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Sample records for polar pollen cell

  1. Knockin' on pollen's door: live cell imaging of early polarization events in germinating Arabidopsis pollen

    Science.gov (United States)

    Vogler, Frank; Konrad, Sebastian S. A.; Sprunck, Stefanie

    2015-01-01

    Pollen tubes are an excellent system for studying the cellular dynamics and complex signaling pathways that coordinate polarized tip growth. Although several signaling mechanisms acting in the tip-growing pollen tube have been described, our knowledge on the subcellular and molecular events during pollen germination and growth site selection at the pollen plasma membrane is rather scarce. To simultaneously track germinating pollen from up to 12 genetically different plants we developed an inexpensive and easy mounting technique, suitable for every standard microscope setup. We performed high magnification live-cell imaging during Arabidopsis pollen activation, germination, and the establishment of pollen tube tip growth by using fluorescent marker lines labeling either the pollen cytoplasm, vesicles, the actin cytoskeleton or the sperm cell nuclei and membranes. Our studies revealed distinctive vesicle and F-actin polarization during pollen activation and characteristic growth kinetics during pollen germination and pollen tube formation. Initially, the germinating Arabidopsis pollen tube grows slowly and forms a uniform roundish bulge, followed by a transition phase with vesicles heavily accumulating at the growth site before switching to rapid tip growth. Furthermore, we found the two sperm cells to be transported into the pollen tube after the phase of rapid tip growth has been initiated. The method presented here is suitable to quantitatively study subcellular events during Arabidopsis pollen germination and growth, and for the detailed analysis of pollen mutants with respect to pollen polarization, bulging, or growth site selection at the pollen plasma membrane. PMID:25954283

  2. Comprehensive analysis of tobacco pollen transcriptome unveils common pathways in polar cell expansion and underlying heterochronic shift during spermatogenesis

    Directory of Open Access Journals (Sweden)

    Hafidh Said

    2012-02-01

    Full Text Available Abstract Background Many flowering plants produce bicellular pollen. The two cells of the pollen grain are destined for separate fates in the male gametophyte, which provides a unique opportunity to study genetic interactions that govern guided single-cell polar expansion of the growing pollen tube and the coordinated control of germ cell division and sperm cell fate specification. We applied the Agilent 44 K tobacco gene chip to conduct the first transcriptomic analysis of the tobacco male gametophyte. In addition, we performed a comparative study of the Arabidopsis root-hair trichoblast transcriptome to evaluate genetic factors and common pathways involved in polarized cell-tip expansion. Results Progression of pollen grains from freshly dehisced anthers to pollen tubes 4 h after germination is accompanied with > 5,161 (14.9% gametophyte-specific expressed probes active in at least one of the developmental stages. In contrast, > 18,821 (54.4% probes were preferentially expressed in the sporophyte. Our comparative approach identified a subset of 104 pollen tube-expressed genes that overlap with root-hair trichoblasts. Reverse genetic analysis of selected candidates demonstrated that Cu/Zn superoxide dismutase 1 (CSD1, a WD-40 containing protein (BP130384, and Replication factor C1 (NtRFC1 are among the central regulators of pollen-tube tip growth. Extension of our analysis beyond the second haploid mitosis enabled identification of an opposing-dynamic accumulation of core regulators of cell proliferation and cell fate determinants in accordance with the progression of the germ cell cycle. Conclusions The current study provides a foundation to isolate conserved regulators of cell tip expansion and those that are unique for pollen tube growth to the female gametophyte. A transcriptomic data set is presented as a benchmark for future functional studies using developing pollen as a model. Our results demonstrated previously unknown functions of

  3. Comprehensive analysis of tobacco pollen transcriptome unveils common pathways in polar cell expansion and underlying heterochronic shift during spermatogenesis

    Science.gov (United States)

    2012-01-01

    Background Many flowering plants produce bicellular pollen. The two cells of the pollen grain are destined for separate fates in the male gametophyte, which provides a unique opportunity to study genetic interactions that govern guided single-cell polar expansion of the growing pollen tube and the coordinated control of germ cell division and sperm cell fate specification. We applied the Agilent 44 K tobacco gene chip to conduct the first transcriptomic analysis of the tobacco male gametophyte. In addition, we performed a comparative study of the Arabidopsis root-hair trichoblast transcriptome to evaluate genetic factors and common pathways involved in polarized cell-tip expansion. Results Progression of pollen grains from freshly dehisced anthers to pollen tubes 4 h after germination is accompanied with > 5,161 (14.9%) gametophyte-specific expressed probes active in at least one of the developmental stages. In contrast, > 18,821 (54.4%) probes were preferentially expressed in the sporophyte. Our comparative approach identified a subset of 104 pollen tube-expressed genes that overlap with root-hair trichoblasts. Reverse genetic analysis of selected candidates demonstrated that Cu/Zn superoxide dismutase 1 (CSD1), a WD-40 containing protein (BP130384), and Replication factor C1 (NtRFC1) are among the central regulators of pollen-tube tip growth. Extension of our analysis beyond the second haploid mitosis enabled identification of an opposing-dynamic accumulation of core regulators of cell proliferation and cell fate determinants in accordance with the progression of the germ cell cycle. Conclusions The current study provides a foundation to isolate conserved regulators of cell tip expansion and those that are unique for pollen tube growth to the female gametophyte. A transcriptomic data set is presented as a benchmark for future functional studies using developing pollen as a model. Our results demonstrated previously unknown functions of certain genes in pollen

  4. The apical actin fringe contributes to localized cell wall deposition and polarized growth in the lily pollen tube.

    Science.gov (United States)

    Rounds, Caleb M; Hepler, Peter K; Winship, Lawrence J

    2014-09-01

    In lily (Lilium formosanum) pollen tubes, pectin, a major component of the cell wall, is delivered through regulated exocytosis. The targeted transport and secretion of the pectin-containing vesicles may be controlled by the cortical actin fringe at the pollen tube apex. Here, we address the role of the actin fringe using three different inhibitors of growth: brefeldin A, latrunculin B, and potassium cyanide. Brefeldin A blocks membrane trafficking and inhibits exocytosis in pollen tubes; it also leads to the degradation of the actin fringe and the formation of an aggregate of filamentous actin at the base of the clear zone. Latrunculin B, which depolymerizes filamentous actin, markedly slows growth but allows focused pectin deposition to continue. Of note, the locus of deposition shifts frequently and correlates with changes in the direction of growth. Finally, potassium cyanide, an electron transport chain inhibitor, briefly stops growth while causing the actin fringe to completely disappear. Pectin deposition continues but lacks focus, instead being delivered in a wide arc across the pollen tube tip. These data support a model in which the actin fringe contributes to the focused secretion of pectin to the apical cell wall and, thus, to the polarized growth of the pollen tube. © 2014 American Society of Plant Biologists. All Rights Reserved.

  5. The Apical Actin Fringe Contributes to Localized Cell Wall Deposition and Polarized Growth in the Lily Pollen Tube1[W][OPEN

    Science.gov (United States)

    Rounds, Caleb M.; Hepler, Peter K.; Winship, Lawrence J.

    2014-01-01

    In lily (Lilium formosanum) pollen tubes, pectin, a major component of the cell wall, is delivered through regulated exocytosis. The targeted transport and secretion of the pectin-containing vesicles may be controlled by the cortical actin fringe at the pollen tube apex. Here, we address the role of the actin fringe using three different inhibitors of growth: brefeldin A, latrunculin B, and potassium cyanide. Brefeldin A blocks membrane trafficking and inhibits exocytosis in pollen tubes; it also leads to the degradation of the actin fringe and the formation of an aggregate of filamentous actin at the base of the clear zone. Latrunculin B, which depolymerizes filamentous actin, markedly slows growth but allows focused pectin deposition to continue. Of note, the locus of deposition shifts frequently and correlates with changes in the direction of growth. Finally, potassium cyanide, an electron transport chain inhibitor, briefly stops growth while causing the actin fringe to completely disappear. Pectin deposition continues but lacks focus, instead being delivered in a wide arc across the pollen tube tip. These data support a model in which the actin fringe contributes to the focused secretion of pectin to the apical cell wall and, thus, to the polarized growth of the pollen tube. PMID:25037212

  6. Nuclear membrane localization during pollen development and apex-focused polarity establishment of SYP124/125 during pollen germination in Arabidopsis thaliana.

    Science.gov (United States)

    Ichikawa, Mie; Iwano, Megumi; Sato, Masa H

    2015-12-01

    Establishment of apex-polarity. Elongation of the pollen tube is a highly coordinated process involving polarized secretion of cell wall and membrane materials to the apical region. We investigated changes in the localization of soluble NSF attachment proteins (SNAREs) in developing pollen grains and the pollen tube for transgenic Arabidopsis expressing pollen-specific plasma-membrane Qa-SNAREs (SYP124, 125 and 131) fused with the green fluorescent protein (GFP). The expression of SYP124 and SYP125 was firstly detected in the microspore nuclear membrane during pollen mitosis II. Although SYP124, 125 and 131 accumulated throughout the cytosol in the mature pollen grain, GFP-SYP124 and GFP-SYP125 were highly concentrated in the apical or subapical regions of the elongating pollen tube with slightly different localization patterns, whereas GFP-SYP131 was uniformly localized to the plasma membrane of the pollen tube. The apex-focused polarity of GFP-SYP125 was established coincident with formation of a Ca(2+) gradient before pollen germination. These results suggest that SNAREs function differentially in the same cells and that at least two distinct membrane transport pathways are involved in the pollen development and the pollen tube germination and elongation.

  7. A cyclic nucleotide-gated channel is essential for polarized tip growth of pollen.

    Science.gov (United States)

    Frietsch, Sabine; Wang, Yong-Fei; Sladek, Chris; Poulsen, Lisbeth R; Romanowsky, Shawn M; Schroeder, Julian I; Harper, Jeffrey F

    2007-09-04

    Ion signals are critical to regulating polarized growth in many cell types, including pollen in plants and neurons in animals. Genetic evidence presented here indicates that pollen tube growth requires cyclic nucleotide-gated channel (CNGC) 18. CNGCs are nonspecific cation channels found in plants and animals and have well established functions in excitatory signal transduction events in animals. In Arabidopsis, male sterility was observed for two cngc18 null mutations. CNGC18 is expressed primarily in pollen, as indicated from a promoter::GUS (beta-glucuronidase) reporter analysis and expression profiling. The underlying cause of sterility was identified as a defect in pollen tube growth, resulting in tubes that were kinky, short, often thin, and unable to grow into the transmitting tract. Expression of a GFP-tagged CNGC18 in mutant pollen provided complementation and evidence for asymmetric localization of CNGC18 to the plasma membrane at the growing tip, starting at the time of pollen grain germination. Heterologous expression of CNGC18 in Escherichia coli resulted in a time- and concentration-dependent accumulation of more Ca2+. Thus, CNGC18 provides a mechanism to directly transduce a cyclic nucleotide (cNMP) signal into an ion flux that can produce a localized signal capable of regulating the pollen tip-growth machinery. These results identify a CNGC that is essential to an organism's life cycle and raise the possibility that CNGCs have a widespread role in regulating cell-growth dynamics in both plant and animals.

  8. Calcium-dependent protein kinases regulate polarized tip growth in pollen tubes.

    Science.gov (United States)

    Myers, Candace; Romanowsky, Shawn M; Barron, Yoshimi D; Garg, Shilpi; Azuse, Corinn L; Curran, Amy; Davis, Ryan M; Hatton, Jasmine; Harmon, Alice C; Harper, Jeffrey F

    2009-08-01

    Calcium signals are critical for the regulation of polarized growth in many eukaryotic cells, including pollen tubes and neurons. In plants, the regulatory pathways that code and decode Ca(2+) signals are poorly understood. In Arabidopsis thaliana, genetic evidence presented here indicates that pollen tube tip growth involves the redundant activity of two Ca(2+)-dependent protein kinases (CPKs), isoforms CPK17 and -34. Both isoforms appear to target to the plasma membrane, as shown by imaging of CPK17-yellow fluorescent protein (YFP) and CPK34-YFP in growing pollen tubes. Segregation analyses from two independent sets of T-DNA insertion mutants indicate that a double disruption of CPK17 and -34 results in an approximately 350-fold reduction in pollen transmission efficiency. The near sterile phenotype of homozygous double mutants could be rescued through pollen expression of a CPK34-YFP fusion. In contrast, a transgene rescue was blocked by mutations engineered to disrupt the Ca(2+)-activation mechanism of CPK34 (CPK34-YFP-E465A,E500A), providing in vivo evidence linking Ca(2+) activation to a biological function of a CPK. While double mutant pollen tubes displayed normal morphology, relative growth rates for the most rapidly growing tubes were reduced by more than three-fold compared with wild type. In addition, while most mutant tubes appeared to grow far enough to reach ovules, the vast majority (>90%) still failed to locate and fertilize ovules. Together, these results provide genetic evidence that CPKs are essential to pollen fitness, and support a mechanistic model in which CPK17 and -34 transduce Ca(2+) signals to increase the rate of pollen tube tip growth and facilitate a response to tropism cues.

  9. Vacuolar CBL-CIPK12 Ca(2+)-sensor-kinase complexes are required for polarized pollen tube growth.

    Science.gov (United States)

    Steinhorst, Leonie; Mähs, Anette; Ischebeck, Till; Zhang, Chunxia; Zhang, Xinxin; Arendt, Sibylle; Schültke, Stefanie; Heilmann, Ingo; Kudla, Jörg

    2015-06-01

    Polarized tip growth is a fundamental process of specialized eukaryotic cells like neuronal axons, fungal hyphae, and plant root hairs and pollen tubes. In pollen tubes, a tip-focused oscillating Ca(2+) gradient governs ions fluxes, vesicle transport, and cytoskeleton dynamics to ensure proper polarized cell growth [1, 2]. While a crucial role of vacuolar Ca(2+) signaling is established for cellular movements like guard cell dynamics [3-5], its contribution to polarized growth remains to be defined. Here we identified the two closely related tonoplast-localized Ca(2+)-sensor proteins CBL2 and CBL3 as crucial regulators of vacuolar dynamics and polarized pollen tube growth. Overexpression of CBL2 or CBL3 in Arabidopsis and tobacco pollen tubes affected vacuolar morphology, pollen germination, and tube growth, but did not alter actin organization, PI(4,5)P2 distribution, or tip-focused Ca(2+) oscillations. Similarly, loss of function of each single Ca(2+) sensor and cbl2/cbl3 double mutants exhibited impaired pollen tube growth in vitro and in vivo. Both Ca(2+) sensors interacted with the kinase CIPK12, which translocated from the cytoplasm to the vacuolar membrane upon this interaction. Also, overexpression of CIPK12 induced severe vacuolar phenotypes, and loss of function of CIPK12 lead to impairment of polar growth. Remarkably, co-expression of CBL2 or CBL3 with CIPK12 resulted in a phosphorylation-dependent, massively enhanced vacuolar inflation and further disruption of polar growth. Together, these findings identify an essential role of the vacuole and vacuolar Ca(2+) signaling for polarized tip growth. We propose that a faithfully balanced activity of Ca(2+)-activated CBL2/3-CIPK12 complexes fulfills fundamental functions to enable the fast growth of pollen tubes in higher plants. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Polar Expansion Dynamics in the Plant Kingdom: A Diverse and Multifunctional Journey on the Path to Pollen Tubes

    Directory of Open Access Journals (Sweden)

    David S. Domozych

    2013-03-01

    Full Text Available Polar expansion is a widespread phenomenon in plants spanning all taxonomic groups from the Charophycean Green Algae to pollen tubes in Angiosperms and Gymnosperms. Current data strongly suggests that many common features are shared amongst cells displaying polar growth mechanics including changes to the structural features of localized regions of the cell wall, mobilization of targeted secretion mechanisms, employment of the actin cytoskeleton for directing secretion and in many cases, endocytosis and coordinated interaction of multiple signal transduction mechanisms prompted by external biotic and abiotic cues. The products of polar expansion perform diverse functions including delivery of male gametes to the egg, absorption, anchorage, adhesion and photo-absorption efficacy. A comparative analysis of polar expansion dynamics is provided with special emphasis on those found in early divergent plants.

  11. Pollen

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Records of past vegetation and climate derived from pollen found in lake and ocean sediments. Parameter keywords describe what was measured in this data set....

  12. Comprehensive cell-specific protein analysis in early and late pollen development from diploid microsporocytes to pollen tube growth.

    Science.gov (United States)

    Ischebeck, Till; Valledor, Luis; Lyon, David; Gingl, Stephanie; Nagler, Matthias; Meijón, Mónica; Egelhofer, Volker; Weckwerth, Wolfram

    2014-01-01

    Pollen development in angiosperms is one of the most important processes controlling plant reproduction and thus productivity. At the same time, pollen development is highly sensitive to environmental fluctuations, including temperature, drought, and nutrition. Therefore, pollen biology is a major focus in applied studies and breeding approaches for improving plant productivity in a globally changing climate. The most accessible developmental stages of pollen are the mature pollen and the pollen tubes, and these are thus most frequently analyzed. To reveal a complete quantitative proteome map, we additionally addressed the very early stages, analyzing eight stages of tobacco pollen development: diploid microsporocytes, meiosis, tetrads, microspores, polarized microspores, bipolar pollen, desiccated pollen, and pollen tubes. A protocol for the isolation of the early stages was established. Proteins were extracted and analyzed by means of a new gel LC-MS fractionation protocol. In total, 3817 protein groups were identified. Quantitative analysis was performed based on peptide count. Exceedingly stage-specific differential protein regulation was observed during the conversion from the sporophytic to the gametophytic proteome. A map of highly specialized functionality for the different stages could be revealed from the metabolic activity and pronounced differentiation of proteasomal and ribosomal protein complex composition up to protective mechanisms such as high levels of heat shock proteins in the very early stages of development.

  13. Comprehensive Cell-specific Protein Analysis in Early and Late Pollen Development from Diploid Microsporocytes to Pollen Tube Growth*

    Science.gov (United States)

    Ischebeck, Till; Valledor, Luis; Lyon, David; Gingl, Stephanie; Nagler, Matthias; Meijón, Mónica; Egelhofer, Volker; Weckwerth, Wolfram

    2014-01-01

    Pollen development in angiosperms is one of the most important processes controlling plant reproduction and thus productivity. At the same time, pollen development is highly sensitive to environmental fluctuations, including temperature, drought, and nutrition. Therefore, pollen biology is a major focus in applied studies and breeding approaches for improving plant productivity in a globally changing climate. The most accessible developmental stages of pollen are the mature pollen and the pollen tubes, and these are thus most frequently analyzed. To reveal a complete quantitative proteome map, we additionally addressed the very early stages, analyzing eight stages of tobacco pollen development: diploid microsporocytes, meiosis, tetrads, microspores, polarized microspores, bipolar pollen, desiccated pollen, and pollen tubes. A protocol for the isolation of the early stages was established. Proteins were extracted and analyzed by means of a new gel LC-MS fractionation protocol. In total, 3817 protein groups were identified. Quantitative analysis was performed based on peptide count. Exceedingly stage-specific differential protein regulation was observed during the conversion from the sporophytic to the gametophytic proteome. A map of highly specialized functionality for the different stages could be revealed from the metabolic activity and pronounced differentiation of proteasomal and ribosomal protein complex composition up to protective mechanisms such as high levels of heat shock proteins in the very early stages of development. PMID:24078888

  14. Proteomics of pollen development and germination.

    Science.gov (United States)

    Dai, Shaojun; Wang, Tai; Yan, Xiufeng; Chen, Sixue

    2007-12-01

    In higher plants, pollen grains represent the vestiges of a highly reduced male gametophyte generation. After germination, the pollen tube delivers the sperm cells by tip-growing to the embryo sac for fertilization. Besides the intrinsic importance for sexual reproduction, pollen development and germination serve as an attractive system to address important questions related to cell division, cell differentiation, polar growth, cell-cell interaction, and cell fate. Recently, pollen functional specification has been well-studied using multidisciplinary approaches. Here, we review recent advances in proteomics of pollen development and germination.

  15. Lost in traffic? The K+ channel of lily pollen, LilKT1, is detected at the endomembranes inside yeast cells, tobacco leaves and lily pollen

    Directory of Open Access Journals (Sweden)

    Minou Jasmin Safiarian

    2015-02-01

    Full Text Available Fertilization in plants relies on fast growth of pollen tubes through the style tissue towards the ovules. This polarized growth depends on influx of ions and water to increase the tube’s volume. K+ inward rectifying channels were detected in many pollen species, with one identified in Arabidopsis. Here, an Arabidopsis AKT1-like channel (LilKT1 was identified from Lilium longiflorum pollen. Complementation of K+ uptake deficient yeast mutants was only successful when the entire LilKT1 C-terminus was replaced by the AKT1 C-terminus. No signals were observed in the plasma membrane (PM of pollen tubes after expression of fluorescence-tagged LilKT1 nor were any LilKT1-derived peptides detectable in the pollen PM by mass spectrometry analysis. In contrast, fluorescent LilKT1 partly co-localized with the lily PM H+ ATPase LilHA2 in the PM of tobacco leaf cells, but exhibited a punctual fluorescence pattern and also sub-plasma membrane localization. Thus, incorporation of LilKT1 into the pollen PM seems tighter controlled than in other cells with still unknown trafficking signals in LilKT1’s C-terminus, resulting in channel densities below detection limits. This highly controlled incorporation might have physiological reasons: an uncontrolled number of K+ inward channels in the pollen PM will give an increased water influx due to the raising cytosolic K+ concentration, and finally, causing the tube to burst.

  16. Visualisation of plastid degradation in sperm cells of wheat pollen.

    Science.gov (United States)

    Primavesi, Lucia F; Wu, Huixia; Mudd, Elisabeth A; Day, Anil; Jones, Huw D

    2017-01-01

    Like most angiosperms, wheat (Triticum aestivum) shows maternal inheritance of plastids. It is thought that this takes place by cytoplasmic stripping at fertilisation rather than the absence of plastids in sperm cells. To determine the fate of plastids during sperm cell development, plastid-targeted green fluorescent protein was used to visualise these organelles in nuclear transgenic wheat lines. Fewer than thirty small 1-2-μm plastids were visible in early uninucleate pollen cells. These dramatically increased to several hundred larger (4 μm) plastids during pollen maturation and went through distinct morphological changes. Only small plastids were visible in generative cells (n = 25) and young sperm cells (n = 9). In mature sperm cells, these green fluorescent protein (GFP)-tagged plastids were absent. This is consistent with maternal inheritance of plastids resulting from their degradation in mature sperm cells in wheat.

  17. Integrins and epithelial cell polarity.

    Science.gov (United States)

    Lee, Jessica L; Streuli, Charles H

    2014-08-01

    Cell polarity is characterised by differences in structure, composition and function between at least two poles of a cell. In epithelial cells, these spatial differences allow for the formation of defined apical and basal membranes. It has been increasingly recognised that cell-matrix interactions and integrins play an essential role in creating epithelial cell polarity, although key gaps in our knowledge remain. This Commentary will discuss the mounting evidence for the role of integrins in polarising epithelial cells. We build a model in which both inside-out signals to polarise basement membrane assembly at the basal surface, and outside-in signals to control microtubule apical-basal orientation and vesicular trafficking are required for establishing and maintaining the orientation of epithelial cell polarity. Finally, we discuss the relevance of the basal integrin polarity axis to cancer. This article is part of a Minifocus on Establishing polarity. © 2014. Published by The Company of Biologists Ltd.

