contractility revealed irregular ventricular contractions with a 2:1, or 3:1 atrial/ventricular conduction ratio, which caused a significant reduction in heart frequency. Recordings of calcium transients with high spatiotemporal resolution using a transgenic calcium indicator line (Tg(cmlc2:gCaMP)s878) and SPIM microscopy confirmed the presence of a severe arrhythmia phenotype. Our results identify popdc2 as a gene important for striated muscle differentiation and cardiac morphogenesis. In addition it is required for the development of the cardiac conduction system. PMID:22290329

  20. Comparison of palatability characteristics of beef gluteus medius and triceps brachii muscles.

    Science.gov (United States)

    King, D A; Wheeler, T L; Shackelford, S D; Koohmaraie, M

    2009-01-01

    The objective of this experiment was to evaluate triceps brachii steaks as a substitute for gluteus medius steaks in foodservice and retail applications, including the effect of aging time and USDA quality grade on the palatability of both muscles. Top sirloin butts (n = 600) and shoulder clod arm roasts (n = 600) representing US Choice and US Select quality grades were selected at 48 h postmortem and aged for 7, 14, 21, 28, 35, or 42 d. Steaks were evaluated using a trained sensory panel, slice shear force, sarcomere length, and Western blotting of desmin measurements. Sarcomere length was measured only on steaks at 14 and 42 d. Triceps brachii and gluteus medius steaks were similar in tenderness rating at 7 and 14 d, but triceps brachii steaks aged longer were more tender (P gluteus medius steaks. Triceps brachii steaks reached ultimate tenderness values by 21 d. Gluteus medius steak tenderness ratings improved through 35 d, and at 42 d were similar to those given to triceps brachii steaks at 21 d. Sarcomere lengths were longer (P gluteus medius (2.09 and 1.58 mum, respectively). Significant increases in desmin degradation were detected through 42 d in both muscles (30.9, 46.3, 50.6, 51.0, 57.6, and 64.1% at d 7, 14, 21, 28, 35, and 42 for gluteus medius and 28.9, 40.8, 49.3, 59.2, 61.8, and 71.9% at d 7, 14, 21, 28, 35, and 42 for triceps brachii). At 14 d, gluteus medius had more (P gluteus medius tenderness variation (r = 0.36) across all aging times, but not at individual aging times. Sarcomere length contributed to variation in slice shear force values of gluteus medius at 14 and 42 d (r = -0.59 and -0.48, respectively). Sarcomere length contributed to triceps brachii tenderness variation at 14 d, but not 42 d (r = 0.44 and -0.12, respectively). Desmin degradation was strongly correlated (r = 0.55) to triceps brachii tenderness ratings pooled across aging times but not at individual aging times. These data indicate that triceps brachii steaks could provide

  1. Differences in Contractile Function of Myofibrils within Human Embryonic Stem Cell-Derived Cardiomyocytes vs. Adult Ventricular Myofibrils Are Related to Distinct Sarcomeric Protein Isoforms

    Directory of Open Access Journals (Sweden)

    Bogdan Iorga

    2018-01-01

    Full Text Available Characterizing the contractile function of human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs is key for advancing their utility for cellular disease models, promoting cell based heart repair, or developing novel pharmacological interventions targeting cardiac diseases. The aim of the present study was to understand whether steady-state and kinetic force parameters of β-myosin heavy chain (βMyHC isoform-expressing myofibrils within human embryonic stem cell-derived cardiomyocytes (hESC-CMs differentiated in vitro resemble those of human ventricular myofibrils (hvMFs isolated from adult donor hearts. Contractile parameters were determined using the same micromechanical method and experimental conditions for both types of myofibrils. We identified isoforms and phosphorylation of main sarcomeric proteins involved in the modulation of force generation of both, chemically demembranated hESC-CMs (d-hESC-CMs and hvMFs. Our results indicate that at saturating Ca2+ concentration, both human-derived contractile systems developed forces with similar rate constants (0.66 and 0.68 s−1, reaching maximum isometric force that was significantly smaller for d-hESC-CMs (42 kPa than for hvMFs (94 kPa. At submaximal Ca2+-activation, where intact cardiomyocytes normally operate, contractile parameters of d-hESC-CMs and hvMFs exhibited differences. Ca2+ sensitivity of force was higher for d-hESC-CMs (pCa50 = 6.04 than for hvMFs (pCa50 = 5.80. At half-maximum activation, the rate constant for force redevelopment was significantly faster for d-hESC-CMs (0.51 s−1 than for hvMFs (0.28 s−1. During myofibril relaxation, kinetics of the slow force decay phase were significantly faster for d-hESC-CMs (0.26 s−1 than for hvMFs (0.21 s−1, while kinetics of the fast force decay were similar and ~20x faster. Protein analysis revealed that hESC-CMs had essentially no cardiac troponin-I, and partially non-ventricular isoforms of some other sarcomeric proteins

  2. Residual force depression following muscle shortening is exaggerated by prior eccentric drop jump exercise.

    Science.gov (United States)

    Dargeviciute, Gintare; Masiulis, Nerijus; Kamandulis, Sigitas; Skurvydas, Albertas; Westerblad, Håkan

    2013-10-15

    We studied the relation between two common force modifications in skeletal muscle: the prolonged force depression induced by unaccustomed eccentric contractions, and the residual force depression (rFD) observed immediately after active shortening. We hypothesized that rFD originates from distortion within the sarcomeres and the extent of rFD: 1) correlates to the force and work performed during the shortening steps, which depend on sarcomeric integrity; and 2) is increased by sarcomeric disorganization induced by eccentric contractions. Nine healthy untrained men (mean age 26 yr) participated in the study. rFD was studied in electrically stimulated knee extensor muscles. rFD was defined as the reduction in isometric torque after active shortening compared with the torque in a purely isometric contraction. Eccentric contractions were performed as 50 repeated drop jumps with active deceleration to 90° knee angle, immediately followed by a maximal upward jump. rFD was assessed before and 5 min to 72 h after drop jumps. The series of drop jumps caused a prolonged force depression, which was about two times larger at 20-Hz than at 50-Hz stimulation. There was a significant correlation between increasing rFD and increasing mechanical work performed during active shortening both before and after drop jumps. In addition, a given rFD was obtained at a markedly lower mechanical work after drop jumps. In conclusion, the extent of rFD correlates to the mechanical work performed during active shortening. A series of eccentric contractions causes a prolonged reduction of isometric force. In addition, eccentric contractions exaggerate rFD, which further decreases muscle performance during dynamic contractions.

  3. Muscle pain

    African Journals Online (AJOL)

    Key Summary Points. • Muscle pain, known as myalgia, can be in one targeted area or across many muscles, occurring with overexertion or overuse of these muscles. • Pain can be classified as acute or chronic pain and further categorized as nociceptive or neuropathic. • Causes of muscle pain include stress, physical ...

  4. Cell division in Apicomplexan parasites is organized by a homolog of the striated rootlet fiber of algal flagella.

    Directory of Open Access Journals (Sweden)

    Maria E Francia

    Full Text Available Apicomplexa are intracellular parasites that cause important human diseases including malaria and toxoplasmosis. During host cell infection new parasites are formed through a budding process that parcels out nuclei and organelles into multiple daughters. Budding is remarkably flexible in output and can produce two to thousands of progeny cells. How genomes and daughters are counted and coordinated is unknown. Apicomplexa evolved from single celled flagellated algae, but with the exception of the gametes, lack flagella. Here we demonstrate that a structure that in the algal ancestor served as the rootlet of the flagellar basal bodies is required for parasite cell division. Parasite striated fiber assemblins (SFA polymerize into a dynamic fiber that emerges from the centrosomes immediately after their duplication. The fiber grows in a polarized fashion and daughter cells form at its distal tip. As the daughter cell is further elaborated it remains physically tethered at its apical end, the conoid and polar ring. Genetic experiments in Toxoplasma gondii demonstrate two essential components of the fiber, TgSFA2 and 3. In the absence of either of these proteins cytokinesis is blocked at its earliest point, the initiation of the daughter microtubule organizing center (MTOC. Mitosis remains unimpeded and mutant cells accumulate numerous nuclei but fail to form daughter cells. The SFA fiber provides a robust spatial and temporal organizer of parasite cell division, a process that appears hard-wired to the centrosome by multiple tethers. Our findings have broader evolutionary implications. We propose that Apicomplexa abandoned flagella for most stages yet retained the organizing principle of the flagellar MTOC. Instead of ensuring appropriate numbers of flagella, the system now positions the apical invasion complexes. This suggests that elements of the invasion apparatus may be derived from flagella or flagellum associated structures.

  5. Methylmercury intoxication and histochemical demonstration of NADPH-diaphorase activity in the striate cortex of adult cats

    Directory of Open Access Journals (Sweden)

    R.B. Oliveira

    1998-09-01

    Full Text Available The effects of methylmercury (MeHg on histochemical demonstration of the NADPH-diaphorase (NADPH-d activity in the striate cortex were studied in 4 adult cats. Two animals were used as control. The contaminated animals received 50 ml milk containing 0.42 µg MeHg and 100 g fish containing 0.03 µg MeHg daily for 2 months. The level of MeHg in area 17 of intoxicated animals was 3.2 µg/g wet weight brain tissue. Two cats were perfused 24 h after the last dose (group 1 and the other animals were perfused 6 months later (group 2. After microtomy, sections were processed for NADPHd histochemistry procedures using the malic enzyme method. Dendritic branch counts were performed from camera lucida drawings for control and intoxicated animals (N = 80. Average, standard deviation and Student t-test were calculated for each data group. The concentrations of mercury (Hg in milk, fish and brain tissue were measured by acid digestion of samples, followed by reduction of total Hg in the digested sample to metallic Hg using stannous chloride followed by atomic fluorescence analysis. Only group 2 revealed a reduction of the neuropil enzyme activity and morphometric analysis showed a reduction in dendritic field area and in the number of distal dendrite branches of the NADPHd neurons in the white matter (P<0.05. These results suggest that NADPHd neurons in the white matter are more vulnerable to the long-term effects of MeHg than NADPHd neurons in the gray matter.

  6. Quantification of birefringence readily measures the level of muscle damage in zebrafish

    International Nuclear Information System (INIS)

    Berger, Joachim; Sztal, Tamar; Currie, Peter D.

    2012-01-01

    Highlights: ► Report of an unbiased quantification of the birefringence of muscle of fish larvae. ► Quantification method readily identifies level of overall muscle damage. ► Compare zebrafish muscle mutants for level of phenotype severity. ► Proposed tool to survey treatments that aim to ameliorate muscular dystrophy. -- Abstract: Muscular dystrophies are a group of genetic disorders that progressively weaken and degenerate muscle. Many zebrafish models for human muscular dystrophies have been generated and analysed, including dystrophin-deficient zebrafish mutants dmd that model Duchenne Muscular Dystrophy. Under polarised light the zebrafish muscle can be detected as a bright area in an otherwise dark background. This light effect, called birefringence, results from the diffraction of polarised light through the pseudo-crystalline array of the muscle sarcomeres. Muscle damage, as seen in zebrafish models for muscular dystrophies, can readily be detected by a reduction in the birefringence. Therefore, birefringence is a very sensitive indicator of overall muscle integrity within larval zebrafish. Unbiased documentation of the birefringence followed by densitometric measurement enables the quantification of the birefringence of zebrafish larvae. Thereby, the overall level of muscle integrity can be detected, allowing the identification and categorisation of zebrafish muscle mutants. In addition, we propose that the establish protocol can be used to analyse treatments aimed at ameliorating dystrophic zebrafish models.

  7. Quantification of birefringence readily measures the level of muscle damage in zebrafish

    Energy Technology Data Exchange (ETDEWEB)

    Berger, Joachim, E-mail: Joachim.Berger@Monash.edu [Australian Regenerative Medicine Institute, EMBL Australia, Monash University, Clayton (Australia); Sztal, Tamar; Currie, Peter D. [Australian Regenerative Medicine Institute, EMBL Australia, Monash University, Clayton (Australia)

    2012-07-13

    Highlights: Black-Right-Pointing-Pointer Report of an unbiased quantification of the birefringence of muscle of fish larvae. Black-Right-Pointing-Pointer Quantification method readily identifies level of overall muscle damage. Black-Right-Pointing-Pointer Compare zebrafish muscle mutants for level of phenotype severity. Black-Right-Pointing-Pointer Proposed tool to survey treatments that aim to ameliorate muscular dystrophy. -- Abstract: Muscular dystrophies are a group of genetic disorders that progressively weaken and degenerate muscle. Many zebrafish models for human muscular dystrophies have been generated and analysed, including dystrophin-deficient zebrafish mutants dmd that model Duchenne Muscular Dystrophy. Under polarised light the zebrafish muscle can be detected as a bright area in an otherwise dark background. This light effect, called birefringence, results from the diffraction of polarised light through the pseudo-crystalline array of the muscle sarcomeres. Muscle damage, as seen in zebrafish models for muscular dystrophies, can readily be detected by a reduction in the birefringence. Therefore, birefringence is a very sensitive indicator of overall muscle integrity within larval zebrafish. Unbiased documentation of the birefringence followed by densitometric measurement enables the quantification of the birefringence of zebrafish larvae. Thereby, the overall level of muscle integrity can be detected, allowing the identification and categorisation of zebrafish muscle mutants. In addition, we propose that the establish protocol can be used to analyse treatments aimed at ameliorating dystrophic zebrafish models.

  8. Fast skeletal muscle troponin activation increases force of mouse fast skeletal muscle and ameliorates weakness due to nebulin-deficiency.

    Directory of Open Access Journals (Sweden)

    Eun-Jeong Lee

    Full Text Available The effect of the fast skeletal muscle troponin activator, CK-2066260, on calcium-induced force development was studied in skinned fast skeletal muscle fibers from wildtype (WT and nebulin deficient (NEB KO mice. Nebulin is a sarcomeric protein that when absent (NEB KO mouse or present at low levels (nemaline myopathy (NM patients with NEB mutations causes muscle weakness. We studied the effect of fast skeletal troponin activation on WT muscle and tested whether it might be a therapeutic mechanism to increase muscle strength in nebulin deficient muscle. We measured tension-pCa relations with and without added CK-2066260. Maximal active tension in NEB KO tibialis cranialis fibers in the absence of CK-2066260 was ∼60% less than in WT fibers, consistent with earlier work. CK-2066260 shifted the tension-calcium relationship leftwards, with the largest relative increase (up to 8-fold at low to intermediate calcium levels. This was a general effect that was present in both WT and NEB KO fiber bundles. At pCa levels above ∼6.0 (i.e., calcium concentrations <1 µM, CK-2066260 increased tension of NEB KO fibers to beyond that of WT fibers. Crossbridge cycling kinetics were studied by measuring k(tr (rate constant of force redevelopment following a rapid shortening/restretch. CK-2066260 greatly increased k(tr at submaximal activation levels in both WT and NEB KO fiber bundles. We also studied the sarcomere length (SL dependence of the CK-2066260 effect (SL 2.1 µm and 2.6 µm and found that in the NEB KO fibers, CK-2066260 had a larger effect on calcium sensitivity at the long SL. We conclude that fast skeletal muscle troponin activation increases force at submaximal activation in both wildtype and NEB KO fiber bundles and, importantly, that this troponin activation is a potential therapeutic mechanism for increasing force in NM and other skeletal muscle diseases with loss of muscle strength.

  9. A novel three-filament model of force generation in eccentric contraction of skeletal muscles.

    Science.gov (United States)

    Schappacher-Tilp, Gudrun; Leonard, Timothy; Desch, Gertrud; Herzog, Walter

    2015-01-01

    We propose and examine a three filament model of skeletal muscle force generation, thereby extending classical cross-bridge models by involving titin-actin interaction upon active force production. In regions with optimal actin-myosin overlap, the model does not alter energy and force predictions of cross-bridge models for isometric contractions. However, in contrast to cross-bridge models, the three filament model accurately predicts history-dependent force generation in half sarcomeres for eccentric and concentric contractions, and predicts the activation-dependent forces for stretches beyond actin-myosin filament overlap.

  10. A novel three-filament model of force generation in eccentric contraction of skeletal muscles.

    Directory of Open Access Journals (Sweden)

    Gudrun Schappacher-Tilp

    Full Text Available We propose and examine a three filament model of skeletal muscle force generation, thereby extending classical cross-bridge models by involving titin-actin interaction upon active force production. In regions with optimal actin-myosin overlap, the model does not alter energy and force predictions of cross-bridge models for isometric contractions. However, in contrast to cross-bridge models, the three filament model accurately predicts history-dependent force generation in half sarcomeres for eccentric and concentric contractions, and predicts the activation-dependent forces for stretches beyond actin-myosin filament overlap.

  11. The emergence of sarcomeric, graded-polarity and spindle-like patterns in bundles of short cytoskeletal polymers and two opposite molecular motors

    International Nuclear Information System (INIS)

    Craig, E M; Dey, S; Mogilner, A

    2011-01-01

    We use linear stability analysis and numerical solutions of partial differential equations to investigate pattern formation in the one-dimensional system of short dynamic polymers and one (plus-end directed) or two (one is plus-end, another minus-end directed) molecular motors. If polymer sliding and motor gliding rates are slow and/or the polymer turnover rate is fast, then the polymer-motor bundle has mixed polarity and homogeneous motor distribution. However, if motor gliding is fast, a sarcomeric pattern with periodic bands of alternating polymer polarity separated by motor aggregates evolves. On the other hand, if polymer sliding is fast, a graded-polarity bundle with motors at the center emerges. In the presence of the second, minus-end directed motor, the sarcomeric pattern is more ubiquitous, while the graded-polarity pattern is destabilized. However, if the minus-end motor is weaker than the plus-end directed one, and/or polymer nucleation is autocatalytic, and/or long polymers are present in the bundle, then a spindle-like architecture with a sorted-out polarity emerges with the plus-end motors at the center and minus-end motors at the edges. We discuss modeling implications for actin-myosin fibers and in vitro and meiotic spindles.

  12. Monoclonal antibodies against muscle actin isoforms: epitope identification and analysis of isoform expression by immunoblot and immunostaining in normal and regenerating skeletal muscle [version 2; referees: 3 approved

    Directory of Open Access Journals (Sweden)

    Christine Chaponnier

    2016-06-01

    Full Text Available Higher vertebrates (mammals and birds express six different highly conserved actin isoforms that can be classified in three subgroups: 1 sarcomeric actins, α-skeletal (α-SKA and α-cardiac (α-CAA, 2 smooth muscle actins (SMAs, α-SMA and γ-SMA, and 3 cytoplasmic actins (CYAs, β-CYA and γ-CYA. The variations among isoactins, in each subgroup, are due to 3-4 amino acid differences located in their acetylated N-decapeptide sequence. The first monoclonal antibody (mAb against an actin isoform (α-SMA was produced and characterized in our laboratory in 1986 (Skalli  et al., 1986 . We have further obtained mAbs against the 5 other isoforms. In this report, we focus on the mAbs anti-α-SKA and anti-α-CAA obtained after immunization of mice with the respective acetylated N-terminal decapeptides using the Repetitive Immunizations at Multiple Sites Strategy (RIMMS. In addition to the identification of their epitope by immunoblotting, we describe the expression of the 2 sarcomeric actins in mature skeletal muscle and during muscle repair after micro-lesions. In particular, we analyze the expression of α-CAA, α-SKA and α-SMA by co-immunostaining in a time course frame during the muscle repair process. Our results indicate that a restricted myocyte population expresses α-CAA and suggest a high capacity of self-regeneration in muscle cells. These antibodies may represent a helpful tool for the follow-up of muscle regeneration and pathological changes.

  13. Altered cross-bridge properties in skeletal muscle dystrophies

    Directory of Open Access Journals (Sweden)

    Aziz eGuellich

    2014-10-01

    Full Text Available Force and motion generated by skeletal muscle ultimately depends on the cyclical interaction of actin with myosin. This mechanical process is regulated by intracellular Ca2+ through the thin filament-associated regulatory proteins i.e.; troponins and tropomyosin. Muscular dystrophies are a group of heterogeneous genetic affections characterized by progressive degeneration and weakness of the skeletal muscle as a consequence of loss of muscle tissue which directly reduces the number of potential myosin cross-bridges involved in force production. Mutations in genes responsible for skeletal muscle dystrophies have been shown to modify the function of contractile proteins and cross-bridge interactions. Altered gene expression or RNA splicing or post-translational modifications of contractile proteins such as those related to oxidative stress, may affect cross-bridge function by modifying key proteins of the excitation-contraction coupling. Micro-architectural change in myofilament is another mechanism of altered cross-bridge performance. In this review, we provide an overview about changes in cross-bridge performance in skeletal muscle dystrophies and discuss their ultimate impacts on striated muscle function.

  14. Relationships between myonuclear domain size and fibre properties in the muscles of Thoroughbred horses.

    Science.gov (United States)

    Kawai, M; Kuwano, A; Hiraga, A; Miyata, H

    2010-11-01

    The myonuclear domain (MND) is the region of cytoplasm governed by a single myonucleus. Myonuclear domain size is an important factor for muscle fibre plasticity because each myonucleus has limitations in the capacity of protein synthesis. Previous studies have demonstrated that differences in MND size exist in different fibre types in several species, including horses. To understand the basic mechanism of muscle plasticity, the relationships between MND size, muscle fibre type population and metabolic properties of skeletal muscles throughout the whole body in Thoroughbred horses were examined. Post mortem samples were taken from 20 muscles in 3 Thoroughbred horses aged 3-5 years of age. Fibre type population was determined on serial cross sections of each muscle sample, stained for monoclonal antibodies to each myosin heavy chain isoform. Oxidative (succinic dehydrogenase; SDH) and glycolytic (phosphofructokinase; PFK) enzyme activities were determined spectrophotometrically in each muscle sample. Furthermore, 30 single fibres were isolated from each muscle under stereomicroscopy and then fibre volume and myonuclear number for a given length analysed under confocal microscopy. The MND size of each single fibre was measured after normalisation of sarcomere length to 2.8 µm by staining with membrane-specific dye. Immunohistochemical staining indicated that soleus, vastus lateralis and gluteus medius muscles had the highest percentage of type I, IIa and IIx muscle fibre, respectively. Biochemical analysis indicated highest activities of SDH and PFK in diaphragm and longissimus lumborum muscles, respectively. MNDs were largest in the splenius muscle and smallest in the soleus and masseter muscles. Myonuclear domain size is significantly related to type I muscle fibre population, but not to SDH activities of the muscles. The MND size of muscle fibre depends on fibre type population rather than mitochondrial enzyme activities. © 2010 EVJ Ltd.

  15. Genome-wide mapping of Sox6 binding sites in skeletal muscle reveals both direct and indirect regulation of muscle terminal differentiation by Sox6

    Directory of Open Access Journals (Sweden)

    An Chung-Il

    2011-10-01

    Full Text Available Abstract Background Sox6 is a multi-faceted transcription factor involved in the terminal differentiation of many different cell types in vertebrates. It has been suggested that in mice as well as in zebrafish Sox6 plays a role in the terminal differentiation of skeletal muscle by suppressing transcription of slow fiber specific genes. In order to understand how Sox6 coordinately regulates the transcription of multiple fiber type specific genes during muscle development, we have performed ChIP-seq analyses to identify Sox6 target genes in mouse fetal myotubes and generated muscle-specific Sox6 knockout (KO mice to determine the Sox6 null muscle phenotype in adult mice. Results We have identified 1,066 Sox6 binding sites using mouse fetal myotubes. The Sox6 binding sites were found to be associated with slow fiber-specific, cardiac, and embryonic isoform genes that are expressed in the sarcomere as well as transcription factor genes known to play roles in muscle development. The concurrently performed RNA polymerase II (Pol II ChIP-seq analysis revealed that 84% of the Sox6 peak-associated genes exhibited little to no binding of Pol II, suggesting that the majority of the Sox6 target genes are transcriptionally inactive. These results indicate that Sox6 directly regulates terminal differentiation of muscle by affecting the expression of sarcomere protein genes as well as indirectly through influencing the expression of transcription factors relevant to muscle development. Gene expression profiling of Sox6 KO skeletal and cardiac muscle revealed a significant increase in the expression of the genes associated with Sox6 binding. In the absence of the Sox6 gene, there was dramatic upregulation of slow fiber-specific, cardiac, and embryonic isoform gene expression in Sox6 KO skeletal muscle and fetal isoform gene expression in Sox6 KO cardiac muscle, thus confirming the role Sox6 plays as a transcriptional suppressor in muscle development

  16. Interaction Between Troponin and Myosin Enhances Contractile Activity of Myosin in Cardiac Muscle

    OpenAIRE

    Schoffstall, Brenda; LaBarbera, Vincent A.; Brunet, Nicolas M.; Gavino, Belinda J.; Herring, Lauren; Heshmati, Sara; Kraft, Brittany H.; Inchausti, Vanessa; Meyer, Nancy L.; Moonoo, Danamarie; Takeda, Aya K.; Chase, Prescott Bryant

    2011-01-01

    Ca2+ signaling in striated muscle cells is critically dependent upon thin filament proteins tropomyosin (Tm) and troponin (Tn) to regulate mechanical output. Using in vitro measurements of contractility, we demonstrate that even in the absence of actin and Tm, human cardiac Tn (cTn) enhances heavy meromyosin MgATPase activity by up to 2.5-fold in solution. In addition, cTn without Tm significantly increases, or superactivates sliding speed of filamentous actin (F-actin) in skeletal motility a...

  17. Effect of gamma irradiation on the microstructure and post-mortem anaerobic metabolism of bovine muscle

    Energy Technology Data Exchange (ETDEWEB)

    Yook, H.-S.; Lee, J.-W.; Lee, K.-H.; Kim, M.-K.; Song, C.-W.; Byun, M.-W. E-mail: mwbyun@nanum.kaeri.re.kr

    2001-05-01

    Experiments were performed to study the effect of gamma irradiation on morphological properties and post-mortem metabolism in bovine M. sternomandibularis with special reference to ultrastructure, shear force, pH and ATP breakdown. The shortening of sarcomere was not observed in gamma-irradiated muscle, however, the disappearance of M-line and of A- and I-bands was perceptible. During cold storage, the destruction of muscle bundles was faster in the gamma-irradiated muscle than in the non-irradiated with a dose-dependent manner. The same is true for the post mortem pH drop and ATP breakdown. So, experimental results confirmed that the anaerobic metabolism and morphological properties are noticeably affected by gamma irradiation in beef.

  18. Stimulus rate dependence of regional cerebral blood flow in human striate cortex, demonstrated by positron emission tomography

    International Nuclear Information System (INIS)

    Fox, P.T.; Raichle, M.E.

    1984-01-01

    The purpose of this investigation was to determine the relationship between the repetition rate of a simple sensory stimulus and regional cerebral blood flow (rCBF) in the human brain. Positron emission tomography (PET), using intravenously administered H 2 ( 15 )O as the diffusible blood-flow tracer, was employed for all CBF measurements. The use of H 2 ( 15 )O with PET allowed eight CBF measurements to be made in rapid sequence under multiple stimulation conditions without removing the subject from the tomograph. Nine normal volunteers each underwent a series of eight H2( 15 )O PET measurements of CBF. Initial and final scans were made during visual deprivation. The six intervening scans were made during visual activation with patterned-flash stimuli given in random order at 1.0-, 3.9-, 7.8-, 15.5-, 33.1-, and 61-Hz repetition rates. The region of greatest rCBF increase was determined. Within this region the rCBF was determined for every test condition and then expressed as the percentage change from the value of the initial unstimulated scan (rCBF% delta). In every subject, striate cortex rCBF% delta varied systematically with stimulus rate. Between 0 and 7.8 Hz, rCBF% delta was a linear function of stimulus repetition rate. The rCBF response peaked at 7.8 Hz and then declined. The rCBF% delta during visual stimulation was significantly greater than that during visual deprivation for every stimulus rate except 1.0 Hz. The anatomical localization of the region of peak rCBF response was determined for every subject to be the mesial occipital lobes along the calcarine fissure, primary visual cortex. Stimulus rate is a significant determinant of rCBF response in the visual cortex. Investigators of brain responses to selective activation procedures should be aware of the potential effects of stimulus rate on rCBF and other measurements of cerebral metabolism

  19. Morphology of the lateral pterygoid muscle associated to the mandibular condyle in the human prenatal stage.

    Science.gov (United States)

    Carranza, Miriam L; Carda, Carmen; Simbrón, Alicia; Quevedo, María C Sánchez; Celaya, Gabriela; de Ferraris, Maria Elsa Gómez

    2006-01-01

    The lateral pterygoid muscle (LPM) inserts at the condyle and the articular disc and plays a central role in mandibular movement via the Temporomandibular Articular Complex. The aim of this study was to examine the association between the morphology of LPM muscular fascicles and the degree of mineralization of the mandibular condyle in the prenatal stage employing structural, ultrastructural and microanalytical evaluation. Sixteen human fetuses at 11-37 weeks of gestation, with no apparent pathology and resulting from spontaneous abortions, were included in the study. Samples from lateral pterygoid muscle and the mandibular condyle were processed for light microscopy and electron microscopy and microanalysis. Desmin immunolabeling (dilution 1: 25 Dako) and alpha sarcomeric actin immunolabeling (dilution 1:50 Dako) employing the avidin-biotin system were used in paraffin embedded samples. Contralateral samples were examine by transmission electron microscopy. Four condyles (at 17-21 weeks of gestation) were used to measure the relative content of calcium and phosphorous employing the X-ray diffraction microanalytical technique. At 11-16 weeks of gestation, the LPM was composed of secondary myotubes associated to satellite cells and nerve fibers. At 18 weeks, the muscle exhibited multiple compact fascicles and the condyle showed a thin, external, subperiostal mineralized layer with few central bone spicules. At 20 weeks, at the site of insertion of the LPM, the bone trabeculae of the condyle contained an electrondense matrix with abundant mineralization nuclei. At 17-21 weeks of gestation no significant variations in the contents of phosphorous and calcium were observed. At 24 weeks, transmission electron calcium and microscopy studies revealed a marked increase in the functional units of the muscle fascicles. Also, at this age muscle fibers exhibited differences in the expression of desmin and alpha sarcomeric actin. At 37 weeks the muscle became multipennate in

  20. The mechanisms of muscle wasting in COPD and heart failure

    Directory of Open Access Journals (Sweden)

    Giorgio Vescovo

    2012-10-01

    Full Text Available Many of the mechanisms leading to skeletal muscle wasting in COPD and heart failure are common to both conditions. These encompass neurohormonal activation and systemic inflammation. The mechanisms leading to muscle dysfunction are both qualitative and quantitative. Qualitative changes comprise the transition from aerobic metabolism and prevalent slow fibers composition toward anaerobic metabolism and fast fibers synthesis. Quantitative changes are mainly linked to muscle loss. These changes occur not only in the major muscles bulks of the body but also in respiratory muscles. The mechanisms leading to muscle wastage include cytokine-triggered skeletal muscle apoptosis and ubiquitin-proteasomeand non-ubiquitin-dependent pathways. The regulation of fiber type involves the growth hormone/insulin-like growth factor 1/calcineurin/transcriptional coactivator PGC1 cascade. The imbalance between protein synthesis and degradation plays an important role. Protein degradation can occur through ubiquitin-dependent and non-ubiquitin-dependent pathways. Very recently, two systems controlling ubiquitin-proteasome activation have been described: FOXO-ubiquitin ligase and NFkB ubiquitin ligase. These are triggered by TNFα and growth hormone/insulin-like growth factor 1. Moreover, apoptosis, which is triggered by tumor necrosis factor α, plays an important role. Another mechanism acting on muscle wastage is malnutrition, with an imbalance between catabolic and anabolic factors toward the catabolic component. Catabolism is also worsened by the activation of the adrenergic system and alteration of the cortisol/DEHA ratio toward cortisol production. Sarcomeric protein oxidation and its consequent contractile impairment can be another cause of skeletal muscle dysfunction in CHF.