  18. Epithelial cell polarity, stem cells and cancer

    DEFF Research Database (Denmark)

    Martin-Belmonte, Fernando; Perez-Moreno, Mirna

    2011-01-01

    After years of extensive scientific discovery much has been learned about the networks that regulate epithelial homeostasis. Loss of expression or functional activity of cell adhesion and cell polarity proteins (including the PAR, crumbs (CRB) and scribble (SCRIB) complexes) is intricately related......, deregulation of adhesion and polarity proteins can cause misoriented cell divisions and increased self-renewal of adult epithelial stem cells. In this Review, we highlight some advances in the understanding of how loss of epithelial cell polarity contributes to tumorigenesis....

  19. Bee Pollen

    Science.gov (United States)

    ... Pollen Extract, Buckwheat Pollen, Extrait de Pollen d’Abeille, Honeybee Pollen, Honey Bee Pollen, Maize Pollen, Pine Pollen, Polen de Abeja, Pollen, Pollen d'Abeille, Pollen d’Abeille de Miel, Pollen de Sarrasin.

  20. Establishment of the male germline and sperm cell movement during pollen germination and tube growth in maize

    OpenAIRE

    Kliwer, Irina; Dresselhaus, Thomas

    2010-01-01

    Two sperm cells are required to achieve double fertilization in flowering plants (angiosperms). In contrast to animals and lower plants such as mosses and ferns, sperm cells of flowering plants (angiosperms) are immobile and are transported to the female gametes (egg and central cell) via the pollen tube. The two sperm cells arise from the generative pollen cell either within the pollen grain or after germination inside the pollen tube. While pollen tube growth and sperm behavior has been int...

  1. Pollen proteomics: from stress physiology to developmental priming.

    Science.gov (United States)

    Chaturvedi, Palak; Ghatak, Arindam; Weckwerth, Wolfram

    2016-06-01

    Pollen development and stress. In angiosperms, pollen or pollen grain (male gametophyte) is a highly reduced two- or three-cell structure which plays a decisive role in plant reproduction. Male gametophyte development takes place in anther locules where diploid sporophytic cells undergo meiotic division followed by two consecutive mitotic processes. A desiccated and metabolically quiescent form of mature pollen is released from the anther which lands on the stigma. Pollen tube growth takes place followed by double fertilization. Apart from its importance in sexual reproduction, pollen is also an interesting model system which integrates fundamental cellular processes like cell division, differentiation, fate determination, polar establishment, cell to cell recognition and communication. Recently, pollen functionality has been studied by multidisciplinary approaches which also include OMICS analyses like transcriptomics, proteomics and metabolomics. Here, we review recent advances in proteomics of pollen development and propose the process of developmental priming playing a key role to guard highly sensitive developmental processes.

  2. Regulation of cell polarity by cell adhesion receptors.

    Science.gov (United States)

    Ebnet, Klaus; Kummer, Daniel; Steinbacher, Tim; Singh, Amrita; Nakayama, Masanori; Matis, Maja

    2017-07-22

    The ability of cells to polarize is an intrinsic property of almost all cells and is required for the devlopment of most multicellular organisms. To develop cell polarity, cells integrate various signals derived from intrinsic as well as extrinsic sources. In the recent years, cell-cell adhesion receptors have turned out as important regulators of cellular polarization. By interacting with conserved cell polarity proteins, they regulate the recruitment of polarity complexes to specific sites of cell-cell adhesion. By initiating intracellular signaling cascades at those sites, they trigger their specific subcellular activation. Not surprisingly, cell-cell adhesion receptors regulate diverse aspects of cell polarity, including apico-basal polarity in epithelial and endothelial cells, front-to-rear polarity in collectively migrating cells, and planar cell polarity during organ development. Here, we review the recent developments highlighting the central roles of cell-cell adhesion molecules in the development of cell polarity. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. Effect of 211At treating pollen and stigma on generative cells and seed setting of rice

    International Nuclear Information System (INIS)

    Jin Jiannan; Mo Shangwu; Liu Ning; Zhou Maolun; Zhang Shuyuan; Chen Fang; Zhang Yizheng; Gao Maoguo

    1998-01-01

    Low specific radioactivity (7.4 kBq/ml) 211 At treating pollen and stigma can obviously affect morphological structures and physiological functions of pollen, stigma and ovule or embryo sac cells, and cause injury. Results showed that because of the radiation effects the seed setting rate of rice was decreased, and the development of some embryos were affected and others became abnormal

  4. Methods to isolate a large amount of generative cells, sperm cells and vegetative nuclei from tomato pollen for omics analysis

    Directory of Open Access Journals (Sweden)

    Yunlong eLu

    2015-06-01

    Full Text Available The development of sperm cells from microspores involves a set of finely regulated molecular and cellular events and the coordination of these events. The mechanisms underlying these events and their interconnections remain a major challenge. Systems analysis of genome-wide molecular networks and functional modules with high-throughput omics approaches is crucial for understanding the mechanisms; however, this study is hindered because of the difficulty in isolating a large amount of cells of different types, especially generative cells (GCs, from the pollen. Here, we optimized the conditions of tomato pollen germination and pollen tube growth to allow for long-term growth of pollen tubes in vitro with sperm cells (SCs generated in the tube. Using this culture system, we developed methods for isolating GCs, SCs and vegetative-cell nuclei (VN from just-germinated tomato pollen grains and growing pollen tubes and their purification by Percoll density gradient centrifugation. The purity and viability of isolated GCs and SCs were confirmed by microscopy examination and fluorescein diacetate staining, respectively, and the integrity of VN was confirmed by propidium iodide staining. We could obtain about 1.5 million GCs and 2.0 million SCs each from 180 mg initiated pollen grains, and 10 million VN from 270 mg initiated pollen grains germinated in vitro in each experiment. These methods provide the necessary preconditions for systematic biology studies of SC development and differentiation in higher plants.

  5. Dynamics of vegetative cytoplasm during generative cell formation and pollen maturation in Arabidopsis thaliana

    Science.gov (United States)

    Kuang, A.; Musgrave, M. E.

    1996-01-01

    Ultrastructural changes of pollen cytoplasm during generative cell formation and pollen maturation in Arabidopsis thaliana were studied. The pollen cytoplasm develops a complicated ultrastructure and changes dramatically during these stages. Lipid droplets increase after generative cell formation and their organization and distribution change with the developmental stage. Starch grains in amyloplasts increase in number and size during generative and sperm cell formation and decrease at pollen maturity. The shape and membrane system of mitochondria change only slightly. Dictyosomes become very prominent, and numerous associated vesicles are observed during and after sperm cell formation. Endoplasmic reticulum appears extensively as stacks during sperm cell formation. Free and polyribosomes are abundant in the cytoplasm at all developmental stages although they appear denser at certain stages and in some areas. In mature pollen, all organelles are randomly distributed throughout the vegetative cytoplasm and numerous small particles appear. Organization and distribution of storage substances and appearance of these small particles during generative and sperm cell formation and pollen maturation are discussed.

  6. Characteristics of T-cell and B-cell immune respoonses to pollen allergens in Bulgarian patients with pollinosis

    Directory of Open Access Journals (Sweden)

    ZHIVKA KOSTOVA

    2015-12-01

    Full Text Available Characterization of the pollen allergens and identification of the main T-cell and B-cell epitopes is of crucial importance, since the clarification of the cellular and molecular mechanisms will allow long-term control of this type allergic diseases and development of safe immunotherapeutic medication. The objective of this study was to identify which pollen allergens are responsible for the T-cell activation in atopic patients with pollinosis and to determine whether the same allergens are responsible for the IgE-mediated reactions. Thirty-seven patients with pollinosis and thirteen non-allergic subjects were recruited. Peripheral blood samples were collected out of the pollen season. T-cell responses (IFN-γ production towards different pollen allergens and levels of pollen specific IgE and IgG in the sera were measured by ELISA. The T-cell reactivity in most patients was directed towards the grass pollen B1, tree pollen I and autumn pollen B5. 70% of the allergic individuals responded to allergens from Lolium perenne, 84% to Dactylis glomerata, 11% to Phleum pratense, 65% to Betula pendula and 70% to Taxus baccata. Elevated serum levels of specific IgE in the allergic patients (p<0.01 were measured against the tree pollen I and autumn pollen, which include widespread deciduous trees (birch, willow, poplar and yew and Artemisia absinthium, respectively. Our results show that the T-cell reactivity and antibody responses may be directed towards different or the same allergens. In addition, we propose that short synthetic peptides, which contain overlapping T- and B-cell epitopes can be used for specific immunotherapy treatment if they lack antibody recognition domains.

  7. Cells of the connective tissue differentiate and migrate into pollen sacs

    Science.gov (United States)

    Iqbal, M. C. M.; Wijesekara, Kolitha B.

    2002-01-01

    In angiosperms, archesporial cells in the anther primordium undergo meiosis to form haploid pollen, the sole occupants of anther sacs. Anther sacs are held together by a matrix of parenchyma cells, the connective tissue. Cells of the connective tissue are not known to differentiate. We report the differentiation of parenchyma cells in the connective tissue of two Gordonia species into pollen-like structures (described as pseudopollen), which migrate into the anther sacs before dehiscence. Pollen and pseudopollen were distinguishable by morphology and staining. Pollen were tricolpate to spherical while pseudopollen were less rigid and transparent with a ribbed surface. Both types were different in size, shape, staining and surface architecture. The ratio of the number of pseudopollen to pollen was 1:3. During ontogeny in the connective tissue, neither cell division nor tetrad formation was observed and hence pseudopollen were presumed to be diploid. Only normal pollen germinated on a germination medium. Fixed preparations in time seemed to indicate that pseudopollen migrate from the connective tissue into the anther sac.

  8. Decreased viability and proliferation of CHANG conjunctival epithelial cells after contact with ultraviolet light-irradiated pollen.

    Science.gov (United States)

    Heidinger, Andrea; Rabensteiner, Dieter Franz; Rabensteiner, Jasmin; Kieslinger, Petra; Horwath-Winter, Jutta; Stabentheiner, Edith; Riedl, Regina; Wedrich, Andreas; Schmut, Otto

    2017-12-27

    Contact with pollen is the major reason for the development of allergic symptoms on the ocular surface leading to a significant increase of allergic diseases worldwide. Environmental changes such as increased ultraviolet (UV) radiation and air pollution are discussed as contributory causes for this increase. We investigated the effect of UV light on the histamine content of pollen and examined if an irradiation of pollen affects the viability and proliferation of conjunctival cells. Alder (Alnus glutinosa) and hazel (Corylus avellana) pollen were irradiated for different time periods with sunlight, UV-A or UV-B light and the histamine content was analysed and compared with non-irradiated pollen. Conjunctival epithelial cells (CHANG cells) were exposed to irradiated and non-irradiated pollen followed by an assessment of cell viability with the colorimetric MTS test and the impedance-based measurement of cell proliferation using the xCELLigence real-time analysis system. UV light irradiation increased the histamine level of alder and hazel pollen in a dose-dependent manner. CHANG cells treated with irradiated pollen induced a statistically significant higher decrease of cell viability than treatment with non-irradiated pollen. Our results indicate that UV light is able to alter pollen thus making them more harmful for conjunctival cells.

  9. Antiproliferative activity and apoptotic effects of Filipendula ulmaria pollen against C26 mice colon tumour cells

    Directory of Open Access Journals (Sweden)

    Mărgăoan Rodica

    2016-06-01

    Full Text Available Honeybee collected pollen exhibits high nutritional and pharmaceutical benefits for the human diet and medicine. Pollen’s antioxidant, anti-ageing, anti-inflammatory, anti-atherosclerosis, and cardioprotective activity, depending on the floral origin, are well known. Recent studies proposed that pollen may also be an excellent cancer-fighting candidate, as pollen harbours high amounts of phenolic substances. In our study, Filipendula ulmaria pollen (bee collected was methanol-water extracted and used to verify its in vitro pharmacological activities on C26 mice cancer tumour cells. Three different concentrations of the extract were tested in antitumour assays. Monitoring was done after 6, 12, 24, and 48 hours. Promising results were obtained for antiproliferative and apoptotic activity of the pollen extracts, with high efficiency for the highest concentration (1 mg/mL. For both activities, time and concentration-dependent effects were observed. Pollen extracts or bee collected pollen has a high potential as an antitumour agent for use in human medicine, because they are both rich in bioactive compounds.

  10. Taking one for the team: self-recognition and cell suicide in pollen.

    Science.gov (United States)

    Wilkins, Katie A; Poulter, Natalie S; Franklin-Tong, Vernonica E

    2014-03-01

    Self-incompatibility (SI) is an important genetically controlled mechanism used by many angiosperms to prevent self-fertilization and inbreeding. A multiallelic S-locus allows discrimination between 'self' (incompatible) pollen from 'nonself' pollen at the pistil. Interaction of matching pollen and pistil S-determinants allows 'self' recognition and triggers rejection of incompatible pollen. The S-determinants for Papaver rhoeas (poppy) are PrsS and PrpS. PrsS is a small secreted protein that acts as a signalling ligand to interact with its cognate pollen S-determinant PrpS, a small novel transmembrane protein. Interaction of PrsS with incompatible pollen stimulates increases in cytosolic free Ca(2+) and involves influx of Ca(2+) and K(+). Data implicate involvement of reactive oxygen species and nitric oxide signalling in the SI response. Downstream targets include the cytoskeleton, a soluble inorganic pyrophosphatase, Pr-p26.1, and a MAP kinase, PrMPK9-1. A major focus for SI-induced signalling is to initiate programmed cell death (PCD). In this review we provide an overview of our understanding of SI, with focus on how the signals and components are integrated, in particular, how reactive oxygen species, nitric oxide, and the actin cytoskeleton feed into a PCD network. We also discuss our recent functional expression of PrpS in Arabidopsis thaliana pollen in the context of understanding how PCD signalling systems may have evolved.

  11. Coronavirus infection of polarized epithelial cells

    NARCIS (Netherlands)

    Rossen, J W; Horzinek, M C; Rottier, P J

    1995-01-01

    Epithelial cells are the first host cells to be infected by incoming c oronaviruses. Recent observations in vitro show that coronaviruses are released from a specific side of these polarized cells, and this polarized release might be important for the spread of the infection in vivo. Mechanisms for

  12. Plant cells which aid in pollen digestion within a beetle's gut.

    Science.gov (United States)

    Rickson, Fred R; Cresti, M; Beach, James H

    1990-03-01

    The peach palm, Bactris gasipaes H.B.K., in Costa Rica, possesses unusual trichomes on the inflorescence epidermal surface. Certain cells of the trichome possess a thick, highly lignified cell wall and are consumed by the beetle Cyclocephala amazona L. before it ingests pollen from the same inflorescence. Chemical analyses show the trichome to possess no nutritive value. The thick-walled trichome cells pass intact through the beetle's digestive system, while ingested pollen is crushed. We suggest that the specialized plant cells function as gastroliths in the beetle's digestive tract.

  13. Actin polymerization drives polar growth in Arabidopsis root hair cells.

    Science.gov (United States)

    Vazquez, Luis Alfredo Bañuelos; Sanchez, Rosana; Hernandez-Barrera, Alejandra; Zepeda-Jazo, Isaac; Sánchez, Federico; Quinto, Carmen; Torres, Luis Cárdenas

    2014-01-01

    In plants, the actin cytoskeleton is a prime regulator of cell polarity, growth, and cytoplasmic streaming. Tip growth, as observed in root hairs, caulonema, and pollen tubes, is governed by many factors, including calcium gradients, exocytosis and endocytosis, reactive oxygen species, and the cytoskeleton. Several studies indicate that the polymerization of G-actin into F-actin also contributes to tip growth. The structure and function of F-actin within the apical dome is variable, ranging from a dense meshwork to sparse single filaments. The presence of multiple F-actin structures in the elongating apices of tip-growing cells suggests that this cytoskeletal array is tightly regulated. We recently reported that sublethal concentrations of fluorescently labeled cytochalasin could be used to visualize the distribution of microfilament plus ends using fluorescence microscopy, and found that the tip region of the growing root hair cells of a legume plant exhibits a clear response to the nodulation factors secreted by Rhizobium. (1) In this current work, we expanded our analysis using confocal microscopy and demonstrated the existence of highly dynamic fluorescent foci along Arabidopsis root hair cells. Furthermore, we show that the strongest fluorescence signal accumulates in the tip dome of the growing root hair and seems to be in close proximity to the apical plasma membrane. Based on these findings, we propose that actin polymerization within the dome of growing root hair cells regulates polar growth.

  14. [Cell polarity in the cardiovascular system].

    Science.gov (United States)

    Haller, C; Kübler, W

    1999-05-01

    The importance of cell polarity as a fundamental biological principle is increasingly recognized in the cardiovascular system. Polar cell mechanisms underlie not only the development of the heart and blood vessels, but also play a major role in the adult organism for polarized endothelial functions such as the separation of the intra- and extravascular compartment and the vectorial exchange of substances between these compartments. Endothelial cells are connected through intercellular junctions which separate the functionally and structurally distinct luminal and abluminal cell surfaces. The luminal plasma membrane is in contact with the blood and takes part in the regulation of hemostasis. The abluminal cell membrane connects the endothelial cell with the basement membrane and modulates blood flow through the release of vasoactive substances. Results from epithelial model systems have shown that the polarized cell phenotype is generated by specific protein sorting and regulated protein trafficking between the trans-Golgi network and the cell surface. The polarized distribution of cell membrane proteins is maintained by anchorage with the cytoskeleton and limitation of lateral diffusion by tight junctions. Disturbances of cell polarity may contribute to the pathogenesis of disease states, including ischemic and radiocontrast-induced acute renal failure and carcinomas. Recent results have demonstrated the importance of cholesterol for protein traffic from the trans-Golgi network to the apical cell membrane. This novel intracellular function of cholesterol could point to a connection between cell polarity and the pathogenesis of arteriosclerosis. The polarity of the endothelium also has to be taken into account when developing gene-therapeutic strategies, since therapeutic success will not only depend on the efficient expression of the desired gene product, but also on its correct cellular location or secretion into the correct extracellular compartment. These

  15. Pectic arabinan side chains are essential for pollen cell wall integrity during pollen development

    NARCIS (Netherlands)

    Cankar, K.; Kortstee, A.J.; Toonen, M.A.J.; Wolters-Arts, M.; Houbein, R.; Mariani, C.; Ulvskov, P.; Jorgensen, B.; Schols, H.A.; Visser, R.G.F.; Trindade, L.M.

    2014-01-01

    Pectin is a complex polysaccharide and an integral part of the primary plant cell wall and middle lamella, contributing to cell wall mechanical strength and cell adhesion. To understand the structure–function relationships of pectin in the cell wall, a set of transgenic potato lines with altered

  16. Pectic arabinan side chains are essential for pollen cell wall integrity during pollen development

    DEFF Research Database (Denmark)

    Cankar, Katarina; Kortstee, Anne; Toonen, Marcel A.J.

    2014-01-01

    Pectin is a complex polysaccharide and an integral part of the primary plant cell wall and middle lamella, contributing to cell wall mechanical strength and cell adhesion. To understand the structure-function relationships of pectin in the cell wall, a set of transgenic potato lines with altered...

  17. Understanding pollen tube growth: the hydrodynamic model versus the cell wall model

    NARCIS (Netherlands)

    Zonia, L.; Munnik, T.

    2011-01-01

    Scientific progress stimulates the evolution of models used to understand and conceptualize biological behaviors. The widely accepted cell wall model of pollen tube growth explains stochastic growth of the apical pectin wall, but fails to explain the mechanism driving oscillations in growth and cell

  18. Symmetry breaking signaling mechanisms during cell polarization

    NARCIS (Netherlands)

    Bruurs, LJM

    2017-01-01

    Breaking of cellular symmetry in order to establish an apico-basal polarity axis initiates de novo formation of cell polarity. However, symmetry breaking provides a formidable challenge from a signaling perspective, because by definition no spatial cues are present to instruct axis establishment.

  19. Elaborate spatial patterning of cell-wall PME and PMEI at the pollen tube tip involves PMEI endocytosis, and reflects the distribution of esterified and de-esterified pectins.

    Science.gov (United States)

    Röckel, Nina; Wolf, Sebastian; Kost, Benedikt; Rausch, Thomas; Greiner, Steffen

    2008-01-01

    In dicots, pectins are the major structural determinant of the cell wall at the pollen tube tip. Recently, immunological studies revealed that esterified pectins are prevalent at the apex of growing pollen tubes, where the cell wall needs to be expandable. In contrast, lateral regions of the cell wall contain mostly de-esterified pectins, which can be cross-linked to rigid gels by Ca(2+) ions. In pollen tubes, several pectin methylesterases (PMEs), enzymes that de-esterify pectins, are co-expressed with different PME inhibitors (PMEIs). This raises the possibility that interactions between PMEs and PMEIs play a key role in the regulation of cell-wall stability at the pollen tube tip. Our data establish that the PME isoform AtPPME1 (At1g69940) and the PMEI isoform AtPMEI2 (At3g17220), which are both specifically expressed in Arabidopsis pollen, physically interact, and that AtPMEI2 inactivates AtPPME1 in vitro. Furthermore, transient expression in tobacco pollen tubes revealed a growth-promoting activity of AtPMEI2, and a growth-inhibiting effect of AtPPME1. Interestingly, AtPPME1:YFP accumulated to similar levels throughout the cell wall of tobacco pollen tubes, including the tip region, whereas AtPMEI2:YFP was exclusively detected at the apex. In contrast to AtPPME1, AtPMEI2 localized to Brefeldin A-induced compartments, and was found in FYVE-induced endosomal aggregates. Our data strongly suggest that the polarized accumulation of PMEI isoforms at the pollen tube apex, which depends at least in part on local PMEI endocytosis at the flanks of the tip, regulates cell-wall stability by locally inhibiting PME activity.

  20. Polarized Cell Division of Chlamydia trachomatis.

    Directory of Open Access Journals (Sweden)

    Yasser Abdelrahman

    2016-08-01

    Full Text Available Bacterial cell division predominantly occurs by a highly conserved process, termed binary fission, that requires the bacterial homologue of tubulin, FtsZ. Other mechanisms of bacterial cell division that are independent of FtsZ are rare. Although the obligate intracellular human pathogen Chlamydia trachomatis, the leading bacterial cause of sexually transmitted infections and trachoma, lacks FtsZ, it has been assumed to divide by binary fission. We show here that Chlamydia divides by a polarized cell division process similar to the budding process of a subset of the Planctomycetes that also lack FtsZ. Prior to cell division, the major outer-membrane protein of Chlamydia is restricted to one pole of the cell, and the nascent daughter cell emerges from this pole by an asymmetric expansion of the membrane. Components of the chlamydial cell division machinery accumulate at the site of polar growth prior to the initiation of asymmetric membrane expansion and inhibitors that disrupt the polarity of C. trachomatis prevent cell division. The polarized cell division of C. trachomatis is the result of the unipolar growth and FtsZ-independent fission of this coccoid organism. This mechanism of cell division has not been documented in other human bacterial pathogens suggesting the potential for developing Chlamydia-specific therapeutic treatments.

  1. Male germ line development in Arabidopsis. duo pollen mutants reveal gametophytic regulators of generative cell cycle progression.