  1. The Burden of Early Phenotypes and the Impact of Left Ventricular Wall Thickness on Hypertrophic Cardiomyopathy Sarcomere Mutation Carriers: Findings from the HCMNet Study

    Science.gov (United States)

    Ho, Carolyn Y.; Day, Sharlene M.; Colan, Steven D.; Russell, Mark W.; Towbin, Jeffrey; Sherrid, Mark V.; Canter, Charles E.; Jefferies, John Lynn; Murphy, Anne M.; Cirino, Allison L.; Abraham, Theodore P.; Taylor, Matthew; Mestroni, Luisa; Bluemke, David A.; Jarolim, Petr; Shi, Ling; Sleeper, Lynn A.; Seidman, Christine E.; Orav, E. John

    2017-01-01

    Importance Sarcomere mutations and left ventricular hypertrophy (LVH) are cardinal features of hypertrophic cardiomyopathy (HCM). However, little is known about the full spectrum of phenotypic manifestations, or how LVH impacts disease expression. Objective Genotyped individuals with or at risk for HCM were studied to: (1) characterize sarcomere mutation carriers (G+), including assessing collective phenotypic burden; and (2) investigate the relationship between left ventricular wall thickness (LVWT) and other disease features. Design Cross-sectional multicenter observational study Setting Network of hypertrophic cardiomyopathy specialty clinical centers (HCMNet). Participants Mutation carriers with LVH (G+/LVH+), without LVH (G+/LVH-), and healthy related controls (G-/LVH-) were enrolled through HCMNet sites. A total of 193 participants were enrolled and underwent study procedures. Analyses were performed on 178 participants (G+/LVH+ n=81, G+/LVH- n=55, G-/LVH- n=42) with mean age 26.9, 20.4 and 17.5 years, respectively. Exposure The primary stratifying variables were the presence of a sarcomere mutation and measures of LV wall thickness. Main Outcomes and Measures Parameters from standardized exercise testing, echocardiography, cardiac magnetic resonance, serum biomarker measurement, and electrocardiography (ECG) were compared across study cohorts. Results G+/LVH- subjects had smaller LV cavity size and higher LV wall thickness (LVWT):diastolic diameter ratio and LV ejection fraction than controls. Phenotypic burden was determined as the cumulative number of 7 traits (ECG changes; decreased LV systolic, diastolic diameter, or septal E′; higher LVWT:diameter ratio; serum troponin, natriuretic peptide levels) in each individual. Mean burden was 4.9 in G+/LVH+, 2.4 in G+/LVH-, and 1.3 in controls (p<0.001). Classification and regression tree analysis identified an LVEDD z-score < -1.85 or the combination of LVEDD z-score ≥ -1.85 and Septal E′ z-score < -0

  2. Intramuscular connective tissue differences in spastic and control muscle: a mechanical and histological study.

    Directory of Open Access Journals (Sweden)

    Marije de Bruin

    Full Text Available Cerebral palsy (CP of the spastic type is a neurological disorder characterized by a velocity-dependent increase in tonic stretch reflexes with exaggerated tendon jerks. Secondary to the spasticity, muscle adaptation is presumed to contribute to limitations in the passive range of joint motion. However, the mechanisms underlying these limitations are unknown. Using biopsies, we compared mechanical as well as histological properties of flexor carpi ulnaris muscle (FCU from CP patients (n = 29 and healthy controls (n = 10. The sarcomere slack length (mean 2.5 µm, SEM 0.05 and slope of the normalized sarcomere length-tension characteristics of spastic fascicle segments and single myofibre segments were not different from those of control muscle. Fibre type distribution also showed no significant differences. Fibre size was significantly smaller (1933 µm2, SEM 190 in spastic muscle than in controls (2572 µm2, SEM 322. However, our statistical analyses indicate that the latter difference is likely to be explained by age, rather than by the affliction. Quantities of endomysial and perimysial networks within biopsies of control and spastic muscle were unchanged with one exception: a significant thickening of the tertiary perimysium (3-fold, i.e. the connective tissue reinforcement of neurovascular tissues penetrating the muscle. Note that this thickening in tertiary perimysium was shown in the majority of CP patients, however a small number of patients (n = 4 out of 23 did not have this feature. These results are taken as indications that enhanced myofascial loads on FCU is one among several factors contributing in a major way to the aetiology of limitation of movement at the wrist in CP and the characteristic wrist position of such patients.

  3. Muscle Lim Protein and myosin binding protein C form a complex regulating muscle differentiation.

    Science.gov (United States)

    Arvanitis, Demetrios A; Vafiadaki, Elizabeth; Papalouka, Vasiliki; Sanoudou, Despina

    2017-12-01

    Muscle Lim Protein (MLP) is a protein with multiple functional roles in striated muscle physiology and pathophysiology. Herein, we demonstrate that MLP directly binds to slow, fast, and cardiac myosin-binding protein C (MyBP-C) during myogenesis, as shown by yeast two-hybrid and a range of protein-protein interaction assays. The minimal interacting domains involve MLP inter-LIM and MyBP-C [C4]. The interaction is sensitive to cytosolic Ca 2+ concentrations changes and to MyBP-C phosphorylation by PKA or CaMKII. Confocal microscopy of differentiating myoblasts showed MLP and MyBP-C colocalization during myoblast differentiation. Suppression of the complex formation with recombinant MyBP-C [C4] peptide overexpression, inhibited myoblast differentiation by 65%. Suppression of both MLP and MyBP-C expression in myoblasts by siRNA revealed negative synergistic effects on differentiation. The MLP/MyBP-C complex modulates the actin activated myosin II ATPase activity in vitro, which could interfere with sarcomerogenesis and myofilaments assembly during differentiation. Our data demonstrate a critical role of the MLP/MyBP-C complex during early myoblast differentiation. Its absence in muscles with mutations or aberrant expression of MLP or MyBP-C could be directly implicated in the development of cardiac and skeletal myopathies. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. The influence of temperature on the distribution and intensity of the reaction product in rat muscle fibers obtained with the histochemical method for myosin ATPase

    DEFF Research Database (Denmark)

    Kirkeby, S; Tuxen, A

    1989-01-01

    The influence of temperature in the incubation medium on the localization and intensity of myosin ATPase was investigated in striated muscles from the rat using a conventional histochemical technique. It was found that the enzyme reaction was temperature-dependent since the activity in some fibers...... was raised and in others was depressed by alteration of the incubation temperature. There was no obvious correlation between the temperature sensitivity of ATPase in the muscle fibers and their activity for succinic dehydrogenase. It is proposed that the histochemical method for myosin ATPase can be used...

  5. Length Tension Function of Puborectalis Muscle: Implications for the Treatment of Fecal Incontinence and Pelvic Floor Disorders

    Science.gov (United States)

    Mittal, Ravinder K; Sheean, Geoff; Padda, Bikram S; Rajasekaran, Mahadevan R

    2014-01-01

    Background/Aims External anal sphincter (EAS) and puborectalis muscle (PRM) play important role in anal continence function. Based on length-tension measurement, we recently reported that the human EAS muscle operates at short sarcomere length under physiological conditions. Goal of our study was to determine if PRM also operates at the short sarcomere length. Methods Length-tension relationship of the PRM muscle was studied in vivo in 10 healthy nullipara women. Length was altered by vaginal distension using custom-designed probes of 5, 10, 15, 20, 25 and 30 mm diameters as well as by distending a polyethylene bag with different volumes of water. Probes were equipped with a reverse perfuse sleeve sensor to measure vaginal pressure (surrogate of PRM tension). PRM electromyogram (EMG) was recorded using wire electrodes. Three-dimensional ultra-sound images were obtained to determine effect of vaginal distension on PRM length. Results Ultrasound images demonstrate distension volume dependent increase in PRM length. Rest and squeeze pressures of vaginal bag increased with the increase in bag volume. Similarly, the change in vaginal pressure, which represents the PRM contraction increased with the increase in the probe size. Increase in probe size was not associated with an increase in EMG activity (a marker of neural drive) of the PRM. Conclusions Probe size dependent increase in PRM contraction pressure, in the presence of constant EMG (neural input) proves that the human PRM operates at short sarcomere length. Surgically adjusting the PRM length may represent a novel strategy to improve treat anal continence and possibly other pelvic floor disorders. PMID:25273124

  6. The Drosophila Z-disc protein Z(210) is an adult muscle isoform of Zasp52, which is required for normal myofibril organization in indirect flight muscles.

    Science.gov (United States)

    Chechenova, Maria B; Bryantsev, Anton L; Cripps, Richard M

    2013-02-08

    The Z-disc is a critical anchoring point for thin filaments as they slide during muscle contraction. Therefore, identifying components of the Z-disc is critical for fully comprehending how myofibrils assemble and function. In the adult Drosophila musculature, the fibrillar indirect flight muscles accumulate a >200 kDa Z-disc protein termed Z(210), the identity of which has to date been unknown. Here, we use mass spectrometry and gene specific knockdown studies, to identify Z(210) as an adult isoform of the Z-disc protein Zasp52. The Zasp52 primary transcript is extensively alternatively spliced, and we describe its splicing pattern in the flight muscles, identifying a new Zasp52 isoform, which is the one recognized by the Z(210) antibody. We also demonstrate that Zasp52 is required for the association of α-actinin with the flight muscle Z-disc, and for normal sarcomere structure. These studies expand our knowledge of Zasp isoforms and their functions in muscle. Given the role of Zasp proteins in mammalian muscle development and disease, our results have relevance to mammalian muscle biology.

  7. Preferential type II muscle fiber damage from plyometric exercise.

    Science.gov (United States)

    Macaluso, Filippo; Isaacs, Ashwin W; Myburgh, Kathryn H

    2012-01-01

    Plyometric training has been successfully used in different sporting contexts. Studies that investigated the effect of plyometric training on muscle morphology are limited, and results are controversial with regard to which muscle fiber type is mainly affected. To analyze the skeletal muscle structural and ultrastructural change induced by an acute bout of plyometric exercise to determine which type of muscle fibers is predominantly damaged. Descriptive laboratory study. Research laboratory. Eight healthy, untrained individuals (age = 22 ± 1 years, height = 179.2 ± 6.4 cm, weight = 78.9 ± 5.9 kg). Participants completed an acute bout of plyometric exercise (10 sets of 10 squat-jumps with a 1-minute rest between sets). Blood samples were collected 9 days and immediately before and 6 hours and 1, 2, and 3 days after the acute intervention. Muscle samples were collected 9 days before and 3 days after the exercise intervention. Blood samples were analyzed for creatine kinase activity. Muscle biopsies were analyzed for damage using fluorescent and electron transmission microscopy. Creatine kinase activity peaked 1 day after the exercise bout (529.0 ± 317.8 U/L). Immunofluorescence revealed sarcolemmal damage in 155 of 1616 fibers analyzed. Mainly fast-twitch fibers were damaged. Within subgroups, 7.6% of type I fibers, 10.3% of type IIa fibers, and 14.3% of type IIx fibers were damaged as assessed by losses in dystrophin staining. Similar damage was prevalent in IIx and IIa fibers. Electron microscopy revealed clearly distinguishable moderate and severe sarcomere damage, with damage quantifiably predominant in type II muscle fibers of both the glycolytic and oxidative subtypes (86% and 84%, respectively, versus only 27% of slow-twitch fibers). We provide direct evidence that a single bout of plyometric exercise affected mainly type II muscle fibers.

  8. Glacially striated, soft sediment surfaces on late Paleozoic tillite at São Luiz do Purunã, PR

    Directory of Open Access Journals (Sweden)

    Ivo Trosdtorf Jr.

    2005-06-01

    Full Text Available Striae and furrows found on the upper surfaces of three stratigraphically superposed decimetric beds of late Paleozoic lodgement tillite of the Itararé Subgroup in the northern Paraná Basin were engraved by ploughing of clasts and possibly also ice protuberances at the base of the glacier, on unconsolidated to partially consolidated sediment. Associated features indicate that the rheology of the bed varied from stiff during lodgement to soft and deformable during ploughing. Poor drainage of meltwater at the glacier-bed interface may have contributed to lower the strength of sediment to deformation. The deformed interval was probably generated during a single glacial phase or advance of a glacier grounding in a marine or lacustrine water body. Changes in the dynamics of the glacier involving slow and fast flow were correlated respectively with alternation of deposition and erosion. The proposed model is analogous to that of lodgement till complexes from the Pleistocene of the northern hemisphere. Retreat of the glacier was probably fast, followed by settling of muds on top of the upper striated and furrowed surface, and progradation of deltaic sands during post-glacial time.Estrias e sulcos encontrados sobre três camadas decimétricas, estratigraficamente superpostas, de tilito de alojamento neopaleozóico do Subgrupo Itararé, na porção norte da Bacia do Paraná, foram formados por aração de clastos e, possivelmente, por protuberâncias de gelo, na base da geleira. Feições associadas indicam que a reologia do sedimento variou de rígido, durante o alojamento, a inconsolidado e deformável durante a aração. A baixa drenagem da água de degelo na interface geleira-substrato pode ter contribuído para reduzir a resistência do sedimento à deformação. A sucessão acima foi gerada provavelmente durante uma única fase glacial ou avanço de geleira sobre corpo de água marinho ou lacustre. Mudanças na dinâmica da geleira envolvendo

  9. Histotopographical study of human periocular elastic fibers using aldehyde-fuchsin staining with special reference to the sleeve and pulley system for extraocular rectus muscles.

    Science.gov (United States)

    Osanai, Hajime; Murakami, Gen; Ohtsuka, Aiji; Suzuki, Daisuke; Nakagawa, Takashi; Tatsumi, Haruyuki

    2009-09-01

    The aim of this study was to investigate the detailed configuration of periocular elastic fibers. Semiserial paraffin sections were made using 40 whole orbital contents from 27 elderly cadavers and stained by the aldehyde-fuchsin method. Periocular tissues were classified into three types according to directions of the elastic fibers, i.e., tissues containing anteroposteriorly running elastic fibers, those with mediolateral fibers, and those with meshwork of fibers. Anteroposterior elastic fiber-dominant tissue was seen in the upper eyelid and newly defined pulley plate for the medial and lateral recti (MR, LR). Mediolateral fibers were predominant in the central part of the inferior rectus pulley. In the pulley plates for the MR and LR, anteroposteriorly running fibers encased the striated muscle. Tenon's capsule and the epimysium of the recti were mediolateral fiber-dominant. However, at the entrance of the muscle terminal where Tenon's capsule reflects and continues to the epimysium, composite elastic fibers provided a meshwork-like skeleton. The elastic mesh was also seen around the lacrimal canaliculi. The pulley for the recti seemed to be composed of two parts--a connective tissue plate encasing the recti and specialized Tenon's capsule at an entrance or porta of the muscle. For both parts, elastic fibers were major functional components. The anteroposterior elastic fibers in the MR and LR pulley plates, especially, seemed to receive anteroposteriorly directed stress and tension from these striated muscles. The elastic interfaces seemed to prevent any concentration of stress that would interfere with periocular striated muscle functions, including hypothetical active pulleys.

  10. Multistability inspired by the oblique, pennate architectures of skeletal muscle

    Science.gov (United States)

    Kidambi, Narayanan; Harne, Ryan L.; Wang, K. W.

    2017-04-01

    Skeletal muscle mechanics exhibit a range of noteworthy characteristics, providing great inspiration for the development of advanced structural and material systems. These characteristics arise from the synergies demonstrated between muscle's constituents across the various length scales. From the macroscale oblique orientation of muscle fibers to the microscale lattice spacing of sarcomeres, muscle takes advantage of geometries and multidimensionality for force generation or length change along a desired axis. Inspired by these behaviors, this research investigates how the incorporation of multidimensionality afforded by oblique, pennate architectures can uncover novel mechanics in structures exhibiting multistability. Experimental investigation of these mechanics is undertaken using specimens of molded silicone rubber with patterned voids, and results reveal tailorable mono-, bi-, and multi-stability under axial displacements by modulation of transverse confinement. If the specimen is considered as an architected material, these results show its ability to generate intriguing, non-monotonic shear stresses. The outcomes would foster the development of novel, advanced mechanical metamaterials that exploit pennation and multidimensionality.

  11. Work Done by Titin Protein Folding Assists Muscle Contraction

    Directory of Open Access Journals (Sweden)

    Jaime Andrés Rivas-Pardo

    2016-02-01

    Full Text Available Current theories of muscle contraction propose that the power stroke of a myosin motor is the sole source of mechanical energy driving the sliding filaments of a contracting muscle. These models exclude titin, the largest protein in the human body, which determines the passive elasticity of muscles. Here, we show that stepwise unfolding/folding of titin immunoglobulin (Ig domains occurs in the elastic I band region of intact myofibrils at physiological sarcomere lengths and forces of 6–8 pN. We use single-molecule techniques to demonstrate that unfolded titin Ig domains undergo a spontaneous stepwise folding contraction at forces below 10 pN, delivering up to 105 zJ of additional contractile energy, which is larger than the mechanical energy delivered by the power stroke of a myosin motor. Thus, it appears inescapable that folding of titin Ig domains is an important, but as yet unrecognized, contributor to the force generated by a contracting muscle.

  12. Ultra structure of the denervated vocal muscle mechanically in hogs (sus scrofa domestica

    Directory of Open Access Journals (Sweden)

    Leão, Henrique Zaquia

    2010-03-01

    Full Text Available Introduction: The literature is not clear in the ultra-structural manifestations of the vocal wrinkles after neural wound. Objective: To verify the alterations that occur in a vocal fold mechanically denervated. Method: In this prospective study, it were utilized 15 hogs of commercial race (Sus scrofa domesticates, with age of 4 to 12 weeks. The animals were distributed in three groups, chosen at random. Everybody was submitted to the denervation of the right vocal fold, with surgical removal of a segment with three centimeters of the recurring right laryngeal nerve. After 45, 90 and 180 days of the operations, it was proceeded the biopsy of the vocal muscles, it was prosecuted the samples for transmission electron microscopy and, for the ultra-structural study, utilized the transmission electron microscopy Philips, model EM208S. Results: The biopsied groups with 45 and 90 days after operation of mechanical denervation, presented disorganization miofibrilar, only vestigial lines Z in many samples, as well like altered mithochondrions presenting limited sizes, and matrix mithocondrial rarefied with rare mithocondrial cristae present. The biopsied group with 180 days after operation of denervation, presented regular sarcomeres, mithocondrions with sizes and regular number with correct positioning between the sarcomerical units. Conclusion: The finds in the ultra-structure of the vocal muscles suggest to re enervation of the muscle being that the muscular mithochondrions were the most sensible structures to the denervated condition, successions by the cytoarchiteture of the miofibrilas; the finds in the ultra-structure of the vocal muscles suggests to reinervation of the muscle in the period of approximately six months.

  13. Dense-body aggregates as plastic structures supporting tension in smooth muscle cells.

    Science.gov (United States)

    Zhang, Jie; Herrera, Ana M; Paré, Peter D; Seow, Chun Y

    2010-11-01

    The wall of hollow organs of vertebrates is a unique structure able to generate active tension and maintain a nearly constant passive stiffness over a large volume range. These properties are predominantly attributable to the smooth muscle cells that line the organ wall. Although smooth muscle is known to possess plasticity (i.e., the ability to adapt to large changes in cell length through structural remodeling of contractile apparatus and cytoskeleton), the detailed structural basis for the plasticity is largely unknown. Dense bodies, one of the most prominent structures in smooth muscle cells, have been regarded as the anchoring sites for actin filaments, similar to the Z-disks in striated muscle. Here, we show that the dense bodies and intermediate filaments formed cable-like structures inside airway smooth muscle cells and were able to adjust the cable length according to cell length and tension. Stretching the muscle cell bundle in the relaxed state caused the cables to straighten, indicating that these intracellular structures were connected to the extracellular matrix and could support passive tension. These plastic structures may be responsible for the ability of smooth muscle to maintain a nearly constant tensile stiffness over a large length range. The finding suggests that the structural plasticity of hollow organs may originate from the dense-body cables within the smooth muscle cells.

  14. Deleting exon 55 from the nebulin gene induces severe muscle weakness in a mouse model for nemaline myopathy.

    Science.gov (United States)

    Ottenheijm, Coen A C; Buck, Danielle; de Winter, Josine M; Ferrara, Claudia; Piroddi, Nicoletta; Tesi, Chiara; Jasper, Jeffrey R; Malik, Fady I; Meng, Hui; Stienen, Ger J M; Beggs, Alan H; Labeit, Siegfried; Poggesi, Corrado; Lawlor, Michael W; Granzier, Henk

    2013-06-01

    Nebulin--a giant sarcomeric protein--plays a pivotal role in skeletal muscle contractility by specifying thin filament length and function. Although mutations in the gene encoding nebulin (NEB) are a frequent cause of nemaline myopathy, the most common non-dystrophic congenital myopathy, the mechanisms by which mutations in NEB cause muscle weakness remain largely unknown. To better understand these mechanisms, we have generated a mouse model in which Neb exon 55 is deleted (Neb(ΔExon55)) to replicate a founder mutation seen frequently in patients with nemaline myopathy with Ashkenazi Jewish heritage. Neb(ΔExon55) mice are born close to Mendelian ratios, but show growth retardation after birth. Electron microscopy studies show nemaline bodies--a hallmark feature of nemaline myopathy--in muscle fibres from Neb(ΔExon55) mice. Western blotting studies with nebulin-specific antibodies reveal reduced nebulin levels in muscle from Neb(ΔExon55) mice, and immunofluorescence confocal microscopy studies with tropomodulin antibodies and phalloidin reveal that thin filament length is significantly reduced. In line with reduced thin filament length, the maximal force generating capacity of permeabilized muscle fibres and single myofibrils is reduced in Neb(ΔExon55) mice with a more pronounced reduction at longer sarcomere lengths. Finally, in Neb(ΔExon55) mice the regulation of contraction is impaired, as evidenced by marked changes in crossbridge cycling kinetics and by a reduction of the calcium sensitivity of force generation. A novel drug that facilitates calcium binding to the thin filament significantly augmented the calcium sensitivity of submaximal force to levels that exceed those observed in untreated control muscle. In conclusion, we have characterized the first nebulin-based nemaline myopathy model, which recapitulates important features of the phenotype observed in patients harbouring this particular mutation, and which has severe muscle weakness caused by

  15. Effects of cross-bridge compliance on the force-velocity relationship and muscle power output.

    Directory of Open Access Journals (Sweden)

    Axel J Fenwick

    Full Text Available Muscles produce force and power by utilizing chemical energy through ATP hydrolysis. During concentric contractions (shortening, muscles generate less force compared to isometric contractions, but consume greater amounts of energy as shortening velocity increases. Conversely, more force is generated and less energy is consumed during eccentric muscle contractions (lengthening. This relationship between force, energy use, and the velocity of contraction has important implications for understanding muscle efficiency, but the molecular mechanisms underlying this behavior remain poorly understood. Here we used spatially-explicit, multi-filament models of Ca2+-regulated force production within a half-sarcomere to simulate how force production, energy utilization, and the number of bound cross-bridges are affected by dynamic changes in sarcomere length. These computational simulations show that cross-bridge binding increased during slow-velocity concentric and eccentric contractions, compared to isometric contractions. Over the full ranges of velocities that we simulated, cross-bridge cycling and energy utilization (i.e. ATPase rates increased during shortening, and decreased during lengthening. These findings are consistent with the Fenn effect, but arise from a complicated relationship between velocity-dependent cross-bridge recruitment and cross-bridge cycling kinetics. We also investigated how force production, power output, and energy utilization varied with cross-bridge and myofilament compliance, which is impossible to address under typical experimental conditions. These important simulations show that increasing cross-bridge compliance resulted in greater cross-bridge binding and ATPase activity, but less force was generated per cross-bridge and throughout the sarcomere. These data indicate that the efficiency of force production decreases in a velocity-dependent manner, and that this behavior is sensitive to cross-bridge compliance. In

  16. Architectural design of the pelvic floor is consistent with muscle functional subspecialization.

    Science.gov (United States)

    Tuttle, Lori J; Nguyen, Olivia T; Cook, Mark S; Alperin, Marianna; Shah, Sameer B; Ward, Samuel R; Lieber, Richard L

    2014-02-01

    Skeletal muscle architecture is the strongest predictor of a muscle's functional capacity. The purpose of this study was to define the architectural properties of the deep muscles of the female pelvic floor (PFMs) to elucidate their structure-function relationships. PFMs coccygeus (C), iliococcygeus (IC), and pubovisceral (PV) were harvested en bloc from ten fixed human cadavers (mean age 85 years, range 55-102). Fundamental architectural parameters of skeletal muscles [physiological cross-sectional area (PCSA), normalized fiber length, and sarcomere length (L(s))] were determined using validated methods. PCSA predicts muscle-force production, and normalized fiber length is related to muscle excursion. These parameters were compared using repeated measures analysis of variance (ANOVA) with post hoc t tests, as appropriate. Significance was set to α = 0.05. PFMs were thinner than expected based on data reported from imaging studies and in vivo palpation. Significant differences in fiber length were observed across PFMs: C = 5.29 ± 0.32 cm, IC = 7.55 ± 0.46 cm, PV = 10.45 ± 0.67 cm (p design shows individual muscles demonstrating differential architecture, corresponding to specialized function in the pelvic floor.

  17. Evaluation of human muscle in vivo by potassium radiometric measuring

    International Nuclear Information System (INIS)

    Sousa, Wanderson de P.

    2000-01-01

    Potassium is an essential element to the human metabolism and is present in all living cells, mainly in the striated muscular fibers. K-40 is one of the natural potassium isotopes with mass percentage of 0,0118% . This isotope emits beta particle and gamma rays with 1460 keV. The energy of K-40 photon and its uniform distribution within the human body allows its in vivo measurement. The objective of this study is to optimize this technique and evaluate the possibility of its medical application in order to quantify muscle increase during recovering procedures. Subjects of both sexes measured until this moment were divided into two groups. Subjects of Group 1 do not exercise routinely and subjects of Group 2 does. In Group 1 the average potassium mass, muscle mass and potassium concentration were (101±16)g of K, (20±3)kg of muscle and (1,3±0,3)g of K/kg of body mass, respectively, while in Group 2 average values were (125±38)g of K, (25±8)kg of muscle and (1,7±0,2)g of K/kg of body mass. The comparison between average values shows a clear difference, which allows to correlate a higher K mass with routine body activity. The technique has shown enough sensitivity for this application. (author)

  18. Effects of yessotoxin (YTX) on the skeletal muscle: an update.

    Science.gov (United States)

    Tubaro, A; Bandi, E; Sosa, S; Soranzo, M R; Giangaspero, A; De Ninis, V; Yasumoto, T; Lorenzon, P

    2008-09-01

    Yessotoxins (YTXs) are algal toxins originally included in the diarrheic toxins. After oral intake, YTXs induce only ultra-structural changes (packages of swollen mitochondria) in cardiac cells. The aim of this study was to investigate the possible effects of YTX on the other contractile striated tissue, the skeletal muscle, in vitro and in vivo. In vitro, in skeletal mouse myotubes, YTX (0.01-1.0 microM) influenced cell excitability in a concentration- and time-dependent way. In the in vivo study, transmission electron microscopy analysis did not reveal any ultrastructural alteration of skeletal muscle after acute (1 mg kg(-1)) or repeated (1 and 2mg kg(-1) day(-1), for 7 days) oral administration of YTX to mice. The observation that effects were detected in vitro but not in vivo supports the hypothesis of a low YTX bioavailability to skeletal muscle after oral intake. Therefore, the results seem to exclude a toxic effect in skeletal muscle when YTX is consumed as a food contaminant.

  19. Power Grid Protection of the Muscle Mitochondrial Reticulum

    Directory of Open Access Journals (Sweden)

    Brian Glancy

    2017-04-01

    Full Text Available Summary: Mitochondrial network connectivity enables rapid communication and distribution of potential energy throughout the cell. However, this connectivity puts the energy conversion system at risk, because damaged elements could jeopardize the entire network. Here, we demonstrate the mechanisms for mitochondrial network protection in heart and skeletal muscle (SKM. We find that the cardiac mitochondrial reticulum is segmented into subnetworks comprising many mitochondria linked through abundant contact sites at highly specific intermitochondrial junctions (IMJs. In both cardiac and SKM subnetworks, a rapid electrical and physical separation of malfunctioning mitochondria occurs, consistent with detachment of IMJs and retraction of elongated mitochondria into condensed structures. Regional mitochondrial subnetworks limit the cellular impact of local dysfunction while the dynamic disconnection of damaged mitochondria allows the remaining mitochondria to resume normal function within seconds. Thus, mitochondrial network security is comprised of both proactive and reactive mechanisms in striated muscle cells. : Network connectivity allows information sharing and distribution but also enables propagation of localized dysfunction. Glancy et al. demonstrate the existence of both proactive and reactive network protection mechanisms designed to minimize the spread of dysfunction throughout the coupled mitochondrial networks in heart and skeletal muscle cells. Keywords: energy distribution, muscle energetics, oxidative phosphorylation, 3D electron microscopy, mitochondrial retraction, mitochondrial dynamics

  20. Genetic architecture of gene expression in ovine skeletal muscle.

    Science.gov (United States)

    Kogelman, Lisette J A; Byrne, Keren; Vuocolo, Tony; Watson-Haigh, Nathan S; Kadarmideen, Haja N; Kijas, James W; Oddy, Hutton V; Gardner, Graham E; Gondro, Cedric; Tellam, Ross L

    2011-12-15

    In livestock populations the genetic contribution to muscling is intensively monitored in the progeny of industry sires and used as a tool in selective breeding programs. The genes and pathways conferring this genetic merit are largely undefined. Genetic variation within a population has potential, amongst other mechanisms, to alter gene expression via cis- or trans-acting mechanisms in a manner that impacts the functional activities of specific pathways that contribute to muscling traits. By integrating sire-based genetic merit information for a muscling trait with progeny-based gene expression data we directly tested the hypothesis that there is genetic structure in the gene expression program in ovine skeletal muscle. The genetic performance of six sires for a well defined muscling trait, longissimus lumborum muscle depth, was measured using extensive progeny testing and expressed as an Estimated Breeding Value by comparison with contemporary sires. Microarray gene expression data were obtained for longissimus lumborum samples taken from forty progeny of the six sires (4-8 progeny/sire). Initial unsupervised hierarchical clustering analysis revealed strong genetic architecture to the gene expression data, which also discriminated the sire-based Estimated Breeding Value for the trait. An integrated systems biology approach was then used to identify the major functional pathways contributing to the genetics of enhanced muscling by using both Estimated Breeding Value weighted gene co-expression network analysis and a differential gene co-expression network analysis. The modules of genes revealed by these analyses were enriched for a number of functional terms summarised as muscle sarcomere organisation and development, protein catabolism (proteosome), RNA processing, mitochondrial function and transcriptional regulation. This study has revealed strong genetic structure in the gene expression program within ovine longissimus lumborum muscle. The balance between

  1. Basal body and striated rootlet changes in primate macular retinal pigmented epithelium after low level diffuse argon laser radiation. Final report 1981-1982

    Energy Technology Data Exchange (ETDEWEB)

    Schuschereba, S.T.; Zwick, H.; Stuck, B.E.; Beatrice, E.S.

    1982-09-01

    Basal bodies or centrioles (BB - microtubule organizing centers) and striated rootlets (SR - bundles of 60 A action-like filaments) have a close association in primate retinal pigmented epithelial (RPE) cells. The frequency of occurrence of these structures was evaluated in the macular RPE after repeated exposure to low level diffuse argon laser radiation (DALR). The awake chaired animal's head was restrained and positioned near the center of the 0.75 m hemisphere which was diffusely irradiated with 514.5 nm laser radiation. The right eye of each subject was occluded during the two-hour exposure session. The first subject received 24 cumulative hours of exposure, the second, 40 hours and the third, 42 hours.

  2. Pregnancy-induced adaptations in intramuscular extracellular matrix of rat pelvic floor muscles.

    Science.gov (United States)

    Alperin, Marianna; Kaddis, Timothy; Pichika, Rajeswari; Esparza, Mary C; Lieber, Richard L

    2016-08-01

    Birth trauma to pelvic floor muscles is a major risk factor for pelvic floor disorders. Intramuscular extracellular matrix determines muscle stiffness, supports contractile component, and shields myofibers from mechanical strain. Our goal was to determine whether pregnancy alters extracellular matrix mechanical and biochemical properties in a rat model, which may provide insights into the pathogenesis of pelvic floor muscle birth injury. To examine whether pregnancy effects were unique to pelvic floor muscles, we also studied a hind limb muscle. Passive mechanical properties of coccygeus, iliocaudalis, pubocaudalis, and tibialis anterior were compared among 3-month old Sprague-Dawley virgin, late-pregnant, and postpartum rats. Muscle tangent stiffness was calculated as the slope of the stress-sarcomere length curve between 2.5 and 4.0 μm, obtained from a stress-relaxation protocol at a bundle level. Elastin and collagen isoform concentrations were quantified by the use of enzyme-linked immunosorbent assay. Enzymatic and glycosylated collagen crosslinks were determined by high-performance liquid chromatography. Data were compared by the use of repeated-measures, 2-way analysis of variance with Tukey post-hoc testing. Correlations between mechanical and biochemical parameters were assessed by linear regressions. Significance was set to P pelvic floor muscle stiffness did not differ from virgins (P > .3). A substantial increase in collagen V in coccygeus and pubocaudalis was observed in late-pregnant, compared with virgin, animals, (P pelvic floor muscles (P pelvic floor muscles, the tibialis anterior was unaltered by pregnancy. In contrast to other pelvic tissues, pelvic floor muscle stiffness increased in pregnancy, returning to prepregnancy state postpartum. This adaptation may shield myofibers from excessive mechanical strain during parturition. Biochemical alterations in pelvic floor muscle extracellular matrix due to pregnancy include increase in collagen V

  3. Pregnancy-induced adaptations in intramuscular extracellular matrix of rat pelvic floor muscles

    Science.gov (United States)

    Alperin, Marianna; Kaddis, Timothy; Pichika, Rajeswari; Esparza, Mary C.; Lieber, Richard L.