    Science.gov (United States)

    Durbarry, Anjusha; Vizir, Igor; Twell, David

    2005-01-01

    Male germ line development in flowering plants is initiated with the formation of the generative cell that is the progenitor of the two sperm cells. While structural features of the generative cell are well documented, genetic programs required for generative cell cycle progression are unknown. We describe two novel Arabidopsis (Arabidopsis thaliana) mutants, duo pollen1 (duo1) and duo pollen2 (duo2), in which generative cell division is blocked, resulting in the formation of bicellular pollen grains at anthesis. duo1 and duo2 map to different chromosomes and act gametophytically in a male-specific manner. Both duo mutants progress normally through the first haploid division at pollen mitosis I (PMI) but fail at distinct stages of the generative cell cycle. Mutant generative cells in duo1 pollen fail to enter mitosis at G2-M transition, whereas mutant generative cells in duo2 enter PMII but arrest at prometaphase. In wild-type plants, generative and sperm nuclei enter S phase soon after inception, implying that male gametic cells follow a simple S to M cycle. Mutant generative nuclei in duo1 complete DNA synthesis but bypass PMII and enter an endocycle during pollen maturation. However, mutant generative nuclei in duo2 arrest in prometaphase of PMII with a 2C DNA content. Our results identify two essential gametophytic loci required for progression through different phases of the generative cell cycle, providing the first evidence to our knowledge for genetic regulators of male germ line development in flowering plants.

  2. Male Germ Line Development in Arabidopsis. duo pollen Mutants Reveal Gametophytic Regulators of Generative Cell Cycle Progression1[w

    Science.gov (United States)

    Durbarry, Anjusha; Vizir, Igor; Twell, David

    2005-01-01

    Male germ line development in flowering plants is initiated with the formation of the generative cell that is the progenitor of the two sperm cells. While structural features of the generative cell are well documented, genetic programs required for generative cell cycle progression are unknown. We describe two novel Arabidopsis (Arabidopsis thaliana) mutants, duo pollen1 (duo1) and duo pollen2 (duo2), in which generative cell division is blocked, resulting in the formation of bicellular pollen grains at anthesis. duo1 and duo2 map to different chromosomes and act gametophytically in a male-specific manner. Both duo mutants progress normally through the first haploid division at pollen mitosis I (PMI) but fail at distinct stages of the generative cell cycle. Mutant generative cells in duo1 pollen fail to enter mitosis at G2-M transition, whereas mutant generative cells in duo2 enter PMII but arrest at prometaphase. In wild-type plants, generative and sperm nuclei enter S phase soon after inception, implying that male gametic cells follow a simple S to M cycle. Mutant generative nuclei in duo1 complete DNA synthesis but bypass PMII and enter an endocycle during pollen maturation. However, mutant generative nuclei in duo2 arrest in prometaphase of PMII with a 2C DNA content. Our results identify two essential gametophytic loci required for progression through different phases of the generative cell cycle, providing the first evidence to our knowledge for genetic regulators of male germ line development in flowering plants. PMID:15618418

  3. Coronaviruses in polarized epithelial cells

    NARCIS (Netherlands)

    Rossen, J. W.; Bekker, C. P.; Voorhout, W. F.; Horzinek, M. C.; van der Ende, A.; Strous, G. J.; Rottier, P. J.

    1995-01-01

    Coronaviruses have a marked tropism for epithelial cells. In this paper the interactions of the porcine transmissible gastroenteritis virus (TGEV) and mouse hepatitis virus (MHV-A59) with epithelial cells are compared. Porcine (LLC-PK1) and murine (mTAL) epithelial cells were grown on permeable

  4. Maize EMBRYO SAC family peptides interact differentially with pollen tubes and fungal cells.

    Science.gov (United States)

    Woriedh, Mayada; Merkl, Rainer; Dresselhaus, Thomas

    2015-08-01

    EMBRYO SAC1-4 (ES1-4) peptides belong to the defensin subgroup of cysteine-rich peptides known to mediate pollen tube burst in Zea mays (maize). ES1-4 are reported here to also be capable of inhibiting germination and growth of the maize fungal pathogens Fusarium graminearum and Ustilago maydis at higher concentrations. Dividing the peptides into smaller pieces showed that a 15-amino-acid peptide located in a highly variable loop region lacking similarity to other defensins or defensin-like peptides binds to maize pollen tube surfaces, causing swelling prior to burst. This peptide fragment and a second conserved neighbouring fragment showed suppression of fungal germination and growth. The two peptides caused swelling of fungal cells, production of reactive oxygen species, and finally the formation of big vacuoles prior to burst at high peptide concentration. Furthermore, peptide fragments were found to bind differently to fungal cells. In necrotrophic F. graminearum, a peptide fragment named ES-d bound only at cell surfaces whereas the peptide ES-c bound at cell surfaces and also accumulated inside cells. Conversely, in biotrophic U. maydis, both peptide fragments accumulated inside cells, but, if applied at higher concentration, ES-c but not ES-d accumulated mainly in vacuoles. Mapping of peptide interaction sites identified amino acids differing in pollen tube burst and fungal response reactions. In summary, these findings indicate that residues targeting pollen tube burst in maize are specific to the ES family, while residues targeting fungal growth are conserved within defensins and defensin-like peptides. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  5. Analysis of exocyst subunit EXO70 family reveals distinct membrane polar domains in Tobacco pollen tubes

    Czech Academy of Sciences Publication Activity Database

    Sekereš, Juraj; Pejchar, Přemysl; Šantrůček, J.; Vukašinović, Nemanja; Žárský, Viktor; Potocký, Martin

    2017-01-01

    Roč. 173, č. 3 (2017), s. 1659-1675 ISSN 0032-0889 R&D Projects: GA ČR GA13-19073S; GA ČR GA15-24711S Grant - others:OPPK(XE) CZ.2.16/3.1.00/21519 Institutional support: RVO:61389030 Keywords : PLASMA-MEMBRANE * ARABIDOPSIS-THALIANA * CELL-MIGRATION Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Plant sciences, botany Impact factor: 6.456, year: 2016

  6. Dissecting the regulation of pollen tube growth by modelling the interplay of hydrodynamics, cell wall and ion dynamics

    Directory of Open Access Journals (Sweden)

    Junli eLiu

    2014-08-01

    Full Text Available Hydrodynamics, cell wall and ion dynamics are all important properties that regulate pollen tube growth. Currently, the two main pollen tube growth models, the cell wall model and the hydrodynamic model do not appear to be reconcilable. Here we develop an integrative model for pollen tube growth and show that our model reproduces key experimental observations: 1 that the hypertonic condition leads to a much longer oscillatory period and that the hypotonic condition halves the oscillatory period; 2 that oscillations in turgor are experimentally undetectable; 3 that increasing the extracellular calcium concentration or decreasing the pH decreases the growth oscillatory amplitude; 4 that knockout of Raba4d, a member of the Rab family of small GTPase proteins, decreases pollen tube length after germination for 24 hours. Using the model generated here, we reveal that 1 when cell wall extensibility is large, pollen tube may sustain growth at different volume changes and maintain relatively stable turgor; 2 turgor increases if cell wall extensibility decreases; 3 increasing turgor due to decrease in osmolarity in the media, although very small, increases volume change . However, increasing turgor due to decrease in cell wall extensibility decreases volume change. In this way regulation of pollen tube growth by turgor is context dependent. By changing the osmolarity in the media, the main regulatory points are extracellular osmolarity for water flow and turgor for the volume encompassed by the cell wall. However, if the viscosity of cell wall changes, the main regulatory points are turgor for water flow and wall extensibility for the volume encompassed by the cell wall. The novel methodology developed here reveals the underlying context-dependent regulatory principle of pollen tube growth.

  7. Microspore embryogenesis: reprogramming cell fate from pollen to embryo development

    NARCIS (Netherlands)

    Hui Li,

    2014-01-01

    Microspore embryogenesis is an expression of plant cell totipotency that leads to the production of haploid embryos. Besides being a widely exploited plant breeding tool, microspore embryogenesis is also a fascinating system that can be used to obtain a deeper mechanistic understanding of plant

  8. A Pollen Coat–Inducible Autoinhibited Ca2+-ATPase Expressed in Stigmatic Papilla Cells Is Required for Compatible Pollination in the Brassicaceae[W

    Science.gov (United States)

    Iwano, Megumi; Igarashi, Motoko; Tarutani, Yoshiaki; Kaothien-Nakayama, Pulla; Nakayama, Hideki; Moriyama, Hideki; Yakabe, Ryo; Entani, Tetsuyuki; Shimosato-Asano, Hiroko; Ueki, Masao; Tamiya, Gen; Takayama, Seiji

    2014-01-01

    In the Brassicaceae, intraspecific non-self pollen (compatible pollen) can germinate and grow into stigmatic papilla cells, while self-pollen or interspecific pollen is rejected at this stage. However, the mechanisms underlying this selective acceptance of compatible pollen remain unclear. Here, using a cell-impermeant calcium indicator, we showed that the compatible pollen coat contains signaling molecules that stimulate Ca2+ export from the papilla cells. Transcriptome analyses of stigmas suggested that autoinhibited Ca2+-ATPase13 (ACA13) was induced after both compatible pollination and compatible pollen coat treatment. A complementation test using a yeast Saccharomyces cerevisiae strain lacking major Ca2+ transport systems suggested that ACA13 indeed functions as an autoinhibited Ca2+ transporter. ACA13 transcription increased in papilla cells and in transmitting tracts after pollination. ACA13 protein localized to the plasma membrane and to vesicles near the Golgi body and accumulated at the pollen tube penetration site after pollination. The stigma of a T-DNA insertion line of ACA13 exhibited reduced Ca2+ export, as well as defects in compatible pollen germination and seed production. These findings suggest that stigmatic ACA13 functions in the export of Ca2+ to the compatible pollen tube, which promotes successful fertilization. PMID:24569769

  9. Acetylesterase-Mediated Deacetylation of Pectin Impairs Cell Elongation, Pollen Germination, and Plant Reproduction

    Energy Technology Data Exchange (ETDEWEB)

    Gou J. Y.; Liu C.; Miller, L. M.; Hou, G.; Yu, X.-H.; Chen, X.-Y.

    2012-01-01

    Pectin is a major component of the primary cell wall of higher plants. Some galacturonyl residues in the backbone of pectinaceous polysaccharides are often O-acetylated at the C-2 or C-3 position, and the resulting acetylesters change dynamically during the growth and development of plants. The processes involve both enzymatic acetylation and deacetylation. Through genomic sequence analysis, we identified a pectin acetylesterase (PAE1) from black cottonwood (Populus trichocarpa). Recombinant Pt PAE1 exhibited preferential activity in releasing the acetate moiety from sugar beet (Beta vulgaris) and potato (Solanum tuberosum) pectin in vitro. Overexpressing Pt PAE1 in tobacco (Nicotiana tabacum) decreased the level of acetyl esters of pectin but not of xylan. Deacetylation engendered differential changes in the composition and/or structure of cell wall polysaccharides that subsequently impaired the cellular elongation of floral styles and filaments, the germination of pollen grains, and the growth of pollen tubes. Consequently, plants overexpressing PAE1 exhibited severe male sterility. Furthermore, in contrast to the conventional view, PAE1-mediated deacetylation substantially lowered the digestibility of pectin. Our data suggest that pectin acetylesterase functions as an important structural regulator in planta by modulating the precise status of pectin acetylation to affect the remodeling and physiochemical properties of the cell wall's polysaccharides, thereby affecting cell extensibility.

  10. Identification of the arabidopsis RAM/MOR signalling network: adding new regulatory players in plant stem cell maintenance and cell polarization

    Science.gov (United States)

    Zermiani, Monica; Begheldo, Maura; Nonis, Alessandro; Palme, Klaus; Mizzi, Luca; Morandini, Piero; Nonis, Alberto; Ruperti, Benedetto

    2015-01-01

    Background and Aims The RAM/MOR signalling network of eukaryotes is a conserved regulatory module involved in co-ordination of stem cell maintenance, cell differentiation and polarity establishment. To date, no such signalling network has been identified in plants. Methods Genes encoding the bona fide core components of the RAM/MOR pathway were identified in Arabidopsis thaliana (arabidopsis) by sequence similarity searches conducted with the known components from other species. The transcriptional network(s) of the arabidopsis RAM/MOR signalling pathway were identified by running in-depth in silico analyses for genes co-regulated with the core components. In situ hybridization was used to confirm tissue-specific expression of selected RAM/MOR genes. Key Results Co-expression data suggested that the arabidopsis RAM/MOR pathway may include genes involved in floral transition, by co-operating with chromatin remodelling and mRNA processing/post-transcriptional gene silencing factors, and genes involved in the regulation of pollen tube polar growth. The RAM/MOR pathway may act upstream of the ROP1 machinery, affecting pollen tube polar growth, based on the co-expression of its components with ROP-GEFs. In silico tissue-specific co-expression data and in situ hybridization experiments suggest that different components of the arabidopsis RAM/MOR are expressed in the shoot apical meristem and inflorescence meristem and may be involved in the fine-tuning of stem cell maintenance and cell differentiation. Conclusions The arabidopsis RAM/MOR pathway may be part of the signalling cascade that converges in pollen tube polarized growth and in fine-tuning stem cell maintenance, differentiation and organ polarity. PMID:26078466

  11. [Additional phragmoplast corrects abnormal cytokinesis in wheat x rye hybrid pollen mother cells].

    Science.gov (United States)

    Gordeeva, E I; Shamina, N V; Dudka, L F; Kovtunenko, V Ia; Bolobolova, E U

    2009-01-01

    The phragmoplast dysfunction in wheat x rye hybrid F1 male meiosis has been described. The pollen mother cells (PMCs) show the phenotype where transition from central spindle fibers (forming a solid bundle) to a phragmoplast (hollow cylinder) is blocked. The blockade suppresses centrifugal movement of the phragmoplast and cell plate formation. The resulting cells occur to be binucleate. Sometimes, the two nuclei join and form one restitution nucleus. PMCs of wheat x rye F1 hybrid N D-144gp 06r. F1 (T. aestivum c. 93-60 T 9 x S. cereale c. Saratovskaya 7) showing this phenotype have an additional phragmoplast at late telophase. This happens like that in the case of immobile phragmoplast formation in meiosis in bicotyledons: the new phragmoplast arises by the aid of microtubules polymerization starting from the spindle poles. The new additional phragmoplast builds a new cell plate and accomplishes cytokinesis.

  12. Mechanisms, safety and efficacy of a B cell epitope-based vaccine for immunotherapy of grass pollen allergy

    Directory of Open Access Journals (Sweden)

    Petra Zieglmayer

    2016-09-01

    Conclusion: The B cell epitope-based recombinant grass pollen allergy vaccine BM32 is well tolerated and few doses are sufficient to suppress immediate allergic reactions as well as allergen-specific T cell responses via a selective induction of allergen-specific IgG antibodies. (ClinicalTrials.gov number, NCT01445002.

  13. Morphogenesis of complex plant cell shapes: the mechanical role of crystalline cellulose in growing pollen tubes.

    Science.gov (United States)

    Aouar, Leila; Chebli, Youssef; Geitmann, Anja

    2010-03-01

    Cellulose is the principal component of the load-bearing system in primary plant cell walls. The great resistance to tensile forces of this polysaccharide and its embedding in matrix components make the cell wall a material similar to a fiber composite. In the rapidly growing pollen tube, the amount of cellulose in the cell wall is untypically low. Therefore, we want to investigate whether the load-bearing function of cellulose is nevertheless important for the architecture of this cell. Enzymatic digestion with cellulase and inhibition of cellulose crystal formation with CGA (1-cyclohexyl-5-(2,3,4,5,6-pentafluorophenoxy)-1lambda4,2,4,6-thiatriazin-3-amine) resulted in the formation of tubes with increased diameter in Solanum chacoense and Lilium orientalis when present during germination. In pre-germinated tubes, application of both agents resulted in the transient arrest of growth accompanied by the formation of an apical swelling indicating a role in the mechanical stabilization of this cellular region. Once growth resumed in the presence of cellulase, however, the cell wall in the newly formed tube showed increased amounts of pectins, possibly to compensate for the reduced amount of cellulose. Scanning electron microscopy of pollen tubes subjected to digestion of matrix polysaccharides revealed the mechanical anisotropy of the cell wall. In both Lilium and Solanum, the angle of highest stability revealed by crack formation was significantly below 45 degrees , an indication that in the mature part of the cell cellulose may not the main stress-bearing component against turgor pressure induced tensile stress in circumferential direction.

  14. Unique and cross-reactive T cell epitope peptides of the major Bahia grass pollen allergen, Pas n 1.

    Science.gov (United States)

    Etto, Tamara; de Boer, Carmela; Prickett, Sara; Gardner, Leanne M; Voskamp, Astrid; Davies, Janet M; O'Hehir, Robyn E; Rolland, Jennifer M

    2012-01-01

    Bahia grass pollen (BaGP) is a major cause of allergic rhinitis. Subcutaneous allergen-specific immunotherapy is effective for grass pollen allergy, but is unsuitable for patients with moderate to severe asthma due to the risk of anaphylaxis. T cell-reactive but IgE nonreactive peptides provide a safer treatment option. This study aimed to identify and characterize dominant CD4(+) T cell epitope peptides of the major BaGP allergen, Pas n 1. Pas n 1-specific T cell lines generated from the peripheral blood of BaGP-allergic subjects were tested for proliferative and cytokine response to overlapping 20-mer Pas n 1 peptides. Cross-reactivity to homologous peptides from Lol p 1 and Cyn d 1 of Ryegrass and Bermuda grass pollen, respectively, was assessed using Pas n 1 peptide-specific T cell clones. MHC class II restriction of Pas n 1 peptide T cell recognition was determined by HLA blocking assays and peptide IgE reactivity tested by dot blotting. Three Pas n 1 peptides showed dominant T cell reactivity; 15 of 18 (83%) patients responded to one or more of these peptides. T cell clones specific for dominant Pas n 1 peptides showed evidence of species-specific T cell reactivity as well as cross-reactivity with other group 1 grass pollen allergens. The dominant Pas n 1 T cell epitope peptides showed HLA binding diversity and were non-IgE reactive. The immunodominant T cell-reactive Pas n 1 peptides are candidates for safe immunotherapy for individuals, including those with asthma, who are allergic to Bahia and possibly other grass pollens. Copyright © 2012 S. Karger AG, Basel.

  15. Analysis of gemini pollen 3 mutant suggests a broad function of AUGMIN in microtubule organization during sexual reproduction in Arabidopsis.

    Science.gov (United States)

    Oh, Sung-Aeong; Jeon, Jien; Park, Hyo-Jin; Grini, Paul Eivind; Twell, David; Park, Soon Ki

    2016-07-01

    In flowering plants, male gametes arise via meiosis of diploid pollen mother cells followed by two rounds of mitotic division. Haploid microspores undergo polar nuclear migration and asymmetric division at pollen mitosis I to segregate the male germline, followed by division of the germ cell to generate a pair of sperm cells. We previously reported two gemini pollen (gem) mutants that produced twin-celled pollen arising from polarity and cytokinesis defects at pollen mitosis I in Arabidopsis. Here, we report an independent mutant, gem3, with a similar division phenotype and severe genetic transmission defects through pollen. Cytological analyses revealed that gem3 disrupts cell division during male meiosis, at pollen mitosis I and during female gametophyte development. We show that gem3 is a hypomorphic allele (aug6-1) of AUGMIN subunit 6, encoding a conserved component in the augmin complex, which mediates microtubule (MT)-dependent MT nucleation in acentrosomal cells. We show that MT arrays are disturbed in gem3/aug6-1 during male meiosis and pollen mitosis I using fluorescent MT-markers. Our results demonstrate a broad role for the augmin complex in MT organization during sexual reproduction, and highlight gem3/aug6-1 mutants as a valuable tool for the investigation of augmin-dependent MT nucleation and dynamics in plant cells. © 2016 The Authors The Plant Journal © 2016 John Wiley & Sons Ltd.

  16. Identification of T-cell epitopes of Lol p 9, a major allergen of ryegrass (Lolium perenne) pollen.

    Science.gov (United States)

    Blaher, B; Suphioglu, C; Knox, R B; Singh, M B; McCluskey, J; Rolland, J M

    1996-07-01

    T-cell recognition of Lol p 9, a major allergen of ryegrass pollen, was investigated by using a T-cell line and T-cell clones generated from the peripheral blood of an atopic donor. The T-cell line reacted with purified Lol p 9, as well as with crude ryegrass pollen extract, but failed to cross-react with Bermuda grass pollen extract. All of six T-cell clones generated from this line proliferated in response to Lol p 9. Epitope mapping was carried out with a panel of 34 overlapping synthetic peptides, which spanned the entire sequence of the Lol p 9 12R isoform. The T-cell line responded to two of the peptides, Lol p 9 (105-116) and Lol p 9 (193-204), whereas reactivity with one or other of these peptides was shown by five T-cell clones. These two peptides contained sequences consistent with motifs previously reported for major histocompatibility complex class II-restricted peptides. HLA antibody blocking studies showed that presentation of peptide Lol p 9 (105-116) to one T-cell clone was HLA-DR-restricted; this clone expressed a T helper cell phenotype (CD3+, CD4+) and the T-cell receptor alpha beta. The identification of immunodominant T-cell epitope(s) on allergens is essential for devising safer and more effective immunotherapy strategies, which can interrupt the chain of events leading to allergic disease.

  17. Lipids in pollen - They are different.

    Science.gov (United States)

    Ischebeck, Till

    2016-09-01

    During evolution, the male gametophyte of Angiosperms has been severely reduced to the pollen grain, consisting of a vegetative cell containing two sperm cells. This vegetative cell has to deliver the sperm cells from the stigma through the style to the ovule. It does so by producing a pollen tube and elongating it to many centimeters in length in some species, requiring vast amounts of fatty acid and membrane lipid synthesis. In order to optimize this polar tip growth, a unique lipid composition in the pollen has evolved. Pollen tubes produce extraplastidial galactolipids and store triacylglycerols in lipid droplets, probably needed as precursors of glycerolipids or for acyl editing. They also possess special sterol and sphingolipid moieties that might together form microdomains in the membranes. The individual lipid classes, the proteins involved in their synthesis as well as the corresponding Arabidopsis knockout mutant phenotypes are discussed in this review. This article is part of a Special Issue entitled: Plant Lipid Biology edited by Kent D. Chapman and Ivo Feussner. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Genome-scale analysis and comparison of gene expression profiles in developing and germinated pollen in Oryza sativa.

    Science.gov (United States)

    Wei, Li Q; Xu, Wen Y; Deng, Zhu Y; Su, Zhen; Xue, Yongbiao; Wang, Tai

    2010-05-28

    Pollen development from the microspore involves a series of coordinated cellular events, and the resulting mature pollen has a specialized function to quickly germinate, produce a polar-growth pollen tube derived from the vegetative cell, and deliver two sperm cells into the embryo sac for double fertilization. The gene expression profiles of developing and germinated pollen have been characterised by use of the eudicot model plant Arabidopsis. Rice, one of the most important cereal crops, has been used as an excellent monocot model. A comprehensive analysis of transcriptome profiles of developing and germinated pollen in rice is important to understand the conserved and diverse mechanism underlying pollen development and germination in eudicots and monocots. We used Affymetrix GeneChip Rice Genome Array to comprehensively analyzed the dynamic changes in the transcriptomes of rice pollen at five sequential developmental stages from microspores to germinated pollen. Among the 51,279 transcripts on the array, we found 25,062 pollen-preferential transcripts, among which 2,203 were development stage-enriched. The diversity of transcripts decreased greatly from microspores to mature and germinated pollen, whereas the number of stage-enriched transcripts displayed a "U-type" change, with the lowest at the bicellular pollen stage; and a transition of overrepresented stage-enriched transcript groups associated with different functional categories, which indicates a shift in gene expression program at the bicellular pollen stage. About 54% of the now-annotated rice F-box protein genes were expressed preferentially in pollen. The transcriptome profile of germinated pollen was significantly and positively correlated with that of mature pollen. Analysis of expression profiles and coexpressed features of the pollen-preferential transcripts related to cell cycle, transcription, the ubiquitin/26S proteasome system, phytohormone signalling, the kinase system and defense

  19. Genome-scale analysis and comparison of gene expression profiles in developing and germinated pollen in Oryza sativa

    Directory of Open Access Journals (Sweden)

    Xue Yongbiao

    2010-05-01

    Full Text Available Abstract Background Pollen development from the microspore involves a series of coordinated cellular events, and the resulting mature pollen has a specialized function to quickly germinate, produce a polar-growth pollen tube derived from the vegetative cell, and deliver two sperm cells into the embryo sac for double fertilization. The gene expression profiles of developing and germinated pollen have been characterised by use of the eudicot model plant Arabidopsis. Rice, one of the most important cereal crops, has been used as an excellent monocot model. A comprehensive analysis of transcriptome profiles of developing and germinated pollen in rice is important to understand the conserved and diverse mechanism underlying pollen development and germination in eudicots and monocots. Results We used Affymetrix GeneChip® Rice Genome Array to comprehensively analyzed the dynamic changes in the transcriptomes of rice pollen at five sequential developmental stages from microspores to germinated pollen. Among the 51,279 transcripts on the array, we found 25,062 pollen-preferential transcripts, among which 2,203 were development stage-enriched. The diversity of transcripts decreased greatly from microspores to mature and germinated pollen, whereas the number of stage-enriched transcripts displayed a "U-type" change, with the lowest at the bicellular pollen stage; and a transition of overrepresented stage-enriched transcript groups associated with different functional categories, which indicates a shift in gene expression program at the bicellular pollen stage. About 54% of the now-annotated rice F-box protein genes were expressed preferentially in pollen. The transcriptome profile of germinated pollen was significantly and positively correlated with that of mature pollen. Analysis of expression profiles and coexpressed features of the pollen-preferential transcripts related to cell cycle, transcription, the ubiquitin/26S proteasome system, phytohormone

  20. Radiation dose-response relationship of micronucleus occurrence in pollen mother cells of tradescantia

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Jin Kyu; Kim, Yeon Ku; Song, Hi Sup [KAERI, Taejon (Korea, Republic of)

    1999-10-01

    This study was carried out to investigate the radiation dose-response of micronucleus frequencies in Tradescantia pollen mother cells. The number of micronuclei increased in the tetrads as a result of chromosome deletion after irradiation. The maximal frequency of micronucleus showed a good dose-response relationship in the range of dose 0{approx}50 cGy. On the basis of the relationship, a dose of 1 cGy resulted in two additional micronuclei in 100 tetrads. The radiation dose-response relationship of micronucleus occurrence is prerequisite to biological monitoring of radiation and can be modified for biological risk assessment of toxicants, and to safety test of water or soil integrity.

  1. Tightly Controlled Expression of bHLH142 Is Essential for Timely Tapetal Programmed Cell Death and Pollen Development in Rice

    Directory of Open Access Journals (Sweden)

    Swee-Suak Ko

    2017-07-01

    Full Text Available Male sterility is important for hybrid seed production. Pollen development is regulated by a complex network. We previously showed that knockout of bHLH142 in rice (Oryza sativa causes pollen sterility by interrupting tapetal programmed cell death (PCD and bHLH142 coordinates with TDR to modulate the expression of EAT1. In this study, we demonstrated that overexpression of bHLH142 (OE142 under the control of the ubiquitin promoter also leads to male sterility in rice by triggering the premature onset of PCD. Protein of bHLH142 was found to accumulate specifically in the OE142 anthers. Overexpression of bHLH142 induced early expression of several key regulatory transcription factors in pollen development. In particular, the upregulation of EAT1 at the early stage of pollen development promoted premature PCD in the OE142 anthers, while its downregulation at the late stage impaired pollen development by suppressing genes involved in pollen wall biosynthesis, ROS scavenging and PCD. Collectively, these events led to male sterility in OE142. Analyses of related mutants further revealed the hierarchy of the pollen development regulatory gene network. Thus, the findings of this study advance our understanding of the central role played by bHLH142 in the regulatory network leading to pollen development in rice and how overexpression of its expression affects pollen development. Exploitation of this novel functionality of bHLH142 may confer a big advantage to hybrid seed production.

  2. The Cysteine Protease CEP1, a Key Executor Involved in Tapetal Programmed Cell Death, Regulates Pollen Development in Arabidopsis[W][OPEN

    Science.gov (United States)

    Zhang, Dandan; Liu, Di; Lv, Xiaomeng; Wang, Ying; Xun, Zhili; Liu, Zhixiong; Li, Fenglan; Lu, Hai

    2014-01-01

    Tapetal programmed cell death (PCD) is a prerequisite for pollen grain development in angiosperms, and cysteine proteases are the most ubiquitous hydrolases involved in plant PCD. We identified a papain-like cysteine protease, CEP1, which is involved in tapetal PCD and pollen development in Arabidopsis thaliana. CEP1 is expressed specifically in the tapetum from stages 5 to 11 of anther development. The CEP1 protein first appears as a proenzyme in precursor protease vesicles and is then transported to the vacuole and transformed into the mature enzyme before rupture of the vacuole. cep1 mutants exhibited aborted tapetal PCD and decreased pollen fertility with abnormal pollen exine. A transcriptomic analysis revealed that 872 genes showed significantly altered expression in the cep1 mutants, and most of them are important for tapetal cell wall organization, tapetal secretory structure formation, and pollen development. CEP1 overexpression caused premature tapetal PCD and pollen infertility. ELISA and quantitative RT-PCR analyses confirmed that the CEP1 expression level showed a strong relationship to the degree of tapetal PCD and pollen fertility. Our results reveal that CEP1 is a crucial executor during tapetal PCD and that proper CEP1 expression is necessary for timely degeneration of tapetal cells and functional pollen formation. PMID:25035401

  3. Pollen types used by Centris (Hemisiella tarsata Smith (1874 (Hymenoptera, Apidae in the provisioning of brood cells in an area of Caatinga

    Directory of Open Access Journals (Sweden)

    Ana Paula Araújo da Cruz

    2015-06-01

    Full Text Available The aim of this study was to identify, by sediment pollen analysis, the plant species used as floral resources for the provisioning of brood cells in Centris (Hemisiella tarsata, in an area of Caatinga, within the municipality of Nova Soure, Bahia State, Brazil. The analysis of pollen contents from three brood cells revealed 11 pollen types, corresponding to four botanical families. Malpighiaceae was represented most, followed by Leguminosae, Ochnaceae, and Solanaceae, the latter two represented by just a single pollen type each. On the basis of the percentages in the samples, it was possible to infer that C. tarsata visited distinct plants, but intensified its pollen collection in species related to Aeschynomene martii and Solanum paniculatumpollen types, which are considered the most important pollen sources in the larval diet of this bee. In addition to the pollen sources, we have also recorded seven pollen types regarded as oil ones, all related to the Malpighiaceae family. The information about the resources for C. tarsata can be of great relevance, in view of the importance of these bees in the pollination of native flora.

  4. Classical cadherins control nucleus and centrosome position and cell polarity.

    Science.gov (United States)

    Dupin, Isabelle; Camand, Emeline; Etienne-Manneville, Sandrine

    2009-06-01

    Control of cell polarity is crucial during tissue morphogenesis and renewal, and depends on spatial cues provided by the extracellular environment. Using micropatterned substrates to impose reproducible cell-cell interactions, we show that in the absence of other polarizing cues, cell-cell contacts are the main regulator of nucleus and centrosome positioning, and intracellular polarized organization. In a variety of cell types, including astrocytes, epithelial cells, and endothelial cells, calcium-dependent cadherin-mediated cell-cell interactions induce nucleus and centrosome off-centering toward cell-cell contacts, and promote orientation of the nucleus-centrosome axis toward free cell edges. Nucleus and centrosome off-centering is controlled by N-cadherin through the regulation of cell interactions with the extracellular matrix, whereas the orientation of the nucleus-centrosome axis is determined by the geometry of N-cadherin-mediated contacts. Our results demonstrate that in addition to the specific function of E-cadherin in regulating baso-apical epithelial polarity, classical cadherins control cell polarization in otherwise nonpolarized cells.

  5. Addition of Phenylboronic Acid to Malus domestica Pollen Tubes Alters Calcium Dynamics, Disrupts Actin Filaments and Affects Cell Wall Architecture.

    Directory of Open Access Journals (Sweden)

    Kefeng Fang

    Full Text Available A key role of boron in plants is to cross-link the cell wall pectic polysaccharide rhamnogalacturonan-II (RG-II through borate diester linkages. Phenylboronic acid (PBA can form the same reversible ester bonds but cannot cross-link two molecules, so can be used as an antagonist to study the function of boron. This study aimed to evaluate the effect of PBA on apple (Malus domestica pollen tube growth and the underlying regulatory mechanism. We observed that PBA caused an inhibition of pollen germination, tube growth and led to pollen tube morphological abnormalities. Fluorescent labeling, coupled with a scanning ion-selective electrode technique, revealed that PBA induced an increase in extracellular Ca2+ influx, thereby elevating the cytosolic Ca2+ concentration [Ca2+]c and disrupting the [Ca2+]c gradient, which is critical for pollen tube growth. Moreover the organization of actin filaments was severely perturbed by the PBA treatment. Immunolocalization studies and fluorescent labeling, together with Fourier-transform infrared analysis (FTIR suggested that PBA caused an increase in the abundance of callose, de-esterified pectins and arabinogalactan proteins (AGPs at the tip. However, it had no effect on the deposition of the wall polymers cellulose. These effects are similar to those of boron deficiency in roots and other organs, indicating that PBA can induce boron deficiency symptoms. The results provide new insights into the roles of boron in pollen tube development, which likely include regulating [Ca2+]c and the formation of the actin cytoskeleton, in addition to the synthesis and assembly of cell wall components.

  6. Retracing the path of planar cell polarity.

    Science.gov (United States)

    Schenkelaars, Quentin; Fierro-Constain, Laura; Renard, Emmanuelle; Borchiellini, Carole

    2016-04-02

    The Planar Cell Polarity pathway (PCP) has been described as the main feature involved in patterning cell orientation in bilaterian tissues. Recently, a similar phenomenon was revealed in cnidarians, in which the inhibition of this pathway results in the absence of cilia orientation in larvae, consequently proving the functional conservation of PCP signaling between Cnidaria and Bilateria. Nevertheless, despite the growing accumulation of databases concerning basal lineages of metazoans, very few information concerning the existence of PCP components have been gathered outside of Bilateria and Cnidaria. Thus, the origin of this module or its prevalence in early emerging metazoans has yet to be elucidated. The present study addresses this question by investigating the genomes and transcriptomes from all poriferan lineages in addition to Trichoplax (Placozoa) and Mnemiopsis (Ctenophora) genomes for the presence of the core components of this pathway. Our results confirm that several PCP components are metazoan innovations. In addition, we show that all members of the PCP pathway, including a bona fide Strabismus ortholog (Van gogh), are retrieved only in one sponge lineage (Homoscleromorpha) out of four. This highly suggests that the full PCP pathway dates back at least to the emergence of homoscleromorph sponges. Consequently, several secondary gene losses would have occurred in the three other poriferan lineages including Amphimedon queenslandica (Demospongiae). Several proteins were not retrieved either in placozoans or ctenophores leading us to discuss the difficulties to predict orthologous proteins in basally branching animals. Finally, we reveal how the study of multigene families may be helpful to unravel the relationships at the base of the metazoan tree. The PCP pathway antedates the radiation of Porifera and may have arisen in the last common ancestor of animals. Oscarella species now appear as key organisms to understand the ancestral function of PCP

  7. Sulfinylated Azadecalins act as functional mimics of a pollen germination stimulant in Arabidopsis pistils

    Science.gov (United States)

    Qin, Yuan; Wysocki, Ronald J; Somogyi, Arpad; Feinstein, Yelena; Franco, Jessica Y; Tsukamoto, Tatsuya; Dunatunga, Damayanthi; Levy, Clara; Smith, Steven; Simpson, Robert; Gang, David; Johnson, Mark A; Palanivelu, Ravishankar

    2011-01-01

    SUMMARY Polarized cell elongation is triggered by small molecule cues during development of diverse organisms. During plant reproduction, pollen interactions with the stigma result in the polar outgrowth of a pollen tube, which delivers sperm cells to the female gametophyte to effect double fertilization. In many plants, pistils stimulate pollen germination. However, in Arabidopsis, the effect of pistils on pollen germination and the pistil factors that stimulate pollen germination remain poorly characterized. Here, we demonstrate that stigma, style, and ovules in Arabidopsis pistils stimulate pollen germination. We isolated an Arabidopsis pistil extract fraction that stimulates Arabidopsis pollen germination, and employed ultrahigh resolution ESI FT-ICR and MS/MS techniques to accurately determine the mass (202.126 daltons) of a compound that is specifically present in this pistil extract fraction. Using the molecular formula (C10H19NOS) and tandem mass spectral fragmentation patterns of the m/z (mass to charge ratio) 202.126 ion, we postulated chemical structures, devised protocols, synthesized N-Methanesulfinyl 1- and 2-azadecalins that are close structural mimics of the m/z 202.126 ion, and showed that they are sufficient to stimulate Arabidopsis pollen germination in vitro (30 µM stimulated ~50% germination) and elicit accession-specific response. Although N-Methanesulfinyl 2-azadecalin stimulated pollen germination in three species of Lineage I of Brassicaceae, it did not induce a germination response in Sisymbrium irio (Lineage II of Brassicaceae) and tobacco, indicating that activity of the compound is not random. Our results show that Arabidopsis pistils promote germination by producing azadecalin-like molecules to ensure rapid fertilization by the appropriate pollen. PMID:21801250

  8. The polarized double cell target of the SMC

    Energy Technology Data Exchange (ETDEWEB)

    Adams, D.; Adeva, B.; Arik, E.; Arvidson, A.; Badelek, B.; Ballintijn, M.K.; Bardin, G.; Baum, G.; Berglund, P.; Betev, L.; Bird, I.G.; Birsa, R.; Bjoerkholm, P.; Bonner, B.E.; Botton, N. de; Boutemeur, M.; Bradamante, F.; Bravar, A.; Bressan, A.; Bueltmann, S.; Burtin, E.; Cavata, C.; Crabb, D.; Cranshaw, J.; Cuhadar, T.; Torre, S. Dalla; Dantzig, R. van; Derro, B.; Deshpande, A.; Dhawan, S.; Dulya, C.; Dyring, A.; Eichblatt, S.; Faivre, J.C.; Fasching, D.; Feinstein, F.; Fernandez, C.; Forthmann, S.; Frois, B.; Gallas, A.; Garzon, J.A.; Gaussiran, T.; Gilly, H.; Giorgi, M.; Goeler, E. von; Goertz, S.; Gracia, G.; Groot, N. de; Perdekamp, M. Grosse; Guelmez, E.; Haft, K.; Harrach, D. von; Hasegawa, T.; Hautle, P.; Hayashi, N.; Heusch, C.A.; Horikawa, N.; Hughes, V.W.; Igo, G.; Ishimoto, S.; Iwata, T.; Kabuss, E.M.; Kageya, T.; Karev, A.; Kessler, H.J.; Ketel, T.J.; Kiryluk, J.; Kishi, A.; Kisselev, Yu.; Klostermann, L.; Kraemer, D.; Krivokhijine, V.; Kroeger, W.; Kurek, K.; Kyynaeraeinen, J.; Lamanna, M.; Landgraf, U.; Layda, T.; Le Goff, J.M.; Lehar, F.; Lesquen, A. de; Lichtenstadt, J.; Lindqvist, T.; Litmaath, M.; Lowe, M.; Magnon, A.; Mallot, G.K.; Marie, F.; Martin, A.; Martino, J.; Matsuda, T.; Mayes, B.; McCarthy, J.S.; Medved, K.; Meyer, W.; Middelkoop, G. van; Miller, D.; Miyachi, Y.; Mori, K.; Moromisato, J.; Nassalski, J.; Naumann, L.; Neganov, B.; Niinikoski, T.O.; Oberski, J.E.J.; Ogawa, A.; Ozben, C.; Parks, D.P.; Pereira, H.; Penzo, A.; Perrot-Kunne, F.; Peshekhonov, D.; Piegaia, R.; Pinsky, L.; Platchkov, S.; Plo, M.; Pose, D.; Postma, H. E-mail: hpostma@dataweb.nl; Pretz, J.; Pussieux, T.; Pyrlik, J.; Raedel, G.; Reyhancan, I.; Reicherz, G.; Rieubland, J.M.; Rijllart, A.; Roberts, J.B.; Rock, S.; Rodriguez, M.; Rondio, E.; Rosado, A.; Roscherr, B.; Sabo, I.; Saborido, J.; Sandacz, A.; Savin, I.; Schiavon, P.; Schiller, A.; Schueler, K.P.; Segel, R.; Seitz, R.; Semertzidis, Y.; Sever, F.; Shanahan, P.; Sichtermann, E.P.; Simeoni, F. [and others

    1999-11-11

    The polarized target of the Spin Muon Collaboration at CERN was used for deep inelastic muon scattering experiments during 1993-1996 with a polarized muon beam to investigate the spin structure of the nucleon. Most of the experiments were carried out with longitudinal target polarization and 190 GeV muons, and some were done with transverse polarization and 100 GeV muons. Protons as well as deuterons were polarized by dynamic nuclear polarization (DNP) in three kinds of solid materials -- butanol, ammonia, and deuterated butanol -- with maximum degrees of polarization of 94%, 91% and 60%, respectively. Considerable attention was paid to the accuracies of the NMR polarization measurements and their analyses, the accuracies achieved were between 2.0% and 3.2%. The SMC target system with two cells of opposite polarizations, each cell 65 cm long and 5 cm in diameter, constitutes the largest polarized target system ever built and facilitates accurate spin asymmetry measurements. The design considerations, construction and performance of the target are reviewed.

  9. The polarized double cell target of the SMC

    CERN Document Server

    Adams, D; Arik, E; Arvidson, A; Badelek, B; Ballintijn, M K; Bardin, G; Baum, G; Berglund, P; Betev, L; Bird, I G; Birsa, R; Björkholm, P; Bonner, B E; De Botton, N R; Boutemeur, M; Bradamante, Franco; Bravar, A; Bressan, A; Bültmann, S; Burtin, E; Cavata, C; Crabb, D; Cranshaw, J; Çuhadar-Dönszelmann, T; Dalla Torre, S; Van Dantzig, R; Derro, B R; Deshpande, A A; Dhawan, S K; Dulya, C M; Dyring, A; Eichblatt, S; Faivre, Jean-Claude; Fasching, D; Feinstein, F; Fernández, C; Forthmann, S; Frois, Bernard; Gallas, A; Garzón, J A; Gaussiran, T; Gilly, H; Giorgi, M A; von Goeler, E; Görtz, S; Gracia, G; De Groot, N; Grosse-Perdekamp, M; Gülmez, E; Haft, K; Von Harrach, D; Hasegawa, T; Hautle, P; Hayashi, N; Heusch, C A; Horikawa, N; Hughes, V W; Igo, G; Ishimoto, S; Iwata, T; Kabuss, E M; Kageya, T; Karev, A G; Kessler, H J; Ketel, T; Kiryluk, J; Kishi, A; Kiselev, Yu F; Klostermann, L; Krämer, Dietrich; Krivokhizhin, V G; Kröger, W; Kurek, K; Kyynäräinen, J; Lamanna, M; Landgraf, U; Layda, T; Le Goff, J M; Lehár, F; de Lesquen, A; Lichtenstadt, J; Lindqvist, T; Litmaath, M; Loewe, M; Magnon, A; Mallot, G K; Marie, F; Martin, A; Martino, J; Matsuda, T; Mayes, B W; McCarthy, J S; Medved, K S; Meyer, W T; Van Middelkoop, G; Miller, D; Miyachi, Y; Mori, K; Moromisato, J H; Nassalski, J P; Naumann, Lutz; Neganov, B S; Niinikoski, T O; Oberski, J; Ogawa, A; Ozben, C; Parks, D P; Pereira, H; Penzo, Aldo L; Perrot-Kunne, F; Peshekhonov, V D; Piegaia, R; Pinsky, L; Platchkov, S K; Pló, M; Pose, D; Postma, H; Pretz, J; Pussieux, T; Pyrlik, J; Rädel, G; Reyhancan, I; Reicherz, G; Rijllart, A; Roberts, J B; Rock, S E; Rodríguez, M; Rondio, Ewa; Rosado, A; Roscherr, B; Sabo, I; Saborido, J; Sandacz, A; Savin, I A; Schiavon, R P; Schiller, A; Schüler, K P; Segel, R E; Seitz, R; Semertzidis, Y K; Sever, F; Shanahan, P; Sichtermann, E P; Simeoni, F; Smirnov, G I; Staude, A; Steinmetz, A; Stiegler, U; Stuhrmann, H B; Szleper, M; Teichert, K M; Tessarotto, F; Thers, D; Tlaczala, W; Trentalange, S; Tripet, A; Ünel, G; Velasco, M; Vogt, J; Voss, Rüdiger; Weinstein, R; Whitten, C; Windmolders, R; Willumeit, R; Wislicki, W; Witzmann, A; Zanetti, A M; Zaremba, K; Zhao, J

    1999-01-01

    The polarized target of the Spin Muon Collaboration at CERN was used for deep inelastic muon scattering experiments during 1993 to 1996 with a polarized muon beam to investigate the spin structure of the nucleon. Most of the experiments were carried out with longitudinal target polarization and 190 GeV muons, and some were done with transverse polarization and 100 GeV muons. Protons as well as deuterons were polarized by dynamic nuclear polarization (DNP) in three kinds of solid materials $-$ butanol, ammonia, and deuterated butanol, with maximum degrees of polarization of 94, 91, and 60 \\%, respectively. Considerable attention was paid to the accuracies of the NMR polarization measurements and their analyses. The achieved accuracies were between 2.0 and 3.2 \\%. The SMC target system with two cells of opposite polarizations, each cell 65 cm long and 5 cm in diameter, constitutes the largest polarized target system ever built and facilitates accurate spin asymmetry measurements. The design considerations, the ...

  10. Polymer photovoltaic cells sensitive to the circular polarization of light

    Energy Technology Data Exchange (ETDEWEB)

    Gilot, Jan; Abbel, Robert; Lakhwani, Girish; Meijer, E.W.; Schenning, Albertus P.H.J.; Meskers, Stefan C.J. [Laboratory of Macromolecular and Organic Chemistry, Eindhoven University of Technology (Netherlands)

    2010-05-25

    Chiral conjugated polymer is used to construct a photovoltaic cell whose response depends on the circular polarization of the incoming light. The selectivity for left and right polarized light as a function of the thickness of the polymer layer is accounted for by modeling of the optical properties of all layers inside the device. (Abstract Copyright [2010], Wiley Periodicals, Inc.)