    2017-01-01

    BACKGROUND Birth trauma to pelvic floor muscles is a major risk factor for pelvic floor disorders. Intramuscular extracellular matrix determines muscle stiffness, supports contractile component, and shields myofibers from mechanical strain. OBJECTIVE Our goal was to determine whether pregnancy alters extracellular matrix mechanical and biochemical properties in a rat model, which may provide insights into the pathogenesis of pelvic floor muscle birth injury. To examine whether pregnancy effects were unique to pelvic floor muscles, we also studied a hind limb muscle. STUDY DESIGN Passive mechanical properties of coccygeus, iliocaudalis, pubocaudalis, and tibialis anterior were compared among 3-month old Sprague–Dawley virgin, late-pregnant, and postpartum rats. Muscle tangent stiffness was calculated as the slope of the stress–sarcomere length curve between 2.5 and 4.0 μm, obtained from a stress-relaxation protocol at a bundle level. Elastin and collagen isoform concentrations were quantified by the use of enzyme-linked immunosorbent assay. Enzymatic and glycosylated collagen crosslinks were determined by high-performance liquid chromatography. Data were compared by the use of repeated-measures, 2-way analysis of variance with Tukey post-hoc testing. Correlations between mechanical and biochemical parameters were assessed by linear regressions. Significance was set to P pelvic floor muscle stiffness did not differ from virgins (P > .3). A substantial increase in collagen V in coccygeus and pubocaudalis was observed in late-pregnant, compared with virgin, animals, (P pelvic floor muscles (P pelvic floor muscles, the tibialis anterior was unaltered by pregnancy. CONCLUSIONS In contrast to other pelvic tissues, pelvic floor muscle stiffness increased in pregnancy, returning to prepregnancy state post-partum. This adaptation may shield myofibers from excessive mechanical strain during parturition. Biochemical alterations in pelvic floor muscle extracellular matrix

  4. Ultrastructural effects on gill, muscle, and gonadal tissues induced in zebrafish (Danio rerio) by a waterborne uranium exposure

    Energy Technology Data Exchange (ETDEWEB)

    Barillet, Sabrina, E-mail: sabrina.barillet@free.fr [Laboratory of Radioecology and Ecotoxicology, IRSN (Institute for Radiological Protection and Nuclear Safety), DEI/SECRE/LRE, Cadarache, Bat 186, BP 3, 13115 St-Paul-Lez-Durance cedex (France); Larno, Valerie, E-mail: valerie.larno@irsn.fr [Laboratory of Radioecology and Ecotoxicology, IRSN (Institute for Radiological Protection and Nuclear Safety), DEI/SECRE/LRE, Cadarache, Bat 186, BP 3, 13115 St-Paul-Lez-Durance cedex (France); Floriani, Magali, E-mail: magali.floriani@irsn.fr [Laboratory of Radioecology and Ecotoxicology, IRSN (Institute for Radiological Protection and Nuclear Safety), DEI/SECRE/LRE, Cadarache, Bat 186, BP 3, 13115 St-Paul-Lez-Durance cedex (France); Devaux, Alain, E-mail: alain.devaux@entpe.fr [INRA, EFPA Department, 54280, Champenoux and Environmental Science Laboratory, ENTPE, 69518 Vaulx en Velin cedex (France); Adam-Guillermin, Christelle, E-mail: christelle.adam-guillermin@irsn.fr [Laboratory of Radioecology and Ecotoxicology, IRSN (Institute for Radiological Protection and Nuclear Safety), DEI/SECRE/LRE, Cadarache, Bat 186, BP 3, 13115 St-Paul-Lez-Durance cedex (France)

    2010-11-01

    Experiments on adult zebrafish (Danio rerio) were conducted to assess histopathological effects induced on gill, muscle, and gonadal tissues after waterborne uranium exposure. Although histopathology is often employed as a tool for the detection and assessment of xenobiotic-mediated effects in aquatic organisms, few studies have been dedicated to the investigation of histopathological consequences of uranium exposure in fish. Results showed that gill tissue architecture was markedly disrupted. Major symptoms were alterations of the secondary lamellae epithelium (from extensive oedema to desquamation), hyperplasia of chloride cells, and breakdown of the pillar cell system. Muscle histology was also affected. Degeneration and disorganization of myofibrillar sarcomeric pattern as well as abnormal localization of mitochondria within muscle and altered endomysial sheaths were observed. Morphological alterations of spermatozoa within the gonadal tissue were also noticed. This study demonstrated that uranium exposure induced a variety of histological impairments in fish, supporting environmental concerns when uranium contaminates aquatic systems.

  5. Muscle biopsy.

    Science.gov (United States)

    Meola, G; Bugiardini, E; Cardani, R

    2012-04-01

    Muscle biopsy is required to provide a definitive diagnosis in many neuromuscular disorders. It can be performed through an open or needle technique under local anesthesia. The major limitations of the needle biopsy technique are the sample size, which is smaller than that obtained with open biopsy, and the impossibility of direct visualization of the sampling site. However, needle biopsy is a less invasive procedure than open biopsy and is particularly indicated for diagnosis of neuromuscular disease in infancy and childhood. The biopsied muscle should be one affected by the disease but not be too weak or too atrophic. Usually, in case of proximal muscle involvement, the quadriceps and the biceps are biopsied, while under suspicion of mitochondrial disorder, the deltoid is preferred. The samples must be immediately frozen or fixed after excision to prevent loss of enzymatic reactivity, DNA depletion or RNA degradation. A battery of stainings is performed on muscle sections from every frozen muscle biopsy arriving in the pathology laboratory. Histological, histochemical, and histoenzymatic stainings are performed to evaluate fiber atrophy, morphological, and structural changes and metabolic disorders. Moreover, immunohistochemistry and Western blotting analysis may be used for expression analysis of muscle proteins to obtain a specific diagnosis. There are myopathies that do not need muscle biopsy since a genetic test performed on a blood sample is enough for definitive diagnosis. Muscle biopsy is a useful technique which can make an enormous contribution in the field of neuromuscular disorders but should be considered and interpreted together with the patient's family and clinical history.

  6. Oxygen dependence of respiration in rat spinotrapezius muscle in situ.

    Science.gov (United States)

    Golub, Aleksander S; Pittman, Roland N

    2012-07-01

    The oxygen dependence of respiration in striated muscle in situ was studied by measuring the rate of decrease of interstitial Po(2) [oxygen disappearance curve (ODC)] following rapid arrest of blood flow by pneumatic tissue compression, which ejected red blood cells from the muscle vessels and made the ODC independent from oxygen bound to hemoglobin. After the contribution of photo-consumption of oxygen by the method was evaluated and accounted for, the corrected ODCs were converted into the Po(2) dependence of oxygen consumption, Vo(2), proportional to the rate of Po(2) decrease. Fitting equations obtained from a model of heterogeneous intracellular Po(2) were applied to recover the parameters describing respiration in muscle fibers, with a predicted sigmoidal shape for the dependence of Vo(2) on Po(2). This curve consists of two regions connected by the point for critical Po(2) of the cell (i.e., Po(2) at the sarcolemma when the center of the cell becomes anoxic). The critical Po(2) was below the Po(2) for half-maximal respiratory rate (P(50)) for the cells. In six muscles at rest, the rate of oxygen consumption was 139 ± 6 nl O(2)/cm(3)·s and mitochondrial P(50) was k = 10.5 ± 0.8 mmHg. The range of Po(2) values inside the muscle fibers was found to be 4-5 mmHg at the critical Po(2). The oxygen dependence of respiration can be studied in thin muscles under different experimental conditions. In resting muscle, the critical Po(2) was substantially lower than the interstitial Po(2) of 53 ± 2 mmHg, a finding that indicates that Vo(2) under this circumstance is independent of oxygen supply and is discordant with the conventional hypothesis of metabolic regulation of the oxygen supply to tissue.

  7. Oxygen dependence of respiration in rat spinotrapezius muscle in situ

    Science.gov (United States)

    Pittman, Roland N.

    2012-01-01

    The oxygen dependence of respiration in striated muscle in situ was studied by measuring the rate of decrease of interstitial Po2 [oxygen disappearance curve (ODC)] following rapid arrest of blood flow by pneumatic tissue compression, which ejected red blood cells from the muscle vessels and made the ODC independent from oxygen bound to hemoglobin. After the contribution of photo-consumption of oxygen by the method was evaluated and accounted for, the corrected ODCs were converted into the Po2 dependence of oxygen consumption, V̇o2, proportional to the rate of Po2 decrease. Fitting equations obtained from a model of heterogeneous intracellular Po2 were applied to recover the parameters describing respiration in muscle fibers, with a predicted sigmoidal shape for the dependence of V̇o2 on Po2. This curve consists of two regions connected by the point for critical Po2 of the cell (i.e., Po2 at the sarcolemma when the center of the cell becomes anoxic). The critical Po2 was below the Po2 for half-maximal respiratory rate (P50) for the cells. In six muscles at rest, the rate of oxygen consumption was 139 ± 6 nl O2/cm3·s and mitochondrial P50 was k = 10.5 ± 0.8 mmHg. The range of Po2 values inside the muscle fibers was found to be 4–5 mmHg at the critical Po2. The oxygen dependence of respiration can be studied in thin muscles under different experimental conditions. In resting muscle, the critical Po2 was substantially lower than the interstitial Po2 of 53 ± 2 mmHg, a finding that indicates that V̇o2 under this circumstance is independent of oxygen supply and is discordant with the conventional hypothesis of metabolic regulation of the oxygen supply to tissue. PMID:22523254

  8. Thick-to-Thin Filament Surface Distance Modulates Cross-Bridge Kinetics in Drosophila Flight Muscle

    Energy Technology Data Exchange (ETDEWEB)

    Tanner, Bertrand C.W.; Farman, Gerrie P.; Irving, Thomas C.; Maughan, David W.; Palmer, Bradley M.; Miller, Mark S. (IIT); (Vermont); (BU)

    2012-09-19

    The demembranated (skinned) muscle fiber preparation is widely used to investigate muscle contraction because the intracellular ionic conditions can be precisely controlled. However, plasma membrane removal results in a loss of osmotic regulation, causing abnormal hydration of the myofilament lattice and its proteins. We investigated the structural and functional consequences of varied myofilament lattice spacing and protein hydration on cross-bridge rates of force development and detachment in Drosophila melanogaster indirect flight muscle, using x-ray diffraction to compare the lattice spacing of dissected, osmotically compressed skinned fibers to native muscle fibers in living flies. Osmolytes of different sizes and exclusion properties (Dextran T-500 and T-10) were used to differentially alter lattice spacing and protein hydration. At in vivo lattice spacing, cross-bridge attachment time (t{sub on}) increased with higher osmotic pressures, consistent with a reduced cross-bridge detachment rate as myofilament protein hydration decreased. In contrast, in the swollen lattice, t{sub on} decreased with higher osmotic pressures. These divergent responses were reconciled using a structural model that predicts t{sub on} varies inversely with thick-to-thin filament surface distance, suggesting that cross-bridge rates of force development and detachment are modulated more by myofilament lattice geometry than protein hydration. Generalizing these findings, our results suggest that cross-bridge cycling rates slow as thick-to-thin filament surface distance decreases with sarcomere lengthening, and likewise, cross-bridge cycling rates increase during sarcomere shortening. Together, these structural changes may provide a mechanism for altering cross-bridge performance throughout a contraction-relaxation cycle.

  9. Respiratory muscle dysfunction in animal models of hypoxic disease: antioxidant therapy goes from strength to strength

    Directory of Open Access Journals (Sweden)

    O'Halloran KD

    2017-07-01

    Full Text Available Ken D O’Halloran,1 Philip Lewis2 1Department of Physiology, School of Medicine, University College Cork, Cork, Ireland; 2Institute and Policlinic for Occupational Medicine, Environmental Medicine and Preventative Research, University Hospital of Cologne, Germany Abstract: The striated muscles of breathing play a critical role in respiratory homeostasis governing blood oxygenation and pH regulation. Upper airway dilator and thoracic pump muscles retain a remarkable capacity for plasticity throughout life, both in health and disease states. Hypoxia, whatever the cause, is a potent driver of respiratory muscle remodeling with evidence of adaptive and maladaptive outcomes for system performance. The pattern, duration, and intensity of hypoxia are key determinants of respiratory muscle structural-, metabolic-, and functional responses and adaptation. Age and sex also influence respiratory muscle tolerance of hypoxia. Redox stress emerges as the principal protagonist driving respiratory muscle malady in rodent models of hypoxic disease. There is a growing body of evidence demonstrating that antioxidant intervention alleviates hypoxia-induced respiratory muscle dysfunction, and that N-acetyl cysteine, approved for use in humans, is highly effective in preventing hypoxia-induced respiratory muscle weakness and fatigue. We posit that oxygen homeostasis is a key driver of respiratory muscle form and function. Hypoxic stress is likely a major contributor to respiratory muscle malaise in diseases of the lungs and respiratory control network. Animal studies provide an evidence base in strong support of the need to explore adjunctive antioxidant therapies for muscle dysfunction in human respiratory disease. Keywords: respiratory muscle, diaphragm, upper airway, hypoxia, antioxidants, N-acetyl-cysteine, OSA, COPD

  10. Expression of a dominant negative CELF protein in vivo leads to altered muscle organization, fiber size, and subtype.

    Directory of Open Access Journals (Sweden)

    Dara S Berger

    2011-04-01

    Full Text Available CUG-BP and ETR-3-like factor (CELF proteins regulate tissue- and developmental stage-specific alternative splicing in striated muscle. We previously demonstrated that heart muscle-specific expression of a nuclear dominant negative CELF protein in transgenic mice (MHC-CELFΔ effectively disrupts endogenous CELF activity in the heart in vivo, resulting in impaired cardiac function. In this study, transgenic mice that express the dominant negative protein under a skeletal muscle-specific promoter (Myo-CELFΔ were generated to investigate the role of CELF-mediated alternative splicing programs in normal skeletal muscle.Myo-CELFΔ mice exhibit modest changes in CELF-mediated alternative splicing in skeletal muscle, accompanied by a reduction of endomysial and perimysial spaces, an increase in fiber size variability, and an increase in slow twitch muscle fibers. Weight gain and mean body weight, total number of muscle fibers, and overall muscle strength were not affected.Although these findings demonstrate that CELF activity contributes to the normal alternative splicing of a subset of muscle transcripts in vivo, the mildness of the effects in Myo-CELFΔ muscles compared to those in MHC-CELFΔ hearts suggests CELF activity may be less determinative for alternative splicing in skeletal muscle than in heart muscle. Nonetheless, even these small changes in CELF-mediated splicing regulation were sufficient to alter muscle organization and muscle fiber properties affected in myotonic dystrophy. This lends further evidence to the hypothesis that dysregulation of CELF-mediated alternative splicing programs may be responsible for the disruption of these properties during muscle pathogenesis.

  11. Cardiac muscle: a miracle of creation.

    Science.gov (United States)

    Seely, S

    1989-09-01

    The paper proposes that energy conversion in muscle is a two-step process, chemical energy being first converted into electrical energy which is then converted into mechanical work. The chemo-electrical transducers are, in effect, minute voltaic cells--more precisely calcium-magnesium cells--with the magnesium electrodes on myosin heads and the calcium electrodes on the C subunits of troponin molecules associated with actin filaments. These cells are established when, after the passage of an action potential, calcium ions are admitted to the sarcomere. In an energy-consuming process, calcium ions are bound to troponin molecules, the energy for the process being supplied by hydrolysis of adenosine triphosphate. The electro-mechanical transducer utilises the electrostatic field established between the oppositely charged electrodes of the voltaic cell. As the two are pulled towards each other, doing mechanical work, energy is supplied by the voltaic cells. In the course of this action, calcium ions go back into solution. The action ceases when, after the passage of an action potential, calcium ions are withdrawn into the sarcoplasmic reticulum.

  12. Defective muscle basement membrane and lack of M-laminin in the dystrophic dy/dy mouse

    DEFF Research Database (Denmark)

    Xu, H; Christmas, P; Wu, X R

    1994-01-01

    -linked Duchenne and Becker muscular dystrophies. We have examined M-laminin expression in mice with autosomal recessive muscular dystrophy caused by the mutation dy. The heavy chain of M-laminin was undetectable in skeletal muscle, heart muscle, and peripheral nerve by immunofluorescence and immunoblotting......M-laminin is a major member of the laminin family of basement membrane proteins. It is prominently expressed in striated muscle and peripheral nerve. M-laminin is deficient in patients with the autosomal recessive Fukuyama congenital muscular dystrophy but is normal in patients with the sex...... tissue from dy/dy mice, suggesting that M-laminin heavy-chain mRNA may be produced at very low levels or is unstable. Information about the chromosomal localization of the M heavy-chain in human and mouse suggests that a mutation in the M-chain gene causes the muscular dystrophy in dy/dy mice. The dy...

  13. Phosphorylation of titan and nebulin in skeletal muscle

    Energy Technology Data Exchange (ETDEWEB)

    Somerville, L.L.

    1986-01-01

    The in vitro and in vivo phosphorylation of skeletal muscle titin and nebulin are examined. It has been proposed that these proteins are the fundamental components of an elastic cytoskeletal lattice within the sarcomere. Determinations of endogenous phosphate in titin and nebulin purified from rabbit back muscle revealed phosphate contents of 3.10 +/- 0.26 mol phosphate/mol titin and 4.63 +/- 0.43 mol phosphate/mol nebulin. Incubation of rabbit back muscle homogenate in the presence of gamma-/sup 32/P ATP resulted in the labeling of both titin and nebulin; labeling was enhanced by the addition of cAMP-dependent protein kinase. Similar results were obtained from the incubation of chemically skinned rabbit psoas fibers in the presence of labeled ATP. A time dependent increase in phosphate incorporation was observed. Purification of titin and nebulin from Xenopus laevis frog gastrocnemius revealed endogenous phosphate contents of 6.15 +/- 0.12 mol phosphate/mol titin and 9.67 +/- 1.5 mol phosphate/mol nebulin. Titin and nebulin labeling after in vivo injection of Xenopus laevis frogs with /sup 32/P-orthophosphate was demonstrated.

  14. Effects of ultrasound and convection cooking to different end point temperatures on cooking characteristics, shear force and sensory properties, composition, and microscopic morphology of beef longissimus and pectoralis muscles.

    Science.gov (United States)

    Pohlman, F W; Dikeman, M E; Zayas, J F; Unruh, J A

    1997-02-01

    Longissimus and pectoralis muscles were removed from 10 steer carcasses at 4 d postmortem, aged for 14 d at 2 degrees C, then assigned to either ultrasound or convection cooking to either 62 degrees C or 70 degrees C internal end point temperature. During cooking, time-temperature profiles and energy consumption were monitored. Ultrasound cooking resulted in greater (P force work to shear muscle samples than convection cooking. The ultrasound treatment also resulted in a reduction (P convection cooking. Electron micrographs indicated that ultrasound-cooked muscles had longer sarcomeres, larger diameter fibers, and more myofibrillar disruption and shattering. Longissimus muscles cooked faster (P < .05) and more (P < .05) energetically efficient, had less (P < .05) total collagen, and were superior (P < .05) in instrumental evaluated texture and sensory tenderness than pectoralis muscles. Cooking to 70 degrees C caused greater (P < .05) moisture and cooking losses, required more (P < .05) time and energy input to cook, and negatively (P < .05) affected instrumental textural and sensory tenderness characteristics. Electron micrographs indicated a shortening of sarcomeres, more deterioration of the banding structure, reduction in fiber diameter, and breakdown of endomysial and perimysial connective tissue at an internal temperature of 70 degrees C vs 62 degrees C. This research identifies ultrasound cooking as a new, rapid, energy-efficient method that may improve some meat textural attributes.

  15. Growth associated protein 43 is expressed in skeletal muscle fibers and is localized in proximity of mitochondria and calcium release units.

    Directory of Open Access Journals (Sweden)

    Simone Guarnieri

    Full Text Available The neuronal Growth Associated Protein 43 (GAP43, also known as B-50 or neuromodulin, is involved in mechanisms controlling pathfinding and branching of neurons during development and regeneration. For many years this protein was classified as neuron-specific, but recent evidences suggest that a GAP43 is expressed in the nervous system not only in neurons, but also in glial cells, and b probably it is present also in other tissues. In particular, its expression was revealed in muscles from patients affected by various myopathies, indicating that GAP43 can no-longer considered only as a neuron-specific molecule. We have investigated the expression and subcellular localization of GAP43 in mouse satellite cells, myotubes, and adult muscle (extensor digitorum longus or EDL using Western blotting, immuno-fluorescence combined to confocal microscopy and electron microscopy. Our in vitro results indicated that GAP43 is indeed expressed in both myoblasts and differentiating myotubes, and its cellular localization changes dramatically during maturation: in myoblasts the localization appeared to be mostly nuclear, whereas with differentiation the protein started to display a sarcomeric-like pattern. In adult fibers, GAP43 expression was evident with the protein labeling forming (in longitudinal views a double cross striation reminiscent of the staining pattern of other organelles, such as calcium release units (CRUs and mitochondria. Double immuno-staining and experiments done in EDL muscles fixed at different sarcomere lengths, allowed us to determine the localization, from the sarcomere Z-line, of GAP43 positive foci, falling between that of CRUs and of mitochondria. Staining of cross sections added a detail to the puzzle: GAP43 labeling formed a reticular pattern surrounding individual myofibrils, but excluding contractile elements. This work leads the way to further investigation about the possible physiological and structural role of GAP43 protein in

  16. Large isoforms of UNC-89 (obscurin are required for muscle cell architecture and optimal calcium release in Caenorhabditis elegans.

    Directory of Open Access Journals (Sweden)

    Patrick M Spooner

    Full Text Available Calcium, a ubiquitous intracellular signaling molecule, controls a diverse array of cellular processes. Consequently, cells have developed strategies to modulate the shape of calcium signals in space and time. The force generating machinery in muscle is regulated by the influx and efflux of calcium ions into the muscle cytoplasm. In order for efficient and effective muscle contraction to occur, calcium needs to be rapidly, accurately and reliably regulated. The mechanisms underlying this highly regulated process are not fully understood. Here, we show that the Caenorhabditis elegans homolog of the giant muscle protein obscurin, UNC-89, is required for normal muscle cell architecture. The large immunoglobulin domain-rich isoforms of UNC-89 are critical for sarcomere and sarcoplasmic reticulum organization. Furthermore, we have found evidence that this structural organization is crucial for excitation-contraction coupling in the body wall muscle, through the coordination of calcium signaling. Thus, our data implicates UNC-89 in maintaining muscle cell architecture and that this precise organization is essential for optimal calcium mobilization and efficient and effective muscle contraction.

  17. Architectural design of the pelvic floor is consistent with muscle functional subspecialization

    Science.gov (United States)

    Tuttle, Lori J.; Nguyen, Olivia T.; Cook, Mark S.; Alperin, Marianna; Shah, Sameer B.; Ward, Samuel R.

    2014-01-01

    Introduction and hypothesis Skeletal muscle architecture is the strongest predictor of a muscle’s functional capacity. The purpose of this study was to define the architectural properties of the deep muscles of the female pelvic floor (PFMs) to elucidate their structure–function relationships. Methods PFMs coccygeus (C), iliococcygeus (IC), and pubovisceral (PV) were harvested en bloc from ten fixed human cadavers (mean age 85 years, range 55–102). Fundamental architectural parameters of skeletal muscles [physiological cross-sectional area (PCSA), normalized fiber length, and sarcomere length (Ls)] were determined using validated methods. PCSA predicts muscle-force production, and normalized fiber length is related to muscle excursion. These parameters were compared using repeated measures analysis of variance (ANOVA) with post hoc t tests, as appropriate. Significance was set to α=0.05. Results PFMs were thinner than expected based on data reported from imaging studies and in vivo palpation. Significant differences in fiber length were observed across PFMs: C=5.29±0.32 cm, IC=7.55±0.46 cm, PV=10.45±0.67 cm (pstabilizer. PV demonstrated the longest fibers, suggesting that it functions to produce large excursions. Conclusions PFM design shows individual muscles demonstrating differential architecture, corresponding to specialized function in the pelvic floor. PMID:23903821

  18. Genomic profiling reveals Pitx2 controls expression of mature extraocular muscle contraction-related genes.

    Science.gov (United States)

    Zhou, Yuefang; Gong, Bendi; Kaminski, Henry J

    2012-04-18

    To assess the influence of the Pitx2 transcription factor on the global gene expression profile of extraocular muscle (EOM) of mice. Mice with a conditional knockout of Pitx2, designated Pitx2(Δflox/Δflox) and their control littermates Pitx2(flox/flox), were used. RNA was isolated from EOM obtained at 3, 6, and 12 weeks of age and processed for microarray-based profiling. Pairwise comparisons were performed between mice of the same age and differentially expressed gene lists were generated. Select genes from the profile were validated using real-time quantitative polymerase chain reaction and protein immunoblot. Ultrastructural analysis was performed to evaluate EOM sarcomeric structure. The number of differentially expressed genes was relatively small. Eleven upregulated and 23 downregulated transcripts were identified common to all three age groups in the Pitx2-deficient extraocular muscle compared with littermate controls. These fell into a range of categories including muscle-specific structural genes, transcription factors, and ion channels. The differentially expressed genes were primarily related to muscle contraction. We verified by protein and ultrastructural analysis that myomesin 2 was expressed in the Pitx2-deficient mice, and this was associated with development of M lines evident in their orbital region. The global transcript expression analysis uncovered that Pitx2 primarily regulates a relatively select number of genes associated with muscle contraction. Pitx2 loss led to the development of M line structures, a feature more typical of other skeletal muscle.

  19. Getting Muscles

    Science.gov (United States)

    ... re thinking about aren't possible for kids. Superheroes, of course, aren't real, and professional athletes ... can make you stronger. Why? Because you're using your muscles when you do it. Eat Strong ...

  20. Muscle cramps

    Science.gov (United States)

    ... the lower leg/calf Back of the thigh (hamstrings) Front of the thigh (quadriceps) Cramps in the ... Names Cramps - muscle Images Chest stretch Groin stretch Hamstring stretch Hip stretch Thigh stretch Triceps stretch References ...

  1. Multi-scale modeling of chemo-mechanical coupling in muscle contraction and applications to cardiac modeling

    Directory of Open Access Journals (Sweden)

    Chapelle Dominique

    2016-01-01

    Full Text Available We propose a muscle chemo-mechanical model by which myosin heads - that can chemically bind to actin, thus creating so-called cross-bridges producing contraction forces in sarcomeres at the subcellular level - are considered as special chemical entities having internal mechanical variables pertaining to the actual geometric configuration. This provides a thermodynamical basis for modeling the complex interplay of chemical and mechanical phenomena at the sarcomere level. The resulting model is in the form of stochastic equations governing the dynamics of these microscopic mechanical variables in a Langevin framework. Equivalently, Fokker-Planck equations can be derived to describe the evolution of the associated probability densities. Under certain assumptions the corresponding moment equations can be closed, thus directly providing access to macroscopic quantities that can be incorporated in the overall constitutive equations of the muscle tissue. The underlying thermodynamical framework also enables the derivation of compatible numerical schemes, in particular in terms of energy balances. These modeling and discretization ingredients can be integrated in a global model of the cardiac system, to represent physiological and pathological phenomena in various medical applications.

  2. Molecular analysis of the muscle protein projectin in Lepidoptera.

    Science.gov (United States)

    Ayme-Southgate, A J; Turner, L; Southgate, R J

    2013-01-01

    Striated muscles of both vertebrates and insects contain a third filament composed of the giant proteins, namely kettin and projectin (insects) and titin (vertebrates). All three proteins have been shown to contain several domains implicated in conferring elasticity, in particular a PEVK segment. In this study, the characterization of the projectin protein in the silkmoth, Bombyx mori L. (Lepidoptera: Bombycidae), and the monarch butterfly, Danaus plexippus L. (Lepidoptera: Nymphalidae), as well as a partial characterization in the Carolina sphinx, Manduca sexta L. (Lepidoptera: Sphingidae), are presented. This study showed that, similar to other insects, projectin's overall modular organization was conserved, but in contrast, the PEVK region had a highly divergent sequence. The analysis of alternative splicing in the PEVK region revealed a small number of possible isoforms and the lack of a flight-muscle specific variant, both characteristics being in sharp contrast with findings from other insects. The possible correlation with difference in flight muscle stiffness and physiology between Lepidoptera and other insect orders is discussed.

  3. Carbonic Anhydrase III Is Expressed in Mouse Skeletal Muscles Independent of Fiber Type-Specific Myofilament Protein Isoforms and Plays a Role in Fatigue Resistance.

    Science.gov (United States)

    Feng, Han-Zhong; Jin, J-P

    2016-01-01

    Carbonic anhydrase III (CAIII) is a metabolic enzyme and a regulator for intracellular pH. CAIII has been reported with high level expression in slow twitch skeletal muscles. Here we demonstrate that CAIII is expressed in multiple slow and fast twitch muscles of adult mouse independent of the expression of myosin isoforms. Expressing similar fast type of myofilament proteins, CAIII-positive tibial anterior (TA) muscle exhibits higher tolerance to fatigue than that of CAIII-negative fast twitch extensor digitorum longus (EDL) muscle in in situ contractility studies. We further studied the muscles of CAIII knockout ( Car3 -KO) mice. The loss of CAIII in soleus and TA muscles in Car3 -KO mice did not change muscle mass, sarcomere protein isoform contents, and the baseline twitch and tetanic contractility as compared with age-matched wild type (WT) controls. On the other hand, Car3 -KO TA muscle showed faster force reduction at the beginning but higher resistance at the end during a fatigue test, followed by slower post fatigue recovery than that of WT TA muscle. Superfused Car3 -KO soleus muscle also had faster total force reduction during fatigue test than that of WT soleus. However, it showed a less elevation of resting tension followed by a better post fatigue recovery under acidotic stress. CAIII was detected in neonatal TA and EDL muscle, downregulated during development, and then re-expressed in adult TA but not EDL muscles. The expression of CAIII in Tnnt1 -KO myopathy mouse soleus muscle that has diminished slow fiber contents due to the loss of slow troponin T remained high. Car3 -KO EDL, TA, and soleus muscles showed no change in the expression of mitochondria biomarker proteins. The data suggest a fiber type independent expression of CAIII with a role in the regulation of intracellular pH in skeletal muscle and may be explored as a target for improving fatigue resistance and for the treatment of TNNT1 myopathies.

  4. Carbonic Anhydrase III Is Expressed in Mouse Skeletal Muscles Independent of Fiber Type-Specific Myofilament Protein Isoforms and Plays a Role in Fatigue Resistance

    Science.gov (United States)

    Feng, Han-Zhong; Jin, J.-P.

    2016-01-01

    Carbonic anhydrase III (CAIII) is a metabolic enzyme and a regulator for intracellular pH. CAIII has been reported with high level expression in slow twitch skeletal muscles. Here we demonstrate that CAIII is expressed in multiple slow and fast twitch muscles of adult mouse independent of the expression of myosin isoforms. Expressing similar fast type of myofilament proteins, CAIII-positive tibial anterior (TA) muscle exhibits higher tolerance to fatigue than that of CAIII-negative fast twitch extensor digitorum longus (EDL) muscle in in situ contractility studies. We further studied the muscles of CAIII knockout (Car3-KO) mice. The loss of CAIII in soleus and TA muscles in Car3-KO mice did not change muscle mass, sarcomere protein isoform contents, and the baseline twitch and tetanic contractility as compared with age-matched wild type (WT) controls. On the other hand, Car3-KO TA muscle showed faster force reduction at the beginning but higher resistance at the end during a fatigue test, followed by slower post fatigue recovery than that of WT TA muscle. Superfused Car3-KO soleus muscle also had faster total force reduction during fatigue test than that of WT soleus. However, it showed a less elevation of resting tension followed by a better post fatigue recovery under acidotic stress. CAIII was detected in neonatal TA and EDL muscle, downregulated during development, and then re-expressed in adult TA but not EDL muscles. The expression of CAIII in Tnnt1-KO myopathy mouse soleus muscle that has diminished slow fiber contents due to the loss of slow troponin T remained high. Car3-KO EDL, TA, and soleus muscles showed no change in the expression of mitochondria biomarker proteins. The data suggest a fiber type independent expression of CAIII with a role in the regulation of intracellular pH in skeletal muscle and may be explored as a target for improving fatigue resistance and for the treatment of TNNT1 myopathies. PMID:28018233

  5. Rbfox-regulated alternative splicing is critical for zebrafish cardiac and skeletal muscle function

    Science.gov (United States)

    Gallagher, Thomas L.; Arribere, Joshua A.; Geurts, Paul A.; Exner, Cameron R. T.; McDonald, Kent L.; Dill, Kariena K.; Marr, Henry L.; Adkar, Shaunak S.; Garnett, Aaron T.; Amacher, Sharon L.; Conboy, John G.