  11. Interference of the Histone Deacetylase Inhibits Pollen Germination and Pollen Tube Growth in Picea wilsonii Mast.

    Directory of Open Access Journals (Sweden)

    Yaning Cui

    Full Text Available Histone deacetylase (HDAC is a crucial component in the regulation of gene expression in various cellular processes in animal and plant cells. HDAC has been reported to play a role in embryogenesis. However, the effect of HDAC on androgamete development remains unclear, especially in gymnosperms. In this study, we used the HDAC inhibitors trichostatin A (TSA and sodium butyrate (NaB to examine the role of HDAC in Picea wilsonii pollen germination and pollen tube elongation. Measurements of the tip-focused Ca2+ gradient revealed that TSA and NaB influenced this gradient. Immunofluorescence showed that actin filaments were disrupted into disorganized fragments. As a result, the vesicle trafficking was disturbed, as determined by FM4-64 labeling. Moreover, the distribution of pectins and callose in cell walls was significantly altered in response to TSA and NaB. Our results suggest that HDAC affects pollen germination and polarized pollen tube growth in Picea wilsonii by affecting the intracellular Ca2+ concentration gradient, actin organization patterns, vesicle trafficking, as well as the deposition and configuration of cell wall components.

  12. EXO70C2 is a key regulatory factor for optimal tip growth of pollen

    KAUST Repository

    Synek, Lukas

    2017-03-30

    The exocyst, an eukaryotic tethering complex, co-regulates targeted exocytosis as an effector of small GTPases in polarized cell growth. In land plants, several exocyst subunits are encoded by double or triple paralogs, culminating in tens of EXO70 paralogs. Out of 23 Arabidopsis EXO70 isoforms, we analyzed seven isoforms expressed in pollen. Genetic and microscopic analyses of single mutants in EXO70A2, C1, C2, F1, H3, H5, and H6 genes revealed that only a loss-of-function EXO70C2 allele resulted in a significant male-specific transmission defect (segregation 40%:51%:9%) due to aberrant pollen tube growth. Mutant pollen tubes grown in vitro exhibited enhanced growth rate and a decreased thickness of the tip cell wall, causing tip bursts. However, exo70C2 pollen tubes could frequently recover and restart their speedy elongation, resulting in a repetitive stop-and-go growth dynamics. A pollen-specific depletion of the closest paralog, EXO70C1, using ami-RNA in the exo70C2 mutant background resulted in a complete pollen-specific transmission defect, suggesting redundant functions of EXO70C1 and EXO70C2. Both EXO70C1 and EXO70C2, GFP-tagged and expressed under their native promoters, localized in the cytoplasm of pollen grains, pollen tubes, and also root trichoblast cells. Expression of EXO70C2-GFP complemented aberrant growth of exo70C2 pollen tubes. The absent EXO70C2 interactions with core exocyst subunits in the yeast two-hybrid assay, cytoplasmic localization, and genetic effect suggest an unconventional EXO70 function possibly as a regulator of exocytosis outside the exocyst complex. In conclusion, EXO70C2 is a novel factor contributing to the regulation of optimal tip growth of Arabidopsis pollen tubes.

  13. Studies on optical pumping cells (OPC) to polarize 3He

    International Nuclear Information System (INIS)

    Hutanu, V.; Rupp, A.

    2004-01-01

    The technique applied at HMI to obtain nuclear-spin-polarized 3 He, used in neutron spin filters (NSFs), is metastability-exchange optical pumping. To prepare efficient NSF, one must highly polarize 3 He nuclei in the optical pumping volume (OPV) and reduce the polarization losses during the compression phase. Great progress has been achieved in reducing of depolarization due to the recent development of both, large polarization preserving piston compressors and long relaxation time filter cells. It is even more important to significantly enhance the 3 He polarization rate during optical pumping in order to increase NSF efficiency. Different cells materials were tested, such as Duran and quartz glass. In order to use the laser light more efficiently and to decrease the risk of 3 He depolarization due to unfavorable reflections, antireflection (AR) coatings were used on cell windows made of quartz glass. They were compared with the ones without coating, made of quartz, Duran and BK7 glass. The comparison of various techniques to mount the windows such as blowing, gluing or molecular diffusion was also conducted. It indicated that the molecular diffusion is the most suitable technique because of a better purity of the gas in the cell and the preservation of the optical flatness of the windows. Cells, for practical reasons each entirely made from the same material (Duran, Quartz glass) with windows mounted using this method, showed the best polarization performance

  14. Llgl1 Connects Cell Polarity with Cell-Cell Adhesion in Embryonic Neural Stem Cells.

    Science.gov (United States)

    Jossin, Yves; Lee, Minhui; Klezovitch, Olga; Kon, Elif; Cossard, Alexia; Lien, Wen-Hui; Fernandez, Tania E; Cooper, Jonathan A; Vasioukhin, Valera

    2017-06-05

    Malformations of the cerebral cortex (MCCs) are devastating developmental disorders. We report here that mice with embryonic neural stem-cell-specific deletion of Llgl1 (Nestin-Cre/Llgl1 fl/fl ), a mammalian ortholog of the Drosophila cell polarity gene lgl, exhibit MCCs resembling severe periventricular heterotopia (PH). Immunohistochemical analyses and live cortical imaging of PH formation revealed that disruption of apical junctional complexes (AJCs) was responsible for PH in Nestin-Cre/Llgl1 fl/fl brains. While it is well known that cell polarity proteins govern the formation of AJCs, the exact mechanisms remain unclear. We show that LLGL1 directly binds to and promotes internalization of N-cadherin, and N-cadherin/LLGL1 interaction is inhibited by atypical protein kinase C-mediated phosphorylation of LLGL1, restricting the accumulation of AJCs to the basolateral-apical boundary. Disruption of the N-cadherin-LLGL1 interaction during cortical development in vivo is sufficient for PH. These findings reveal a mechanism responsible for the physical and functional connection between cell polarity and cell-cell adhesion machineries in mammalian cells. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. The Major Birch Pollen Allergen Bet v 1 Induces Different Responses in Dendritic Cells of Birch Pollen Allergic and Healthy Individuals

    Science.gov (United States)

    Smole, Ursula; Radauer, Christian; Lengger, Nina; Svoboda, Martin; Rigby, Neil; Bublin, Merima; Gaier, Sonja; Hoffmann-Sommergruber, Karin; Jensen-Jarolim, Erika; Mechtcheriakova, Diana; Breiteneder, Heimo

    2015-01-01

    Dendritic cells play a fundamental role in shaping the immune response to allergens. The events that lead to allergic sensitization or tolerance induction during the interaction of the major birch pollen allergen Bet v 1 and dendritic cells are not very well studied. Here, we analyzed the uptake of Bet v 1 and the cross-reactive celery allergen Api g 1 by immature monocyte-derived dendritic cells (iMoDCs) of allergic and normal donors. In addition, we characterized the allergen-triggered intracellular signaling and transcriptional events. Uptake kinetics, competitive binding, and internalization pathways of labeled allergens by iMoDCs were visualized by live-cell imaging. Surface-bound IgE was detected by immunofluorescence microscopy and flow cytometry. Allergen- and IgE-induced gene expression of early growth response genes and Th1 and Th2 related cytokines and chemokines were analyzed by real-time PCR. Phosporylation of signaling kinases was analyzed by Western blot. Internalization of Bet v 1 by iMoDCs of both donor groups, likely by receptor-mediated caveolar endocytosis, followed similar kinetics. Bet v 1 outcompeted Api g 1 in cell surface binding and uptake. MoDCs of allergic and healthy donors displayed surface-bound IgE and showed a pronounced upregulation of Th2 cytokine- and NFκB-dependent genes upon non-specific Fcε receptor cross-linking. In contrast to these IgE-mediated responses, Bet v 1-stimulation increased transcript levels of the Th2 cytokines IL-4 and IL-13 but not of NFκB-related genes in MoDCs of BP allergic donors. Cells of healthy donors were either unresponsive or showed elevated mRNA levels of Th1-promoting chemokines. Moreover, Bet v 1 was able to induce Erk1/2 and p38 MAPK activation in BP allergics but only a slight p38 activation in normal donors. In conclusion, our data indicate that Bet v 1 favors the activation of a Th2 program only in DCs of BP allergic individuals. PMID:25635684

  16. The major birch pollen allergen Bet v 1 induces different responses in dendritic cells of birch pollen allergic and healthy individuals.

    Directory of Open Access Journals (Sweden)

    Ursula Smole

    Full Text Available Dendritic cells play a fundamental role in shaping the immune response to allergens. The events that lead to allergic sensitization or tolerance induction during the interaction of the major birch pollen allergen Bet v 1 and dendritic cells are not very well studied. Here, we analyzed the uptake of Bet v 1 and the cross-reactive celery allergen Api g 1 by immature monocyte-derived dendritic cells (iMoDCs of allergic and normal donors. In addition, we characterized the allergen-triggered intracellular signaling and transcriptional events. Uptake kinetics, competitive binding, and internalization pathways of labeled allergens by iMoDCs were visualized by live-cell imaging. Surface-bound IgE was detected by immunofluorescence microscopy and flow cytometry. Allergen- and IgE-induced gene expression of early growth response genes and Th1 and Th2 related cytokines and chemokines were analyzed by real-time PCR. Phosporylation of signaling kinases was analyzed by Western blot. Internalization of Bet v 1 by iMoDCs of both donor groups, likely by receptor-mediated caveolar endocytosis, followed similar kinetics. Bet v 1 outcompeted Api g 1 in cell surface binding and uptake. MoDCs of allergic and healthy donors displayed surface-bound IgE and showed a pronounced upregulation of Th2 cytokine- and NFκB-dependent genes upon non-specific Fcε receptor cross-linking. In contrast to these IgE-mediated responses, Bet v 1-stimulation increased transcript levels of the Th2 cytokines IL-4 and IL-13 but not of NFκB-related genes in MoDCs of BP allergic donors. Cells of healthy donors were either unresponsive or showed elevated mRNA levels of Th1-promoting chemokines. Moreover, Bet v 1 was able to induce Erk1/2 and p38 MAPK activation in BP allergics but only a slight p38 activation in normal donors. In conclusion, our data indicate that Bet v 1 favors the activation of a Th2 program only in DCs of BP allergic individuals.

  17. Synthetic spatially graded Rac activation drives cell polarization and movement.

    Science.gov (United States)

    Lin, Benjamin; Holmes, William R; Wang, C Joanne; Ueno, Tasuku; Harwell, Andrew; Edelstein-Keshet, Leah; Inoue, Takanari; Levchenko, Andre

    2012-12-26

    Migrating cells possess intracellular gradients of active Rho GTPases, which serve as central hubs in transducing signals from extracellular receptors to cytoskeletal and adhesive machinery. However, it is unknown whether shallow exogenously induced intracellular gradients of Rho GTPases are sufficient to drive cell polarity and motility. Here, we use microfluidic control to generate gradients of a small molecule and thereby directly induce linear gradients of active, endogenous Rac without activation of chemotactic receptors. Gradients as low as 15% were sufficient not only to trigger cell migration up the chemical gradient but to induce both cell polarization and repolarization. Cellular response times were inversely proportional to the steepness of Rac inducer gradient in agreement with a mathematical model, suggesting a function for chemoattractant gradient amplification upstream of Rac. Increases in activated Rac levels beyond a well-defined threshold augmented polarization and decreased sensitivity to the imposed gradient. The threshold was governed by initial cell polarity and PI3K activity, supporting a role for both in defining responsiveness to Rac activation. Our results reveal that Rac can serve as a starting point in defining cell polarity. Furthermore, our methodology may serve as a template to investigate processes regulated by intracellular signaling gradients.

  18. Blended learning fitting algorithm for polarization curves of fuel cells

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Fengxiang; Ji, Guangji; Zhang, Chuansheng [School of Automotive Studies of Tongji University, Shanghai 201804 (China); Zhou, Su [School of Automotive Studies of Tongji University, Shanghai 201804 (China); CDHK of Tongji University, Shanghai 200092 (China); Sundmacher, Kai [Max-Planck-Institute for Dynamics of Complex Technical Systems, Magdeburg 39106 (Germany)

    2009-07-15

    Fuel cell polarization curves, characterized by nonlinear models and the parameters of which are time-consuming to be identified, can represent fuel cell performance but will alter as the fuel cell degrades. For getting the information on degradation in time, a less time-consuming and an easily programmed algorithm, based on blended learning technique and linear least square estimation (LSE), is proposed to fit polarization curves obtained from the fuel cell systems. Simulations show that the proposed algorithm, compared with classical nonlinear LSE algorithms, converges much faster, features better extrapolation and less average quadratic error, and is easy to be programmed by C language. Therefore, the algorithm is a good option not only for fitting the polarization curves but also for implementation in embedded systems. (author)

  19. Defective planar cell polarity in polycystic kidney disease.

    Science.gov (United States)

    Fischer, Evelyne; Legue, Emilie; Doyen, Antonia; Nato, Faridabano; Nicolas, Jean-François; Torres, Vicente; Yaniv, Moshe; Pontoglio, Marco

    2006-01-01

    Morphogenesis involves coordinated proliferation, differentiation and spatial distribution of cells. We show that lengthening of renal tubules is associated with mitotic orientation of cells along the tubule axis, demonstrating intrinsic planar cell polarization, and we demonstrate that mitotic orientations are significantly distorted in rodent polycystic kidney models. These results suggest that oriented cell division dictates the maintenance of constant tubule diameter during tubular lengthening and that defects in this process trigger renal tubular enlargement and cyst formation.

  20. In vitro cytotoxicity effects of date palm (Phoenix dactylifera L.) pollen on neonate mouse spermatogonial stem cells.

    Science.gov (United States)

    Mahaldashtian, Maryam; Makoolati, Zohreh; Ghorbanian, Mohamad Taghi; Naghdi, Majid; Kouhpayeh, Seyed Amin

    2015-01-01

    There is a fast growing tendency in the use of herbal remedies in developing countries. One of the traditional medicines used for male infertility treatment is date palm (Phoenix dactylifera) pollen (DPP). Isolated spermatogonial stem cells and sertoli cells using enzymatic digestion were grown in Dulbecco's modified Eagle's medium supplemented with 4% foetal bovine serum in the absence or presence of 0.06, 0.25 and 0.62 mg/mL concentrations of aqueous extract of DPP for 2 weeks. The assessment of mean number of the whole cells and the living cells showed that there were no significant differences between the mean viability percentage and proliferation rate between control and experimental groups (P>0.05). As there are no cytotoxicity effects of DPP in our cultural system, this system can be utilised for the enrichment or differentiation of these cells in clinical applications, cell replacement therapy, tissue regeneration and tissue engineering applications.

  1. Unique stigmatic hairs and pollen-tube growth within the stigmatic cell wall in the early-divergent angiosperm family Hydatellaceae

    Science.gov (United States)

    Prychid, Christina J.; Sokoloff, Dmitry D.; Remizowa, Margarita V.; Tuckett, Renee E.; Yadav, Shrirang R.; Rudall, Paula J.

    2011-01-01

    Background and Aims The ultrastructure of the pollen tubes and the unusual multicellular stigmatic hairs of Trithuria, the sole genus of Hydatellaceae, are described in the context of comparative studies of stigmatic and transmitting tissue in other early-divergent angiosperms. Methods Scanning and transmission electron microscopy and immunocytochemistry are used to study the structure and composition of both mature and immature stigmatic hair cells and pollen-tube growth in Trithuria. Key Results Trithuria possesses a dry-type stigma. Pollen tubes grow within the cell walls of the long multicellular stigmatic hairs. Immunocytochemistry results suggest that arabinogalactan proteins are involved in attracting the pollen tubes through the stigmatic cuticle. Most tubes grow along the hair axis towards its base, but some grow towards the hair apex, suggesting that pollen tubes are guided by both physical constraints such as microfibril orientation and the presence of binding factors such as unesterified pectins and adhesive proteins. Conclusions The presence of a dry-type stigma in Trithuria supports the hypothesis that this condition is ancestral in angiosperms. Each multicellular stigmatic hair of Hydatellaceae is morphologically homologous with a stigmatic papilla of other angiosperms, but functions as an independent stigma and style. This unusual combination of factors makes Hydatellaceae a useful model for comparative studies of pollen-tube growth in early angiosperms. PMID:21320877

  2. Microinjection of Guanine Nucleotide Analogues into Lily Pollen Tubes Results in Isodiametric Tip Expansion

    Czech Academy of Sciences Publication Activity Database

    Eliáš, M.; Cvrčková, F.; Obermeyer, G.; Žárský, Viktor

    2001-01-01

    Roč. 3, - (2001), s. 489-492 ISSN 1435-8603 R&D Projects: GA AV ČR IAA5038907; GA ČR GA206/99/1138 Institutional research plan: CEZ:AV0Z5038910 Keywords : pollen tube * G-proteins * cell polarity Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.828, year: 2001

  3. Oral immunotherapy for pollen allergy using T-cell epitope-containing egg white derived from genetically manipulated chickens.

    Directory of Open Access Journals (Sweden)

    Yoshinori Kawabe

    Full Text Available Peptide immunotherapy using T-cell epitopes is expected to be an effective treatment for allergic diseases such as Japanese cedar (Cryptomeria japonica; Cj pollinosis. To develop a treatment for pollen allergy by inducing oral tolerance, we generated genetically manipulated (GM chickens by retroviral gene transduction, to produce a fusion protein of chicken egg white lysozyme and a peptide derived from seven dominant human T-cell epitopes of Japanese cedar pollen allergens (cLys-7crp. The transgene sequence was detected in all chickens transduced with the retroviral vector. Transduction efficiency in blood cells correlated to transgene expression. Western blot analysis revealed that cLys-7crp was expressed in the egg white of GM hens. Mice induced to develop allergic rhinitis by Cj pollinosis were fed with cLys-7crp-containing egg white produced by GM chickens. Total and Cj allergen (Cry j 1-specific IgE levels were significantly decreased in allergic mice fed with cLys-7crp-containing egg white compared with allergic mice fed with normal egg white. These results suggest that oral administration of T-cell epitope-containing egg white derived from GM chickens is effective for the induction of immune tolerance as an allergy therapy.

  4. Oral Immunotherapy for Pollen Allergy Using T-Cell Epitope-Containing Egg White Derived from Genetically Manipulated Chickens

    Science.gov (United States)

    Kawabe, Yoshinori; Hayashida, Yuuki; Numata, Kensaku; Harada, Shota; Hayashida, Yoshifumi; Ito, Akira; Kamihira, Masamichi

    2012-01-01

    Peptide immunotherapy using T-cell epitopes is expected to be an effective treatment for allergic diseases such as Japanese cedar (Cryptomeria japonica; Cj) pollinosis. To develop a treatment for pollen allergy by inducing oral tolerance, we generated genetically manipulated (GM) chickens by retroviral gene transduction, to produce a fusion protein of chicken egg white lysozyme and a peptide derived from seven dominant human T-cell epitopes of Japanese cedar pollen allergens (cLys-7crp). The transgene sequence was detected in all chickens transduced with the retroviral vector. Transduction efficiency in blood cells correlated to transgene expression. Western blot analysis revealed that cLys-7crp was expressed in the egg white of GM hens. Mice induced to develop allergic rhinitis by Cj pollinosis were fed with cLys-7crp-containing egg white produced by GM chickens. Total and Cj allergen (Cry j 1)-specific IgE levels were significantly decreased in allergic mice fed with cLys-7crp-containing egg white compared with allergic mice fed with normal egg white. These results suggest that oral administration of T-cell epitope-containing egg white derived from GM chickens is effective for the induction of immune tolerance as an allergy therapy. PMID:23144766

  5. FijiWingsPolarity: An open source toolkit for semi-automated detection of cell polarity.

    Science.gov (United States)

    Dobens, Leonard L; Shipman, Anna; Axelrod, Jeffrey D

    2017-12-22

    Epithelial cells are defined by apical-basal and planar cell polarity (PCP) signaling, the latter of which establishes an orthogonal plane of polarity in the epithelial sheet. PCP signaling is required for normal cell migration, differentiation, stem cell generation and tissue repair, and defects in PCP have been associated with developmental abnormalities, neuropathologies and cancers. While the molecular mechanism of PCP is incompletely understood, the deepest insights have come from Drosophila, where PCP is manifest in hairs and bristles across the adult cuticle and organization of the ommatidia in the eye. Fly wing cells are marked by actin-rich trichome structures produced at the distal edge of each cell in the developing wing epithelium and in a mature wing the trichomes orient collectively in the distal direction. Genetic screens have identified key PCP signaling pathway components that disrupt trichome orientation, which has been measured manually in a tedious and error prone process. Here we describe a set of image processing and pattern-recognition macros that can quantify trichome arrangements in micrographs and mark these directly by color, arrow or colored arrow to indicate trichome location, length and orientation. Nearest neighbor calculations are made to exploit local differences in orientation to better and more reliably detect and highlight local defects in trichome polarity. We demonstrate the use of these tools on trichomes in adult wing preps and on actin-rich developing trichomes in pupal wing epithelia stained with phalloidin. FijiWingsPolarity is freely available and will be of interest to a broad community of fly geneticists studying the effect of gene function on PCP.

  6. Direct contact between dendritic cells and bronchial epithelial cells inhibits T cell recall responses towards mite and pollen allergen extracts in vitro

    DEFF Research Database (Denmark)

    Papazian, Dick; Wagtmann, Valery R; Hansen, Soren

    2015-01-01

    Background: Airway epithelial cells (AECs) form a polarized barrier along the respiratory tract. They are the first point of contact with airborne antigens and are able to instruct resident immune cells to mount appropriate immune responses by either soluble or contact-dependent mechanisms....... Objective: We hypothesize that a healthy, polarized epithelial cell layer inhibits inflammatory responses toward allergens to uphold homeostasis. Methods: Using an in vitro co-culture model of the airway epithelium, where a polarized cell layer of bronchial epithelial cells can interact with dendritic cells...... cell recall responses towards Bet v 1, Phl p 5 and Der p 1 in vitro, suggesting that AECs-DC contact in vivo constitute a key element in mucosal homeostasis. This article is protected by copyright. All rights reserved....

  7. Honey bees preferentially consume freshly-stored pollen.

    Science.gov (United States)

    Carroll, Mark J; Brown, Nicholas; Goodall, Craig; Downs, Alexandra M; Sheenan, Timothy H; Anderson, Kirk E

    2017-01-01

    Honey bees (Apis mellifera) collect and store both honey and pollen in preserved forms. Pollen storage involves the addition of honey or nectar and oral secretions to pollen granules. It is controversial whether the duration of pollen storage alters the palatability or nutritive value of the pollen storage medium. We examined how bees utilize different-aged stored pollen during an extended pollen flow. The deposition of pollen into wax cells and subsequent consumption were monitored daily on 18 brood frames from 6 colonies over an 8d observation period. Despite a greater abundance of older stored pollen cells on brood frames, bees showed a marked preference for the consumption of freshly-stored pollen. Two to four day-old pollen cell contents were significantly more likely to be consumed, while pollen cell contents more than seven days old were eaten at much lower rates. Similar experiments that controlled for cell abundance and spatial effects using cage assays yielded the same result. One day-old stored pollen was consumed approximately three times more often than 10d-old stored pollen, and two times more often than 5d-old stored pollen. These consumption preferences for freshly-stored pollen occurred despite a lack of clear developmental advantages. Young adult workers reared for 7 days on 1d-, 5d-, or 10d-old stored pollen showed no difference in body mass, stored pollen consumption, hindgut fecal material accumulation, or hypopharyngeal gland (HPG) protein titers, suggesting that different-aged pollen stores did not vary in their nutritional value to adult bees. These findings are inconsistent with the hypothesis promoting a period of microbially-mediated, "beebread maturation" that results in greater palatability or nutritive value for aged pollen stores. Rather, stored pollen that is not eaten in the first few days accumulates as excess stores preserved in a less preferred, but nutritionally-similar state.