    2012-01-01

    Rbfox RNA binding proteins are implicated as regulators of phylogenetically-conserved alternative splicing events important for muscle function. To investigate the function of rbfox genes, we used morpholino-mediated knockdown of muscle-expressed rbfox1l and rbfox2 in zebrafish embryos. Single and double morphant embryos exhibited changes in splicing of overlapping sets of bioinformatically-predicted rbfox target exons, many of which exhibit a muscle-enriched splicing pattern that is conserved in vertebrates. Thus, conservation of intronic Rbfox binding motifs is a good predictor of Rbfox-regulated alternative splicing. Morphology and development of single morphant embryos was strikingly normal; however, muscle development in double morphants was severely disrupted. Defects in cardiac muscle were marked by reduced heart rate and in skeletal muscle by complete paralysis. The predominance of wavy myofibers and abnormal thick and thin filaments in skeletal muscle revealed that myofibril assembly is defective and disorganized in double morphants. Ultra-structural analysis revealed that although sarcomeres with electron dense M- and Z-bands are present in muscle fibers of rbfox1l/rbox2 morphants, they are substantially reduced in number and alignment. Importantly, splicing changes and morphological defects were rescued by expression of morpholino-resistant rbfox cDNA. Additionally, a target-blocking MO complementary to a single UGCAUG motif adjacent to an rbfox target exon of fxr1 inhibited inclusion in a similar manner to rbfox knockdown, providing evidence that Rbfox regulates the splicing of target exons via direct binding to intronic regulatory motifs. We conclude that Rbfox proteins regulate an alternative splicing program essential for vertebrate heart and skeletal muscle function. PMID:21925157

  6. Rbfox-regulated alternative splicing is critical for zebrafish cardiac and skeletal muscle functions.

    Science.gov (United States)

    Gallagher, Thomas L; Arribere, Joshua A; Geurts, Paul A; Exner, Cameron R T; McDonald, Kent L; Dill, Kariena K; Marr, Henry L; Adkar, Shaunak S; Garnett, Aaron T; Amacher, Sharon L; Conboy, John G

    2011-11-15

    Rbfox RNA binding proteins are implicated as regulators of phylogenetically-conserved alternative splicing events important for muscle function. To investigate the function of rbfox genes, we used morpholino-mediated knockdown of muscle-expressed rbfox1l and rbfox2 in zebrafish embryos. Single and double morphant embryos exhibited changes in splicing of overlapping sets of bioinformatically-predicted rbfox target exons, many of which exhibit a muscle-enriched splicing pattern that is conserved in vertebrates. Thus, conservation of intronic Rbfox binding motifs is a good predictor of Rbfox-regulated alternative splicing. Morphology and development of single morphant embryos were strikingly normal; however, muscle development in double morphants was severely disrupted. Defects in cardiac muscle were marked by reduced heart rate and in skeletal muscle by complete paralysis. The predominance of wavy myofibers and abnormal thick and thin filaments in skeletal muscle revealed that myofibril assembly is defective and disorganized in double morphants. Ultra-structural analysis revealed that although sarcomeres with electron dense M- and Z-bands are present in muscle fibers of rbfox1l/rbox2 morphants, they are substantially reduced in number and alignment. Importantly, splicing changes and morphological defects were rescued by expression of morpholino-resistant rbfox cDNA. Additionally, a target-blocking MO complementary to a single UGCAUG motif adjacent to an rbfox target exon of fxr1 inhibited inclusion in a similar manner to rbfox knockdown, providing evidence that Rbfox regulates the splicing of target exons via direct binding to intronic regulatory motifs. We conclude that Rbfox proteins regulate an alternative splicing program essential for vertebrate heart and skeletal muscle functions. Published by Elsevier Inc.

  7. Diaphragm Muscle Adaptation to Sustained Hypoxia: Lessons from Animal Models with Relevance to High Altitude and Chronic Respiratory Diseases

    Directory of Open Access Journals (Sweden)

    Philip Lewis

    2016-12-01

    Full Text Available The diaphragm is the primary inspiratory pump muscle of breathing. Notwithstanding its critical role in pulmonary ventilation, the diaphragm like other striated muscles is malleable in response to physiological and pathophysiological stressors, with potential implications for the maintenance of respiratory homeostasis. This review considers hypoxic adaptation of the diaphragm muscle, with a focus on functional, structural, and metabolic remodeling relevant to conditions such as high altitude and chronic respiratory disease. On the basis of emerging data in animal models, we posit that hypoxia is a significant driver of respiratory muscle plasticity, with evidence suggestive of both compensatory and deleterious adaptations in conditions of sustained exposure to low oxygen. Cellular strategies driving diaphragm remodeling during exposure to sustained hypoxia appear to confer hypoxic tolerance at the expense of peak force-generating capacity, a key functional parameter that correlates with patient morbidity and mortality. Changes include, but are not limited to: redox-dependent activation of hypoxia-inducible factor (HIF and MAP kinases; time-dependent carbonylation of key metabolic and functional proteins; decreased mitochondrial respiration; activation of atrophic signaling and increased proteolysis; and, altered functional performance. Diaphragm muscle weakness may be a signature effect of sustained hypoxic exposure. We discuss the putative role of reactive oxygen species as mediators of both advantageous and disadvantageous adaptations of diaphragm muscle to sustained hypoxia, and the role of antioxidants in mitigating adverse effects of chronic hypoxic stress on respiratory muscle function.

  8. Reconstruction of the muscle system in Antygomonas sp. (Kinorhyncha, Cyclorhagida) by means of phalloidin labeling and cLSM.

    Science.gov (United States)

    Müller, Monika C M; Schmidt-Rhaesa, Andreas

    2003-05-01

    In the present investigation the entire muscle system of the cyclorhagid kinorhynch Antygomonas sp. was three-dimensionally reconstructed from whole mounts by means of FITC-phalloidin labeling and confocal scanning microscopy. With this technique, which proved to be especially useful for microscopically small species, we wanted to reinvestigate and supplement the findings obtained by histological and electron microscopical methods. The organization of the major muscle systems can be summarized as follows: 1) All muscle fibers, apart from the intestinal ones, the spine, and the mouth cone muscles, show a cross-striated pattern; 2) Dorsal longitudinal muscle fibers as well as segmentally arranged dorsoventral fibers occur from segment III to XIII; 3) Diagonal muscle fibers are located laterally in segments III to X; 4) Two rings of circular fibers are present in segment II, forming the closing apparatus in Cyclorhagida. Further circular muscles are present in segment I, forming the mouth cone and the eversible introvert, and in the pharyngeal bulb. Copyright 2003 Wiley-Liss, Inc.

  9. Target-derived trophic effect on skeletal muscle innervation in senescent mice.

    Science.gov (United States)

    Messi, Maria Laura; Delbono, Osvaldo

    2003-02-15

    In the present work, we tested the hypothesis that target-derived insulin-like growth factor-1 (IGF-1) prevents alterations in neuromuscular innervation in aging mammals. To explore this hypothesis, we studied senescent wild-type mice as a model of deficient IGF-1 secretion and signaling and S1S2 transgenic mice as a tool to investigate the role of sustained overexpression of IGF-1 in striated muscle in neuromuscular innervation. The analysis of the nerve terminal in extensor digitorum longus muscles from senescent mice showed that the decrease in the percentage of cholinesterase-stained zones (CSZ) exhibiting nerve terminal branching, number of nerve branches at the CSZ, and nerve branch points was partially or completely reversed by sustained overexpression of IGF-1 in skeletal muscle. Target-derived IGF-1 also prevented age-related decreases in the postterminal alpha-bungarotoxin immunostained area, as well as the reduction in the number and length of postsynaptic folds, and area and density of postsynaptic folds studied with electron microscopy. Overexpression of IGF-1 in skeletal muscle may account for the lack of age-dependent switch in muscle fiber type composition recorded in senescent mice. In summary, the use of the S1S2 IGF-1 transgenic mouse model allowed us to provide morphological evidence for the role of target-derived IGF-1 in spinal cord motor neurons in senescent mice.

  10. Generation of a vascularized organoid using skeletal muscle as the inductive source.

    LENUS (Irish Health Repository)

    Messina, Aurora

    2005-09-01

    The technology required for creating an in vivo microenvironment and a neovasculature that can grow with and service new tissue is lacking, precluding the possibility of engineering complex three-dimensional organs. We have shown that when an arterio-venous (AV) loop is constructed in vivo in the rat groin, and placed inside a semisealed chamber, an extensive functional vasculature is generated. To test whether this unusually angiogenic environment supports the survival and growth of implanted tissue or cells, we inserted various preparations of rat and human skeletal muscle. We show that after 6 weeks incubation of muscle tissue, the chamber filled with predominantly well-vascularized recipient-derived adipose tissue, but some new donor-derived skeletal muscle and connective tissue were also evident. When primary cultured myoblasts were inserted into the chamber with the AV loop, they converted to mature striated muscle fibers. Furthermore, we identify novel adipogenesis-inducing properties of skeletal muscle. This represents the first report of a specific three-dimensional tissue grown on its own vascular supply.

  11. Physical activity counteracts tumor cell growth in colon carcinoma C26-injected muscles: an interim report

    Directory of Open Access Journals (Sweden)

    Charlotte Hiroux

    2016-06-01

    Full Text Available Skeletal muscle tissue is a rare site of tumor metastasis but is the main target of the degenerative processes occurring in cancer-associated cachexia syndrome. Beneficial effects of physical activity in counteracting cancer-related muscle wasting have been described in the last decades. Recently it has been shown that, in tumor xeno-transplanted mouse models, physical activity is able to directly affect tumor growth by modulating inflammatory responses in the tumor mass microenvironment. Here, we investigated the effect of physical activity on tumor cell growth in colon carcinoma C26 cells injected tibialis anterior muscles of BALB/c mice. Histological analyses revealed that 4 days of voluntary wheel running significantly counteracts tumor cell growth in C26-injected muscles compared to the non-injected sedentary controls. Since striated skeletal muscle tissue is the site of voluntary contraction, our results confirm that physical activity can also directly counteract tumor cell growth in a metabolically active tissue that is usually not a target for metastasis.

  12. Membrane-stabilizing copolymers confer marked protection to dystrophic skeletal muscle in vivo

    Directory of Open Access Journals (Sweden)

    Evelyne M Houang

    Full Text Available Duchenne muscular dystrophy (DMD is a fatal disease of striated muscle deterioration. A unique therapeutic approach for DMD is the use of synthetic membrane stabilizers to protect the fragile dystrophic sarcolemma against contraction-induced mechanical stress. Block copolymer-based membrane stabilizer poloxamer 188 (P188 has been shown to protect the dystrophic myocardium. In comparison, the ability of synthetic membrane stabilizers to protect fragile DMD skeletal muscles has been less clear. Because cardiac and skeletal muscles have distinct structural and functional features, including differences in the mechanism of activation, variance in sarcolemma phospholipid composition, and differences in the magnitude and types of forces generated, we speculated that optimized membrane stabilization could be inherently different. Our objective here is to use principles of pharmacodynamics to evaluate membrane stabilization therapy for DMD skeletal muscles. Results show a dramatic differential effect of membrane stabilization by optimization of pharmacodynamic-guided route of poloxamer delivery. Data show that subcutaneous P188 delivery, but not intravascular or intraperitoneal routes, conferred significant protection to dystrophic limb skeletal muscles undergoing mechanical stress in vivo. In addition, structure-function examination of synthetic membrane stabilizers further underscores the importance of copolymer composition, molecular weight, and dosage in optimization of poloxamer pharmacodynamics in vivo.

  13. Serotonin as an integrator of leech behavior and muscle mechanical performance.

    Science.gov (United States)

    Gerry, Shannon P; Daigle, Amanda J; Feilich, Kara L; Liao, Jessica; Oston, Azzara L; Ellerby, David J

    2012-08-01

    The obliquely striated muscle in the leech body wall has a broad functional repertoire; it provides power for both locomotion and suction feeding. It also operates over an unusually high strain range, undergoing up to threefold changes in length. Serotonin (5-HT) may support this functional flexibility, integrating behavior and biomechanics. It can act centrally, promoting motor outputs that drive body wall movements, and peripherally, modulating the mechanical properties of body wall muscle. During isometric contractions 5-HT enhances active force production and reduces resting muscle tone. We therefore hypothesized that 5-HT would increase net work output during the cyclical contractions associated with locomotion and feeding. Longitudinal strains measured during swimming, crawling and feeding were applied to body wall muscle in vitro with the timing and duration of stimulation selected to maximize net work output. The net work output during all simulated behaviors significantly increased in the presence of 100μM 5-HT relative to the 5-HT-free control condition. Without 5-HT the muscle strips could not achieve a net positive work output during simulated swimming. The decrease in passive tension associated with 5-HT may also be important in reducing muscle antagonist work during longitudinal muscle lengthening. The behavioral and mechanical effects of 5-HT during locomotion are clearly complementary, promoting particular behaviors and enhancing muscle performance during those behaviors. Although 5-HT can enhance muscle mechanical performance during simulated feeding, low in vivo activity in serotonergic neurons during feeding may mean that its mechanical role during this behavior is less important than during locomotion. Copyright © 2012 Elsevier GmbH. All rights reserved.

  14. Sub-cellular localisation of fukutin related protein in different cell lines and in the muscle of patients with MDC1C and LGMD2I

    DEFF Research Database (Denmark)

    Torelli, Silvia; Brown, Susan C; Brockington, Martin

    2005-01-01

    MDC1C and LGMD2I are two allelic forms of muscular dystrophies caused by mutations in the gene encoding for fukutin related protein (FKRP). FKRP encodes for a putative glycosyltransferase, the precise function of which is unknown. However, the marked reduction of alpha-dystroglycan glycosylation...... and in transverse sections of normal skeletal and cardiac muscle, endogenous FKRP surrounded the myonuclei. This localisation was unaffected in the skeletal muscle of patients with MDC1C and LGMD2I carrying various FKRP mutations. These observations imply a specific role for FKRP during striated muscle, neuronal...... and glial development and suggest that protein mis-localisation is not a common mechanism of disease in FKRP-related dystrophies....

  15. Replication in cultured C2C12 muscle cells correlates with the neuroinvasiveness of California serogroup bunyaviruses.

    Science.gov (United States)

    Griot, C; Pekosz, A; Davidson, R; Stillmock, K; Hoek, M; Lukac, D; Schmeidler, D; Cobbinah, I; Gonzalez-Scarano, F; Nathanson, N

    1994-06-01

    The neuroinvasiveness of California serogroup bunyaviruses is determined by the ability of the virus to replicate in striated muscle after peripheral inoculation of mice. Neuroinvasiveness was mapped to the medium (M) RNA segment of the virus, which encodes the viral glycoproteins, when reassortants were made between La Crosse/original virus, a neuroinvasive isolate, and Tahyna-181/57 virus, a nonneuroinvasive clone. We have tested the murine muscle cell line C2C12 as a surrogate for myotropism and have found that there is a slight, but reproducible difference in the replication of virus clones bearing the M RNA segment of La Crosse/original virus compared to clones bearing the M RNA segment of Tahyna-181/57 virus, as determined by viral titer, antigen expression, and plaque formation.

  16. The effect of transient, moderate dietary phosphorus deprivation on phosphorus metabolism, muscle content of different phosphorus-containing compounds, and muscle function in dairy cows.

    Science.gov (United States)

    Grünberg, W; Scherpenisse, P; Dobbelaar, P; Idink, M J; Wijnberg, I D

    2015-08-01

    decline in muscle tissue P content. Electromyographic examination revealed increased occurrence of pathological spontaneous activity in striated muscles after 2 wk of dietary P depletion in several cows, which could be suggestive of neuromuscular membrane instability. No effect on heart muscle activity was identified electrocardiographically. These results suggest that counter-regulatory mechanisms were sufficient to maintain normal muscle tissue P content during transient and moderate P deprivation. Muscle function was not grossly affected, although the increased occurrence of pathological spontaneous activity suggests that subclinical neuropathy or myopathy, or both, may have occurred with ongoing P deprivation. The results presented here indicate that plasma [Pi] is unsuitable for assessing muscle tissue P content in cattle. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  17. Skeletal muscle expression of the adhesion-GPCR CD97: CD97 deletion induces an abnormal structure of the sarcoplasmatic reticulum but does not impair skeletal muscle function.

    Directory of Open Access Journals (Sweden)

    Tatiana Zyryanova

    Full Text Available CD97 is a widely expressed adhesion class G-protein-coupled receptor (aGPCR. Here, we investigated the presence of CD97 in normal and malignant human skeletal muscle as well as the ultrastructural and functional consequences of CD97 deficiency in mice. In normal human skeletal muscle, CD97 was expressed at the peripheral sarcolemma of all myofibers, as revealed by immunostaining of tissue sections and surface labeling of single myocytes using flow cytometry. In muscle cross-sections, an intracellular polygonal, honeycomb-like CD97-staining pattern, typical for molecules located in the T-tubule or sarcoplasmatic reticulum (SR, was additionally found. CD97 co-localized with SR Ca2+-ATPase (SERCA, a constituent of the longitudinal SR, but not with the receptors for dihydropyridine (DHPR or ryanodine (RYR, located in the T-tubule and terminal SR, respectively. Intracellular expression of CD97 was higher in slow-twitch compared to most fast-twitch myofibers. In rhabdomyosarcomas, CD97 was strongly upregulated and in part more N-glycosylated compared to normal skeletal muscle. All tumors were strongly CD97-positive, independent of the underlying histological subtype, suggesting high sensitivity of CD97 for this tumor. Ultrastructural analysis of murine skeletal myofibers confirmed the location of CD97 in the SR. CD97 knock-out mice had a dilated SR, resulting in a partial increase in triad diameter yet not affecting the T-tubule, sarcomeric, and mitochondrial structure. Despite these obvious ultrastructural changes, intracellular Ca2+ release from single myofibers, force generation and fatigability of isolated soleus muscles, and wheel-running capacity of mice were not affected by the lack of CD97. We conclude that CD97 is located in the SR and at the peripheral sarcolemma of human and murine skeletal muscle, where its absence affects the structure of the SR without impairing skeletal muscle function.

  18. Early effects of muscle atrophy on shoulder joint development in infants with unilateral birth brachial plexus injury.

    Science.gov (United States)

    Van Gelein Vitringa, Valerie M; Jaspers, Richard; Mullender, Margriet; Ouwerkerk, Willem J; Van Der Sluijs, Johannes A

    2011-02-01

    Shoulder deformities in children with a birth brachial plexus injury (BBPI) are caused by muscle imbalances; however, the underlying mechanisms are unclear. The aim of this study was to assess the early interactions between shoulder muscles and shoulder joint development. In a retrospective magnetic resonance imaging (MRI) study of 36 infants (21 males, 15 females) younger than 12 months (mean 4.8 mo) with unilateral BBPI, volumes and thicknesses of standardized segments of the infraspinatus, subscapularis, and deltoid muscles were measured in both shoulders and expressed as ratios of pathological/unaffected side. The relation between muscle ratios and humeral head subluxation, passive external rotation, glenoid version, and deformity was analysed. Compared with the unaffected side, the muscles of the affected side were of significantly smaller volume and thickness. The subscapularis was the most severely affected muscle, its volume being only 64% (SD 21%) and its thickness only 79% (SD 23%) of the corresponding values on the unaffected side (p atrophy and, possibly, by a reduction in the number of sarcomeres in series. These effects are strongly related to shoulder joint subluxation. © The Authors. Journal compilation © Mac Keith Press 2010.

  19. De novo assembly and comprehensive characterization of the skeletal muscle transcriptomes of the European anchovy (Engraulis encrasicolus).

    Science.gov (United States)

    Eldem, Vahap; Zararsız, Gökmen; Erkan, Melike; Bakir, Yakup

    2015-04-01

    European anchovy has considerable economic and ecological importance due to its high reproduction capacity and growth rate. As one of the largest source of wild marine protein, an increasing muscle mass strength has a major contribution to this growth rate during transition from subadult to adult stage. In the present study, using Illumina sequencing technology (HiSeq2000) accompanied with appropriate bioinformatics softwares; we have sequenced, assembled and annotated the transcriptome of wild subadult and adult anchovy muscles. A total of 131,081,776 high-quality reads were assembled into 125,506 contigs with an average length of 709.35 bp and N50 length of 1159 bp. Functional annotations of assembled contigs have been summarized according to 3325 GO terms, 3370 PFAM domains and 378 predicted KEGG metabolic pathways. About 11% of all contigs had at least one type of SSR motif in their sequences. According to the sequence homology analysis by BlasTN it was concluded that the assembled contigs include 16 skeletal muscle-expressed miRNAs, 14 ncRNAs and most of sarcomeric/myofibrillar genes. We hope that the sequence information regarding the muscle transcriptome of anchovy can provide some insight into the understanding of genome-wide transcriptome profile of teleost muscle tissue and give useful information in fish muscle development. Copyright © 2015 Elsevier B.V. All rights reserved.

  20. Catch-slip bonds can be dispensable for motor force regulation during skeletal muscle contraction.

    Science.gov (United States)

    Dong, Chenling; Chen, Bin

    2015-07-01

    It is intriguing how multiple molecular motors can perform coordinated and synchronous functions, which is essential in various cellular processes. Recent studies on skeletal muscle might have shed light on this issue, where rather precise motor force regulation was partly attributed to the specific stochastic features of a single attached myosin motor. Though attached motors can randomly detach from actin filaments either through an adenosine triphosphate (ATP) hydrolysis cycle or through "catch-slip bond" breaking, their respective contribution in motor force regulation has not been clarified. Here, through simulating a mechanical model of sarcomere with a coupled Monte Carlo method and finite element method, we find that the stochastic features of an ATP hydrolysis cycle can be sufficient while those of catch-slip bonds can be dispensable for motor force regulation.

  1. Comparative study of the effects of the Ga-As (904 nm, 150 mW) laser and the pulsed ultrasound of 1 MHz in inflammation of tibialis muscle of Wistar rats

    Energy Technology Data Exchange (ETDEWEB)

    Pinto, Marcus Vinicius de Mello; Rocha, Lamara Laguardia Valente; Santos, Helio Ricardo dos; Silva, Andre Luis dos Santos; Barbosa, Luis Guilherme; Reis, Joao Batista Alves dos [Centro Universitario de Caratinga, MG (Brazil)]. E-mail; orofacial_1@hotmail.com; Costa, Daniel Almeida da [Faculdade de Minas, Belo Horizonte, MG (Brazil); Bernardo-Filho, Mario [Universidade do Estado do Rio de Janeiro (UERJ), RJ (Brazil). Inst. de Biologia Roberto Alcantara Gomes. Dept. de Biofisica e Biometria

    2008-12-15

    This paper aims to compare the therapeutic effect of the laser As-Ga of 904 nm and pulsed Ultrasound of 1 MHz applied in striated skeletal muscle of inflamed rats. The animals received an intramuscular injection of bupivacaine hydrochloride in tibialis muscle in order to induce the inflammatory process, and after 24 hours, the time was considered 0 for the initiation of therapy, using a laser and ultrasound. Samples collected the muscles of the animals were stained with Hematoxylin-Eosin and histological sections of the groups used for the analysis of the muscle tissue in relation to reducing the inflammatory process, comparing the results of the two therapies used. In this study it is suggested that both treatment with laser as with ultrasound can act as anti-inflammatory. However, the laser seems to have anti-inflammatory effect for all periods observed, while the ultrasound was only able to induce declining inflammatory response to seven days. (author)

  2. Park7 expression influences myotube size and myosin expression in muscle.

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    Hui Yu

    Full Text Available Callipyge sheep exhibit postnatal muscle hypertrophy due to the up-regulation of DLK1 and/or RTL1. The up-regulation of PARK7 was identified in hypertrophied muscles by microarray analysis and further validated by quantitative PCR. The expression of PARK7 in hypertrophied muscle of callipyge lambs was confirmed to be up-regulated at the protein level. PARK7 was previously identified to positively regulate PI3K/AKT pathway by suppressing the phosphatase activity of PTEN in mouse fibroblasts. The purpose of this study was to investigate the effects of PARK7 in muscle growth and protein accretion in response to IGF1. Primary myoblasts isolated from Park7 (+/+ and Park7 (-/- mice were used to examine the effect of differential expression of Park7. The Park7 (+/+ myotubes had significantly larger diameters and more total sarcomeric myosin expression than Park7 (-/- myotubes. IGF1 treatment increased the mRNA abundance of Myh4, Myh7 and Myh8 between 20-40% in Park7 (+/+ myotubes relative to Park7 (-/-. The level of AKT phosphorylation was increased in Park7 (+/+ myotubes at all levels of IGF1 supplementation. After removal of IGF1, the Park7 (+/+ myotubes maintained higher AKT phosphorylation through 3 hours. PARK7 positively regulates the PI3K/AKT pathway by inhibition of PTEN phosphatase activity in skeletal muscle. The increased PARK7 expression can increase protein synthesis and result in myotube hypertrophy. These results support the hypothesis that elevated expression of PARK7 in callipyge muscle would increase levels of AKT activity to cause hypertrophy in response to the normal IGF1 signaling in rapidly growing lambs. Increasing expression of PARK7 could be a novel mechanism to increase protein accretion and muscle growth in livestock or help improve muscle mass with disease or aging.

  3. Park7 expression influences myotube size and myosin expression in muscle.

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    Yu, Hui; Waddell, Jolena N; Kuang, Shihuan; Bidwell, Christopher A

    2014-01-01

    Callipyge sheep exhibit postnatal muscle hypertrophy due to the up-regulation of DLK1 and/or RTL1. The up-regulation of PARK7 was identified in hypertrophied muscles by microarray analysis and further validated by quantitative PCR. The expression of PARK7 in hypertrophied muscle of callipyge lambs was confirmed to be up-regulated at the protein level. PARK7 was previously identified to positively regulate PI3K/AKT pathway by suppressing the phosphatase activity of PTEN in mouse fibroblasts. The purpose of this study was to investigate the effects of PARK7 in muscle growth and protein accretion in response to IGF1. Primary myoblasts isolated from Park7 (+/+) and Park7 (-/-) mice were used to examine the effect of differential expression of Park7. The Park7 (+/+) myotubes had significantly larger diameters and more total sarcomeric myosin expression than Park7 (-/-) myotubes. IGF1 treatment increased the mRNA abundance of Myh4, Myh7 and Myh8 between 20-40% in Park7 (+/+) myotubes relative to Park7 (-/-). The level of AKT phosphorylation was increased in Park7 (+/+) myotubes at all levels of IGF1 supplementation. After removal of IGF1, the Park7 (+/+) myotubes maintained higher AKT phosphorylation through 3 hours. PARK7 positively regulates the PI3K/AKT pathway by inhibition of PTEN phosphatase activity in skeletal muscle. The increased PARK7 expression can increase protein synthesis and result in myotube hypertrophy. These results support the hypothesis that elevated expression of PARK7 in callipyge muscle would increase levels of AKT activity to cause hypertrophy in response to the normal IGF1 signaling in rapidly growing lambs. Increasing expression of PARK7 could be a novel mechanism to increase protein accretion and muscle growth in livestock or help improve muscle mass with disease or aging.

  4. Energy capture and storage in asymmetrically multistable modular structures inspired by skeletal muscle

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    Kidambi, Narayanan; Harne, Ryan L.; Wang, K. W.

    2017-08-01

    The remarkable versatility and adaptability of skeletal muscle that arises from the assembly of its nanoscale cross-bridges into micro-scale assemblies known as sarcomeres provides great inspiration for the development of advanced adaptive structures and material systems. Motivated by the capability of cross-bridges to capture elastic strain energy to improve the energetic efficiency of sudden movements and repeated motions, and by models of cross-bridge power stroke motions and sarcomere contractile behaviors that incorporate asymmetric, bistable potential energy landscapes, this research develops and studies modular mechanical structures that trap and store energy in higher-energy configurations. Modules exhibiting tailorable asymmetric bistability are first designed and fabricated, revealing how geometric parameters influence the asymmetry of the resulting double-well energy landscapes. These experimentally-observed characteristics are then investigated with numerical and analytical methods to characterize the dynamics of asymmetrically bistable modules. The assembly of such modules into greater structures generates complex, multi-well energy landscapes with stable system configurations exhibiting different quantities of stored elastic potential energy. Dynamic analyses illustrate the ability of these structures to capture a portion of the initial kinetic energy due to impulsive excitations as recoverable strain potential energy, and reveal how stiffness parameters, damping, and the presence of thermal noise in micro- and nano-scale applications influence energy capture behaviors. The insights gained could foster the development of advanced structural/material systems inspired by skeletal muscle, including actuators that effectively capture, store, and release energy, as well as adaptive, robust, and reusable armors and protective devices.

  5. Dynamic Strength of Titin's Z-Disk End

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    Veronika Kollár

    2010-01-01

    Full Text Available Titin is a giant filamentous protein traversing the half sarcomere of striated muscle with putative functions as diverse as providing structural template, generating elastic response, and sensing and relaying mechanical information. The Z-disk region of titin, which corresponds to the N-terminal end of the molecule, has been thought to be a hot spot for mechanosensing while also serving as anchorage for its sarcomeric attachment. Understanding the mechanics of titin's Z-disk region, particularly under the effect of binding proteins, is of great interest. Here we briefly review recent findings on the structure, molecular associations, and mechanics of titin's Z-disk region. In addition, we report experimental results on the dynamic strength of titin's Z1Z2 domains measured by nanomechanical manipulation of the chemical dimer of a recombinant protein fragment.

  6. O-GlcNAcylation, contractile protein modifications and calcium affinity in skeletal muscle

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    Caroline eCieniewski-Bernard

    2014-10-01

    Full Text Available Phosphorylation is recognized as being one of the major post-translational modification involved in the fine modulation of all if not all cellular processes including skeletal muscle contractile activity. However, phosphorylation presents a dynamic and highly regulated interplay with an atypical glycosylation on intracellular proteins, the O-GlcNAcylation. Akin to phosphorylation, O-GlcNAcylation is also involved in the physiopathology of several acquired diseases, such as muscle insulin resistance in diabetes or muscle atrophy. Recent data underline that the interplay between phosphorylation and O-GlcNAcylation acts as a modulator of skeletal muscle contractile activity. In particular, the O-GlcNAcylation level of the phosphoprotein MLC2 seems to be crucial in the modulation of the calcium activation properties, and should be responsible of changes in calcium properties observed in atrophy. Moreover, since several key structural proteins are O-GlcNAc-modified, and because of the localization of the enzymes involved in the O-GlcNAcylation/de-O-GlcNAcylation process to the nodal Z disk, a role of O-GlcNAcylation in the modulation of the sarcomeric structure should be considered.

  7. Interactome Mapping Reveals Important Pathways in Skeletal Muscle Development of Pigs

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    Jianhua Cao

    2014-11-01

    Full Text Available The regulatory relationship and connectivity among genes involved in myogenesis and hypertrophy of skeletal muscle in pigs still remain large challenges. Presentation of gene interactions is a potential way to understand the mechanisms of developmental events in skeletal muscle. In this study, genome-wide transcripts and miRNA profiling was determined for Landrace pigs at four time points using microarray chips. A comprehensive method integrating gene ontology annotation and interactome network mapping was conducted to analyze the biological patterns and interaction modules of muscle development events based on differentially expressed genes and miRNAs. Our results showed that in total 484 genes and 34 miRNAs were detected for the duration from embryonic stage to adult in pigs, which composed two linear expression patterns with consensus changes. Moreover, the gene ontology analysis also disclosed that there were three typical biological events i.e., microstructure assembly of sarcomere at early embryonic stage, myofibril formation at later embryonic stage and function establishments of myoblast cells at postnatal stage. The interactome mappings of different time points also found the down-regulated trend of gene expression existed across the whole duration, which brought a possibility to introduce the myogenesis related miRNAs into the interactome regulatory networks of skeletal muscle in pigs.