  8. High Throughput Method to Quantify Anterior-Posterior Polarity of T-Cells and Epithelial Cells

    Directory of Open Access Journals (Sweden)

    Susan J. Marriott

    2011-11-01

    Full Text Available The virologic synapse (VS, which is formed between a virus-infected and uninfected cell, plays a central role in the transmission of certain viruses, such as HIV and HTLV-1. During VS formation, HTLV-1-infected T-cells polarize cellular and viral proteins toward the uninfected T-cell. This polarization resembles anterior-posterior cell polarity induced by immunological synapse (IS formation, which is more extensively characterized than VS formation and occurs when a T-cell interacts with an antigen-presenting cell. One measure of cell polarity induced by both IS or VS formation is the repositioning of the microtubule organizing center (MTOC relative to the contact point with the interacting cell. Here we describe an automated, high throughput system to score repositioning of the MTOC and thereby cell polarity establishment. The method rapidly and accurately calculates the angle between the MTOC and the IS for thousands of cells. We also show that the system can be adapted to score anterior-posterior polarity establishment of epithelial cells. This general approach represents a significant advancement over manual cell polarity scoring, which is subject to experimenter bias and requires more time and effort to evaluate large numbers of cells.

  9. Planar Cell Polarity Controls Pancreatic Beta Cell Differentiation and Glucose Homeostasis

    DEFF Research Database (Denmark)

    Cortijo, Cedric; Gouzi, Mathieu; Tissir, Fadel

    2012-01-01

    Planar cell polarity (PCP) refers to the collective orientation of cells within the epithelial plane. We show that progenitor cells forming the ducts of the embryonic pancreas express PCP proteins and exhibit an active PCP pathway. Planar polarity proteins are acquired at embryonic day 11.5 synch...... that tridimensional organization and collective communication of cells are needed in the pancreatic epithelium in order to generate appropriate numbers of endocrine cells....

  10. Massively Parallelized Pollen Tube Guidance and Mechanical Measurements on a Lab-on-a-Chip Platform.

    Directory of Open Access Journals (Sweden)

    Naveen Shamsudhin

    Full Text Available Pollen tubes are used as a model in the study of plant morphogenesis, cellular differentiation, cell wall biochemistry, biomechanics, and intra- and intercellular signaling. For a "systems-understanding" of the bio-chemo-mechanics of tip-polarized growth in pollen tubes, the need for a versatile, experimental assay platform for quantitative data collection and analysis is critical. We introduce a Lab-on-a-Chip (LoC concept for high-throughput pollen germination and pollen tube guidance for parallelized optical and mechanical measurements. The LoC localizes a large number of growing pollen tubes on a single plane of focus with unidirectional tip-growth, enabling high-resolution quantitative microscopy. This species-independent LoC platform can be integrated with micro-/nano-indentation systems, such as the cellular force microscope (CFM or the atomic force microscope (AFM, allowing for rapid measurements of cell wall stiffness of growing tubes. As a demonstrative example, we show the growth and directional guidance of hundreds of lily (Lilium longiflorum and Arabidopsis (Arabidopsis thaliana pollen tubes on a single LoC microscopy slide. Combining the LoC with the CFM, we characterized the cell wall stiffness of lily pollen tubes. Using the stiffness statistics and finite-element-method (FEM-based approaches, we computed an effective range of the linear elastic moduli of the cell wall spanning the variability space of physiological parameters including internal turgor, cell wall thickness, and tube diameter. We propose the LoC device as a versatile and high-throughput phenomics platform for plant reproductive and development biology using the pollen tube as a model.

  11. Massively Parallelized Pollen Tube Guidance and Mechanical Measurements on a Lab-on-a-Chip Platform.

    Science.gov (United States)

    Shamsudhin, Naveen; Laeubli, Nino; Atakan, Huseyin Baris; Vogler, Hannes; Hu, Chengzhi; Haeberle, Walter; Sebastian, Abu; Grossniklaus, Ueli; Nelson, Bradley J

    2016-01-01

    Pollen tubes are used as a model in the study of plant morphogenesis, cellular differentiation, cell wall biochemistry, biomechanics, and intra- and intercellular signaling. For a "systems-understanding" of the bio-chemo-mechanics of tip-polarized growth in pollen tubes, the need for a versatile, experimental assay platform for quantitative data collection and analysis is critical. We introduce a Lab-on-a-Chip (LoC) concept for high-throughput pollen germination and pollen tube guidance for parallelized optical and mechanical measurements. The LoC localizes a large number of growing pollen tubes on a single plane of focus with unidirectional tip-growth, enabling high-resolution quantitative microscopy. This species-independent LoC platform can be integrated with micro-/nano-indentation systems, such as the cellular force microscope (CFM) or the atomic force microscope (AFM), allowing for rapid measurements of cell wall stiffness of growing tubes. As a demonstrative example, we show the growth and directional guidance of hundreds of lily (Lilium longiflorum) and Arabidopsis (Arabidopsis thaliana) pollen tubes on a single LoC microscopy slide. Combining the LoC with the CFM, we characterized the cell wall stiffness of lily pollen tubes. Using the stiffness statistics and finite-element-method (FEM)-based approaches, we computed an effective range of the linear elastic moduli of the cell wall spanning the variability space of physiological parameters including internal turgor, cell wall thickness, and tube diameter. We propose the LoC device as a versatile and high-throughput phenomics platform for plant reproductive and development biology using the pollen tube as a model.

  12. Massively Parallelized Pollen Tube Guidance and Mechanical Measurements on a Lab-on-a-Chip Platform

    Science.gov (United States)

    Laeubli, Nino; Atakan, Huseyin Baris; Vogler, Hannes; Hu, Chengzhi; Haeberle, Walter; Sebastian, Abu; Grossniklaus, Ueli; Nelson, Bradley J.

    2016-01-01

    Pollen tubes are used as a model in the study of plant morphogenesis, cellular differentiation, cell wall biochemistry, biomechanics, and intra- and intercellular signaling. For a “systems-understanding” of the bio-chemo-mechanics of tip-polarized growth in pollen tubes, the need for a versatile, experimental assay platform for quantitative data collection and analysis is critical. We introduce a Lab-on-a-Chip (LoC) concept for high-throughput pollen germination and pollen tube guidance for parallelized optical and mechanical measurements. The LoC localizes a large number of growing pollen tubes on a single plane of focus with unidirectional tip-growth, enabling high-resolution quantitative microscopy. This species-independent LoC platform can be integrated with micro-/nano-indentation systems, such as the cellular force microscope (CFM) or the atomic force microscope (AFM), allowing for rapid measurements of cell wall stiffness of growing tubes. As a demonstrative example, we show the growth and directional guidance of hundreds of lily (Lilium longiflorum) and Arabidopsis (Arabidopsis thaliana) pollen tubes on a single LoC microscopy slide. Combining the LoC with the CFM, we characterized the cell wall stiffness of lily pollen tubes. Using the stiffness statistics and finite-element-method (FEM)-based approaches, we computed an effective range of the linear elastic moduli of the cell wall spanning the variability space of physiological parameters including internal turgor, cell wall thickness, and tube diameter. We propose the LoC device as a versatile and high-throughput phenomics platform for plant reproductive and development biology using the pollen tube as a model. PMID:27977748

  13. Influence of Electric Fields and Conductivity on Pollen Tube Growth assessed via Electrical Lab-on-Chip

    Science.gov (United States)

    Agudelo, Carlos; Packirisamy, Muthukumaran; Geitmann, Anja

    2016-01-01

    Pollen tubes are polarly growing plant cells that are able to rapidly respond to a combination of chemical, mechanical, and electrical cues. This behavioural feature allows them to invade the flower pistil and deliver the sperm cells in highly targeted manner to receptive ovules in order to accomplish fertilization. How signals are perceived and processed in the pollen tube is still poorly understood. Evidence for electrical guidance in particular is vague and highly contradictory. To generate reproducible experimental conditions for the investigation of the effect of electric fields on pollen tube growth we developed an Electrical Lab-on-Chip (ELoC). Pollen from the species Camellia displayed differential sensitivity to electric fields depending on whether the entire cell or only its growing tip was exposed. The response to DC fields was dramatically higher than that to AC fields of the same strength. However, AC fields were found to restore and even promote pollen growth. Surprisingly, the pollen tube response correlated with the conductivity of the growth medium under different AC frequencies—consistent with the notion that the effect of the field on pollen tube growth may be mediated via its effect on the motion of ions. PMID:26804186

  14. Influence of Electric Fields and Conductivity on Pollen Tube Growth assessed via Electrical Lab-on-Chip.

    Science.gov (United States)

    Agudelo, Carlos; Packirisamy, Muthukumaran; Geitmann, Anja

    2016-01-25

    Pollen tubes are polarly growing plant cells that are able to rapidly respond to a combination of chemical, mechanical, and electrical cues. This behavioural feature allows them to invade the flower pistil and deliver the sperm cells in highly targeted manner to receptive ovules in order to accomplish fertilization. How signals are perceived and processed in the pollen tube is still poorly understood. Evidence for electrical guidance in particular is vague and highly contradictory. To generate reproducible experimental conditions for the investigation of the effect of electric fields on pollen tube growth we developed an Electrical Lab-on-Chip (ELoC). Pollen from the species Camellia displayed differential sensitivity to electric fields depending on whether the entire cell or only its growing tip was exposed. The response to DC fields was dramatically higher than that to AC fields of the same strength. However, AC fields were found to restore and even promote pollen growth. Surprisingly, the pollen tube response correlated with the conductivity of the growth medium under different AC frequencies--consistent with the notion that the effect of the field on pollen tube growth may be mediated via its effect on the motion of ions.

  15. Structure of polarization-resolved conoscopic patterns of planar oriented liquid crystal cells

    Science.gov (United States)

    Kiselev, A. D.; Vovk, R. G.

    2010-05-01

    The geometry of distributions of the polarization of light in conoscopic patterns of planar oriented nematic and cholesteric liquid crystal (LC) cells is described in terms of the polarization singularities including C-points (points of circular polarization) and L lines (lines of linear polarization). Conditions for the formation of polarization singularities ( C-points) in an ensemble of conoscopic patterns parametrized by the polarization azimuth and ellipticity of the incident light wave have been studied. A characteristic feature of these conditions is selectivity with respect to the polarization parameters of the incident light wave. The polarization azimuth and ellipticity are determining parameters for nematic and cholesteric LC cells, respectively.

  16. Pollen performance, cell number, and physiological state in the early-divergent angiosperm Annona cherimola Mill. (Annonaceae) are related to environmental conditions during the final stages of pollen development.

    Science.gov (United States)

    Lora, J; Herrero, M; Hormaza, J I

    2012-09-01

    Pollen performance is an important determinant for fertilization success, but high variability in pollen behavior both between and within species occurs in different years and under varying environmental conditions. Annona cherimola, an early-divergent angiosperm, is a species that releases a variable ratio of bicellular and tricellular hydrated pollen at anther dehiscence depending on temperature. The presence of both bi- and tricellular types of pollen is an uncommon characteristic in angiosperms and makes Annona cherimola an interesting model to study the effect of varying environmental conditions on subsequent pollen performance during the final stages of pollen development. In this work, we study the influence of changes in temperature and humidity during the final stages of pollen development on subsequent pollen performance, evaluating pollen germination, presence of carbohydrates, number of nuclei, and water content. At 25 °C, which is the average field temperature during the flowering period of this species, pollen had a viability of 60-70 %, starch hydrolyzed just prior to shedding, and pollen mitosis II was taking place, resulting in a mixture of bi- and tricellular pollen. This activity may be related to the pollen retaining 70 % water content at shedding. Temperatures above 30 °C resulted in a decrease in pollen germination, whereas lower temperatures did not have a clear influence on pollen germination, although they did have a clear effect on starch hydrolysis. On the other hand, slightly higher dehydration accelerated mitosis II, whereas strong dehydration arrested starch hydrolysis and reduced pollen germination. These results show a significant influence of environmental conditions on myriad pollen characteristics during the final stages of pollen development modifying subsequent pollen behavior and contributing to our understanding of the variability observed in pollen tube performance.

  17. Pollen development in Annona cherimola Mill. (Annonaceae. Implications for the evolution of aggregated pollen

    Directory of Open Access Journals (Sweden)

    Hormaza Jose I

    2009-10-01

    Full Text Available Abstract Background In most flowering plants, pollen is dispersed as monads. However, aggregated pollen shedding in groups of four or more pollen grains has arisen independently several times during angiosperm evolution. The reasons behind this phenomenon are largely unknown. In this study, we followed pollen development in Annona cherimola, a basal angiosperm species that releases pollen in groups of four, to investigate how pollen ontogeny may explain the rise and establishment of this character. We followed pollen development using immunolocalization and cytochemical characterization of changes occurring from anther differentiation to pollen dehiscence. Results Our results show that, following tetrad formation, a delay in the dissolution of the pollen mother cell wall and tapetal chamber is a key event that holds the four microspores together in a confined tapetal chamber, allowing them to rotate and then bind through the aperture sites through small pectin bridges, followed by joint sporopollenin deposition. Conclusion Pollen grouping could be the result of relatively minor ontogenetic changes beneficial for pollen transfer or/and protection from desiccation. Comparison of these events with those recorded in the recent pollen developmental mutants in Arabidopsis indicates that several failures during tetrad dissolution may convert to a common recurring phenotype that has evolved independently several times, whenever this grouping conferred advantages for pollen transfer.

  18. Pollen development in Annona cherimola Mill. (Annonaceae). Implications for the evolution of aggregated pollen

    Science.gov (United States)

    Lora, Jorge; Testillano, Pilar S; Risueño, Maria C; Hormaza, Jose I; Herrero, Maria

    2009-01-01

    Background In most flowering plants, pollen is dispersed as monads. However, aggregated pollen shedding in groups of four or more pollen grains has arisen independently several times during angiosperm evolution. The reasons behind this phenomenon are largely unknown. In this study, we followed pollen development in Annona cherimola, a basal angiosperm species that releases pollen in groups of four, to investigate how pollen ontogeny may explain the rise and establishment of this character. We followed pollen development using immunolocalization and cytochemical characterization of changes occurring from anther differentiation to pollen dehiscence. Results Our results show that, following tetrad formation, a delay in the dissolution of the pollen mother cell wall and tapetal chamber is a key event that holds the four microspores together in a confined tapetal chamber, allowing them to rotate and then bind through the aperture sites through small pectin bridges, followed by joint sporopollenin deposition. Conclusion Pollen grouping could be the result of relatively minor ontogenetic changes beneficial for pollen transfer or/and protection from desiccation. Comparison of these events with those recorded in the recent pollen developmental mutants in Arabidopsis indicates that several failures during tetrad dissolution may convert to a common recurring phenotype that has evolved independently several times, whenever this grouping conferred advantages for pollen transfer. PMID:19874617

  19. Cell polarity signaling in the plasticity of cancer cell invasiveness

    Czech Academy of Sciences Publication Activity Database

    Gandalovičová, A.; Vomastek, Tomáš; Rosel, D.; Brábek, J.

    2016-01-01

    Roč. 7, č. 18 (2016), s. 25022-25049 ISSN 1949-2553 R&D Projects: GA ČR GA13-06405S; GA MŠk(CZ) ED1.1.00/02.0109 Institutional support: RVO:61388971 Keywords : polarity * invasion * plasticity Subject RIV: EE - Microbiology, Virology Impact factor: 5.168, year: 2016

  20. Prion infection of epithelial Rov cells is a polarized event.

    Science.gov (United States)

    Paquet, Sophie; Sabuncu, Elifsu; Delaunay, Jean-Louis; Laude, Hubert; Vilette, Didier

    2004-07-01

    During prion infections, the cellular glycosylphosphatidylinositol-anchored glycoprotein PrP is converted into a conformational isoform. This abnormal conformer is thought to recruit and convert the normal cellular PrP into a likeness of itself and is proposed to be the infectious agent. We investigated the distribution of the PrP protein on the surface of Rov cells, an epithelial cell line highly permissive to prion multiplication, and we found that PrP is primarily expressed on the apical side. We further show that prion transmission to Rov cells is much more efficient if infectivity contacts the apical side, indicating that the apical and basolateral sides of Rov cells are not equally competent for prion infection and adding prions to the list of the conventional infectious agents (viruses and bacteria) that infect epithelial cells in a polarized manner. These data raise the possibility that apically expressed PrP may be involved in this polarized process of infection. This would add further support for a crucial role of PrP at the cell surface in prion infection of target cells.

  1. Centrosome polarization in T cells: a task for formins

    Directory of Open Access Journals (Sweden)

    Laura eAndrés-Delgado

    2013-07-01

    Full Text Available T-cell antigen receptor (TCR engagement triggers the rapid reorientation of the centrosome, which is associated with the secretory machinery, towards the immunological synapse (IS for polarized protein trafficking. Recent evidence indicates that upon TCR triggering the INF2 formin, together with the formins DIA1 and FMNL1, promotes the formation of a specialized array of stable detyrosinated MTs that breaks the symmetrical organization of the T-cell microtubule (MT cytoskeleton. The detyrosinated MT array and TCR-induced tyrosine phosphorylation should coincide for centrosome polarization. We propose that the pushing forces produced by the detyrosinated MT array, which modify the position of the centrosome, in concert with Src kinase dependent TCR signaling, which provide the reference frame with respect to which the centrosome reorients, result in the repositioning of the centrosome to the IS.

  2. Heme and non-heme iron transporters in non-polarized and polarized cells

    Directory of Open Access Journals (Sweden)

    Yasui Yumiko

    2010-06-01

    Full Text Available Abstract Background Heme and non-heme iron from diet, and recycled iron from hemoglobin are important products of the synthesis of iron-containing molecules. In excess, iron is potentially toxic because it can produce reactive oxygen species through the Fenton reaction. Humans can absorb, transport, store, and recycle iron without an excretory system to remove excess iron. Two candidate heme transporters and two iron transporters have been reported thus far. Heme incorporated into cells is degraded by heme oxygenases (HOs, and the iron product is reutilized by the body. To specify the processes of heme uptake and degradation, and the reutilization of iron, we determined the subcellular localizations of these transporters and HOs. Results In this study, we analyzed the subcellular localizations of 2 isoenzymes of HOs, 4 isoforms of divalent metal transporter 1 (DMT1, and 2 candidate heme transporters--heme carrier protein 1 (HCP1 and heme responsive gene-1 (HRG-1--in non-polarized and polarized cells. In non-polarized cells, HCP1, HRG-1, and DMT1A-I are located in the plasma membrane. In polarized cells, they show distinct localizations: HCP1 and DMT1A-I are located in the apical membrane, whereas HRG-1 is located in the basolateral membrane and lysosome. 16Leu at DMT1A-I N-terminal cytosolic domain was found to be crucial for plasma membrane localization. HOs are located in smooth endoplasmic reticulum and colocalize with NADPH-cytochrome P450 reductase. Conclusions HCP1 and DMT1A-I are localized to the apical membrane, and HRG-1 to the basolateral membrane and lysosome. These findings suggest that HCP1 and DMT1A-I have functions in the uptake of dietary heme and non-heme iron. HRG-1 can transport endocytosed heme from the lysosome into the cytosol. These localization studies support a model in which cytosolic heme can be degraded by HOs, and the resulting iron is exported into tissue fluids via the iron transporter ferroportin 1, which is

  3. Polarization-resolved angular patterns of nematic liquid crystal cells: Topological events driven by incident light polarization

    Science.gov (United States)

    Kiselev, Alexei D.; Vovk, Roman G.; Egorov, Roman I.; Chigrinov, Vladimir G.

    2008-09-01

    We study the angular structure of polarization of light transmitted through a nematic liquid crystal (NLC) cell by analyzing the polarization state as a function of the incidence angles and the polarization of the incident wave. The polarization-resolved angular (conoscopic) patterns emerging after the NLC cell illuminated by the convergent light beam are described in terms of the polarization singularities such as C points (points of circular polarization) and L lines (lines of linear polarization). For the homeotropically aligned cell, the Stokes polarimetry technique is used to measure the polarization resolved conoscopic patterns at different values of the ellipticity of the incident light, γell(inc) , impinging onto the cell. Using the exact analytical expressions for the transfer matrix we show that variations of the ellipticity, γell(inc) , induce transformations of the angular pattern exhibiting the effect of avoided L -line crossings and characterized by topological events such as creation and annihilation of the C points. The predictions of the theory are found to be in good agreement with the experimental results.

  4. Appearance of differentiated cells derived from polar body nuclei in the silkworm, Bombyx mori

    Directory of Open Access Journals (Sweden)

    Hiroki eSakai

    2013-09-01

    Full Text Available AbstractIn Bombyx mori, polar body nuclei are observed until 9h after egg lying, however, the fate of polar body nuclei remains unclear. To examine the fate of polar body nuclei, we employed a mutation of serosal cell pigmentation, pink-eyed white egg (pe. The heterozygous pe/+pe females produced black serosal cells in white eggs, while pe/pe females did not produce black serosal cells in white eggs. These results suggest that the appearance of black serosal cells in white eggs depends on the genotype (pe/ +pe of the mother. Because the polar body nuclei had +pe genes in the white eggs laid by a pe/ +pe female, polar body nuclei participate in development and differentiate into functional cell (serosal cells. Analyses of serosal cells pigmentation indicated that approximately 30% of the eggs contained polar-body-nucleus-derived cells. These results demonstrate that polar-body-nucleus-derived cells appeared at a high frequency under natural conditions. Approximately 80% of polar-body-nucleus-derived cells appeared near the anterior pole and the dorsal side, which is opposite to where embryogenesis occurs. The number of cells derived from the polar body nuclei was very low. Approximately 26 % of these eggs contained only one black serosal cell. PCR-based analysis revealed that the polar-body-nucleus-derived cells disappeared in late embryonic stages (stage 25. Overall, polar-body-nuclei-derived cells were unlikely to contribute to embryos.

  5. Gametophytic Pollen Tube Guidance: Attractant Peptides, Gametic Controls, and Receptors.

    Science.gov (United States)

    Higashiyama, Tetsuya; Yang, Wei-Cai

    2017-01-01

    Pollen tube guidance in flowering plants is a unique and critical process for successful sexual reproduction. The pollen tube that grows from pollen, which is the male gametophyte, precisely navigates to the embryo sac, which is the female gametophyte, within the pistil. Recent advances have clarified the molecular framework of gametophytic pollen tube guidance. Multiple species-specific attractant peptides are secreted from synergid cells, the proper development and function of which are regulated by female gametes. Multiple receptor-like kinases on the pollen tube tip are involved in sensing species-specific attractant peptides. In this Update article, recent progress in our understanding of the mechanism of gametophytic pollen tube guidance is reviewed, including attraction by synergid cells, control of pollen tube guidance by female gametes, and directional growth of the pollen tube by directional cue sensing. Future directions in the study of pollen tube guidance also are discussed. © 2017 American Society of Plant Biologists. All Rights Reserved.

  6. De novo post-pollen mitosis II tobacco pollen tube transcriptome

    OpenAIRE

    Hafidh, S. (Said); Breznenová, K. (Katarína); Honys, D. (David)

    2012-01-01

    In our previous study we applied the Agilent 44K tobacco gene chip to introduce and analyze the tobacco male gametophyte transcriptome in mature pollen and 4h pollen tubes. Here we extended our analysis post-pollen mitosis II (PMII) by including a new data set obtained from more advanced stage of the ongoing progamic phase – pollen tubes cultivated in vitro for 24 h. Pollen mitosis II marks key events in the control of male gametophyte development, the production of two sperm cells. In bicell...