  8. Calpain 3 is important for muscle regeneration: Evidence from patients with limb girdle muscular dystrophies

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    Hauerslev Simon

    2012-03-01

    Full Text Available Abstract Background Limb girdle muscular dystrophy (LGMD type 2A is caused by mutations in the CAPN3 gene and complete lack of functional calpain 3 leads to the most severe muscle wasting. Calpain 3 is suggested to be involved in maturation of contractile elements after muscle degeneration. The aim of this study was to investigate how mutations in the four functional domains of calpain 3 affect muscle regeneration. Methods We studied muscle regeneration in 22 patients with LGMD2A with calpain 3 deficiency, in five patients with LGMD2I, with a secondary reduction in calpain 3, and in five patients with Becker muscular dystrophy (BMD with normal calpain 3 levels. Regeneration was assessed by using the developmental markers neonatal myosin heavy chain (nMHC, vimentin, MyoD and myogenin and counting internally nucleated fibers. Results We found that the recent regeneration as determined by the number of nMHC/vimentin-positive fibers was greatly diminished in severely affected LGMD2A patients compared to similarly affected patients with LGMD2I and BMD. Whorled fibers, a sign of aberrant regeneration, was highly elevated in patients with a complete lack of calpain 3 compared to patients with residual calpain 3. Regeneration is not affected by location of the mutation in the CAPN3 gene. Conclusions Our findings suggest that calpain 3 is needed for the regenerative process probably during sarcomere remodeling as the complete lack of functional calpain 3 leads to the most severe phenotypes.

  9. Titin isoform variance and length dependence of activation in skinned bovine cardiac muscle.

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    Fukuda, Norio; Wu, Yiming; Farman, Gerrie; Irving, Thomas C; Granzier, Henk

    2003-11-15

    We have explored the role of the giant elastic protein titin in the Frank-Starling mechanism of the heart by measuring the sarcomere length (SL) dependence of activation in skinned cardiac muscles with different titin-based passive stiffness characteristics. We studied muscle from the bovine left ventricle (BLV), which expresses a high level of a stiff titin isoform, and muscle from the bovine left atrium (BLA), which expresses more compliant titin isoforms. Passive tension was also varied in each muscle type by manipulating the pre-history of stretch prior to activation. We found that the SL-dependent increases in Ca2+ sensitivity and maximal Ca2+-activated tension were markedly more pronounced when titin-based passive tension was high. Small-angle X-ray diffraction experiments revealed that the SL dependence of reduction of interfilament lattice spacing is greater in BLV than in BLA and that the lattice spacing is coupled with titin-based passive tension. These results support the notion that titin-based passive tension promotes actomyosin interaction by reducing the lattice spacing. This work indicates that titin may be a factor involved in the Frank-Starling mechanism of the heart by promoting actomyosin interaction in response to stretch.

  10. C. elegans PAT-9 is a nuclear zinc finger protein critical for the assembly of muscle attachments

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    Liu Qian

    2012-05-01

    Full Text Available Abstract Background Caenorhabditis elegans sarcomeres have been studied extensively utilizing both forward and reverse genetic techniques to provide insight into muscle development and the mechanisms behind muscle contraction. A previous genetic screen investigating early muscle development produced 13 independent mutant genes exhibiting a Pat (paralyzed and arrested elongation at the two-fold length of embryonic development muscle phenotype. This study reports the identification and characterization of one of those genes, pat-9. Results Positional cloning, reverse genetics, and plasmid rescue experiments were used to identify the predicted C. elegans gene T27B1.2 (recently named ztf-19 as the pat-9 gene. Analysis of pat-9 showed it is expressed early in development and within body wall muscle lineages, consistent with a role in muscle development and producing a Pat phenotype. However, unlike most of the other known Pat gene family members, which encode structural components of muscle attachment sites, PAT-9 is an exclusively nuclear protein. Analysis of the predicted PAT-9 amino acid sequence identified one putative nuclear localization domain and three C2H2 zinc finger domains. Both immunocytochemistry and PAT-9::GFP fusion expression confirm that PAT-9 is primarily a nuclear protein and chromatin immunoprecipitation (ChIP experiments showed that PAT-9 is present on certain gene promoters. Conclusions We have shown that the T27B1.2 gene is pat-9. Considering the Pat-9 mutant phenotype shows severely disrupted muscle attachment sites despite PAT-9 being a nuclear zinc finger protein and not a structural component of muscle attachment sites, we propose that PAT-9 likely functions in the regulation of gene expression for some necessary structural or regulatory component(s of the muscle attachment sites.

  11. Relationship between physical exercise, muscle damage and delayed-onset muscle soreness

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    Denis Foschini

    2007-03-01

    Full Text Available The objective of the present study was to investigate the relationship between physical exercise involving muscle damage and delayed-onset muscle soreness (DOMS. A literature review of national and international periodicals was carried out. Muscle structures (membranes, Z-line, sarcomeres, T tubules and myofi brils can become damaged as a result of an imposed mechanical overload. Of greatest note are exercises requiring strength, particularly when muscular action is eccentric. Damage to skeletal musculature can be analyzed by direct methods (muscle biopsy or magnetic resonance or by indirect methods (maximum voluntary movement, subjective pain perception scales, analysis of enzyme and protein concentrations in blood. Creatine kinase (CK, lactate dehydrogenase (LDH, myosin heavy chain fragments, troponin-I and myoglobin can be used as indirect markers of muscle damage. Both DOMS and muscle damage can be infl uenced by the type of activity, with emphasis on eccentric muscle movements, type of exercise, velocity of the movement, interval period between series, the level of individual fi tness, this last primarily affecting beginners. When myotrauma occurs, muscle damage repair is initiated by leukocytes migrating to the injured area, although, the histamines, prostaglandins, kinins and K+ produced by neutrophils and macrophages stimulate free nerve endings in the muscle, causing the DOMS. Despite this apparent relationship between muscle damage and DOMS, it is not possible toestablish a linear relationship between these two variables, since published data are divergent. RESUMO O objetivo desse estudo foi investigar as relações do exercício físico com o dano muscular e dor muscular de início tardio (DMIT. Para tanto, foi realizada uma revisão de literatura de periódicos nacionais e internacionais. O dano muscular pode ocorrer em estruturas musculares (membranas, linha Z, sarcolema, túbulos T e miofi brilas em função da sobrecarga mec

  12. An elderly-onset limb girdle muscular dystrophy type 1B (LGMD1B) with pseudo-hypertrophy of paraspinal muscles.

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    Furuta, Mitsuru; Sumi-Akamaru, Hisae; Takahashi, Masanori P; Hayashi, Yukiko K; Nishino, Ichizo; Mochizuki, Hideki

    2016-09-01

    Mutations in LMNA, encoding A-type lamins, lead to diverse disorders, collectively called "laminopathies," which affect the striated muscle, cardiac muscle, adipose tissue, skin, peripheral nerve, and premature aging. We describe a patient with limb-girdle muscular dystrophy type 1B (LGMD1B) carrying a heterozygous p.Arg377His mutation in LMNA, in whom skeletal muscle symptom onset was at the age of 65 years. Her weakness started at the erector spinae muscles, which showed marked pseudo-hypertrophy even at the age of 72 years. Her first episode of syncope was at 44 years; however, aberrant cardiac conduction was not revealed until 60 years. The p.Arg377His mutation has been previously reported in several familial LMNA-associated myopathies, most of which showed muscle weakness before the 6th decade. This is the first report of pseudo-hypertrophy of paravertebral muscles in LMNA-associated myopathies. The pseudo-hypertrophy of paravertebral muscles and the elderly-onset of muscle weakness make this case unique and reportable. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Reduced nuclear translocation of serum response factor is associated with skeletal muscle atrophy in a cigarette smoke-induced mouse model of COPD

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    Ma R

    2017-02-01

    Full Text Available Ran Ma, Xuefang Gong, Hua Jiang, Chunyi Lin, Yuqin Chen, Xiaoming Xu, Chenting Zhang, Jian Wang, Wenju Lu, Nanshan ZhongGuangzhou Institute of Respiratory Disease, State Key Laboratory of Respiratory Diseases, The 1st Affiliated Hospital, Guangzhou Medical University, Guangzhou, Guangdong, People’s Republic of ChinaAbstract: Skeletal muscle atrophy and dysfunction are common complications in the chronic obstructive pulmonary disease (COPD. However, the underlying molecular mechanism remains elusive. Serum response factor (SRF is a transcription factor which is critical in myocyte differentiation and growth. In this study, we established a mouse COPD model induced by cigarette smoking (CS exposure for 24 weeks, with apparent pathophysiological changes, including increased airway resistance, enlarged alveoli, and skeletal muscle atrophy. Levels of upstream regulators of SRF, striated muscle activator of Rho signaling (STARS, and ras homolog gene family, member A (RhoA were decreased in quadriceps muscle of COPD mice. Meanwhile, the nucleic location of SRF was diminished along with its cytoplasmic accumulation. There was a downregulation of the target muscle-specific gene, Igf1. These results suggest that the CS is one of the major cause for COPD pathogenesis, which induces the COPD-associated skeletal muscle atrophy which is closely related to decreasing SRF nucleic translocation, consequently downregulating the SRF target genes involved in muscle growth and nutrition. The STARS/RhoA signaling pathway might contribute to this course by impacting SRF subcellular distribution. Keywords: SRF, chronic obstructive pulmonary disease, skeletal muscle atrophy, cigarette smoking

  14. Nebulin-like protein in the earthworm Eisenia foetida. Immunocytochemical electron microscopic study and western blot analysis of several muscle cell types.

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    Royuela, M; Fraile, B; Paniagua, R

    1997-07-01

    Nebulin is a giant protein (500-900 kDa), which has been reported only in the skeletal muscle (not in cardiac muscle) of vertebrates. The possible presence and distribution of nebulin-like proteins in obliquely striated muscles (body wall and inner muscular layer of the pseudoheart) and smooth muscle (outer muscular layer of the pseudoheart) from the earthworm Eisenia foetida have been examined by means of Western blotting analysis and immunoelectron microscopy, using antibodies against mouse nebulin. The results were compared with those obtained in skeletal, cardiac and smooth muscles of the mouse. In the mouse, immunoreaction to nebulin was observed only in the skeletal muscle and extended along the length of the thin filament. In the earthworm, immunoreaction to a nebulin-like protein was found in the muscle of the body wall and the inner muscular layer of the pseudoheart, but not in the outer muscular layer of the pseudoheart. By electron microscopy, immunolabeling to this protein was observed along the whole length of the thin filament. Western blotting analysis of this nebulin-like protein showed a single band at an estimated molecular mass between 350 and 450 kDa that is slightly lower than that of mouse skeletal muscle nebulin.

  15. A comparative study of aging of the elastic fiber system of the diaphragm and the rectus abdominis muscles in rats

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    Rodrigues C.J.

    2000-01-01

    Full Text Available In the present study the age-related changes of the striated muscle elastic fiber system were investigated in the diaphragm and rectus abdominis muscles of 1-, 4-, 8- and 18-month-old rats. The activation patterns of these muscles differ in that the diaphragm is regularly mobilized tens of times every minute during the entire life of the animal whereas the rectus abdominis, although mobilized in respiration, is much less and more irregularly activated. The elastic fibers were stained by the Verhoeff technique for mature elastic fibers. Weigert stain was used to stain mature and elaunin elastic fibers, and Weigert-oxone to stain mature, elaunin and oxytalan elastic fibers. The density of mature and elaunin elastic fibers showed a progressive increase with age, whereas the amount of oxytalan elastic fibers decreased in both diaphragm and rectus abdominis muscles and their muscular fascias. These age-related quantitative and structural changes of the elastic fiber system may reduce the viscoelastic properties of the diaphragm and rectus abdominis muscles, which may compromise the transmission of tensile muscle strength to the tendons and may affect maximum total strength.

  16. Muscle channelopathies.

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    Statland, Jeffrey; Phillips, Lauren; Trivedi, Jaya R

    2014-08-01

    Skeletal muscle channelopathies are rare heterogeneous diseases with marked genotypic and phenotypic variability. Despite advances in understanding of the molecular pathology of these disorders, the diverse phenotypic manifestations remain a challenge in diagnosis and therapeutics. These disorders can cause lifetime disability and affect quality of life. There is no treatment of these disorders approved by the US Food and Drug Administration at this time. Recognition and treatment of symptoms might reduce morbidity and improve quality of life. This article summarizes the clinical manifestations, diagnostic studies, pathophysiology, and treatment options in nondystrophic myotonia, congenital myasthenic syndrome, and periodic paralyses. Copyright © 2014 Elsevier Inc. All rights reserved.

  17. Overexpression of miRNA-9 Generates Muscle Hypercontraction Through Translational Repression of Troponin-T in Drosophila melanogaster Indirect Flight Muscles

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    Prasanna Katti

    2017-10-01

    Full Text Available MicroRNAs (miRNAs are small noncoding endogenous RNAs, typically 21–23 nucleotides long, that regulate gene expression, usually post-transcriptionally, by binding to the 3′-UTR of target mRNA, thus blocking translation. The expression of several miRNAs is significantly altered during cardiac hypertrophy, myocardial ischemia, fibrosis, heart failure, and other cardiac myopathies. Recent studies have implicated miRNA-9 (miR-9 in myocardial hypertrophy. However, a detailed mechanism remains obscure. In this study, we have addressed the roles of miR-9 in muscle development and function using a genetically tractable model system, the indirect flight muscles (IFMs of Drosophila melanogaster. Bioinformatics analysis identified 135 potential miR-9a targets, of which 27 genes were associated with Drosophila muscle development. Troponin-T (TnT was identified as major structural gene target of miR-9a. We show that flies overexpressing miR-9a in the IFMs have abnormal wing position and are flightless. These flies also exhibit a loss of muscle integrity and sarcomeric organization causing an abnormal muscle condition known as “hypercontraction.” Additionally, miR-9a overexpression resulted in the reduction of TnT protein levels while transcript levels were unaffected. Furthermore, muscle abnormalities associated with miR-9a overexpression were completely rescued by overexpression of TnT transgenes which lacked the miR-9a binding site. These findings indicate that miR-9a interacts with the 3′-UTR of the TnT mRNA and downregulates the TnT protein levels by translational repression. The reduction in TnT levels leads to a cooperative downregulation of other thin filament structural proteins. Our findings have implications for understanding the cellular pathophysiology of cardiomyopathies associated with miR-9 overexpression.

  18. Acute effects of stretching exercise on the soleus muscle of female aged rats.

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    Zotz, Talita Gnoato; Capriglione, Luiz Guilherme A; Zotz, Rafael; Noronha, Lucia; Viola De Azevedo, Marina Louise; Fiuza Martins, Hilana Rickli; Silveira Gomes, Anna Raquel

    2016-01-01

    It has been shown that stretching exercises can improve the flexibility and independence of the elderly. However, although these exercises commonly constitute training programs, the morphological adaptations induced by stretching exercises in aged skeletal muscle are still unclear. To assess the acute effects of passive mechanical static stretching on the morphology, sarcomerogenesis and modulation of important components of the extracellular matrix of the soleus muscle of aged female rats. Fifteen old female rats with 26 months were divided into two groups: stretching (n=8, SG) and control (n=7, CG): The stretching protocol consisted of 4 repetitions each of 1 min with 30s interval between sets. Stretching was performed on the left soleus muscle, 3 times a week for 1 week. After three sessions, the rats were anesthetized to remove the left soleus muscle, and then euthanized. The following analyses were carried out: muscle fiber cross-sectional area and serial sarcomere number; immunohistochemistry for the quantification of collagen I, III and TGFβ-1. a decrease in muscle fiber cross-sectional area of the SG was observed when compared to the CG (p=0.0001, Kruskal-Wallis); the percentage of type I collagen was significantly lower in the SG when compared to the CG (p=0.01, Kruskal-Wallis), as well as the percentage of TGFβ-1 (p=0.04, Kruskal-Wallis); collagen III was significantly higher in the SG than in the CG (7.06±6.88% vs 4.92±5.30%, p=0.01, Kruskal-Wallis). Although the acute stretching induced muscle hypotrophy, an antifibrotic action was detected. Copyright © 2015 Elsevier GmbH. All rights reserved.

  19. Shaping leg muscles in Drosophila: role of ladybird, a conserved regulator of appendicular myogenesis.

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    Tariq Maqbool

    Full Text Available Legs are locomotor appendages used by a variety of evolutionarily distant vertebrates and invertebrates. The primary biological leg function, locomotion, requires the formation of a specialised appendicular musculature. Here we report evidence that ladybird, an orthologue of the Lbx1 gene recognised as a hallmark of appendicular myogenesis in vertebrates, is expressed in leg myoblasts, and regulates the shape, ultrastructure and functional properties of leg muscles in Drosophila. Ladybird expression is progressively activated in myoblasts associated with the imaginal leg disc and precedes that of the founder cell marker dumbfounded. The RNAi-mediated attenuation of ladybird expression alters properties of developing myotubes, impairing their ability to grow and interact with the internal tendons and epithelial attachment sites. It also affects sarcomeric ultrastructure, resulting in reduced leg muscle performance and impaired mobility in surviving flies. The over-expression of ladybird also results in an abnormal pattern of dorsally located leg muscles, indicating different requirements for ladybird in dorsal versus ventral muscles. This differential effect is consistent with the higher level of Ladybird in ventrally located myoblasts and with positive ladybird regulation by extrinsic Wingless signalling from the ventral epithelium. In addition, ladybird expression correlates with that of FGF receptor Heartless and the read-out of FGF signalling downstream of FGF. FGF signals regulate the number of leg disc associated myoblasts and are able to accelerate myogenic differentiation by activating ladybird, leading to ectopic muscle fibre formation. A key role for ladybird in leg myogenesis is further supported by its capacity to repress vestigial and to down-regulate the vestigial-governed flight muscle developmental programme. Thus in Drosophila like in vertebrates, appendicular muscles develop from a specialised pool of myoblasts expressing

  20. Identification of a conserved set of upregulated genes in mouse skeletal muscle hypertrophy and regrowth

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    Chaillou, Thomas; Jackson, Janna R.; England, Jonathan H.; Kirby, Tyler J.; Richards-White, Jena; Esser, Karyn A.; Dupont-Versteegden, Esther E.

    2014-01-01

    The purpose of this study was to compare the gene expression profile of mouse skeletal muscle undergoing two forms of growth (hypertrophy and regrowth) with the goal of identifying a conserved set of differentially expressed genes. Expression profiling by microarray was performed on the plantaris muscle subjected to 1, 3, 5, 7, 10, and 14 days of hypertrophy or regrowth following 2 wk of hind-limb suspension. We identified 97 differentially expressed genes (≥2-fold increase or ≥50% decrease compared with control muscle) that were conserved during the two forms of muscle growth. The vast majority (∼90%) of the differentially expressed genes was upregulated and occurred at a single time point (64 out of 86 genes), which most often was on the first day of the time course. Microarray analysis from the conserved upregulated genes showed a set of genes related to contractile apparatus and stress response at day 1, including three genes involved in mechanotransduction and four genes encoding heat shock proteins. Our analysis further identified three cell cycle-related genes at day and several genes associated with extracellular matrix (ECM) at both days 3 and 10. In conclusion, we have identified a core set of genes commonly upregulated in two forms of muscle growth that could play a role in the maintenance of sarcomere stability, ECM remodeling, cell proliferation, fast-to-slow fiber type transition, and the regulation of skeletal muscle growth. These findings suggest conserved regulatory mechanisms involved in the adaptation of skeletal muscle to increased mechanical loading. PMID:25554798

  1. Identification of a conserved set of upregulated genes in mouse skeletal muscle hypertrophy and regrowth.

    Science.gov (United States)

    Chaillou, Thomas; Jackson, Janna R; England, Jonathan H; Kirby, Tyler J; Richards-White, Jena; Esser, Karyn A; Dupont-Versteegden, Esther E; McCarthy, John J

    2015-01-01

    The purpose of this study was to compare the gene expression profile of mouse skeletal muscle undergoing two forms of growth (hypertrophy and regrowth) with the goal of identifying a conserved set of differentially expressed genes. Expression profiling by microarray was performed on the plantaris muscle subjected to 1, 3, 5, 7, 10, and 14 days of hypertrophy or regrowth following 2 wk of hind-limb suspension. We identified 97 differentially expressed genes (≥2-fold increase or ≥50% decrease compared with control muscle) that were conserved during the two forms of muscle growth. The vast majority (∼90%) of the differentially expressed genes was upregulated and occurred at a single time point (64 out of 86 genes), which most often was on the first day of the time course. Microarray analysis from the conserved upregulated genes showed a set of genes related to contractile apparatus and stress response at day 1, including three genes involved in mechanotransduction and four genes encoding heat shock proteins. Our analysis further identified three cell cycle-related genes at day and several genes associated with extracellular matrix (ECM) at both days 3 and 10. In conclusion, we have identified a core set of genes commonly upregulated in two forms of muscle growth that could play a role in the maintenance of sarcomere stability, ECM remodeling, cell proliferation, fast-to-slow fiber type transition, and the regulation of skeletal muscle growth. These findings suggest conserved regulatory mechanisms involved in the adaptation of skeletal muscle to increased mechanical loading. Copyright © 2015 the American Physiological Society.

  2. Effect of muscle length on cross-bridge kinetics in intact cardiac trabeculae at body temperature.

    Science.gov (United States)

    Milani-Nejad, Nima; Xu, Ying; Davis, Jonathan P; Campbell, Kenneth S; Janssen, Paul M L

    2013-01-01

    Dynamic force generation in cardiac muscle, which determines cardiac pumping activity, depends on both the number of sarcomeric cross-bridges and on their cycling kinetics. The Frank-Starling mechanism dictates that cardiac force development increases with increasing cardiac muscle length (corresponding to increased ventricular volume). It is, however, unclear to what extent this increase in cardiac muscle length affects the rate of cross-bridge cycling. Previous studies using permeabilized cardiac preparations, sub-physiological temperatures, or both have obtained conflicting results. Here, we developed a protocol that allowed us to reliably and reproducibly measure the rate of tension redevelopment (k(tr); which depends on the rate of cross-bridge cycling) in intact trabeculae at body temperature. Using K(+) contractures to induce a tonic level of force, we showed the k(tr) was slower in rabbit muscle (which contains predominantly β myosin) than in rat muscle (which contains predominantly α myosin). Analyses of k(tr) in rat muscle at optimal length (L(opt)) and 90% of optimal length (L(90)) revealed that k(tr) was significantly slower at L(opt) (27.7 ± 3.3 and 27.8 ± 3.0 s(-1) in duplicate analyses) than at L(90) (45.1 ± 7.6 and 47.5 ± 9.2 s(-1)). We therefore show that k(tr) can be measured in intact rat and rabbit cardiac trabeculae, and that the k(tr) decreases when muscles are stretched to their optimal length under near-physiological conditions, indicating that the Frank-Starling mechanism not only increases force but also affects cross-bridge cycling kinetics.

  3. Shaping leg muscles in Drosophila: role of ladybird, a conserved regulator of appendicular myogenesis.

    Science.gov (United States)

    Maqbool, Tariq; Soler, Cedric; Jagla, Teresa; Daczewska, Malgorzata; Lodha, Neha; Palliyil, Sudhir; VijayRaghavan, K; Jagla, Krzysztof

    2006-12-27

    Legs are locomotor appendages used by a variety of evolutionarily distant vertebrates and invertebrates. The primary biological leg function, locomotion, requires the formation of a specialised appendicular musculature. Here we report evidence that ladybird, an orthologue of the Lbx1 gene recognised as a hallmark of appendicular myogenesis in vertebrates, is expressed in leg myoblasts, and regulates the shape, ultrastructure and functional properties of leg muscles in Drosophila. Ladybird expression is progressively activated in myoblasts associated with the imaginal leg disc and precedes that of the founder cell marker dumbfounded. The RNAi-mediated attenuation of ladybird expression alters properties of developing myotubes, impairing their ability to grow and interact with the internal tendons and epithelial attachment sites. It also affects sarcomeric ultrastructure, resulting in reduced leg muscle performance and impaired mobility in surviving flies. The over-expression of ladybird also results in an abnormal pattern of dorsally located leg muscles, indicating different requirements for ladybird in dorsal versus ventral muscles. This differential effect is consistent with the higher level of Ladybird in ventrally located myoblasts and with positive ladybird regulation by extrinsic Wingless signalling from the ventral epithelium. In addition, ladybird expression correlates with that of FGF receptor Heartless and the read-out of FGF signalling downstream of FGF. FGF signals regulate the number of leg disc associated myoblasts and are able to accelerate myogenic differentiation by activating ladybird, leading to ectopic muscle fibre formation. A key role for ladybird in leg myogenesis is further supported by its capacity to repress vestigial and to down-regulate the vestigial-governed flight muscle developmental programme. Thus in Drosophila like in vertebrates, appendicular muscles develop from a specialised pool of myoblasts expressing ladybird/Lbx1. The

  4. Troponin T nuclear localization and its role in aging skeletal muscle.

    Science.gov (United States)

    Zhang, Tan; Birbrair, Alexander; Wang, Zhong-Min; Taylor, Jackson; Messi, María Laura; Delbono, Osvaldo

    2013-04-01

    Troponin T (TnT) is known to mediate the interaction between Tn complex and tropomyosin (Tm), which is essential for calcium-activated striated muscle contraction. This regulatory function takes place in the myoplasm, where TnT binds Tm. However, recent findings of troponin I and Tm nuclear translocation in Drosophila and mammalian cells imply other roles for the Tn-Tm complex. We hypothesized that TnT plays a nonclassical role through nuclear translocation. Immunoblotting with different antibodies targeting the NH2- or COOH-terminal region uncovered a pool of fast skeletal muscle TnT3 localized in the nuclear fraction of mouse skeletal muscle as either an intact or fragmented protein. Construction of TnT3-DsRed fusion proteins led to the further observation that TnT3 fragments are closely related to nucleolus and RNA polymerase activity, suggesting a role for TnT3 in regulating transcription. Functionally, overexpression of TnT3 fragments produced significant defects in nuclear shape and caused high levels of apoptosis. Interestingly, nuclear TnT3 and its fragments were highly regulated by aging, thus creating a possible link between the deleterious effects of TnT3 and sarcopenia. We propose that changes in nuclear TnT3 and its fragments cause the number of myonuclei to decrease with age, contributing to muscle damage and wasting.

  5. MyoScreen, a High-Throughput Phenotypic Screening Platform Enabling Muscle Drug Discovery.

    Science.gov (United States)

    Young, Joanne; Margaron, Yoran; Fernandes, Mathieu; Duchemin-Pelletier, Eve; Michaud, Joris; Flaender, Mélanie; Lorintiu, Oana; Degot, Sébastien; Poydenot, Pauline

    2018-03-01

    Despite the need for more effective drug treatments to address muscle atrophy and disease, physiologically accurate in vitro screening models and higher information content preclinical assays that aid in the discovery and development of novel therapies are lacking. To this end, MyoScreen was developed: a robust and versatile high-throughput high-content screening (HT/HCS) platform that integrates a physiologically and pharmacologically relevant micropatterned human primary skeletal muscle model with a panel of pertinent phenotypic and functional assays. MyoScreen myotubes form aligned, striated myofibers, and they show nerve-independent accumulation of acetylcholine receptors (AChRs), excitation-contraction coupling (ECC) properties characteristic of adult skeletal muscle and contraction in response to chemical stimulation. Reproducibility and sensitivity of the fully automated MyoScreen platform are highlighted in assays that quantitatively measure myogenesis, hypertrophy and atrophy, AChR clusterization, and intracellular calcium release dynamics, as well as integrating contractility data. A primary screen of 2560 compounds to identify stimulators of myofiber regeneration and repair, followed by further biological characterization of two hits, validates MyoScreen for the discovery and testing of novel therapeutics. MyoScreen is an improvement of current in vitro muscle models, enabling a more predictive screening strategy for preclinical selection of the most efficacious new chemical entities earlier in the discovery pipeline process.

  6. Alterations in Muscle Mass and Contractile Phenotype in Response to Unloading Models: Role of Transcriptional/Pretranslational Mechanisms

    Directory of Open Access Journals (Sweden)

    Kenneth M Baldwin

    2013-10-01

    Full Text Available Skeletal muscle is the largest organ system in mammalian organisms providing postural control and movement patterns of varying intensity. Through evolution, skeletal muscle fibers have evolved into three phenotype clusters defined as a muscle unit which consists of all muscle fibers innervated by a single motoneuron linking varying numbers of fibers of similar phenotype. This fundamental organization of the motor unit reflects the fact that there is a remarkable interdependence of gene regulation between the motoneurons and the muscle mainly via activity-dependent mechanisms. These fiber types can be classified via the primary type of myosin heavy chain (MHC gene expressed in the motor unit. Four MHC gene encoded proteins have been identified in striated muscle: slow type I MHC and three fast MHC types, IIa, IIx, and IIb. These MHCs dictate the intrinsic contraction speed of the myofiber with the type I generating the slowest and IIb the fastest contractile speed. Over the last ~35 years, a large body of knowledge suggests that altered loading state cause both fiber atrophy/wasting and a slow to fast shift in the contractile phenotype in the target muscle(s. Hence, this review will examine findings from three different animal models of unloading: 1 space flight (SF, i.e., microgravity; 2 hindlimb suspension (HS, a procedure that chronically eliminates weight bearing of the lower limbs; and 3 spinal cord isolation (SI, a surgical procedure that eliminates neural activation of the motoneurons and associated muscles while maintaining neurotrophic motoneuron-muscle connectivity. The collective findings demonstrate: 1 all three models show a similar pattern of fiber atrophy with differences mainly in the magnitude and kinetics of alteration; 2 transcriptional/pretranslational processes play a major role in both the atrophy process and phenotype shifts; and 3 signaling pathways impacting these alterations appear to be similar in each of the models

  7. Structural and molecular characterization of Kudoa quraishii n. sp. from the trunk muscle of the Indian mackerel Rastrelliger kanagurta (Perciforme, Scombridae) in Saudi Arabia coasts.

    Science.gov (United States)

    Mansour, Lamjed; Harrath, Abdel Halim; Abd-Elkader, Omar H; Alwasel, Saleh; Abdel-Baki, Abdel-Azeem S; Al Omar, Suliman Y

    2014-04-01

    A new Myxozoa, Kudoa quraishii n. sp., is reported in the striated muscle of the Indian mackerel Rastrelliger kanagurta from the Red Sea and the Arabian Gulf in Saudi Arabia. Mean prevalence of infection is about 20% and varies between localities. The parasite develops whitish and oval or rounded pseudocysts of 0.2-3 mm in the striated muscles of the body. Pseudocysts are filled with mature spores. Myxospores are quadrate in shape in apical view with rounded edges and ovoid in side view. Each spore is formed by four equal shell valves and four symmetrical polar capsules. Polar capsules are pyriform in apical view and drop-like in side view. Myxospore measurements in micrometers are 6.14 (5.9-6.34) in width, 5.48 (5.3-5.71) in thickness, and 4.27 (4.1-4.42) in length. Polar capsule measurements in apical view in micrometers are 2.08 (1.88-2.28) and 1.31 (1.10-1.52) length by width. Molecular analysis based on SSU rDNA gene shows closest association with K. amamiensis and K. kenti with respectively 98 and 97.2% of similarities.

  8. Changes in muscle cell metabolism and mechanotransduction are associated with myopathic phenotype in a mouse model of collagen VI deficiency.

    Directory of Open Access Journals (Sweden)

    Sara De Palma

    Full Text Available This study identifies metabolic and protein phenotypic alterations in gastrocnemius, tibialis anterior and diaphragm muscles of Col6a1(-/- mice, a model of human collagen VI myopathies. All three muscles of Col6a1(-/- mice show some common changes in proteins involved in metabolism, resulting in decreased glycolysis and in changes of the TCA cycle fluxes. These changes lead to a different fate of α-ketoglutarate, with production of anabolic substrates in gastrocnemius and tibialis anterior, and with lipotoxicity in diaphragm. The metabolic changes are associated with changes of proteins involved in mechanotransduction at the myotendineous junction/costameric/sarcomeric level (TN-C, FAK, ROCK1, troponin I fast and in energy metabolism (aldolase, enolase 3, triose phosphate isomerase, creatine kinase, adenylate kinase 1, parvalbumin, IDH1 and FASN. Together, these change may explain Ca(2+ deregulation, impaired force development, increased muscle-relaxation-time and fiber damage found in the mouse model as well as in patients. The severity of these changes differs in the three muscles (gastrocnemiusmuscle morphology.

  9. Effect of South African beef production systems on post-mortem muscle energy status and meat quality.

    Science.gov (United States)

    Frylinck, L; Strydom, P E; Webb, E C; du Toit, E

    2013-04-01

    Post-slaughter muscle energy metabolism meat colour of South African production systems were compared; steers (n=182) of Nguni, Simmental Brahman crossbreds were reared on pasture until A-, AB-, or B-age, in feedlot until A-AB-age. After exsanguination carcasses were electrically stimulated (400 V for 15 s). M. longissimus dorsi muscle energy samples were taken at 1, 2, 4 and 20 h. Post-mortem samples for meat quality studies were taken at 1, 7 and 14 days post-mortem. Production systems affected muscle glycogen, glucose, glucose-6-P, lactic acid, ATP, creatine-P glycolytic potential (P0.5) water holding capacity, drip loss, and Warner Bratzler shear force. Muscle energy only affected muscle contraction of the A-age-pasture system (shortest sarcomere length of 1.66 μm vs 1.75 μm highest WBS of 6 kg vs 5 kg 7 days post-mortem). Copyright © 2012 Elsevier Ltd. All rights reserved.

  10. Changes in Muscle Cell Metabolism and Mechanotransduction Are Associated with Myopathic Phenotype in a Mouse Model of Collagen VI Deficiency

    Science.gov (United States)

    De Palma, Sara; Leone, Roberta; Grumati, Paolo; Vasso, Michele; Polishchuk, Roman; Capitanio, Daniele; Braghetta, Paola; Bernardi, Paolo; Bonaldo, Paolo; Gelfi, Cecilia

    2013-01-01

    This study identifies metabolic and protein phenotypic alterations in gastrocnemius, tibialis anterior and diaphragm muscles of Col6a1−/− mice, a model of human collagen VI myopathies. All three muscles of Col6a1−/− mice show some common changes in proteins involved in metabolism, resulting in decreased glycolysis and in changes of the TCA cycle fluxes. These changes lead to a different fate of α-ketoglutarate, with production of anabolic substrates in gastrocnemius and tibialis anterior, and with lipotoxicity in diaphragm. The metabolic changes are associated with changes of proteins involved in mechanotransduction at the myotendineous junction/costameric/sarcomeric level (TN-C, FAK, ROCK1, troponin I fast) and in energy metabolism (aldolase, enolase 3, triose phosphate isomerase, creatine kinase, adenylate kinase 1, parvalbumin, IDH1 and FASN). Together, these change may explain Ca2+ deregulation, impaired force development, increased muscle-relaxation-time and fiber damage found in the mouse model as well as in patients. The severity of these changes differs in the three muscles (gastrocnemiusmass-to-tendon (myotendineous junction) ratio and to muscle morphology. PMID:23437220

  11. Transgenic overexpression of LARGE induces α-dystroglycan hyperglycosylation in skeletal and cardiac muscle.

    Directory of Open Access Journals (Sweden)

    Martin Brockington

    2010-12-01

    Full Text Available LARGE is one of seven putative or demonstrated glycosyltransferase enzymes defective in a common group of muscular dystrophies with reduced glycosylation of α-dystroglycan. Overexpression of LARGE induces hyperglycosylation of α-dystroglycan in both wild type and in cells from dystroglycanopathy patients, irrespective of their primary gene defect, restoring functional glycosylation. Viral delivery of LARGE to skeletal muscle in animal models of dystroglycanopathy has identical effects in vivo, suggesting that the restoration of functional glycosylation could have therapeutic applications in these disorders. Pharmacological strategies to upregulate Large expression are also being explored.In order to asses the safety and efficacy of long term LARGE over-expression in vivo, we have generated four mouse lines expressing a human LARGE transgene. On observation, LARGE transgenic mice were indistinguishable from the wild type littermates. Tissue analysis from young mice of all four lines showed a variable pattern of transgene expression: highest in skeletal and cardiac muscles, and lower in brain, kidney and liver. Transgene expression in striated muscles correlated with α-dystroglycan hyperglycosylation, as determined by immunoreactivity to antibody IIH6 and increased laminin binding on an overlay assay. Other components of the dystroglycan complex and extracellular matrix ligands were normally expressed, and general muscle histology was indistinguishable from wild type controls. Further detailed muscle physiological analysis demonstrated a loss of force in response to eccentric exercise in the older, but not in the younger mice, suggesting this deficit developed over time. However this remained a subclinical feature as no pathology was observed in older mice in any muscles including the diaphragm, which is sensitive to mechanical load-induced damage.This work shows that potential therapies in the dystroglycanopathies based on LARGE upregulation

  12. Growth and the regulation of myotomal muscle mass in teleost fish.

    Science.gov (United States)

    Johnston, Ian A; Bower, Neil I; Macqueen, Daniel J

    2011-05-15

    Teleost muscle first arises in early embryonic life and its development is driven by molecules present in the egg yolk and modulated by environmental stimuli including temperature and oxygen. Several populations of myogenic precursor cells reside in the embryonic somite and external cell layer and contribute to muscle fibres in embryo, larval, juvenile and adult stages. Many signalling proteins and transcription factors essential for these events are known. In all cases, myogenesis involves myoblast proliferation, migration, fusion and terminal differentiation. Maturation of the embryonic muscle is associated with motor innervation and the development of a scaffold of connective tissue and complex myotomal architecture needed to generate swimming behaviour. Adult muscle is a heterogeneous tissue composed of several cell types that interact to affect growth patterns. The development of capillary and lymphatic circulations and extramuscular organs--notably the gastrointestinal, endocrine, neuroendocrine and immune systems--serves to increase information exchange between tissues and with the external environment, adding to the complexity of growth regulation. Teleosts often exhibit an indeterminate growth pattern, with body size and muscle mass increasing until mortality or senescence occurs. The dramatic increase in myotomal muscle mass between embryo and adult requires the continuous production of muscle fibres until 40-50% of the maximum body length is reached. Sarcomeric proteins can be mobilised as a source of amino acids for energy metabolism by other tissues and for gonad generation, requiring the dynamic regulation of muscle mass throughout the life cycle. The metabolic and contractile phenotypes of muscle fibres also show significant plasticity with respect to environmental conditions, migration and spawning. Many genes regulating muscle growth are found as multiple copies as a result of paralogue retention following whole-genome duplication events in teleost

  13. Identification of an FHL1 protein complex containing gamma-actin and non-muscle myosin IIB by analysis of protein-protein interactions.

    Directory of Open Access Journals (Sweden)

    Lili Wang

    Full Text Available FHL1 is multifunctional and serves as a modular protein binding interface to mediate protein-protein interactions. In skeletal muscle, FHL1 is involved in sarcomere assembly, differentiation, growth, and biomechanical stress. Muscle abnormalities may play a major role in congenital clubfoot (CCF deformity during fetal development. Thus, identifying the interactions of FHL1 could provide important new insights into its functional role in both skeletal muscle development and CCF pathogenesis. Using proteins derived from rat L6GNR4 myoblastocytes, we detected FHL1 interacting proteins by immunoprecipitation. Samples were analyzed by liquid chromatography mass spectrometry (LC-MS. Dynamic gene expression of FHL1 was studied. Additionally, the expression of the possible interacting proteins gamma-actin and non-muscle myosin IIB, which were isolated from the lower limbs of E14, E15, E17, E18, E20 rat embryos or from adult skeletal muscle was analyzed. Potential interacting proteins isolated from E17 lower limbs were verified by immunoprecipitation, and co-localization in adult gastrocnemius muscle was visualized by fluorescence microscopy. FHL1 expression was associated with skeletal muscle differentiation. E17 was found to be the critical time-point for skeletal muscle differentiation in the lower limbs of rat embryos. We also identified gamma-actin and non-muscle myosin IIB as potential binding partners of FHL1, and both were expressed in adult skeletal muscle. We then demonstrated that FHL1 exists as part of a complex, which binds gamma-actin and non-muscle myosin IIB.

  14. Conservation and divergence between cytoplasmic and muscle-specific actin capping proteins: insights from the crystal structure of cytoplasmic Cap32/34 from Dictyostelium discoideum

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    Eckert Christian

    2012-06-01

    Full Text Available Abstract Background Capping protein (CP, also known as CapZ in muscle cells and Cap32/34 in Dictyostelium discoideum, plays a major role in regulating actin filament dynamics. CP is a ubiquitously expressed heterodimer comprising an α- and β-subunit. It tightly binds to the fast growing end of actin filaments, thereby functioning as a “cap” by blocking the addition and loss of actin subunits. Vertebrates contain two somatic variants of CP, one being primarily found at the cell periphery of non-muscle tissues while the other is mainly localized at the Z-discs of skeletal muscles. Results To elucidate structural and functional differences between cytoplasmic and sarcomercic CP variants, we have solved the atomic structure of Cap32/34 (32 = β- and 34 = α-subunit from the cellular slime mold Dictyostelium at 2.2 Å resolution and compared it to that of chicken muscle CapZ. The two homologs display a similar overall arrangement including the attached α-subunit C-terminus (α-tentacle and the flexible β-tentacle. Nevertheless, the structures exhibit marked differences suggesting considerable structural flexibility within the α-subunit. In the α-subunit we observed a bending motion of the β-sheet region located opposite to the position of the C-terminal β-tentacle towards the antiparallel helices that interconnect the heterodimer. Recently, a two domain twisting attributed mainly to the β-subunit has been reported. At the hinge of these two domains Cap32/34 contains an elongated and highly flexible loop, which has been reported to be important for the interaction of cytoplasmic CP with actin and might contribute to the more dynamic actin-binding of cytoplasmic compared to sarcomeric CP (CapZ. Conclusions The structure of Cap32/34 from Dictyostelium discoideum allowed a detailed analysis and comparison between the cytoplasmic and sarcomeric variants of CP. Significant structural flexibility could particularly be found within the

  15. Composição de fibras musculares esqueléticas de eqüinos jovens da raça Brasileiro de Hipismo Composition of skeletal muscle fibers of young Brasileiro de Hipismo horse breed

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    F.H.F. D’Angelis

    2006-08-01

    Full Text Available The aim of this study was to typify the skeletal striated fibers of the gluteus medius muscle of young Brasileiro de Hipismo (BH horses by means of histochemical analysis with m-ATPase and NADH-TR according to the sex and the biopsy depth. It was observed that the frequency (F;% and the relative cross sectional area (RCSA;% of the fibers type IIX were greater than the fibers type IIA, which F and RCSA were greater than the fibers type I. The comparison between sex and muscles depht, showed no significant difference in F and RCSA in the three types of fibers. The results of morphometry showed that the gluteus medius muscle has greater glycolitic metabolism and anaerobic capacity because of the presence of large proportion of type IIX fibers. This may be justified by the genetic influence of Thoroughbred in the formation of Brasileiro de Hipismo breed.

  16. Aging Enhances Indirect Flight Muscle Fiber Performance yet Decreases Flight Ability in Drosophila

    Energy Technology Data Exchange (ETDEWEB)

    Miller, Mark S.; Lekkas, Panagiotis; Braddock, Joan M.; Farman, Gerrie P.; Ballif, Bryan A.; Irving, Thomas C.; Maughan, David W.; Vigoreaux, Jim O. (IIT); (Vermont)

    2008-10-02

    We investigated the effects of aging on Drosophila melanogaster indirect flight muscle from the whole organism to the actomyosin cross-bridge. Median-aged (49-day-old) flies were flight impaired, had normal myofilament number and packing, barely longer sarcomeres, and slight mitochondrial deterioration compared with young (3-day-old) flies. Old (56-day-old) flies were unable to beat their wings, had deteriorated ultrastructure with severe mitochondrial damage, and their skinned fibers failed to activate with calcium. Small-amplitude sinusoidal length perturbation analysis showed median-aged indirect flight muscle fibers developed greater than twice the isometric force and power output of young fibers, yet cross-bridge kinetics were similar. Large increases in elastic and viscous moduli amplitude under active, passive, and rigor conditions suggest that median-aged fibers become stiffer longitudinally. Small-angle x-ray diffraction indicates that myosin heads move increasingly toward the thin filament with age, accounting for the increased transverse stiffness via cross-bridge formation. We propose that the observed protein composition changes in the connecting filaments, which anchor the thick filaments to the Z-disk, produce compensatory increases in longitudinal stiffness, isometric tension, power and actomyosin interaction in aging indirect flight muscle. We also speculate that a lack of MgATP due to damaged mitochondria accounts for the decreased flight performance.

  17. Titin-based stiffening of muscle fibers in Ehlers-Danlos Syndrome

    Energy Technology Data Exchange (ETDEWEB)

    Ottenheijm, Coen A.C.; Voermans, Nicol C.; Hudson, Bryan D.; Irving, Thomas; Stienen, Ger J.M.; van Engelen, Baziel G.; Granzier, Henk (IIT); (Radboud); (Ariz); (Vrije)

    2012-05-09

    Tenascin-X (TNX) is an extracellular matrix glycoprotein whose absence leads to Ehlers-Danlos Syndrome (EDS). TNX-deficient EDS patients present with joint hypermobility and muscle weakness attributable to increased compliance of the extracellular matrix. We hypothesized that in response to the increased compliance of the extracellular matrix in TNX-deficient EDS patients, intracellular adaptations take place in the elastic properties of the giant muscle protein titin. We performed extensive single muscle fiber mechanical studies to determine active and passive properties in TNX-deficient EDS patients. Gel-electrophoresis, Western blotting, and microarray studies were used to evaluate titin expression and phosphorylation. X-ray diffraction was used to measure myofilament lattice spacing. Passive tension of muscle fibers from TNX-deficient EDS patients was markedly increased. Myofilament extraction experiments indicated that the increased passive tension is attributable to changes in the properties of the sarcomeric protein titin. Transcript and protein data indicated no changes in titin isoform expression. Instead, differences in posttranslational modifications within titin's elastic region were found. In patients, active tension was not different at maximal activation level, but at submaximal activation level it was augmented attributable to increased calcium sensitivity. This increased calcium sensitivity might be attributable to stiffer titin molecules. In response to the increased compliance of the extracellular matrix in muscle of TNX-deficient EDS patients, a marked intracellular stiffening occurs of the giant protein titin. The stiffening of titin partly compensates for the muscle weakness in these patients by augmenting submaximal active tension generation.

  18. Ultrastructural pathological study on skeletal muscle injury in rabbit after a high-dose radiation

    International Nuclear Information System (INIS)

    Sun Wei; Ni Xinchu; Sun Suping; Cai Leiming; Yu Jingping; Wang Jian; Nie Bin; Sun Zhiqiang; Ni Xinye; Cao Xiufeng

    2012-01-01

    Objective: To establish a rabbit model of radiation-induced skeletal muscle injury in order to study the ultrastructural pathological changes and underlying mechanism. Methods: 28 New Zealand rabbits were randomly divided into 2 groups with 16 rabbits in experimental group and 12 rabbits in control group. The experimental rabbits were irradiated on hip with a single dose of 80 Gy of 9 MeV electrons from a linear accelerator. 1 month and 6 months after irradiation the pathological changes were respectively observed under light microscope and electron microscope. Results: One month after irradiation, the morphologic changes including degeneration, necrosis of muscle cells, and hemorrhage between the muscle cells were observed under light microscope and the swelling of myofibrillae, blurring of light and shade band, vacuolar degeneration of mitochondria and amorphous areas of necrosis were observed under electron microscope. Six months after irradiation, the morphologic changes of nucleolus chips, fibrous connective tissue, thickening of vascular wall and vascular congestion between the muscle cells and the amorphous areas of necrosis in the experimental group were much more serious than those of 1 month after irradiation. In addition, the myofilaments were lost in degeneration areas and the sarcomere became shorten. Observation with electron microscope showed that the mitochondrial size and its morphological changes were varied and the amounts of collagen between myofibrillaes were increased 6 months after irradiation. Conclusions: A rabbit model of high-dose irradiated skeleton muscle injury was successfully established with a single dose of 80 Gy of 9 MeV electrons from a linear accelerator. The degeneration and necrosis of muscle cells may be promoted by mitochondrial and vascular injury, degeneration of vessel and nerve fiber. (authors)

  19. Imaging of muscle injuries

    Energy Technology Data Exchange (ETDEWEB)

    El-Khoury, G.Y. [Iowa Univ., Iowa City, IA (United States). Dept. of Radiology; Brandser, E.A. [Iowa Univ., Iowa City, IA (United States). Dept. of Radiology; Kathol, M.H. [Iowa Univ., Iowa City, IA (United States). Dept. of Radiology; Tearse, D.S. [Iowa Univ., Iowa City, IA (United States). Dept. of Orthopaedic Surgery; Callaghan, J.J. [Iowa Univ., Iowa City, IA (United States). Dept. of Orthopaedic Surgery

    1996-01-01

    Although skeletal muscle is the single largest tissue in the body, there is little written about it in the radiologic literature. Indirect muscle injuries, also called strains or tears, are common in athletics, and knowing the morphology and physiology of the muscle-tendon unit is the key to the understanding of these injuries. Eccentric muscle activation produces more tension within the muscle tan when it is activated concentrically, making it more susceptible to tearing. Injuries involving the muscle belly tend to occur near the myotendinous junction. In adolescents, the weakest link in the muscle-tendon-bone complex is the apophysis. Traditionally, plain radiography has been the main diagnostic modality for evaluation of these injuries; however, with the advent of MRI it has become much easier to diagnose injuries primarily affecting the soft tissues. This article reviews the anatomy and physiology of the muscle-tendon unit as they relate to indirect muscle injuries. Examples of common muscle injuries are illustrated. (orig.)

  20. Meat quality of broiler breast fillets with white striping and woody breast muscle myopathies.

    Science.gov (United States)

    Tijare, V V; Yang, F L; Kuttappan, V A; Alvarado, C Z; Coon, C N; Owens, C M

    2016-09-01

    The global poultry industry has been faced with emerging broiler breast meat quality issues including conditions known as white striping (WS, white striations parallel to muscle fibers) and woody breast (WB, hardness of raw fillet). Experiments were conducted to evaluate effects of WS and WB hardness on meat quality traits in broiler breast fillets. In Exp. 1, birds were processed at approximately 9 wk of age and deboned at 4 h postmortem (PM); in Exp. 2, birds were processed at approximately 6 and 9 wk of age and deboned at 2 h PM. Fillets were categorized as: normal for both white striping and woody breast (NORM); moderate for white striping and mild for woody breast (MILD); severe for white striping and mild for woody breast (WS); severe for woody breast and moderate for white striping (WB); or severe for both white striping and woody breast (BOTH). Sarcomere length, gravimetric fragmentation index, marination uptake, cook loss, and Meullenet-Owens razor shear energy (MORSE) values on non-marinated and marinated fillets were assessed. Sarcomeres tended to be longer (P = 0.07) with increasing severity of WS and WB in both experiments and gravimetric fragmentation index did not differ (P > 0.05) among categories. Marinade uptake decreased (P  0.05) in non-marinated fillets, the marinated BOTH fillets had greater MORSE values (P  0.05) among categories of marinated breasts. At 9 wk, WS and BOTH were higher (P white striping and woody breast, individually or in combination, negatively impact meat quality, especially water holding capacity attributes such as marinade uptake and cook loss. © 2016 Poultry Science Association Inc.

  1. Engineering Skeletal Muscle Repair

    OpenAIRE

    Juhas, Mark; Bursac, Nenad

    2013-01-01

    Healthy skeletal muscle has a remarkable capacity for regeneration. Even at a mature age, muscle tissue can undergo a robust rebuilding process that involves the formation of new muscle cells and extracellular matrix and the re-establishment of vascular and neural networks. Understanding and reverse-engineering components of this process is essential for our ability to restore loss of muscle mass and function in cases where the natural ability of muscle for self-repair is exhausted or impaire...

  2. Interplay between ROS and Antioxidants during Ischemia-Reperfusion Injuries in Cardiac and Skeletal Muscle

    Directory of Open Access Journals (Sweden)

    Tingyang Zhou

    2018-01-01

    Full Text Available Ischemia reperfusion (IR, present in myocardial infarction or extremity injuries, is a major clinical issue and leads to substantial tissue damage. Molecular mechanisms underlying IR injury in striated muscles involve the production of reactive oxygen species (ROS. Excessive ROS accumulation results in cellular oxidative stress, mitochondrial dysfunction, and initiation of cell death by activation of the mitochondrial permeability transition pore. Elevated ROS levels can also decrease myofibrillar Ca2+ sensitivity, thereby compromising muscle contractile function. Low levels of ROS can act as signaling molecules involved in the protective pathways of ischemic preconditioning (IPC. By scavenging ROS, antioxidant therapies aim to prevent IR injuries with positive treatment outcomes. Novel therapies such as postconditioning and pharmacological interventions that target IPC pathways hold great potential in attenuating IR injuries. Factors such as aging and diabetes could have a significant impact on the severity of IR injuries. The current paper aims to provide a comprehensive review on the multifaceted roles of ROS in IR injuries, with a focus on cardiac and skeletal muscle, as well as recent advancement in ROS-related therapies.

  3. Mesodermal origin of median fin mesenchyme and tail muscle in amphibian larvae.

    Science.gov (United States)

    Taniguchi, Yuka; Kurth, Thomas; Medeiros, Daniel Meulemans; Tazaki, Akira; Ramm, Robert; Epperlein, Hans-Henning

    2015-06-18

    Mesenchyme is an embryonic precursor tissue that generates a range of structures in vertebrates including cartilage, bone, muscle, kidney, and the erythropoietic system. Mesenchyme originates from both mesoderm and the neural crest, an ectodermal cell population, via an epithelial to mesenchymal transition (EMT). Because ectodermal and mesodermal mesenchyme can form in close proximity and give rise to similar derivatives, the embryonic origin of many mesenchyme-derived tissues is still unclear. Recent work using genetic lineage tracing methods have upended classical ideas about the contributions of mesodermal mesenchyme and neural crest to particular structures. Using similar strategies in the Mexican axolotl (Ambystoma mexicanum), and the South African clawed toad (Xenopus laevis), we traced the origins of fin mesenchyme and tail muscle in amphibians. Here we present evidence that fin mesenchyme and striated tail muscle in both animals are derived solely from mesoderm and not from neural crest. In the context of recent work in zebrafish, our experiments suggest that trunk neural crest cells in the last common ancestor of tetrapods and ray-finned fish lacked the ability to form ectomesenchyme and its derivatives.

  4. Impaired cell surface expression of HLA-B antigens on mesenchymal stem cells and muscle cell progenitors

    DEFF Research Database (Denmark)

    Isa, Adiba; Nehlin, Jan; Sabir, Hardee Jawad

    2010-01-01

    HLA class-I expression is weak in embryonic stem cells but increases rapidly during lineage progression. It is unknown whether all three classical HLA class-I antigens follow the same developmental program. In the present study, we investigated allele-specific expression of HLA-A, -B, and -C...... at the mRNA and protein levels on human mesenchymal stem cells from bone marrow and adipose tissue as well as striated muscle satellite cells and lymphocytes. Using multicolour flow cytometry, we found high cell surface expression of HLA-A on all stem cells and PBMC examined. Surprisingly, HLA-B was either...... undetectable or very weakly expressed on all stem cells protecting them from complement-dependent cytotoxicity (CDC) using relevant human anti-B and anti-Cw sera. IFNgamma stimulation for 48-72 h was required to induce full HLA-B protein expression. Quantitative real-time RT-PCR showed that IFNgamma induced...

  5. Dynamic gene expression in fish muscle during recovery growth induced by a fasting-refeeding schedule

    Directory of Open Access Journals (Sweden)

    Esquerré Diane

    2007-11-01

    Full Text Available Abstract Background Recovery growth is a phase of rapid growth that is triggered by adequate refeeding of animals following a period of weight loss caused by starvation. In this study, to obtain more information on the system-wide integration of recovery growth in muscle, we undertook a time-course analysis of transcript expression in trout subjected to a food deprivation-refeeding sequence. For this purpose complex targets produced from muscle of trout fasted for one month and from muscle of trout fasted for one month and then refed for 4, 7, 11 and 36 days were hybridized to cDNA microarrays containing 9023 clones. Results Significance analysis of microarrays (SAM and temporal expression profiling led to the segregation of differentially expressed genes into four major clusters. One cluster comprising 1020 genes with high expression in muscle from fasted animals included a large set of genes involved in protein catabolism. A second cluster that included approximately 550 genes with transient induction 4 to 11 days post-refeeding was dominated by genes involved in transcription, ribosomal biogenesis, translation, chaperone activity, mitochondrial production of ATP and cell division. A third cluster that contained 480 genes that were up-regulated 7 to 36 days post-refeeding was enriched with genes involved in reticulum and Golgi dynamics and with genes indicative of myofiber and muscle remodelling such as genes encoding sarcomeric proteins and matrix compounds. Finally, a fourth cluster of 200 genes overexpressed only in 36-day refed trout muscle contained genes with function in carbohydrate metabolism and lipid biosynthesis. Remarkably, among the genes induced were several transcriptional regulators which might be important for the gene-specific transcriptional adaptations that underlie muscle recovery. Conclusion Our study is the first demonstration of a coordinated expression of functionally related genes during muscle recovery growth

  6. RNA sequencing reveals a slow to fast muscle fiber type transition after olanzapine infusion in rats.

    Directory of Open Access Journals (Sweden)

    Christopher J Lynch

    Full Text Available Second generation antipsychotics (SGAs, like olanzapine, exhibit acute metabolic side effects leading to metabolic inflexibility, hyperglycemia, adiposity and diabetes. Understanding how SGAs affect the skeletal muscle transcriptome could elucidate approaches for mitigating these side effects. Male Sprague-Dawley rats were infused intravenously with vehicle or olanzapine for 24h using a dose leading to a mild hyperglycemia. RNA-Seq was performed on gastrocnemius muscle, followed by alignment of the data with the Rat Genome Assembly 5.0. Olanzapine altered expression of 1347 out of 26407 genes. Genes encoding skeletal muscle fiber-type specific sarcomeric, ion channel, glycolytic, O2- and Ca2+-handling, TCA cycle, vascularization and lipid oxidation proteins and pathways, along with NADH shuttles and LDH isoforms were affected. Bioinformatics analyses indicate that olanzapine decreased the expression of slower and more oxidative fiber type genes (e.g., type 1, while up regulating those for the most glycolytic and least metabolically flexible, fast twitch fiber type, IIb. Protein turnover genes, necessary to bring about transition, were also up regulated. Potential upstream regulators were also identified. Olanzapine appears to be rapidly affecting the muscle transcriptome to bring about a change to a fast-glycolytic fiber type. Such fiber types are more susceptible than slow muscle to atrophy, and such transitions are observed in chronic metabolic diseases. Thus these effects could contribute to the altered body composition and metabolic disease olanzapine causes. A potential interventional strategy is implicated because aerobic exercise, in contrast to resistance exercise, can oppose such slow to fast fiber transitions.

  7. Myo/Nog cells: targets for preventing the accumulation of skeletal muscle-like cells in the human lens.

    Directory of Open Access Journals (Sweden)

    Jacquelyn Gerhart

    Full Text Available Posterior capsule opacification (PCO is a vision impairing condition that arises in some patients following cataract surgery. The fibrotic form of PCO is caused by myofibroblasts that may emerge in the lens years after surgery. In the chick embryo lens, myofibroblasts are derived from Myo/Nog cells that are identified by their expression of the skeletal muscle specific transcription factor MyoD, the bone morphogenetic protein inhibitor Noggin, and the epitope recognized by the G8 monoclonal antibody. The goal of this study was to test the hypothesis that depletion of Myo/Nog cells will prevent the accumulation of myofibroblasts in human lens tissue. Myo/Nog cells were present in anterior, equatorial and bow regions of the human lens, cornea and ciliary processes. In anterior lens tissue removed by capsulorhexis, Myo/Nog cells had synthesized myofibroblast and skeletal muscle proteins, including vimentin, MyoD and sarcomeric myosin. Alpha smooth muscle actin (α-SMA was detected in a subpopulation of Myo/Nog cells. Areas of the capsule denuded of epithelial cells were surrounded by Myo/Nog cells. Some of these cell free areas contained a wrinkle in the capsule. Depletion of Myo/Nog cells eliminated cells expressing skeletal muscle proteins in 5-day cultures but did not affect cells immunoreactive for beaded filament proteins that accumulate in differentiating lens epithelial cells. Transforming growth factor-betas 1 and 2 that mediate an epithelial-mesenchymal transition, did not induce the expression of skeletal muscle proteins in lens cells following Myo/Nog cell depletion. This study demonstrates that Myo/Nog cells in anterior lens tissue removed from cataract patients have undergone a partial differentiation to skeletal muscle. Myo/Nog cells appear to be the source of skeletal muscle-like cells in explants of human lens tissue. Targeting Myo/Nog cells with the G8 antibody during cataract surgery may reduce the incidence of PCO.

  8. High resolution X-ray diffraction of muscle using undulator radiation from the Tristan main ring at KEK

    Energy Technology Data Exchange (ETDEWEB)

    Wakabayashi, Katsuzo [Osaka Univ., Toyonaka (Japan). Faculty of Engineering Science; Yagi, Naoto

    1997-04-01

    Results are reported of static and time-resolved X-ray diffraction studies on muscle fibers using a hard X-ray undulator installed in the Tristan main ring at KEK, as an innovative source of synchrotron radiation more intense and better collimated than that available with the Photon Factory bending-magnet beamline. The low divergence of the source made it possible to obtain high-quality diffraction patterns from static state of muscles, clearly resolving, with an angular resolution of ca. 700 nm, closely spaced diffraction peaks arising from the two symmetrical halves of the thick filaments centered on the M lines in a sarcomere. The detailed analysis of the meridional pattern lead to a more precise modeling of the complicated molecular packing of myosin molecules and C-proteins in the thick filaments. Time-resolved experiments using a focusing mirror aimed to prove crossbridge behaviors in muscle fibers by collecting X-ray diffraction data at a 185 {mu}s time resolution. When sinusoidal length changes at 500 Hz with an amplitude of 0.3% of the muscle length were applied to a bundle of several single fibers during active contraction, the intensity of the 14.5 nm meridional reflection changed out of phase with the tension change during the oscillating length change, contrasting to the response in the rigor muscle. The high time-resolved experiments provided an important insight to the molecular mechanism of force generation in muscle. These studies yield a preview of the expected gains for muscle studies from the more widespread use of undulator radiation at third-generation synchrotron source. (author)

  9. Remobilização por alongamento estático cíclico em músculo sóleo de ratos imobilizados em encurtamento Remobilization by cycle static stretching in soleus muscle of rats immobilized in shortening position

    Directory of Open Access Journals (Sweden)

    Evandro Augusto Boian Konno

    2008-04-01

    Full Text Available A fibra muscular tem grandes propriedades plásticas, respondendo a diferentes estímulos com o aumento ou diminuição de sua massa, comprimento e número de sarcômeros em série. O objetivo deste estudo foi analisar os efeitos do alongamento passivo estático em 3 séries diárias de 30 s e da remobilização livre, por um período de duas semanas, no músculo sóleo de ratos imobilizado em posição de encurtamento. Para isso, foram utilizados 12 ratos (Wistar divididos em 2 grupos: G1 (n=6 - músculo sóleo esquerdo (MSE imobilizado e solto na gaiola (GIS; G2 (n=6 - MSE imobilizado e alongado diariamente (GIA. Foram comparadas as variações encontradas entre o MSE e o MSD (direito de cada grupo. As variáveis foram: peso muscular, comprimento muscular, número de sarcômeros em série e comprimento de sarcômeros. Os resultados das variáveis analisadas, comparando o MSE com o MSD foram: peso muscular: GIS=-23,16%, (p=0,0007, GIA-32,43% (p=0,0008; comprimento muscular: GIS=-5,47% (p=0,0120; GIA=-9,99% (p=0,0034; número de sarcômeros em série: GIS=-15,42% (p=0,0047; GIA=-8,08% (p=0,0008; comprimento do sarcômero: GIS=11,16% (p=0,0142; GIA=-1,92% (p=0,3783. Através desses resultados, pode-se concluir que nem o alongamento, nem a remobilização livre promovem a restauração na estrutura dos músculos esqueléticos submetidos à imobilização prolongada.Muscle fibers have great plastic properties, responding to different stimuli with their weight increase or reduction, length and serial sarcomere number. The aim of this study was to analyze the static passive stretching in 3 daily bouts of 30 s and the free remobilization effects for two weeks, in the soleus muscle of rats immobilized in shortening position. For this purpose, 12 rats (Wistar were used and divided in 2 groups: G1 (n=6 - left soleus muscle (LSM immobilized and free in the cage (GIF; G2 (n=6 - LSM immobilized and stretched daily (GIS. The variations found between the LSM

  10. Healthy Muscles Matter

    Science.gov (United States)

    ... do. Exercising, getting enough rest, and eating a balanced diet will help to keep your muscles healthy for ... keep your muscles in good health. Eating a balanced diet will help manage your weight and provide a ...

  11. The length-force behavior and operating length range of squid muscle vary as a function of position in the mantle wall.