  7. Pollen analyses for pollination research, unacetolyzed pollen

    Directory of Open Access Journals (Sweden)

    Gretchen D. Jones

    2012-10-01

    Full Text Available Pollinators feed on the pollen, nectar, and other plant exudates that are associated with flowers. As a result of this feeding activity, pollen becomes attached to them. Analysis of this pollen can reveal what they eat, their dispersal patterns in and around cropping systems, and their role in pollination. However, finding pollen on and or in a pollinator depends on the technique used to recover pollen. Two very easy techniques are described in detail that have been used to recover pollen from a variety of pollinators including beneficial and harmful insects, spiders, bats, and other pollinators. These techniques can be used to recover pollen from internal tissues (gut, alimentary canal, crop, etc., external tissues (proboscis, legs, eyes, etc., or both. By using the proper technique, better pollen recovery can be made and thus better data can be obtained about the pollinators, the foods they eat, the plants they pollinate, their migration routes and source zones.

  8. Pollen performance, cell number, and physiological state in the early-divergent angiosperm Annona cherimola Mill. (Annonaceae) are related to environmental conditions during the final stages of pollen development

    OpenAIRE

    Lora, Jorge; Herrero Romero, María; Hormaza Urroz, José Ignacio

    2012-01-01

    Pollen performance is an important determinant for fertilization success, but high variability in pollen behavior both between and within species occurs in different years and under varying environmental conditions. Annona cherimola, an early-divergent angiosperm, is a species that releases a variable ratio of bicellular and tricellular hydrated pollen at anther dehiscence depending on temperature. The presence of both bi- and tricellular types of pollen is an uncommon characteristic in angio...

  9. Characterization of the human T cell response to rye grass pollen allergens Lol p 1 and Lol p 5.

    Science.gov (United States)

    Burton, M D; Papalia, L; Eusebius, N P; O'Hehir, R E; Rolland, J M

    2002-12-01

    Knowledge of dominant T cell epitopes of major allergens recognized by allergic individuals is required to improve efficacy and safety of allergen immunotherapy. Rye grass pollen (RGP) is the most important source of seasonal aeroallergens in temperate climates and Lol p 1 and Lol p 5 are the two major IgE-reactive allergens. This study aimed to characterize the T cell response to these allergens using a large panel of RGP-sensitive individuals. Short-term RGP-specific T cell lines (TCL) were generated from 38 RGP-sensitive subjects and stimulated with Lol p 1 and/or Lol p 5 allergens and synthetic 20-mer peptides. Proliferative responses were determined by 3H-thymidine uptake and IL-5 and IFN-gamma in culture supernatants analysed by ELISA. Of 17 subjects tested for reactivity to both allergens 16 (94%) responded to Lol p 1 and/or Lol p 5, establishing these as major T cell-reactive allergens. Sites of T cell reactivity were spread throughout the allergen molecules but regions of high reactivity were found. For Lol p 1 these spanned residues 19-38, 109-128, 154-173, 190-209, and for Lol p 5 37-56, 100-119, 145-164, 154-173, 190-209, 217-236 and 226-245. IL-5 and IFN-gamma were produced by T cells cultured with proliferation-inducing peptides. T cell responses to RGP major allergens have been extensively characterized, providing fundamental information for developing T cell-targeted immunotherapy for RGP allergy.

  10. T-helper 17 cell polarization in pulmonary arterial hypertension.

    Science.gov (United States)

    Hautefort, Aurélie; Girerd, Barbara; Montani, David; Cohen-Kaminsky, Sylvia; Price, Laura; Lambrecht, Bart N; Humbert, Marc; Perros, Frédéric

    2015-06-01

    Inflammation may contribute to the pathobiology of pulmonary arterial hypertension (PAH). Deciphering the PAH fingerprint on the inflammation orchestrated by dendritic cells (DCs) and T cells, key driver and effector cells, respectively, of the immune system, may allow the identification of immunopathologic approaches to PAH management. Using flow cytometry, we performed immunophenotyping of monocyte-derived DCs (MoDCs) and circulating lymphocytes from patients with idiopathic PAH and control subjects. With the same technique, we performed cytokine profiling of both populations following stimulation, coculture, or both. We tested the immunomodulatory effects of a glucocorticoid (dexamethasone [Dex]) on this immunophenotype and cytokine profile. Using an epigenetic approach, we confirmed the immune polarization in blood DNA of patients with PAH. The profile of membrane costimulatory molecules of PAH MoDCs was similar to that of control subjects. However, PAH MoDCs retained higher levels of the T-cell activating molecules CD86 and CD40 after Dex pretreatment than did control MoDCs. This was associated with an increased expression of IL-12p40 and a reduced migration toward chemokine (C-C motif) ligand 21. Moreover, both with and without Dex, PAH MoDCs induced a higher activation and proliferation of CD4+ T cells, associated with a reduced expression of IL-4 (T helper 2 response) and a higher expression of IL-17 (T helper 17 response). Purified PAH CD4+ T cells expressed a higher level of IL-17 after activation than did those of control subjects. Lastly, there was significant hypomethylation of the IL-17 promoter in the PAH blood DNA as compared with the control blood. We have highlighted T helper 17 cell immune polarization in patients with PAH, as has been previously demonstrated in other chronic inflammatory and autoimmune conditions.

  11. The first cell-fate decision of mouse preimplantation embryo development: integrating cell position and polarity.

    Science.gov (United States)

    Mihajlović, Aleksandar I; Bruce, Alexander W

    2017-11-01

    During the first cell-fate decision of mouse preimplantation embryo development, a population of outer-residing polar cells is segregated from a second population of inner apolar cells to form two distinct cell lineages: the trophectoderm and the inner cell mass (ICM), respectively. Historically, two models have been proposed to explain how the initial differences between these two cell populations originate and ultimately define them as the two stated early blastocyst stage cell lineages. The 'positional' model proposes that cells acquire distinct fates based on differences in their relative position within the developing embryo, while the 'polarity' model proposes that the differences driving the lineage segregation arise as a consequence of the differential inheritance of factors, which exhibit polarized subcellular localizations, upon asymmetric cell divisions. Although these two models have traditionally been considered separately, a growing body of evidence, collected over recent years, suggests the existence of a large degree of compatibility. Accordingly, the main aim of this review is to summarize the major historical and more contemporarily identified events that define the first cell-fate decision and to place them in the context of both the originally proposed positional and polarity models, thus highlighting their functional complementarity in describing distinct aspects of the developmental programme underpinning the first cell-fate decision in mouse embryogenesis. © 2017 The Authors.

  12. A Review of the Effects of Major Atmospheric Pollutants on Pollen Grains, Pollen Content, and Allergenicity

    Science.gov (United States)

    Sénéchal, Hélène; Visez, Nicolas; Charpin, Denis; Shahali, Youcef; Peltre, Gabriel; Biolley, Jean-Philippe; Lhuissier, Franck; Couderc, Rémy; Yamada, Ohri; Malrat-Domenge, Audrey; Pham-Thi, Nhân; Poncet, Pascal; Sutra, Jean-Pierre

    2015-01-01

    This review summarizes the available data related to the effects of air pollution on pollen grains from different plant species. Several studies carried out either on in situ harvested pollen or on pollen exposed in different places more or less polluted are presented and discussed. The different experimental procedures used to monitor the impact of pollution on pollen grains and on various produced external or internal subparticles are listed. Physicochemical and biological effects of artificial pollution (gaseous and particulate) on pollen from different plants, in different laboratory conditions, are considered. The effects of polluted pollen grains, subparticles, and derived aeroallergens in animal models, in in vitro cell culture, on healthy human and allergic patients are described. Combined effects of atmospheric pollutants and pollen grains-derived biological material on allergic population are specifically discussed. Within the notion of “polluen,” some methodological biases are underlined and research tracks in this field are proposed. PMID:26819967

  13. A Review of the Effects of Major Atmospheric Pollutants on Pollen Grains, Pollen Content, and Allergenicity

    Directory of Open Access Journals (Sweden)

    Hélène Sénéchal

    2015-01-01

    Full Text Available This review summarizes the available data related to the effects of air pollution on pollen grains from different plant species. Several studies carried out either on in situ harvested pollen or on pollen exposed in different places more or less polluted are presented and discussed. The different experimental procedures used to monitor the impact of pollution on pollen grains and on various produced external or internal subparticles are listed. Physicochemical and biological effects of artificial pollution (gaseous and particulate on pollen from different plants, in different laboratory conditions, are considered. The effects of polluted pollen grains, subparticles, and derived aeroallergens in animal models, in in vitro cell culture, on healthy human and allergic patients are described. Combined effects of atmospheric pollutants and pollen grains-derived biological material on allergic population are specifically discussed. Within the notion of “polluen,” some methodological biases are underlined and research tracks in this field are proposed.

  14. Cell polarity, cell adhesion, and spermatogenesis: role of cytoskeletons [version 1; referees: 2 approved

    Directory of Open Access Journals (Sweden)

    Linxi Li

    2017-08-01

    Full Text Available In the rat testis, studies have shown that cell polarity, in particular spermatid polarity, to support spermatogenesis is conferred by the coordinated efforts of the Par-, Crumbs-, and Scribble-based polarity complexes in the seminiferous epithelium. Furthermore, planar cell polarity (PCP is conferred by PCP proteins such as Van Gogh-like 2 (Vangl2 in the testis. On the other hand, cell junctions at the Sertoli cell–spermatid (steps 8–19 interface are exclusively supported by adhesion protein complexes (for example, α6β1-integrin-laminin-α3,β3,γ3 and nectin-3-afadin at the actin-rich apical ectoplasmic specialization (ES since the apical ES is the only anchoring device in step 8–19 spermatids. For cell junctions at the Sertoli cell–cell interface, they are supported by adhesion complexes at the actin-based basal ES (for example, N-cadherin-β-catenin and nectin-2-afadin, tight junction (occludin-ZO-1 and claudin 11-ZO-1, and gap junction (connexin 43-plakophilin-2 and also intermediate filament-based desmosome (for example, desmoglein-2-desmocollin-2. In short, the testis-specific actin-rich anchoring device known as ES is crucial to support spermatid and Sertoli cell adhesion. Accumulating evidence has shown that the Par-, Crumbs-, and Scribble-based polarity complexes and the PCP Vangl2 are working in concert with actin- or microtubule-based cytoskeletons (or both and these polarity (or PCP protein complexes exert their effects through changes in the organization of the cytoskeletal elements across the seminiferous epithelium of adult rat testes. As such, there is an intimate relationship between cell polarity, cell adhesion, and cytoskeletal function in the testis. Herein, we critically evaluate these recent findings based on studies on different animal models. We also suggest some crucial future studies to be performed.

  15. Developmental evolution of flowering plant pollen tube cell walls: callose synthase (CalS gene expression patterns

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    Abercrombie Jason M

    2011-07-01

    Full Text Available Abstract Background A number of innovations underlie the origin of rapid reproductive cycles in angiosperms. A critical early step involved the modification of an ancestrally short and slow-growing pollen tube for faster and longer distance transport of sperm to egg. Associated with this shift are the predominantly callose (1,3-β-glucan walls and septae (callose plugs of angiosperm pollen tubes. Callose synthesis is mediated by callose synthase (CalS. Of 12 CalS gene family members in Arabidopsis, only one (CalS5 has been directly linked to pollen tube callose. CalS5 orthologues are present in several monocot and eudicot genomes, but little is known about the evolutionary origin of CalS5 or what its ancestral function may have been. Results We investigated expression of CalS in pollen and pollen tubes of selected non-flowering seed plants (gymnosperms and angiosperms within lineages that diverged below the monocot/eudicot node. First, we determined the nearly full length coding sequence of a CalS5 orthologue from Cabomba caroliniana (CcCalS5 (Nymphaeales. Semi-quantitative RT-PCR demonstrated low CcCalS5 expression within several vegetative tissues, but strong expression in mature pollen. CalS transcripts were detected in pollen tubes of several species within Nymphaeales and Austrobaileyales, and comparative analyses with a phylogenetically diverse group of sequenced genomes indicated homology to CalS5. We also report in silico evidence of a putative CalS5 orthologue from Amborella. Among gymnosperms, CalS5 transcripts were recovered from germinating pollen of Gnetum and Ginkgo, but a novel CalS paralog was instead amplified from germinating pollen of Pinus taeda. Conclusion The finding that CalS5 is the predominant callose synthase in pollen tubes of both early-diverging and model system angiosperms is an indicator of the homology of their novel callosic pollen tube walls and callose plugs. The data suggest that CalS5 had transient expression

  16. Role of the epithelial cell-specific clathrin adaptor complex AP-1B in cell polarity

    Science.gov (United States)

    Fölsch, Heike

    2015-01-01

    Epithelial cells are important for organ development and function. To this end, they polarize their plasma membrane into biochemically and physically distinct membrane domains. The apical membrane faces the luminal site of an organ and the basolateral domain is in contact with the basement membrane and neighboring cells. To establish and maintain this polarity it is important that newly synthesized and endocytic cargos are correctly sorted according to their final destinations at either membrane. Sorting takes place at one of 2 major sorting stations in the cells, the trans-Golgi network (TGN) and recycling endosomes (REs). Polarized sorting may involve epithelial cell-specific sorting adaptors like the AP-1B clathrin adaptor complex. AP-1B facilitates basolateral sorting from REs. This review will discuss various aspects of basolateral sorting in epithelial cells with a special emphasis on AP-1B. PMID:27057418

  17. Pollen features of hazelnut (Corylus avellana L. from different habitats

    Directory of Open Access Journals (Sweden)

    Natalia Nikolaieva

    2014-04-01

    Full Text Available The aim of this work was to study general morphological characteristics of pollen grains of Corylus avellana L. Seven samples of pollen were investigated. Samples were collected from different habitats in Ukraine – from botanical gardens (Kyiv, Kamianets-Podilskyi and natural habitats (Kyiv region, Kamianets-Podilskyi, and Sumy region. We studied such morphological traits of pollen grains as length of polar and equatorial axes, diameter of pores, and shape of the pollen grain (elongation index. Analysis of morphological characters of pollen was carried out using electron microscope. Comparison of data was performed with the data of the base polleninfo.org. During research the differences in these parameters were marked. Pollen grains of C. avellana are generally isopolar, from suboblate to oblate or oblate-spheroidal, and contain 3 pores. The article contains an attempt to explain the size variations noted for the pollen collected from different habitats.

  18. Topological events in polarization resolved angular patterns of nematic liquid crystal cells at varying ellipticity of incident wave

    OpenAIRE

    Kiselev, Alexei D.; Vovk, Roman G.

    2008-01-01

    We study the angular structure of polarization of light transmitted through a nematic liquid crystal (NLC) cell by analyzing the polarization state as a function of the incidence angles and the polarization of the incident wave. The polarization resolved angular patterns emerging after the NLC cell illuminated by the convergent light beam are described in terms of the polarization singularities such as C-points (points of circular polarization) and L-lines (lines of linear polarization). For ...

  19. The cell polarity determinant CDC42 controls division symmetry to block leukemia cell differentiation.

    Science.gov (United States)

    Mizukawa, Benjamin; O'Brien, Eric; Moreira, Daniel C; Wunderlich, Mark; Hochstetler, Cindy L; Duan, Xin; Liu, Wei; Orr, Emily; Grimes, H Leighton; Mulloy, James C; Zheng, Yi

    2017-09-14

    As a central regulator of cell polarity, the activity of CDC42 GTPase is tightly controlled in maintaining normal hematopoietic stem and progenitor cell (HSC/P) functions. We found that transformation of HSC/P to acute myeloid leukemia (AML) is associated with increased CDC42 expression and activity in leukemia cells. In a mouse model of AML, the loss of Cdc42 abrogates MLL-AF9 -induced AML development. Furthermore, genetic ablation of CDC42 in both murine and human MLL-AF9 (MA9) cells decreased survival and induced differentiation of the clonogenic leukemia-initiating cells. We show that MLL-AF9 leukemia cells maintain cell polarity in the context of elevated Cdc42-guanosine triphosphate activity, similar to nonmalignant, young HSC/Ps. The loss of Cdc42 resulted in a shift to depolarized AML cells that is associated with a decrease in the frequency of symmetric and asymmetric cell divisions producing daughter cells capable of self-renewal. Importantly, we demonstrate that inducible CDC42 suppression in primary human AML cells blocks leukemia progression in a xenograft model. Thus, CDC42 loss suppresses AML cell polarity and division asymmetry, and CDC42 constitutes a useful target to alter leukemia-initiating cell fate for differentiation therapy. © 2017 by The American Society of Hematology.

  20. In vitro effects of date palm (Phoenix dactylifera L.) pollen on colonization of neonate mouse spermatogonial stem cells.

    Science.gov (United States)

    Mahaldashtian, Maryam; Naghdi, Majid; Ghorbanian, Mohamad Taghi; Makoolati, Zohreh; Movahedin, Mansoureh; Mohamadi, Seyedeh Momeneh

    2016-06-20

    Date palm (Phoenix dactylifera L.) pollen (DPP) is widely used as a folk remedy for male infertility treatment, and has well known medicinal effects. This study aimed to determine the in vitro effects of DPP on the efficiency of neonate mouse spermatogonial stem cells (SSCs) proliferation. Sertoli and SSCs were isolated from 6 to 10-days-old mouse testes, and their identity was confirmed using immunocytochemistry against cytokeratin for sertoli cells and PLZF, Oct-4 and CDH-1 for SSCs. Isolated testicular cells were cultured in the absence or presence of 0.06, 0.25 and 0.62mg/ml concentrations of DPP aqueous extract for 2 weeks. The number and diameter of SSC colonies were assessed during third, 7th, 9th and 14th day of culture, and the expression of the Mvh, GFRα-1 and Oct-4 was evaluated using quantitative PCR at the end of the culture period. The significance of the data was analyzed using ANOVA and paired samples t-test and Tukey and Bonferroni test as post hoc tests at the level of p≤0.05. Pattern assay of colony formation showed that SSCs numbers increased in the present of 0.62mg/ml concentration of DPP extract with higher slop relative to other groups (P <0.05). Colony diameters had no significant difference between groups in 3th, 7th, 9th and 14th days after culture. The Mvh and Oct-4 genes expression had no significant difference between groups, while GFRα1 expression was increased significantly in cells treated with 0.06mg/ml concentration relative to other groups (P<0.05). It seems that co-culture of SSCs with sertoli sells in the presence of low doses of DPP can increase SSCs proliferation and keep their stemness state, while higher concentrations can differentiate the treated cells. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  1. Phosphatidylinositol 4-phosphate 5-kinases 1 and 2 are involved in the regulation of vacuole morphology during Arabidopsis thaliana pollen development.

    Science.gov (United States)

    Ugalde, José-Manuel; Rodriguez-Furlán, Cecilia; Rycke, Riet De; Norambuena, Lorena; Friml, Jiří; León, Gabriel; Tejos, Ricardo

    2016-09-01

    The pollen grains arise after meiosis of pollen mother cells within the anthers. A series of complex structural changes follows, generating mature pollen grains capable of performing the double fertilization of the female megasporophyte. Several signaling molecules, including hormones and lipids, have been involved in the regulation and appropriate control of pollen development. Phosphatidylinositol 4-phophate 5-kinases (PIP5K), which catalyze the biosynthesis of the phosphoinositide PtdIns(4,5)P2, are important for tip polar growth of root hairs and pollen tubes, embryo development, vegetative plant growth, and responses to the environment. Here, we report a role of PIP5Ks during microgametogenesis. PIP5K1 and PIP5K2 are expressed during early stages of pollen development and their transcriptional activity respond to auxin in pollen grains. Early male gametophytic lethality to certain grade was observed in both pip5k1(-/-) and pip5k2(-/-) single mutants. The number of pip5k mutant alleles is directly related to the frequency of aborted pollen grains suggesting the two genes are involved in the same function. Indeed PIP5K1 and PIP5K2 are functionally redundant since homozygous double mutants did not render viable pollen grains. The loss of function of PIP5K1 and PIP5K2results in defects in vacuole morphology in pollen at the later stages and epidermal root cells. Our results show that PIP5K1, PIP5K2 and phosphoinositide signaling are important cues for early developmental stages and vacuole formation during microgametogenesis. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  2. Targeting Cell Polarity Machinery to Exhaust Breast Cancer Stem Cells

    Science.gov (United States)

    2017-10-01

    since, in the absence of such knowledge, the development of effective therapeutic interventions to target CSCs and prevent cancer progression and...yes) (2) Presentations: a. 2016 Keystone Symposia- Stem Cells & Cancer, Breckenridge, “Epigenetic regulation promotes obesity related breast

  3. Pyrrolizidine alkaloids in pollen and pollen products.

    Science.gov (United States)

    Kempf, Michael; Heil, Sandra; Hasslauer, Iris; Schmidt, Lukas; von der Ohe, Katharina; Theuring, Claudine; Reinhard, Annika; Schreier, Peter; Beuerle, Till

    2010-02-01

    Recently, 1,2-dehydropyrrolizidine alkaloid (PA) ester alkaloids, found predominantly as their N-oxides (PANOs, pyrrolizidine N-oxides), have been reported in both honey and in pollen obtained directly from PA plants and pollen loads collected by bees, raising the possibility of health risks for consumers of these products. We confirm these findings in regard to floral pollen, using pollen collected directly from flowers of the known PA plants Senecio jacobaea, S. vernalis, Echium vulgare and pollinia of Phalaenopsis hybrids, and we extend analyses of 1,2-unsaturated PAs and 1,2-unsaturated PANOs to include bee-pollen products currently being sold in supermarkets and on the Internet as food supplements. PA content of floral pollen ranged from 0.5 to 5 mg/g. The highest values were observed in pollen obtained from Senecio species. Up to 95% of the PAs are found as PANOs. Detailed studies with S. vernalis revealed unique PA patterns in pollen and flowers. While seneciphylline was the most prominent PA in S. vernalis pollen, the flowers were dominated by senecionine. To analyze trace amounts of 1,2-unsaturated PAs in pollen products, our previously elaborated method consisting of strong cation exchange-SPE, two reduction steps followed by silylation and subsequent capillary high-resolution GC-MS using SIM mode was applied. In total, 55 commercially available pollen products were analyzed. Seventeen (31%) samples contained 1,2-unsaturated PAs in the range from 1.08 to 16.35 microg/g, calculated as retronecine equivalents. The 1,2-unsaturated PA content of pollen products is expressed in terms of a single sum parameter and no background information such as foraged plants, pollen analysis, etc. was needed to analyze the samples. The detection limit of overall procedure and the reliable quantitation limit were 0.003 and 0.01 microg/g, respectively.

  4. Targeting Cell Polarity Machinery to Exhaust Breast Cancer Stem Cells

    Science.gov (United States)

    2016-10-01

    stably express miR-200c (pCDH- miR200c) and MCF7 cells with knock- down of miR-200c (pZIP-miR200c) (Months 1-2) Completed! We have successfully...established BT549-pCDH-miR200c and MCF7 - pZIP-miR200c and examined the protein expression levels as described in subtask 2 (Fig. 1). Subtask 2...Determine expression levels of PKCζ and phospho-NUMB (p- NUMB), by re-expressing PKCζ in BT549-pCDH-miR200c cells and knocking-down PKCζ in MCF7

  5. Angular structure of light polarization and singularities in transmittance of nematic liquid crystal cells

    Science.gov (United States)

    Kiselev, Alexei D.; Vovk, Roman G.; Buinyi, Igor O.; Soskin, Marat S.