    Science.gov (United States)

    Thompson, Joseph T; Shelton, Ryan M; Kier, William M

    2014-06-15

    Hollow cylindrical muscular organs are widespread in animals and are effective in providing support for locomotion and movement, yet are subject to significant non-uniformities in circumferential muscle strain. During contraction of the mantle of squid, the circular muscle fibers along the inner (lumen) surface of the mantle experience circumferential strains 1.3 to 1.6 times greater than fibers along the outer surface of the mantle. This transmural gradient of strain may require the circular muscle fibers near the inner and outer surfaces of the mantle to operate in different regions of the length-tension curve during a given mantle contraction cycle. We tested the hypothesis that circular muscle contractile properties vary transmurally in the mantle of the Atlantic longfin squid, Doryteuthis pealeii. We found that both the length-twitch force and length-tetanic force relationships of the obliquely striated, central mitochondria-poor (CMP) circular muscle fibers varied with radial position in the mantle wall. CMP circular fibers near the inner surface of the mantle produced higher force relative to maximum isometric tetanic force, P0, at all points along the ascending limb of the length-tension curve than CMP circular fibers near the outer surface of the mantle. The mean ± s.d. maximum isometric tetanic stresses at L₀ (the preparation length that produced the maximum isometric tetanic force) of 212 ± 105 and 290 ± 166 kN m(-2) for the fibers from the outer and inner surfaces of the mantle, respectively, did not differ significantly (P=0.29). The mean twitch:tetanus ratios for the outer and inner preparations, 0.60 ± 0.085 and 0.58 ± 0.10, respectively, did not differ significantly (P=0.67). The circular fibers did not exhibit length-dependent changes in contraction kinetics when given a twitch stimulus. As the stimulation frequency increased, L₀ was approximately 1.06 times longer than LTW, the mean preparation length that yielded maximum isometric twitch

  12. Recoil after severing reveals stress fiber contraction mechanisms.

    Science.gov (United States)

    Stachowiak, Matthew R; O'Shaughnessy, Ben

    2009-07-22

    Stress fibers are cellular contractile actomyosin machines central to wound healing, shear stress response, and other processes. Contraction mechanisms have been difficult to establish because stress fibers in cultured cells typically exert isometric tension and present little kinetic activity. In a recent study, living cell stress fibers were severed with laser nanoscissors and recoiled several mum over approximately 5 s. We developed a quantitative model of stress fibers based on known components and available structural information suggesting periodic sarcomeric organization similar to striated muscle. The model was applied to the severing assay and compared to the observed recoil. We conclude that the sarcomere force-length relation is similar to that of muscle with two distinct regions on the ascending limb and that substantial external drag forces act on the recoiling fiber corresponding to effective cytosolic viscosity approximately 10(4) times that of water. This may originate from both nonspecific and specific interactions. The model predicts highly nonuniform contraction with caps of collapsed sarcomeres growing at the severed ends. A directly measurable signature of external drag is that cap length and recoil distance increase at intermediate times as t(1/2). The severing data is consistent with this prediction.

  13. Disassociation of insulin action and Akt/FOXO signaling in skeletal muscle of older Akt-deficient mice

    Science.gov (United States)

    Merrell, Erin; Cinquino, Nicholas; Gaugler, Megan; Ng, Lily

    2012-01-01

    The purpose of the present study was to determine the effect of Akt gene ablation on Akt/Forkhead Box O (FOXO) signaling and atrogene expression. This was accomplished by studying wild-type (WT) and isoform-specific Akt knockout (Akt1−/− and Akt2−/−) mice. The ability of insulin to promote Akt phosphorylation on Ser473 was significantly lower in extensor digitorum longus (EDL) and soleus muscles from Akt1−/− and Akt2−/− mice compared with WT mice. Total Akt1 protein levels were significantly lower in EDL muscles of Akt2−/− mice compared with WT mice, a process that appears to be posttranscriptionally regulated as Akt1 mRNA levels were unchanged. The loss of Akt1 protein in EDL muscles of Akt2−/− mice does not appear to be due to insulin resistance because 4 mo of a high-fat diet failed to reduce Akt1 protein levels in muscles of WT mice. Although FOXO3a phosphorylation and atrogin-1 expression were unaltered in muscles of Akt1−/− and Akt2−/− mice, the expression of the atrogenes Bnip3 and gabarapl were significantly elevated in muscles of both Akt1 and Akt2 knockout mice. Finally, the expression of striated activator of Rho signaling was significantly increased in muscles of Akt2−/− mice compared with Akt1−/− and WT mice. Our results demonstrate that the ablation of Akt isoforms disassociates insulin action and Akt/FOXO signaling to atrogenes. PMID:23100026

  14. Obturator internus muscle strains

    Directory of Open Access Journals (Sweden)

    Caoimhe Byrne, MB BCh, BAO

    2017-03-01

    Full Text Available We report 2 cases of obturator internus muscle strains. The injuries occurred in young male athletes involved in kicking sports. Case 1 details an acute obturator internus muscle strain with associated adductor longus strain. Case 2 details an overuse injury of the bilateral obturator internus muscles. In each case, magnetic resonance imaging played a crucial role in accurate diagnosis.

  15. the sternalis muscle

    African Journals Online (AJOL)

    2009-08-17

    Aug 17, 2009 ... scan of the chest wall was performed to gain clarity of the mam- mographic findings (Figs 1a, 1b and 2). The CT scan demonstrated a flattened band of muscle density lying anterior to the medial margin of the pectoralis muscle. This structure was separated from the underlying pectoralis muscle by a thin ...

  16. Architectural properties of the neuromuscular compartments in selected forearm skeletal muscles.

    Science.gov (United States)

    Liu, An-Tang; Liu, Ben-Li; Lu, Li-Xuan; Chen, Gang; Yu, Da-Zhi; Zhu, Lie; Guo, Rong; Dang, Rui-Shan; Jiang, Hua

    2014-07-01

    The purposes f this study were to (i) explore the possibility of splitting the selected forearm muscles into separate compartments in human subjects; (ii) quantify the architectural properties of each neuromuscular compartment; and (iii) discuss the implication of these properties in split tendon transfer procedures. Twenty upper limbs from 10 fresh human cadavers were used in this study. Ten limbs of five cadavers were used for intramuscular nerve study by modified Sihler's staining technique, which confirmed the neuromuscular compartments. The other 10 limbs were included for architectural analysis of neuromuscular compartments. The architectural features of the compartments including muscle weight, muscle length, fiber length, pennation angle, and sarcomere length were determined. Physiological cross-sectional area and fiber length/muscle length ratio were calculated. Five of the selected forearm muscles were ideal candidates for splitting, including flexor carpi ulnaris, flexor carpi radials, extensor carpi radialis brevis, extensor carpi ulnaris and pronator teres. The humeral head of pronator teres contained the longest fiber length (6.23 ± 0.31 cm), and the radial compartment of extensor carpi ulnaris contained the shortest (2.90 ± 0.28 cm). The ulnar compartment of flexor carpi ulnaris had the largest physiological cross-sectional area (5.17 ± 0.59 cm(2)), and the ulnar head of pronator teres had the smallest (0.67 ± 0.06 cm(2)). Fiber length/muscle length ratios of the neuromuscular compartments were relatively low (average 0.27 ± 0.09, range 0.18-0.39) except for the ulnar head of pronator teres, which had the highest one (0.72 ± 0.05). Using modified Sihler's technique, this research demonstrated that each compartment of these selected forearm muscles has its own neurovascular supply after being split along its central tendon. Data of the architectural properties of each neuromuscular compartment provide insight into the 'design' of their

  17. The Structural Biology of Muscle: Spatial and Temporal Aspects

    Science.gov (United States)

    Holmes, Kenneth C.

    Understanding muscle contraction has resulted from the synergy of a ­number of approaches for which structure has provided an integrating framework. Nearly 60 years ago interference and phase contrast light microscopy established the sliding filament model of muscle contraction. A little later H.E. Huxley exploited electron microscopy to visualize the macromolecular architecture of the sarcomere: the thick (myosin) and thin (actin) filaments with connecting myosin cross-bridges. These observations allowed him to outline a structural basis for muscle contraction: a rowing-like progression of the myosin cross-bridges along the actin filament. X-ray fibre diffraction from insect flight muscle first demonstrated that the cross-bridges could indeed take up two configurations that might represent the ends of an active stroke. Later intense X-ray synchrotron radiation allowed the recording of the movements of the cross-bridges during a contraction with high precision. In 1993 Rayments's group ushered in a much more detailed understanding of myosin function by solving the structure of the myosin cross-bridge by X-ray crystallography. It showed that the cross-bridge consists of a large catalytic domain, often called the motor domain, containing the ATP binding site and the actin binding site. At the C-terminus of the motor domain is a long lever arm. The catalytic mechanism is similar to the G-proteins: the active site contains a P-loop and switch 1 and switch 2 elements. The lever arm was later found in two different conformations (so called pre-power-stroke and post-rigor) showing how switch 2 movement is coupled to a swing of the lever arm. Further crystallographic studies coupled with high resolution em reconstructions of decorated actin (the rigor complex) showed how ATP binding sequesters switch 1 thereby opening the large cleft in the motor domain and breaking the strong binding to actin. Conversely, the strong binding to actin causes a movement of switch 1 with

  18. Fiber-type susceptibility to eccentric contraction-induced damage of hindlimb-unloaded rat AL muscles

    Science.gov (United States)

    Vijayan, K.; Thompson, J. L.; Norenberg, K. M.; Fitts, R. H.; Riley, D. A.

    2001-01-01

    Slow oxidative (SO) fibers of the adductor longus (AL) were predominantly damaged during voluntary reloading of hindlimb unloaded (HU) rats and appeared explainable by preferential SO fiber recruitment. The present study assessed damage after eliminating the variable of voluntary recruitment by tetanically activating all fibers in situ through the motor nerve while applying eccentric (lengthening) or isometric contractions. Muscles were aldehyde fixed and resin embedded, and semithin sections were cut. Sarcomere lesions were quantified in toluidine blue-stained sections. Fibers were typed in serial sections immunostained with antifast myosin and antitotal myosin (which highlights slow fibers). Both isometric and eccentric paradigms caused fatigue. Lesions occurred only in eccentrically contracted control and HU muscles. Fatigue did not cause lesions. HU increased damage because lesioned- fiber percentages within fiber types and lesion sizes were greater than control. Fast oxidative glycolytic (FOG) fibers were predominantly damaged. In no case did damaged SO fibers predominate. Thus, when FOG, SO, and hybrid fibers are actively lengthened in chronically unloaded muscle, FOG fibers are intrinsically more susceptible to damage than SO fibers. Damaged hybrid-fiber proportions ranged between these extremes.

  19. Flight capacity of Bactrocera dorsalis (Diptera: Tephritidae) adult females based on flight mill studies and flight muscle ultrastructure.

    Science.gov (United States)

    Chen, Min; Chen, Peng; Ye, Hui; Yuan, Ruiling; Wang, Xiaowei; Xu, Jin

    2015-01-01

    The oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), is considered a major economic threat in many regions worldwide. To better comprehend flight capacity of B. dorsalis and its physiological basis, a computer-monitored flight mill was used to study flight capacity of B. dorsalis adult females of various ages, and the changes of its flight muscle ultrastructures were studied by transmission electron microscopy. The flight capacity (both speed and distance) changed significantly with age of B. dorsalis female adults, peaking at about 15 d; the myofibril diameter of the flight muscle of test insects at 15-d old was the longest, up to 1.56 µm, the sarcomere length at 15-d old was the shortest, averaging at 1.37 µm, volume content of mitochondria of flight muscle at 15-d old reached the peak, it was 32.64%. This study provides the important scientific data for better revealing long-distance movement mechanism of B. dorsalis. © The Author 2015. Published by Oxford University Press on behalf of the Entomological Society of America.

  20. Lipolysis in Skeletal Muscle

    DEFF Research Database (Denmark)

    Serup, Annette Karen Lundbeck

    of AMPK in regulation of lipid handling and lipolysis in the basal non-contracting state and during muscle contractions in skeletal muscle. To evaluate the role of AMPK, we measured protein expression and phosphorylation as well as gene expression of proteins important for regulation of lipid handling...... and lipolysis in skeletal muscle from wildtype mice and mice overexpressing a kinase dead AMPKα2 construct (AMPKα2 KD) in the basal non-contracting state and during in situ stimulated muscle contractions. We found, that IMTG levels were ~50% lower in AMPKα2 KD in the basal resting state, explained by a lower....... IMTG was in wildtype mice reduced with ~50% after muscle contractions with no effect of contractions in AMPKα2 KD mice. Concomitantly, ATGL was phosphorylated at ser406 and HSL on ser565 with muscle contractions in an AMPK dependent manner, suggesting that these sites actives lipolysis during muscle...

  1. Fine-tuning of substrate architecture and surface chemistry promotes muscle tissue development.

    Science.gov (United States)

    Guex, A G; Kocher, F M; Fortunato, G; Körner, E; Hegemann, D; Carrel, T P; Tevaearai, H T; Giraud, M N

    2012-04-01

    Tissue engineering has been increasingly brought to the scientific spotlight in response to the tremendous demand for regeneration, restoration or substitution of skeletal or cardiac muscle after traumatic injury, tumour ablation or myocardial infarction. In vitro generation of a highly organized and contractile muscle tissue, however, crucially depends on an appropriate design of the cell culture substrate. The present work evaluated the impact of substrate properties, in particular morphology, chemical surface composition and mechanical properties, on muscle cell fate. To this end, aligned and randomly oriented micron (3.3±0.8 μm) or nano (237±98 nm) scaled fibrous poly(ε-caprolactone) non-wovens were processed by electrospinning. A nanometer-thick oxygen functional hydrocarbon coating was deposited by a radio frequency plasma process. C2C12 muscle cells were grown on pure and as-functionalized substrates and analysed for viability, proliferation, spatial orientation, differentiation and contractility. Cell orientation has been shown to depend strongly on substrate architecture, being most pronounced on micron-scaled parallel-oriented fibres. Oxygen functional hydrocarbons, representing stable, non-immunogenic surface groups, were identified as strong triggers for myotube differentiation. Accordingly, the highest myotube density (28±15% of total substrate area), sarcomeric striation and contractility were found on plasma-coated substrates. The current study highlights the manifold material characteristics to be addressed during the substrate design process and provides insight into processes to improve bio-interfaces. Copyright © 2012 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  2. Effect of Pulsatile Electric Field on Cultured Muscle Cells in Vitro

    Directory of Open Access Journals (Sweden)

    Shigehiro Hashimoto

    2012-02-01

    Full Text Available An effect of an electric field on proliferation and on differentiation of cultured muscle cells has been studied in vitro. C2C12 (the mouse myoblast cell line originated with the cross-striated muscle of C3H mouse was exposed to electric stimuli. In the first experiment, the adhered cells were exposed to the electric field between two electrodes made of platinum wire of 0.2 mm diameter dipped in the medium at 37 degrees Celsius for 72 hours. The electric pulses at a period of one second with a pulse width of 0.1 second were generated with a function generator. Variation was made on the pulse amplitude < 12 V. The number of adhered cells was counted after exposure to electric stimulation. In the second experiment, the cells were cultivated for 96 hours without electric stimulation in an incubator, after electric stimulation of 0.1 V for 72 hours. After incubation, the movement of myotubes was observed with electric stimulation at a period of one second with a pulse width of one millisecond of 30 V. The experimental results show that cells adhere and proliferate under electric pulses lower than 8 V, and that differentiation accelerates with the electric pulses of 0.1 V.

  3. Shared gene structures and clusters of mutually exclusive spliced exons within the metazoan muscle myosin heavy chain genes.

    Directory of Open Access Journals (Sweden)

    Martin Kollmar

    Full Text Available Multicellular animals possess two to three different types of muscle tissues. Striated muscles have considerable ultrastructural similarity and contain a core set of proteins including the muscle myosin heavy chain (Mhc protein. The ATPase activity of this myosin motor protein largely dictates muscle performance at the molecular level. Two different solutions to adjusting myosin properties to different muscle subtypes have been identified so far: Vertebrates and nematodes contain many independent differentially expressed Mhc genes while arthropods have single Mhc genes with clusters of mutually exclusive spliced exons (MXEs. The availability of hundreds of metazoan genomes now allowed us to study whether the ancient bilateria already contained MXEs, how MXE complexity subsequently evolved, and whether additional scenarios to control contractile properties in different muscles could be proposed, By reconstructing the Mhc genes from 116 metazoans we showed that all intron positions within the motor domain coding regions are conserved in all bilateria analysed. The last common ancestor of the bilateria already contained a cluster of MXEs coding for part of the loop-2 actin-binding sequence. Subsequently the protostomes and later the arthropods gained many further clusters while MXEs got completely lost independently in several branches (vertebrates and nematodes and species (for example the annelid Helobdella robusta and the salmon louse Lepeophtheirus salmonis. Several bilateria have been found to encode multiple Mhc genes that might all or in part contain clusters of MXEs. Notable examples are a cluster of six tandemly arrayed Mhc genes, of which two contain MXEs, in the owl limpet Lottia gigantea and four Mhc genes with three encoding MXEs in the predatory mite Metaseiulus occidentalis. Our analysis showed that similar solutions to provide different myosin isoforms (multiple genes or clusters of MXEs or both have independently been developed

  4. Progressive Structural Defects in Canine Centronuclear Myopathy Indicate a Role for HACD1 in Maintaining Skeletal Muscle Membrane Systems.

    Science.gov (United States)

    Walmsley, Gemma L; Blot, Stéphane; Venner, Kerrie; Sewry, Caroline; Laporte, Jocelyn; Blondelle, Jordan; Barthélémy, Inès; Maurer, Marie; Blanchard-Gutton, Nicolas; Pilot-Storck, Fanny; Tiret, Laurent; Piercy, Richard J

    2017-02-01

    Mutations in HACD1/PTPLA cause recessive congenital myopathies in humans and dogs. Hydroxyacyl-coA dehydratases are required for elongation of very long chain fatty acids, and HACD1 has a role in early myogenesis, but the functions of this striated muscle-specific enzyme in more differentiated skeletal muscle remain unknown. Canine HACD1 deficiency is histopathologically classified as a centronuclear myopathy (CNM). We investigated the hypothesis that muscle from HACD1-deficient dogs has membrane abnormalities in common with CNMs with different genetic causes. We found progressive changes in tubuloreticular and sarcolemmal membranes and mislocalized triads and mitochondria in skeletal muscle from animals deficient in HACD1. Furthermore, comparable membranous abnormalities in cultured HACD1-deficient myotubes provide additional evidence that these defects are a primary consequence of altered HACD1 expression. Our novel findings, including T-tubule dilatation and disorganization, associated with defects in this additional CNM-associated gene provide a definitive pathophysiologic link with these disorders, confirm that dogs deficient in HACD1 are relevant models, and strengthen the evidence for a unifying pathogenesis in CNMs via defective membrane trafficking and excitation-contraction coupling in muscle. These results build on previous work by determining further functional roles of HACD1 in muscle and provide new insight into the pathology and pathogenetic mechanisms of HACD1 CNM. Consequently, alterations in membrane properties associated with HACD1 mutations should be investigated in humans with related phenotypes. Copyright © 2017 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.

  5. Myocardial fibrosis in patients with symptomatic obstructive hypertrophic cardiomyopathy: correlation with echocardiographic measurements, sarcomeric genotypes, and pro-left ventricular hypertrophy polymorphisms involving the renin-angiotensin-aldosterone system.

    Science.gov (United States)

    Blauwet, Lori A; Ackerman, Michael J; Edwards, William D; Riehle, Darren L; Ommen, Steve R

    2009-01-01

    Hypertrophic cardiomyopathy (HCM) is a heterogeneous disorder of the cardiac sarcomere, resulting in myocyte hypertrophy and disarray, interstitial fibrosis, and cardiac dysfunction. Our aim was to determine whether the amount of fibrosis in HCM correlates with echocardiographic measures of diastolic dysfunction, presence of HCM-susceptibility mutations, or polymorphisms in the renin-angiotensin-aldosterone system (RAAS). Surgical specimens from patients with obstructive HCM undergoing septal myectomy at the Mayo Clinic (2001-2004) were examined and compared with autopsy-derived tissues from age- and sex-matched normal controls. Digital image analysis was used to quantitate the fibrosis in representative microscopic sections. Genotyping was performed for myofilament-HCM using polymerase chain reaction, high-performance liquid chromatography, and direct DNA sequencing. RAAS polymorphism status was similarly established. The study included 59 HCM cases and 44 controls. Patients with HCM exhibited more fibrosis (mean 17%, range 3-45%) than controls (mean 8%, range 3-17%) (P or =1 C-encoding allele in CYP11B2-encoded aldosterone synthase. Patients with HCM undergoing septal myectomy had significantly more myocardial interstitial fibrosis than controls. The amount of fibrosis in HCM patients correlated with degree of septal hypertrophy and left ventricular systolic and diastolic function. Notably, neither mutations in cardiac myofilament proteins or polymorphisms in RAAS exhibited strong associations with severity of myocardial fibrosis.

  6. The impact of prolonged hyperinsulinaemia on glucose transport in equine skeletal muscle and digital lamellae.

    Science.gov (United States)

    de Laat, M A; Clement, C K; Sillence, M N; McGowan, C M; Pollitt, C C; Lacombe, V A

    2015-07-01

    An increased incidence of metabolic disease in horses has led to heightened recognition of the pathological consequences of insulin resistance. Laminitis, failure of the weightbearing digital lamellae, is an important consequence. Altered trafficking of specialised glucose transporters (GLUTs), responsible for glucose uptake, is central to the dysregulation of glucose metabolism and may play a role in the pathophysiology of laminitis. We hypothesised that prolonged hyperinsulinaemia alters the regulation of glucose transport in insulin-sensitive tissue and digital lamellae. Our objectives were to compare the relative protein expression of major GLUT isoforms in striated muscle and digital lamellae in healthy horses and during marked and moderate hyperinsulinaemia. Randomised, controlled study. Prolonged hyperinsulinaemia and lamellar damage were induced by a prolonged euglycaemic-hyperinsulinaemic clamp or a prolonged glucose infusion, and results were compared with those of electrolyte-treated control animals. Protein expression of GLUTs was examined with immunoblotting. Lamellar tissue contained more GLUT1 protein than skeletal muscle (P = 0.002) and less GLUT4 than the heart (P = 0.037). During marked hyperinsulinaemia and acute laminitis (induced by the prolonged euglycaemic-hyperinsulinaemic clamp), GLUT1 protein expression was decreased in skeletal muscle (P = 0.029) but unchanged in the lamellae, while novel GLUTs (8 and 12) were increased in the lamellae (P = 0.03) but not in skeletal muscle. However, moderate hyperinsulinaemia and subclinical laminitis (induced by the prolonged glucose infusion) did not cause differential GLUT protein expression in the lamellae compared with control horses. The results suggest that lamellar tissue functions independently of insulin and that insulin resistance may not be an essential component of the aetiology of laminitis. Marked differences in GLUT expression exist between insulin-sensitive and insulin-independent tissues

  7. Bion 11 Spaceflight Project: Effect of Weightlessness on Single Muscle Fiber Function in Rhesus Monkeys

    Science.gov (United States)

    Fitts, Robert H.; Romatowski, Janell G.; Widrick, Jeffrey J.; DeLaCruz, Lourdes

    1999-01-01

    Although it is well known that microgravity induces considerable limb muscle atrophy, little is known about how weightlessness alters cell function. In this study, we investigated how weightlessness altered the functional properties of single fast and slow striated muscle fibers. Physiological studies were carried out to test the hypothesis that microgravity causes fiber atrophy, a decreased peak force (Newtons), tension (Newtons/cross-sectional area) and power, an elevated peak rate of tension development (dp/dt), and an increased maximal shortening velocity (V(sub o)) in the slow type I fiber, while changes in the fast-twitch fiber are restricted to atrophy and a reduced peak force. For each fiber, we determined the peak force (P(sub o)), V(sub o), dp/dt, the force-velocity relationship, peak power, the power-force relationship, the force-pCa relationship, and fiber stiffness. Biochemical studies were carried out to assess the effects of weightlessness on the enzyme and substrate profile of the fast- and slow-twitch fibers. We predicted that microgravity would increase resting muscle glycogen and glycolytic metabolism in the slow fiber type, while the fast-twitch fiber enzyme profile would be unaltered. The increased muscle glycogen would in part result from an elevated hexokinase and glycogen synthase. The enzymes selected for study represent markers for mitochondrial function (citrate synthase and 0-hydroxyacyl-CoA dehydrogenase), glycolysis (Phosphofructokinase and lactate dehydrogenase), and fatty acid transport (Carnitine acetyl transferase). The substrates analyzed will include glycogen, lactate, adenosine triphosphate, and phosphocreatine.

  8. Efeitos da corrida em esteira em músculos sóleos de ratos encurtados por imobilização Effects of running on treadmill in soleus muscles of rats shortened by immobilization

    Directory of Open Access Journals (Sweden)

    Luiz Henrique Natali

    2008-12-01

    Full Text Available O objetivo deste trabalho foi verificar as adaptações de peso e comprimento do músculo sóleo de ratos Wistar machos, além da estimativa do total de sarcômeros em série e comprimento médio dos sarcômeros, quando submetidos a um processo de remobilização em esteira. Foram utilizados 18 ratos (Wistar, divididos em três grupos: GC - músculo sóleo esquerdo (MSE imobilizado e remobilizado solto; G10 - MSE imobilizado e remobilizado em velocidade de 10m/min; e G12 (n = 6 - MSE imobilizado e remobilizado em velocidade de 12m/min. Os resultados mostraram as seguintes variações, peso muscular: GC -22,35% (p = 0,0089, G10 -12,52% (p = 0,0623, G12 -12,07%, (p = 0,0004; comprimento muscular: GC -5,47% (p = 0,0120, G10 -3,31% (p = 0,2868, G12 0,41% (p = 0,8987; estimativa de sarcômeros em série: GC -15,42% (p = 0,0047, G10 -10,87% (p = 0,0193, G12 -4,97 (p = 0,2409; comprimento de sarcômeros GC 11,16% (p = 0,0142, G10 9,31% (p = 0,1270, G12 5,58% (p = 0,1327. Conclui-se que G12 obteve maior eficácia após o período de imobilização, pois apresentou maior semelhança com o membro não imobilizado.The aim of this work was to compare weight and length adaptations of the soleus muscle of male Wistar rats as well as estimation of the total number of serial sarcomere and mean sarcomere length, when they are submitted to remobilization on treadmill. 18 Wistar male rats were used and divided in the three following groups: CG - left soleus (LS muscle immobilized and remobilized free in the cage; G10 - LS muscle immobilized and remobilized in speed of 10 m/min on treadmill; and G12 - LS muscle immobilized and remobilized in speed of 12 m/min. The right muscles (RS of each animal were used for comparison. The results showed the following variations, muscular weigh: CG -22.35% (p = 0.0089, G10 -12.52% (p = 0.0623, G12 -12.07%, (p = 0.0004; muscle length: CG -5.47% (p = 0.0120, G10 -3.31% (p = 0.2868, G12 0.41% (p = 0.8987; estimation of number of

  9. The hamstring muscle complex.

    Science.gov (United States)

    van der Made, A D; Wieldraaijer, T; Kerkhoffs, G M; Kleipool, R P; Engebretsen, L; van Dijk, C N; Golanó, P

    2015-07-01

    The anatomical appearance of the hamstring muscle complex was studied to provide hypotheses for the hamstring injury pattern and to provide reference values of origin dimensions, muscle length, tendon length, musculotendinous junction (MTJ) length as well as width and length of a tendinous inscription in the semitendinosus muscle known as the raphe. Fifty-six hamstring muscle groups were dissected in prone position from 29 human cadaveric specimens with a median age of 71.5 (range 45-98). Data pertaining to origin dimensions, muscle length, tendon length, MTJ length and length as well as width of the raphe were collected. Besides these data, we also encountered interesting findings that might lead to a better understanding of the hamstring injury pattern. These include overlapping proximal and distal tendons of both the long head of the biceps femoris muscle and the semimembranosus muscle (SM), a twist in the proximal SM tendon and a tendinous inscription (raphe) in the semitendinosus muscle present in 96 % of specimens. No obvious hypothesis can be provided purely based on either muscle length, tendon length or MTJ length. However, it is possible that overlapping proximal and distal tendons as well as muscle architecture leading to a resultant force not in line with the tendon predispose to muscle injury, whereas the presence of a raphe might plays a role in protecting the muscle against gross injury. Apart from these architectural characteristics that may contribute to a better understanding of the hamstring injury pattern, the provided reference values complement current knowledge on surgically relevant hamstring anatomy. IV.

  10. GRAF1 deficiency blunts sarcolemmal injury repair and exacerbates cardiac and skeletal muscle pathology in dystrophin-deficient mice.

    Science.gov (United States)

    Lenhart, Kaitlin C; O'Neill, Thomas J; Cheng, Zhaokang; Dee, Rachel; Demonbreun, Alexis R; Li, Jianbin; Xiao, Xiao; McNally, Elizabeth M; Mack, Christopher P; Taylor, Joan M

    2015-01-01

    The plasma membranes of striated muscle cells are particularly susceptible to rupture as they endure significant mechanical stress and strain during muscle contraction, and studies have shown that defects in membrane repair can contribute to the progression of muscular dystrophy. The synaptotagmin-related protein, dysferlin, has been implicated in mediating rapid membrane repair through its ability to direct intracellular vesicles to sites of membrane injury. However, further work is required to identify the precise molecular mechanisms that govern dysferlin targeting and membrane repair. We previously showed that the bin-amphiphysin-Rvs (BAR)-pleckstrin homology (PH) domain containing Rho-GAP GTPase regulator associated with focal adhesion kinase-1 (GRAF1) was dynamically recruited to the tips of fusing myoblasts wherein it promoted membrane merging by facilitating ferlin-dependent capturing of intracellular vesicles. Because acute membrane repair responses involve similar vesicle trafficking complexes/events and because our prior studies in GRAF1-deficient tadpoles revealed a putative role for GRAF1 in maintaining muscle membrane integrity, we postulated that GRAF1 might also play an important role in facilitating dysferlin-dependent plasma membrane repair. We used an in vitro laser-injury model to test whether GRAF1 was necessary for efficient muscle membrane repair. We also generated dystrophin/GRAF1 doubledeficient mice by breeding mdx mice with GRAF1 hypomorphic mice. Evans blue dye uptake and extensive morphometric analyses were used to assess sarcolemmal integrity and related pathologies in cardiac and skeletal muscles isolated from these mice. Herein, we show that GRAF1 is dynamically recruited to damaged skeletal and cardiac muscle plasma membranes and that GRAF1-depleted muscle cells have reduced membrane healing abilities. Moreover, we show that dystrophin depletion exacerbated muscle damage in GRAF1-deficient mice and that mice with dystrophin/GRAF1

  11. MUSCLE INJURIES IN ATHLETES.

    Science.gov (United States)

    Barroso, Guilherme Campos; Thiele, Edilson Schwansee

    2011-01-01

    This article had the aim of demonstrating the physiology, diagnosis and treatment of muscle injuries, focusing on athletes and their demands and expectations. Muscle injuries are among the most common complaints in orthopedic practice, occurring both among athletes and among non-athletes. These injuries present a challenge for specialists, due to the slow recovery, during which time athletes are unable to take part in training and competitions, and due to frequent sequelae and recurrences of the injuries. Most muscle injuries (between 10% and 55% of all injuries) occur during sports activities. The muscles most commonly affected are the ischiotibial, quadriceps and gastrocnemius. These muscles go across two joints and are more subject to acceleration and deceleration forces. The treatment for muscle injuries varies from conservative treatment to surgery. New procedures are being used, like the hyperbaric chamber and the use of growth factors. However, there is still a high rate of injury recurrence. Muscle injury continues to be a topic of much controversy. New treatments are being researched and developed, but prevention through muscle strengthening, stretching exercises and muscle balance continues to be the best "treatment".

  12. An artificial muscle computer

    Science.gov (United States)

    Marc O'Brien, Benjamin; Alexander Anderson, Iain

    2013-03-01

    We have built an artificial muscle computer based on Wolfram's "2, 3" Turing machine architecture, the simplest known universal Turing machine. Our computer uses artificial muscles for its instruction set, output buffers, and memory write and addressing mechanisms. The computer is very slow and large (0.15 Hz, ˜1 m3); however by using only 13 artificial muscle relays, it is capable of solving any computable problem given sufficient memory, time, and reliability. The development of this computer shows that artificial muscles can think—paving the way for soft robots with reflexes like those seen in nature.

  13. Accessory piriformis muscle

    Directory of Open Access Journals (Sweden)

    Sedat Develi

    2017-03-01

    Full Text Available Piriformis muscle originates from facies pelvica of sacrum and inserts on the trochanter major. It is one of the lateral rotator muscles of the hip and a landmark point in the gluteal region since n. ischiadicus descends to the thigh by passing close to the muscle. This contiguity may be associated with the irritation of the nerve which is known as piriformis syndrome. A rare anatomic variation of the muscle which observed on 74 years old male cadaver is discussed in this case report. [Cukurova Med J 2017; 42(1.000: 182-183

  14. Uncoordinated transcription and compromised muscle function in the lmna-null mouse model of Emery- Emery-Dreyfuss muscular dystrophy.

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    Viola F Gnocchi

    2011-02-01

    Full Text Available LMNA encodes both lamin A and C: major components of the nuclear lamina. Mutations in LMNA underlie a range of tissue-specific degenerative diseases, including those that affect skeletal muscle, such as autosomal-Emery-Dreifuss muscular dystrophy (A-EDMD and limb girdle muscular dystrophy 1B. Here, we examine the morphology and transcriptional activity of myonuclei, the structure of the myotendinous junction and the muscle contraction dynamics in the lmna-null mouse model of A-EDMD. We found that there were fewer myonuclei in lmna-null mice, of which ∼50% had morphological abnormalities. Assaying transcriptional activity by examining acetylated histone H3 and PABPN1 levels indicated that there was a lack of coordinated transcription between myonuclei lacking lamin A/C. Myonuclei with abnormal morphology and transcriptional activity were distributed along the length of the myofibre, but accumulated at the myotendinous junction. Indeed, in addition to the presence of abnormal myonuclei, the structure of the myotendinous junction was perturbed, with disorganised sarcomeres and reduced interdigitation with the tendon, together with lipid and collagen deposition. Functionally, muscle contraction became severely affected within weeks of birth, with specific force generation dropping as low as ∼65% and ∼27% of control values in the extensor digitorum longus and soleus muscles respectively. These observations illustrate the importance of lamin A/C for correct myonuclear function, which likely acts synergistically with myotendinous junction disorganisation in the development of A-EDMD, and the consequential reduction in force generation and muscle wasting.

  15. Muscle wasting and the temporal gene expression pattern in a novel rat intensive care unit model

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    Llano-Diez Monica

    2011-12-01

    Full Text Available Abstract Background Acute quadriplegic myopathy (AQM or critical illness myopathy (CIM is frequently observed in intensive care unit (ICU patients. To elucidate duration-dependent effects of the ICU intervention on molecular and functional networks that control the muscle wasting and weakness associated with AQM, a gene expression profile was analyzed at time points varying from 6 hours to 14 days in a unique experimental rat model mimicking ICU conditions, i.e., post-synaptically paralyzed, mechanically ventilated and extensively monitored animals. Results During the observation period, 1583 genes were significantly up- or down-regulated by factors of two or greater. A significant temporal gene expression pattern was constructed at short (6 h-4 days, intermediate (5-8 days and long (9-14 days durations. A striking early and maintained up-regulation (6 h-14d of muscle atrogenes (muscle ring-finger 1/tripartite motif-containing 63 and F-box protein 32/atrogin-1 was observed, followed by an up-regulation of the proteolytic systems at intermediate and long durations (5-14d. Oxidative stress response genes and genes that take part in amino acid catabolism, cell cycle arrest, apoptosis, muscle development, and protein synthesis together with myogenic factors were significantly up-regulated from 5 to 14 days. At 9-14 d, genes involved in immune response and the caspase cascade were up-regulated. At 5-14d, genes related to contractile (myosin heavy chain and myosin binding protein C, regulatory (troponin, tropomyosin, developmental, caveolin-3, extracellular matrix, glycolysis/gluconeogenesis, cytoskeleton/sarcomere regulation and mitochondrial proteins were down-regulated. An activation of genes related to muscle growth and new muscle fiber formation (increase of myogenic factors and JunB and down-regulation of myostatin and up-regulation of genes that code protein synthesis and translation factors were found from 5 to 14 days. Conclusions Novel

  16. Ischemia causes muscle fatigue

    Science.gov (United States)

    Murthy, G.; Hargens, A. R.; Lehman, S.; Rempel, D. M.

    2001-01-01

    The purpose of this investigation was to determine whether ischemia, which reduces oxygenation in the extensor carpi radialis (ECR) muscle, causes a reduction in muscle force production. In eight subjects, muscle oxygenation (TO2) of the right ECR was measured noninvasively and continuously using near infrared spectroscopy (NIRS) while muscle twitch force was elicited by transcutaneous electrical stimulation (1 Hz, 0.1 ms). Baseline measurements of blood volume, muscle oxygenation and twitch force were recorded continuously, then a tourniquet on the upper arm was inflated to one of five different pressure levels: 20, 40, 60 mm Hg (randomized order) and diastolic (69 +/- 9.8 mm Hg) and systolic (106 +/- 12.8 mm Hg) blood pressures. Each pressure level was maintained for 3-5 min, and was followed by a recovery period sufficient to allow measurements to return to baseline. For each respective tourniquet pressure level, mean TO2 decreased from resting baseline (100% TO2) to 99 +/- 1.2% (SEM), 96 +/- 1.9%, 93 +/- 2.8%, 90 +/- 2.5%, and 86 +/- 2.7%, and mean twitch force decreased from resting baseline (100% force) to 99 +/- 0.7% (SEM), 96 +/- 2.7%, 93 +/- 3.1%, 88 +/- 3.2%, and 86 +/- 2.6%. Muscle oxygenation and twitch force at 60 mm Hg tourniquet compression and above were significantly lower (P muscle oxygenation (r = 0.78, P muscle oxygenation causes decreased muscle force production in the forearm extensor muscle. Thus, ischemia associated with a modest decline in TO2 causes muscle fatigue.

  17. Physical Rehabilitation Improves Muscle Function Following Volumetric Muscle Loss Injury

    Science.gov (United States)

    2014-12-19

    muscle , but it did so without significant morphological adaptations (e.g., no hypertrophy and hyperplasia). Wheel running up-regulated metabolic genes...been shown to foster regeneration of injured muscle [5,32,33] and promote hypertrophy (i.e., increased protein synthesis or muscle weight) in muscle ...remaining muscle tissue. Strengthening of synergist muscles can partially compensate for the loss of function due to VML injury. Compensatory hypertrophy

  18. Muscles, exercise and obesity

    DEFF Research Database (Denmark)

    Pedersen, Bente K; Febbraio, Mark A

    2012-01-01

    of yet unidentified factors, secreted from muscle cells, which may influence cancer cell growth and pancreas function. Many proteins produced by skeletal muscle are dependent upon contraction; therefore, physical inactivity probably leads to an altered myokine response, which could provide a potential...

  19. Proteomics of Skeletal Muscle

    DEFF Research Database (Denmark)

    Deshmukh, Atul

    2016-01-01

    Skeletal muscle is the largest tissue in the human body and plays an important role in locomotion and whole body metabolism. It accounts for ~80% of insulin stimulated glucose disposal. Skeletal muscle insulin resistance, a primary feature of Type 2 diabetes, is caused by a decreased ability of m...

  20. The hamstring muscle complex

    NARCIS (Netherlands)

    van der Made, A. D.; Wieldraaijer, T.; Kerkhoffs, G. M.; Kleipool, R. P.; Engebretsen, L.; van Dijk, C. N.; Golanó, P.

    2015-01-01

    The anatomical appearance of the hamstring muscle complex was studied to provide hypotheses for the hamstring injury pattern and to provide reference values of origin dimensions, muscle length, tendon length, musculotendinous junction (MTJ) length as well as width and length of a tendinous

  1. Muscle phosphorylase kinase deficiency

    DEFF Research Database (Denmark)

    Preisler, N; Orngreen, M C; Echaniz-Laguna, A

    2012-01-01

    To examine metabolism during exercise in 2 patients with muscle phosphorylase kinase (PHK) deficiency and to further define the phenotype of this rare glycogen storage disease (GSD).......To examine metabolism during exercise in 2 patients with muscle phosphorylase kinase (PHK) deficiency and to further define the phenotype of this rare glycogen storage disease (GSD)....

  2. Force Transmission in the Actin Cytoskeleton

    Science.gov (United States)

    Gardel, Margaret

    2012-02-01

    The ability of cells to sense and generate mechanical forces is essential to numerous aspects of their physiology, including adhesion, migration, division and differentiation. To a large degree, cellular tension is regulated by the transmission of myosin II-generated forces through the filamentous actin (F-actin) cytoskeleton. While transmission of myosin-generated stresses from the molecular to cellular length scale is well understood in the context of highly organized sarcomeres found in striated muscle, non-muscle and smooth muscle cells contain a wide variety of bundles and networks lacking sarcomeric organization. I will describe the in vitro and in vivo approaches we use to study force transmission in such disordered actomyosin assemblies. Our in vivo results are showing that highly organized stress fibers contribute surprisingly little to the overall extent of cellular tension as compared to disordered actomyosin meshworks. Our in vitro results are demonstrating the mechanisms of symmetry breaking in disordered actomyosin bundles that facilitate the formation of contractile bundles with well-defined ``contractile elements.'' These results provide insight into the self-organization of actomyosin cytoskeleton in non-muscle cells that regulate and maintain cellular tension.

  3. [Diabetic muscle infarction].

    Science.gov (United States)

    ter Bals, Edske; van der Woude, Henk-Jan; Smets, Yves F C

    2013-01-01

    Diabetic muscle infarction is a rare complication of diabetes mellitus that typically presents in the thigh; microvascular abnormalities may play a role. A 32-year-old female presented at the outpatient clinic with a painful, swollen thigh. She had suffered from type 1 diabetes for 22 years. The patient was admitted to the nephrology ward for further evaluation. Deep-venous thrombosis and abscess were excluded with echography. After additional investigations - MRI and a biopsy of skin, muscle and fascia - the diagnosis diabetic muscle infarction was made. The patient was treated with bed rest and analgesics. With hindsight, the muscle biopsy was not actually required in reaching a diagnosis. The diagnosis 'diabetic muscle infarction' is made on the basis of clinical presentation in combination with MRI findings. The treatment consists of bed rest and analgesics.

  4. Pharmacological Blockade of Cannabinoid CB1 Receptors in Diet-Induced Obesity Regulates Mitochondrial Dihydrolipoamide Dehydrogenase in Muscle.

    Directory of Open Access Journals (Sweden)

    Sergio Arrabal

    Full Text Available Cannabinoid CB1 receptors peripherally modulate energy metabolism. Here, we investigated the role of CB1 receptors in the expression of glucose/pyruvate/tricarboxylic acid (TCA metabolism in rat abdominal muscle. Dihydrolipoamide dehydrogenase (DLD, a flavoprotein component (E3 of α-ketoacid dehydrogenase complexes with diaphorase activity in mitochondria, was specifically analyzed. After assessing the effectiveness of the CB1 receptor antagonist AM251 (3 mg kg(-1, 14 days on food intake and body weight, we could identified seven key enzymes from either glycolytic pathway or TCA cycle--regulated by both diet and CB1 receptor activity--through comprehensive proteomic approaches involving two-dimensional electrophoresis and MALDI-TOF/LC-ESI trap mass spectrometry. These enzymes were glucose 6-phosphate isomerase (GPI, triosephosphate isomerase (TPI, enolase (Eno3, lactate dehydrogenase (LDHa, glyoxalase-1 (Glo1 and the mitochondrial DLD, whose expressions were modified by AM251 in hypercaloric diet-induced obesity. Specifically, AM251 blocked high-carbohydrate diet (HCD-induced expression of GPI, TPI, Eno3 and LDHa, suggesting a down-regulation of glucose/pyruvate/lactate pathways under glucose availability. AM251 reversed the HCD-inhibited expression of Glo1 and DLD in the muscle, and the DLD and CB1 receptor expression in the mitochondrial fraction. Interestingly, we identified the presence of CB1 receptors at the membrane of striate muscle mitochondria. DLD over-expression was confirmed in muscle of CB1-/- mice. AM251 increased the pyruvate dehydrogenase and glutathione reductase activity in C2C12 myotubes, and the diaphorase/oxidative activity in the mitochondria fraction. These results indicated an up-regulation of methylglyoxal and TCA cycle activity. Findings suggest that CB1 receptors in muscle modulate glucose/pyruvate/lactate pathways and mitochondrial oxidative activity by targeting DLD.

  5. Pharmacological Blockade of Cannabinoid CB1 Receptors in Diet-Induced Obesity Regulates Mitochondrial Dihydrolipoamide Dehydrogenase in Muscle.

    Science.gov (United States)

    Arrabal, Sergio; Lucena, Miguel Angel; Canduela, Miren Josune; Ramos-Uriarte, Almudena; Rivera, Patricia; Serrano, Antonia; Pavón, Francisco Javier; Decara, Juan; Vargas, Antonio; Baixeras, Elena; Martín-Rufián, Mercedes; Márquez, Javier; Fernández-Llébrez, Pedro; De Roos, Baukje; Grandes, Pedro; Rodríguez de Fonseca, Fernando; Suárez, Juan

    2015-01-01

    Cannabinoid CB1 receptors peripherally modulate energy metabolism. Here, we investigated the role of CB1 receptors in the expression of glucose/pyruvate/tricarboxylic acid (TCA) metabolism in rat abdominal muscle. Dihydrolipoamide dehydrogenase (DLD), a flavoprotein component (E3) of α-ketoacid dehydrogenase complexes with diaphorase activity in mitochondria, was specifically analyzed. After assessing the effectiveness of the CB1 receptor antagonist AM251 (3 mg kg(-1), 14 days) on food intake and body weight, we could identified seven key enzymes from either glycolytic pathway or TCA cycle--regulated by both diet and CB1 receptor activity--through comprehensive proteomic approaches involving two-dimensional electrophoresis and MALDI-TOF/LC-ESI trap mass spectrometry. These enzymes were glucose 6-phosphate isomerase (GPI), triosephosphate isomerase (TPI), enolase (Eno3), lactate dehydrogenase (LDHa), glyoxalase-1 (Glo1) and the mitochondrial DLD, whose expressions were modified by AM251 in hypercaloric diet-induced obesity. Specifically, AM251 blocked high-carbohydrate diet (HCD)-induced expression of GPI, TPI, Eno3 and LDHa, suggesting a down-regulation of glucose/pyruvate/lactate pathways under glucose availability. AM251 reversed the HCD-inhibited expression of Glo1 and DLD in the muscle, and the DLD and CB1 receptor expression in the mitochondrial fraction. Interestingly, we identified the presence of CB1 receptors at the membrane of striate muscle mitochondria. DLD over-expression was confirmed in muscle of CB1-/- mice. AM251 increased the pyruvate dehydrogenase and glutathione reductase activity in C2C12 myotubes, and the diaphorase/oxidative activity in the mitochondria fraction. These results indicated an up-regulation of methylglyoxal and TCA cycle activity. Findings suggest that CB1 receptors in muscle modulate glucose/pyruvate/lactate pathways and mitochondrial oxidative activity by targeting DLD.

  6. A muscle model for hybrid muscle activation

    Directory of Open Access Journals (Sweden)

    Klauer Christian

    2015-09-01

    Full Text Available To develop model-based control strategies for Functional Electrical Stimulation (FES in order to support weak voluntary muscle contractions, a hybrid model for describing joint motions induced by concurrent voluntary-and FES induced muscle activation is proposed. It is based on a Hammerstein model – as commonly used in feedback controlled FES – and exemplarily applied to describe the shoulder abduction joint angle. Main component of a Hammerstein muscle model is usually a static input nonlinearity depending on the stimulation intensity. To additionally incorporate voluntary contributions, we extended the static non-linearity by a second input describing the intensity of the voluntary contribution that is estimated by electromyography (EMG measurements – even during active FES. An Artificial Neural Network (ANN is used to describe the static input non-linearity. The output of the ANN drives a second-order linear dynamical system that describes the combined muscle activation and joint angle dynamics. The tunable parameters are adapted to the individual subject by a system identification approach using previously recorded I/O-data. The model has been validated in two healthy subjects yielding RMS values for the joint angle error of 3.56° and 3.44°, respectively.

  7. Automated multiscale morphometry of muscle disease from second harmonic generation microscopy using tensor-based image processing.

    Science.gov (United States)

    Garbe, Christoph S; Buttgereit, Andreas; Schürmann, Sebastian; Friedrich, Oliver

    2012-01-01

    Practically, all chronic diseases are characterized by tissue remodeling that alters organ and cellular function through changes to normal organ architecture. Some morphometric alterations become irreversible and account for disease progression even on cellular levels. Early diagnostics to categorize tissue alterations, as well as monitoring progression or remission of disturbed cytoarchitecture upon treatment in the same individual, are a new emerging field. They strongly challenge spatial resolution and require advanced imaging techniques and strategies for detecting morphological changes. We use a combined second harmonic generation (SHG) microscopy and automated image processing approach to quantify morphology in an animal model of inherited Duchenne muscular dystrophy (mdx mouse) with age. Multiphoton XYZ image stacks from tissue slices reveal vast morphological deviation in muscles from old mdx mice at different scales of cytoskeleton architecture: cell calibers are irregular, myofibrils within cells are twisted, and sarcomere lattice disruptions (detected as "verniers") are larger in number compared to samples from healthy mice. In young mdx mice, such alterations are only minor. The boundary-tensor approach, adapted and optimized for SHG data, is a suitable approach to allow quick quantitative morphometry in whole tissue slices. The overall detection performance of the automated algorithm compares very well with manual "by eye" detection, the latter being time consuming and prone to subjective errors. Our algorithm outperfoms manual detection by time with similar reliability. This approach will be an important prerequisite for the implementation of a clinical image databases to diagnose and monitor specific morphological alterations in chronic (muscle) diseases. © 2011 IEEE

  8. The intimate nature of oculomotor muscles contracture A natureza íntima da contratura do músculo oculomotor

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    Carlos Ramos de Souza-Dias

    2010-04-01

    Full Text Available The author makes comments about the shortening and loss of elasticity of the oculomotor muscle that remains slack for some time (contracture, by means of a reasoning based on the Hooke´s law and on the papers carried out to demonstrate that a muscle that remains relaxed for some time suffers a shortening due to loss of sarcomeres on the longitudinal direction and the increase of the cross-sectional area due to the increase of collagen tissue in the perimysium and the endomysium.O autor procura demonstrar a razão da perda de elasticidade e do encurtamento do músculo oculomotor que permanece relaxado durante certo tempo (contratura, mediante raciocínio baseado na lei de Hooke e nos trabalhos que demonstram que o músculo oculomotor que permanece frouxo por algum tempo sofre encurtamento devido à perda de sarcômeros no sentido longitudinal e ao aumento da área da secção transversa, devida ao aumento do tecido colágeno do perimísio e do endomísio.

  9. Proteome Changes in biceps femoris Muscle of Iranian One-Humped Camel and Their Effect on Meat Quality Traits

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    Mohammad-Javad Varidi

    2016-01-01

    Full Text Available In this study physicochemical and quality traits of biceps femoris and longissimus thoracis muscles of male and female Iranian one-humped camel were determined during 14 days of refrigeration storage. Analysis of variance of the results showed that only shear force and temperature were affected by the gender (p<0.05. Anatomical location of the muscle influenced the meat properties except for drip loss (p<0.05. Also, except for cooking loss, ageing influenced the physicochemical and quality properties of meat; during 14 days of storage, proteolysis resulted in an increase of L* and b* values, drip loss and myofibrillar fragmentation index, and the decrease of a* value, expressed juice, shear force and sarcomere length. Proteome changes (myofi brillar proteins during storage were investigated. Gel analysis revealed that 19 protein spots were signifi cantly changed during 24, 72 and 168 h post-mortem. Fifteen spots were identified by MALDI-TOF/TOF mass spectrometer. Correlation analysis revealed significant correlations of actin, troponin T, capping protein, heat shock proteins (HSP and desmin with physicochemical and quality properties of meat (p<0.05. Actin might be a potential protein marker for colour, tenderness and water-holding capacity, and HSP27 and desmin are good candidate markers for colour and tenderness, respectively.

  10. Gene expression changes of single skeletal muscle fibers in response to modulation of the mitochondrial calcium uniporter (MCU

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    Francesco Chemello

    2015-09-01

    Full Text Available The mitochondrial calcium uniporter (MCU gene codifies for the inner mitochondrial membrane (IMM channel responsible for mitochondrial Ca2+ uptake. Cytosolic Ca2+ transients are involved in sarcomere contraction through cycles of release and storage in the sarcoplasmic reticulum. In addition cytosolic Ca2+ regulates various signaling cascades that eventually lead to gene expression reprogramming. Mitochondria are strategically placed in close contact with the ER/SR, thus cytosolic Ca2+ transients elicit large increases in the [Ca2+] of the mitochondrial matrix ([Ca2+]mt. Mitochondrial Ca2+ uptake regulates energy production and cell survival. In addition, we recently showed that MCU-dependent mitochondrial Ca2+ uptake controls skeletal muscle trophism. In the same report, we dissected the effects of MCU-dependent mitochondrial Ca2+ uptake on gene expression through microarray gene expression analysis upon modulation of MCU expression by in vivo AAV infection. Analyses were performed on single skeletal muscle fibers at two time points (7 and 14 days post-AAV injection. Raw and normalized data are available on the GEO database (http://www.ncbi.nlm.nih.gov/geo/ (GSE60931.

  11. Respiratory Muscle Plasticity

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    Gransee, Heather M.; Mantilla, Carlos B.; Sieck, Gary C.

    2014-01-01

    Muscle plasticity is defined as the ability of a given muscle to alter its structural and functional properties in accordance with the environmental conditions imposed on it. As such, respiratory muscle is in a constant state of remodeling, and the basis of muscle’s plasticity is its ability to change protein expression and resultant protein balance in response to varying environmental conditions. Here, we will describe the changes of respiratory muscle imposed by extrinsic changes in mechanical load, activity, and innervation. Although there is a large body of literature on the structural and functional plasticity of respiratory muscles, we are only beginning to understand the molecular-scale protein changes that contribute to protein balance. We will give an overview of key mechanisms regulating protein synthesis and protein degradation, as well as the complex interactions between them. We suggest future application of a systems biology approach that would develop a mathematical model of protein balance and greatly improve treatments in a variety of clinical settings related to maintaining both muscle mass and optimal contractile function of respiratory muscles. PMID:23798306

  12. The musculature of coleoid cephalopod arms and tentacles

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    William McKee Kier

    2016-02-01

    Full Text Available The regeneration of coleoid cephalopod arms and tentacles is a common occurrence, recognized since Aristotle. The complexity of the arrangement of the muscle and connective tissues of these appendages make them of great interest for research on regeneration. They lack rigid skeletal elements and consist of a three-dimensional array of muscle fibers, relying on a type of skeletal support system called a muscular hydrostat. Support and movement in the arms and tentacles depends on the fact that muscle tissue resists volume change. The basic principle of function is straightforward; because the volume of the appendage is essentially constant, a decrease in one dimension must result in an increase in another dimension. Since the muscle fibers are arranged in three mutually perpendicular directions, all three dimensions can be actively controlled and thus a remarkable diversity of movements and deformations can be produced. In the arms and tentacles of coleoids, three main muscle orientations are observed: 1 transverse muscle fibers arranged in planes perpendicular to the longitudinal axis; 2 longitudinal muscle fibers typically arranged in bundles parallel to the longitudinal axis; and 3 helical or obliquely arranged layers of muscle fibers, arranged in both right- and left-handed helixes. By selective activation of these muscle groups, elongation, shortening, bending, torsion and stiffening of the appendage can be produced. The predominant muscle fiber type is obliquely striated. Cross-striated fibers are found only in the transverse muscle mass of the prey capture tentacles of squid and cuttlefish. These fibers have unusually short myofilaments and sarcomeres, generating the high shortening velocity required for rapid elongation of the tentacles. It is likely that coleoid cephalopods use ultrastructural modifications rather than tissue-specific myosin isoforms to tune contraction velocities.

  13. The Musculature of Coleoid Cephalopod Arms and Tentacles

    Science.gov (United States)

    Kier, William M.

    2016-01-01

    The regeneration of coleoid cephalopod arms and tentacles is a common occurrence, recognized since Aristotle. The complexity of the arrangement of the muscle and connective tissues of these appendages make them of great interest for research on regeneration. They lack rigid skeletal elements and consist of a three-dimensional array of muscle fibers, relying on a type of skeletal support system called a muscular hydrostat. Support and movement in the arms and tentacles depends on the fact that muscle tissue resists volume change. The basic principle of function is straightforward; because the volume of the appendage is essentially constant, a decrease in one dimension must result in an increase in another dimension. Since the muscle fibers are arranged in three mutually perpendicular directions, all three dimensions can be actively controlled and thus a remarkable diversity of movements and deformations can be produced. In the arms and tentacles of coleoids, three main muscle orientations are observed: (1) transverse muscle fibers arranged in planes perpendicular to the longitudinal axis; (2) longitudinal muscle fibers typically arranged in bundles parallel to the longitudinal axis; and (3) helical or obliquely arranged layers of muscle fibers, arranged in both right- and left-handed helixes. By selective activation of these muscle groups, elongation, shortening, bending, torsion and stiffening of the appendage can be produced. The predominant muscle fiber type is obliquely striated. Cross-striated fibers are found only in the transverse muscle mass of the prey capture tentacles of squid and cuttlefish. These fibers have unusually short myofilaments and sarcomeres, generating the high shortening velocity required for rapid elongation of the tentacles. It is likely that coleoid cephalopods use ultrastructural modifications rather than tissue-specific myosin isoforms to tune contraction velocities. PMID:26925401

  14. Long-term insulin-like growth factor-I expression in skeletal muscles attenuates the enhanced in vitro proliferation ability of the resident satellite cells in transgenic mice

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    Chakravarthy, M. V.; Fiorotto, M. L.; Schwartz, R. J.; Booth, F. W.

    2001-01-01

    Insulin-like growth factor-I (IGF-I) overexpression for 1-month in mouse skeletal muscle increases satellite cell proliferation potential. However, it is unknown whether this beneficial enhancement by IGF-I expression would persist over a longer-term duration in aged mice. This is an important issue to address if a prolonged course of IGF-I is to be used clinically in muscle-wasting conditions where satellite cells may become limiting. Using the IGF-I transgenic (IGF-I Tg) mouse that selectively expresses the IGF-I transgene in striated muscles, we found that 18-months of continuous IGF-I overexpression led to a loss in the enhanced in vitro proliferative capacity of satellite cells from Tg skeletal muscles. Also 18-month-old IGF-I Tg satellite cells lost the enhanced BrdU incorporation, greater pRb and Akt phosphorylations, and decreased p27(Kip1) levels initially observed in cells from 1-month-old IGF-I Tg mice. The levels of those biochemical markers reverted to similar values seen in the 18-months WT littermates. These findings, therefore, suggest that there is no further beneficial effect on enhancing satellite cell proliferation ability with persistent long-term expression of IGF-I in skeletal muscles of these transgenic mice.

  15. Infection of the muscle tissue of the filter-feeding cichlid, Chaetobranchopsis orbicularis Steindachner, 1875, by Kudoa orbicularis (Myxozoa: Multivalvulidae on Marajó Island in the Brazilian Amazon region

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    J.L. Sindeaux-Neto

    Full Text Available ABSTRACT This study describes aspects of infections caused by the myxosporidian Kudoa orbicularis in filter-feeding cichlids, Chaetobranchopsis orbicularis, caught in the Arari River in the municipality of Cachoeira do Arari, on Marajó Island, Pará, Brazil. The parasite forms pseudocysts scattered throughout the striated epaxial and hypaxial muscles. Samples embedded in paraffin were analyzed histologically using hematoxylin-eosin, Gömöri, Ziehl-Neelsen, and Giemsa staining. Necropsy of the C. orbicularis specimens revealed that 100% (50/50 were infected with K. orbicularis. The specimens presented grossly abnormal muscle texture, resulting in extensive inconsistencies and weakness. Progressive softening of the muscles was observed during necropsy, indicating the rapid enzymatic autolysis of the tissue. The parasite found in the muscle tissue of C. orbicularis was identified as K. orbicularis, with clinical signs of disease being observed in the fish. The necropsy revealed extensive damage to the host organism, with well-established fibrocystic infections in the muscle fibers, associated with post mortem myoliquefaction.

  16. Muscle Bioenergetic Considerations for Intrinsic Laryngeal Skeletal Muscle Physiology

    Science.gov (United States)

    Sandage, Mary J.; Smith, Audrey G.

    2017-01-01

    Purpose: Intrinsic laryngeal skeletal muscle bioenergetics, the means by which muscles produce fuel for muscle metabolism, is an understudied aspect of laryngeal physiology with direct implications for voice habilitation and rehabilitation. The purpose of this review is to describe bioenergetic pathways identified in limb skeletal muscle and…

  17. Human muscle LIM protein dimerizes along the actin cytoskeleton and cross-links actin filaments.

    Science.gov (United States)

    Hoffmann, Céline; Moreau, Flora; Moes, Michèle; Luthold, Carole; Dieterle, Monika; Goretti, Emeline; Neumann, Katrin; Steinmetz, André; Thomas, Clément

    2014-08-01

    The muscle LIM protein (MLP) is a nucleocytoplasmic shuttling protein playing important roles in the regulation of myocyte remodeling and adaptation to hypertrophic stimuli. Missense mutations in human MLP or its ablation in transgenic mice promotes cardiomyopathy and heart failure. The exact function(s) of MLP in the cytoplasmic compartment and the underlying molecular mechanisms remain largely unknown. Here, we provide evidence that MLP autonomously binds to, stabilizes, and bundles actin filaments (AFs) independently of calcium and pH. Using total internal reflection fluorescence microscopy, we have shown how MLP cross-links actin filaments into both unipolar and mixed-polarity bundles. Quantitative analysis of the actin cytoskeleton configuration confirmed that MLP substantially promotes actin bundling in live myoblasts. In addition, bimolecular fluorescence complementation (BiFC) assays revealed MLP self-association. Remarkably, BiFC complexes mostly localize along actin filament-rich structures, such as stress fibers and sarcomeres, supporting a functional link between MLP self-association and actin cross-linking. Finally, we have demonstrated that MLP self-associates through its N-terminal LIM domain, whereas it binds to AFs through its C-terminal LIM domain. Together our data support that MLP contributes to the maintenance of cardiomyocyte cytoarchitecture by a mechanism involving its self-association and actin filament cross-linking. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  18. Painful unilateral temporalis muscle enlargement: reactive masticatory muscle hypertrophy.

    Science.gov (United States)

    Katsetos, Christos D; Bianchi, Michael A; Jaffery, Fizza; Koutzaki, Sirma; Zarella, Mark; Slater, Robert

    2014-06-01

    An instance of isolated unilateral temporalis muscle hypertrophy (reactive masticatory muscle hypertrophy with fiber type 1 predominance) confirmed by muscle biopsy with histochemical fiber typing and image analysis in a 62 year-old man is reported. The patient presented with bruxism and a painful swelling of the temple. Absence of asymmetry or other abnormalities of the craniofacial skeleton was confirmed by magnetic resonance imaging and cephalometric analyses. The patient achieved symptomatic improvement only after undergoing botulinum toxin injections. Muscle biopsy is key in the diagnosis of reactive masticatory muscle hypertrophy and its distinction from masticatory muscle myopathy (hypertrophic branchial myopathy) and other non-reactive causes of painful asymmetric temporalis muscle enlargement.

  19. Site directed mutagenesis of Drosophila flightin disrupts phosphorylation and impairs flight muscle structure and mechanics.

    Science.gov (United States)

    Barton, Byron; Ayer, Gretchen; Maughan, David W; Vigoreaux, Jim O

    2007-01-01

    Flightin is a myosin rod binding protein that in Drosophila melanogaster is expressed exclusively in the asynchronous indirect flight muscles (IFM). Hyperphosphorylation of flightin coincides with the completion of myofibril assembly and precedes the emergence of flight competency in young adults. To investigate the role of flightin phosphorylation in vivo we generated three flightin null (fln(0)) Drosophila strains that express a mutant flightin transgene with two (Thr158, Ser 162), three (Ser139, Ser141, Ser145) or all five potential phosphorylation sites mutated to alanines. These amino acid substitutions result in lower than normal levels of flightin accumulation and transgenic strains that are unable to beat their wings. On two dimensional gels of IFM proteins, the transgenic strain with five mutant sites (fln(5STA)) is devoid of all phosphovariants, the transgenic strain with two mutant sites (fln(2TSA)) expresses only the two least acidic of the nine phosphovariants, and the transgenic strain with three mutant sites (fln(3SA)) expresses all nine phosphovariants, as the wild-type strain. These results suggest that phosphorylation of Thr158 and/or Ser162 is necessary for subsequent phosphorylation of other sites. All three transgenic strains show normal, albeit long, IFM sarcomeres in newly eclosed adults. In contrast, sarcomeres in fully mature fln(5STA) and fln(2TSA) adults show extensive breakdown while those in fln(3SA) are not as disordered. The fiber hypercontraction phenotype that characterizes fln(0) is fully evident in fln(5STA) and fln(2TSA) but partially rescued in fln(3SA). Mechanics on skinned fibers from newly eclosed flies show alterations in viscous modulus for fln(5STA) and fln(2TSA) that result in a significant reduction in oscillatory power output. Expression of fln(5STA) and fln(2TSA), but not fln(3SA), in a wild-type (fln(+)/fln(+)) background resulted in a dominant negative effect manifested as flight impairments and hypercontracted IFM

  20. Muscle Disorders - Multiple Languages

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    ... Health Information Translations Spanish (español) Expand Section Muscle Disorders: MedlinePlus Health Topic - English ... Health Information Translations Characters not displaying correctly on this page? See language display issues . Return to the MedlinePlus Health Information ...