    2007-06-01

    We study the angular structure of polarization of light transmitted through a nematic liquid crystal (NLC) cell by analyzing the polarization state as a function of the incidence angles. Our theoretical results are obtained by evaluating the Stokes parameters that characterize the polarization state of plane waves propagating through the NLC layer at varying direction of incidence. Using the Stokes polarimetry technique we carried out the measurements of the polarization resolved conoscopic patterns emerging after the homeotropically aligned NLC cell illuminated by the convergent light beam. The resulting polarization resolved angular patterns are described both theoretically and experimentally in terms of the polarization singularities such as C-points (points of circular polarization) and L-lines (lines of linear polarization). When the ellipticity of the incident light varies, the angular patterns are found to undergo transformations involving the processes of creation and annihilation of the C-points.

  6. Muscle Stem Cell Fate Is Controlled by the Cell-Polarity Protein Scrib

    Directory of Open Access Journals (Sweden)

    Yusuke Ono

    2015-02-01

    Full Text Available Satellite cells are resident skeletal muscle stem cells that supply myonuclei for homeostasis, hypertrophy, and repair in adult muscle. Scrib is one of the major cell-polarity proteins, acting as a potent tumor suppressor in epithelial cells. Here, we show that Scrib also controls satellite-cell-fate decisions in adult mice. Scrib is undetectable in quiescent cells but becomes expressed during activation. Scrib is asymmetrically distributed in dividing daughter cells, with robust accumulation in cells committed to myogenic differentiation. Low Scrib expression is associated with the proliferative state and preventing self-renewal, whereas high Scrib levels reduce satellite cell proliferation. Satellite-cell-specific knockout of Scrib in mice causes a drastic and insurmountable defect in muscle regeneration. Thus, Scrib is a regulator of tissue stem cells, controlling population expansion and self-renewal with Scrib expression dynamics directing satellite cell fate.

  7. Receptor-like kinases as surface regulators for RAC/ROP-mediated pollen tube growth and interaction with the pistil

    Science.gov (United States)

    Zou, Yanjiao; Aggarwal, Mini; Zheng, Wen-Guang; Wu, Hen-Ming; Cheung, Alice Y.

    2011-01-01

    Background RAC/ROPs are RHO-type GTPases and are known to play diverse signalling roles in plants. Cytoplasmic RAC/ROPs are recruited to the cell membrane and activated in response to extracellular signals perceived and mediated by cell surface-located signalling assemblies, transducing the signals to regulate cellular processes. More than any other cell types in plants, pollen tubes depend on continuous interactions with an extracellular environment produced by their surrounding tissues as they grow within the female organ pistil to deliver sperm to the female gametophyte for fertilization. Scope We review studies on pollen tube growth that provide compelling evidence indicating that RAC/ROPs are crucial for regulating the cellular processes that underlie the polarized cell growth process. Efforts to identify cell surface regulators that mediate extracellular signals also point to RAC/ROPs being the molecular switches targeted by growth-regulating female factors for modulation to mediate pollination and fertilization. We discuss a large volume of work spanning more than two decades on a family of pollen-specific receptor kinases and some recent studies on members of the FERONIA family of receptor-like kinases (RLKs). Significance The research described shows the crucial roles that two RLK families play in transducing signals from growth regulatory factors to the RAC/ROP switch at the pollen tube apex to mediate and target pollen tube growth to the female gametophyte and signal its disintegration to achieve fertilization once inside the female chamber. PMID:22476487

  8. Optimization of conditions for in vitro pollen germination and pollen ...

    African Journals Online (AJOL)

    Pollen germination and pollen tube growth might have a significant effect on fruit and seed production. This study was conducted to investigate the best medium for pollen germination and pollen tube growth of date palm male. Significant differences in percentages of pollen germination and pollen tube growth were ...

  9. Polarity in plant asymmetric cell division: Division orientation and cell fate differentiation.

    Science.gov (United States)

    Shao, Wanchen; Dong, Juan

    2016-11-01

    Asymmetric cell division (ACD) is universally required for the development of multicellular organisms. Unlike animal cells, plant cells have a rigid cellulosic extracellular matrix, the cell wall, which provides physical support and forms communication routes. This fundamental difference leads to some unique mechanisms in plants for generating asymmetries during cell division. However, plants also utilize intrinsically polarized proteins to regulate asymmetric signaling and cell division, a strategy similar to the differentiation mechanism found in animals. Current progress suggests that common regulatory modes, i.e. protein spontaneous clustering and cytoskeleton reorganization, underlie protein polarization in both animal and plant cells. Despite these commonalities, it is important to note that intrinsic mechanisms in plants are heavily influenced by extrinsic cues. To control physical asymmetry in cell division, although our understanding is fragmentary thus far, plants might have evolved novel polarization strategies to orientate cell division plane. Recent studies also suggest that the phytohormone auxin, one of the most pivotal small molecules in plant development, regulates ACD in plants. Copyright © 2016. Published by Elsevier Inc.

  10. Emerging roles for microtubules in angiosperm pollen tube growth highlight new research cues

    Directory of Open Access Journals (Sweden)

    Alessandra eMoscatelli

    2015-02-01

    Full Text Available In plants, actin filaments have an important role in organelle movement and cytoplasmic streaming. Otherwise microtubules have a role in restricting organelles to specific areas of the cell and in maintaining organelle morphology. In somatic plant cells, microtubules also participate in cell division and morphogenesis, allowing cells to take their definitive shape in order to perform specific functions. In the latter case, microtubules influence assembly of the cell wall, controlling the delivery of enzymes involved in cellulose synthesis and of wall modulation material to the proper sites.In angiosperm pollen tubes, organelle movement is generally attributed to the acto-myosin system, the main role of which is in distributing organelles in the cytoplasm and in carrying secretory vesicles to the apex for polarized growth. Recent data on membrane trafficking suggests a role of microtubules in fine delivery and repositioning of vesicles to sustain pollen tube growth. This review examines the role of microtubules in secretion and endocytosis, highlighting new research cues regarding cell wall construction and pollen tube-pistil crosstalk, that help unravel the role of microtubules in polarized growth.

  11. Human Leukocyte Antigen-G and Regulatory T Cells during Specific Immunotherapy for Pollen Allergy

    DEFF Research Database (Denmark)

    Sørensen, Anja Elaine; Johnsen, Claus R; Dalgaard, Louise Torp

    2013-01-01

    Background: TH2-biased immune responses are important in allergy pathogenesis. Mechanisms of allergen-specific immunotherapy (SIT) might include the induction of regulatory T cells (Tregs) and immunoglobulin (Ig) G4 blocking antibodies, a reduction in the number of effector cells, and skewing...

  12. Planar cell polarity breaks bilateral symmetry by controlling ciliary positioning.

    Science.gov (United States)

    Song, Hai; Hu, Jianxin; Chen, Wen; Elliott, Gene; Andre, Philipp; Gao, Bo; Yang, Yingzi

    2010-07-15

    Defining the three body axes is a central event of vertebrate morphogenesis. Establishment of left-right (L-R) asymmetry in development follows the determination of dorsal-ventral and anterior-posterior (A-P) body axes, although the molecular mechanism underlying precise L-R symmetry breaking in reference to the other two axes is still poorly understood. Here, by removing both Vangl1 and Vangl2, the two mouse homologues of a Drosophila core planar cell polarity (PCP) gene Van Gogh (Vang), we reveal a previously unrecognized function of PCP in the initial breaking of lateral symmetry. The leftward nodal flow across the posterior notochord (PNC) has been identified as the earliest event in the de novo formation of L-R asymmetry. We show that PCP is essential in interpreting the A-P patterning information and linking it to L-R asymmetry. In the absence of Vangl1 and Vangl2, cilia are positioned randomly around the centre of the PNC cells and nodal flow is turbulent, which results in disrupted L-R asymmetry. PCP in mouse, unlike what has been implicated in other vertebrate species, is not required for ciliogenesis, cilium motility, Sonic hedgehog (Shh) signalling or apical docking of basal bodies in ciliated tracheal epithelial cells. Our data suggest that PCP acts earlier than the unidirectional nodal flow during bilateral symmetry breaking in vertebrates and provide insight into the functional mechanism of PCP in organizing the vertebrate tissues in development.

  13. Mechanistic Framework for Establishment, Maintenance, and Alteration of Cell Polarity in Plants

    Directory of Open Access Journals (Sweden)

    Pankaj Dhonukshe

    2012-01-01

    Full Text Available Cell polarity establishment, maintenance, and alteration are central to the developmental and response programs of nearly all organisms and are often implicated in abnormalities ranging from patterning defects to cancer. By residing at the distinct plasma membrane domains polar cargoes mark the identities of those domains, and execute localized functions. Polar cargoes are recruited to the specialized membrane domains by directional secretion and/or directional endocytic recycling. In plants, auxin efflux carrier PIN proteins display polar localizations in various cell types and play major roles in directional cell-to-cell transport of signaling molecule auxin that is vital for plant patterning and response programs. Recent advanced microscopy studies applied to single cells in intact plants reveal subcellular PIN dynamics. They uncover the PIN polarity generation mechanism and identified important roles of AGC kinases for polar PIN localization. AGC kinase family members PINOID, WAG1, and WAG2, belonging to the AGC-3 subclass predominantly influence the polar localization of PINs. The emerging mechanism for AGC-3 kinases action suggests that kinases phosphorylate PINs mainly at the plasma membrane after initial symmetric PIN secretion for eventual PIN internalization and PIN sorting into distinct ARF-GEF-regulated polar recycling pathways. Thus phosphorylation status directs PIN translocation to different cell sides. Based on these findings a mechanistic framework evolves that suggests existence of cell side-specific recycling pathways in plants and implicates AGC3 kinases for differential PIN recruitment among them for eventual PIN polarity establishment, maintenance, and alteration.

  14. Dystroglycan is required for polarizing the epithelial cells and the oocyte in Drosophila

    DEFF Research Database (Denmark)

    Deng, Wu-Min; Schneider, Martina; Frock, Richard

    2003-01-01

    , and plays a role in linking the ECM to the actin cytoskeleton; however, how these interactions are regulated and their basic cellular functions are poorly understood. Using mosaic analysis and RNAi in the model organism Drosophila melanogaster, we show that Dystroglycan is required cell......-autonomously for cellular polarity in two different cell types, the epithelial cells (apicobasal polarity) and the oocyte (anteroposterior polarity). Loss of Dystroglycan function in follicle and disc epithelia results in expansion of apical markers to the basal side of cells and overexpression results in a reduced apical...... localization of these same markers. In Dystroglycan germline clones early oocyte polarity markers fail to be localized to the posterior, and oocyte cortical F-actin organization is abnormal. Dystroglycan is also required non-cell-autonomously to organize the planar polarity of basal actin in follicle cells...

  15. Sequential development of apical-basal and planar polarities in aggregating epitheliomuscular cells of Hydra.

    Science.gov (United States)

    Seybold, Anna; Salvenmoser, Willi; Hobmayer, Bert

    2016-04-01

    Apical-basal and planar cell polarities are hallmarks of metazoan epithelia required to separate internal and external environments and to regulate trans- and intracellular transport, cytoskeletal organization, and morphogenesis. Mechanisms of cell polarization have been intensively studied in bilaterian model organisms, particularly in early embryos and cultured cells, while cell polarity in pre-bilaterian tissues is poorly understood. Here, we have studied apical-basal and planar polarization in regenerating (aggregating) clusters of epitheliomuscular cells of Hydra, a simple representative of the ancestral, pre-bilaterian phylum Cnidaria. Immediately after dissociation, single epitheliomuscular cells do not exhibit cellular polarity, but they polarize de novo during aggregation. Reestablishment of the Hydra-specific epithelial bilayer is a result of short-range cell sorting. In the early phase of aggregation, apical-basal polarization starts with an enlargement of the epithelial apical-basal diameter and by the development of belt-like apical septate junctions. Specification of the basal pole of epithelial cells occurs shortly later and is linked to synthesis of mesoglea, development of hemidesmosome-like junctions, and formation of desmosome-like junctions connecting the basal myonemes of neighbouring cells. Planar polarization starts, while apical-basal polarization is already ongoing. It is executed gradually starting with cell-autonomous formation, parallelization, and condensation of myonemes at the basal end of each epithelial cell and continuing with a final planar alignment of epitheliomuscular cells at the tissue level. Our findings reveal that epithelial polarization in Hydra aggregates occurs in defined steps well accessible by histological and ultrastructural techniques and they will provide a basis for future molecular studies. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Pollen reference collection digitization

    NARCIS (Netherlands)

    Ercan, F.E.Z.|info:eu-repo/dai/nl/41250085X; Donders, T.H.|info:eu-repo/dai/nl/290469872; Bijl, P.K.|info:eu-repo/dai/nl/314028110; Wagner, F.|info:eu-repo/dai/nl/173870783

    2016-01-01

    The extensive Utrecht University pollen reference collection holds thousands of pollen samples of many species and genera from all over the world and has been a basis for the widely-used North West European Pollen Flora. These samples are fixed on glass slides for microscopy use, but the aging

  17. Phenotype and polarization of autologous T cells by biomaterial-treated dendritic cells.

    Science.gov (United States)

    Park, Jaehyung; Gerber, Michael H; Babensee, Julia E

    2015-01-01

    Given the central role of dendritic cells (DCs) in directing T-cell phenotypes, the ability of biomaterial-treated DCs to dictate autologous T-cell phenotype was investigated. In this study, we demonstrate that differentially biomaterial-treated DCs differentially directed autologous T-cell phenotype and polarization, depending on the biomaterial used to pretreat the DCs. Immature DCs (iDCs) were derived from human peripheral blood monocytes and treated with biomaterial films of alginate, agarose, chitosan, hyaluronic acid, or 75:25 poly(lactic-co-glycolic acid) (PLGA), followed by co-culture of these biomaterial-treated DCs and autologous T cells. When autologous T cells were co-cultured with DCs treated with biomaterial film/antigen (ovalbumin, OVA) combinations, different biomaterial films induced differential levels of T-cell marker (CD4, CD8, CD25, CD69) expression, as well as differential cytokine profiles [interferon (IFN)-γ, interleukin (IL)-12p70, IL-10, IL-4] in the polarization of T helper (Th) types. Dendritic cells treated with agarose films/OVA induced CD4+CD25+FoxP3+ (T regulatory cells) expression, comparable to untreated iDCs, on autologous T cells in the DC-T co-culture system. Furthermore, in this co-culture, agarose treatment induced release of IL-12p70 and IL-10 at higher levels as compared with DC treatment with other biomaterial films/OVA, suggesting Th1 and Th2 polarization, respectively. Dendritic cells treated with PLGA film/OVA treatment induced release of IFN-γ at higher levels compared with that observed for co-cultures with iDCs or DCs treated with all other biomaterial films. These results indicate that DC treatment with different biomaterial films has potential as a tool for immunomodulation by directing autologous T-cell responses. © 2014 Wiley Periodicals, Inc.

  18. Pollen morphology of Cornus mas L. and Cornus sanguinea L.

    OpenAIRE

    Karlıoğlu Kılıç, Nurgül; Tuttu, Gamze

    2017-01-01

    Pollen morphology of Cornus mas L. and Cornus sanguinea L.Abstract: The pollen morphology of 2 species of the genus Cornus L. distributed in Turkey was studied with light and scanning electron microscope. The pollen of genus Cornus is 3-colporate. Pollen of the Cornus mas is spheroidal and Cornus sanguinea is subprolate. Size varies of Cornus mas pollen with the polar axis ranging from 21.54 to 27.36 μm and equatorial axis from 21.31 to 26.72 μm. Size varies of Cornus sanguinea pollen with po...

  19. A polarized cell model for Chikungunya virus infection: entry and egress of virus occurs at the apical domain of polarized cells.

    Directory of Open Access Journals (Sweden)

    Pei Jin Lim

    2014-02-01

    Full Text Available Chikungunya virus (CHIKV has resulted in several outbreaks in the past six decades. The clinical symptoms of Chikungunya infection include fever, skin rash, arthralgia, and an increasing incidence of encephalitis. The re-emergence of CHIKV with more severe pathogenesis highlights its potential threat on our human health. In this study, polarized HBMEC, polarized Vero C1008 and non-polarized Vero cells grown on cell culture inserts were infected with CHIKV apically or basolaterally. Plaque assays, viral binding assays and immunofluorescence assays demonstrated apical entry and release of CHIKV in polarized HBMEC and Vero C1008. Drug treatment studies were performed to elucidate both host cell and viral factors involved in the sorting and release of CHIKV at the apical domain of polarized cells. Disruption of host cell myosin II, microtubule and microfilament networks did not disrupt the polarized release of CHIKV. However, treatment with tunicamycin resulted in a bi-directional release of CHIKV, suggesting that N-glycans of CHIKV envelope glycoproteins could serve as apical sorting signals.

  20. The keratin-binding protein Albatross regulates polarization of epithelial cells

    OpenAIRE

    Sugimoto, Masahiko; Inoko, Akihito; Shiromizu, Takashi; Nakayama, Masanori; Zou, Peng; Yonemura, Shigenobu; Hayashi, Yuko; Izawa, Ichiro; Sasoh, Mikio; Uji, Yukitaka; Kaibuchi, Kozo; Kiyono, Tohru; Inagaki, Masaki

    2008-01-01

    The keratin intermediate filament network is abundant in epithelial cells, but its function in the establishment and maintenance of cell polarity is unclear. Here, we show that Albatross complexes with Par3 to regulate formation of the apical junctional complex (AJC) and maintain lateral membrane identity. In nonpolarized epithelial cells, Albatross localizes with keratin filaments, whereas in polarized epithelial cells, Albatross is primarily localized in the vicinity of the AJC. Knockdown o...

  1. She's the boss: signaling in pollen tube reception.

    Science.gov (United States)

    Kessler, Sharon A; Grossniklaus, Ueli

    2011-10-01

    In angiosperms, the sperm cells are carried within the pollen tubes (male gametophytes) to the female gametophyte so that double fertilization can occur. The female gametophyte exerts control over the male, with specialized cells known as synergids guiding the pollen tubes and controlling their behavior when they enter the female gametophyte so that the sperm cells can be delivered to the egg and central cell. Upon pollen tube arrival at the ovule, signal transduction cascades mediated by receptor-like kinases are initiated in both the synergid and the tip of the pollen tube, leading to synergid cell death and pollen tube rupture. In this review, we discuss the role of these receptors and of newly discovered members of the pollen tube reception pathway. Copyright © 2011 Elsevier Ltd. All rights reserved.

  2. Horse chestnut pollen quality

    Directory of Open Access Journals (Sweden)

    Ćalić Dušica

    2017-01-01

    Full Text Available Pollen quality of horse chestnut, expressed as pollen productivity, viability and germination was studied. Anthers of horse chestnut genotypes had pollen production from 3.66 to 5.06 x 103 pollen grains per anther, depending of genotype. Also, pollen of horse chestnut Ah1-Ah4 genotypes showed different viability (from 56 to 68%, after staining with fluorescein diacetate. Pollen germination of Ah1-Ah4 genotypes varied from 50-66% on basic medium. Inclusion of polyethylene glycol-PEG from 10%, 15% and 20% v/w increased pollen germination. The best results were achieved on medium with the largest PEG concentration. On these medium 76-91% pollen grains were germinated, depending of genotype. The best pollen quality, for all tested parameters, had genotype Ah2. Knowledge about morphology, production, viability, in vitro germination, tube growth as well as pollen: ovule ratio can be of great importance for future pollen biology studies. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 173015

  3. Flotillins are involved in the polarization of primitive and mature hematopoietic cells.

    Directory of Open Access Journals (Sweden)

    Lawrence Rajendran

    Full Text Available BACKGROUND: Migration of mature and immature leukocytes in response to chemokines is not only essential during inflammation and host defense, but also during development of the hematopoietic system. Many molecules implicated in migratory polarity show uniform cellular distribution under non-activated conditions, but acquire a polarized localization upon exposure to migratory cues. METHODOLOGY/PRINCIPAL FINDINGS: Here, we present evidence that raft-associated endocytic proteins (flotillins are pre-assembled in lymphoid, myeloid and primitive hematopoietic cells and accumulate in the uropod during migration. Furthermore, flotillins display a polarized distribution during immunological synapse formation. Employing the membrane lipid-order sensitive probe Laurdan, we show that flotillin accumulation in the immunological synapse is concomittant with membrane ordering in these regions. CONCLUSIONS: Together with the observation that flotillin polarization does not occur in other polarized cell types such as polarized epithelial cells, our results suggest a specific role for flotillins in hematopoietic cell polarization. Based on our results, we propose that in hematopoietic cells, flotillins provide intrinsic cues that govern segregation of certain microdomain-associated molecules during immune cell polarization.

  4. Polarization resolved conoscopic patterns in nematic cells: effects induced by the incident light ellipticity

    Science.gov (United States)

    Buinyi, Igor O.; Soskin, Marat S.; Vovk, Roman G.

    2008-05-01

    Topological structure of the polarization resolved conoscopic patterns, calculated theoretically and measured experimentally for nematic liquid crystal (NLC) cells, is described in terms of polarization singularities, saddle points and bifurcation lines. The parametric dynamics of the topological network, induced by the variation of the incident light ellipticity, is analyzed for the nematic cells with uniform and non-uniform director configuration. Different stages of similar dynamics are observed for homeotropically oriented NLC cell. Non-uniform director configuration within the cell results in broken central symmentry in the arrangement of the topological network. Main features of the experimentally obtained polarization resolved conoscopic patterns are the same to the theoretically predicted ones.

  5. Topological structure in polarization resolved conoscopic patterns for nematic liquid crystal cells

    Science.gov (United States)

    Buinyi, Igor O.; Denisenko, Vladimir G.; Soskin, Marat S.

    2009-01-01

    We investigate the polarization structure of coherent light, produced by a convergent light beam transmitted through nematic liquid crystal (NLC) cells with different director configurations. Employing solutions to the transmission problem for the case when plane wave propagates through an anisotropic layer, we analyze the arrangement of the topological elements, such as polarization singularities (C points with circular polarization and L lines with linear polarization), saddle points and extrema of polarization azimuth. We observe transformations of the topological structure under the variation of the incident light ellipticity and represent it by corresponding trajectories of topological elements in three-dimensional space. For the cells with uniform and non-uniform director configuration we describe the processes of creation/annihilation of C point pairs, which can be controlled precisely in the case of the cell with non-uniform director. Our experimental measurements for the homeotropically oriented NLC cells are in good agreement with the theoretical predictions.

  6. Monitoring single-channel water permeability in polarized cells.

    Science.gov (United States)

    Erokhova, Liudmila; Horner, Andreas; Kügler, Philipp; Pohl, Peter

    2011-11-18

    So far the determination of unitary permeability (p(f)) of water channels that are expressed in polarized cells is subject to large errors because the opening of a single water channel does not noticeably increase the water permeability of a membrane patch above the background. That is, in contrast to the patch clamp technique, where the single ion channel conductance may be derived from a single experiment, two experiments separated in time and/or space are required to obtain the single-channel water permeability p(f) as a function of the incremental water permeability (P(f,c)) and the number (n) of water channels that contributed to P(f,c). Although the unitary conductance of ion channels is measured in the native environment of the channel, p(f) is so far derived from reconstituted channels or channels expressed in oocytes. To determine the p(f) of channels from live epithelial monolayers, we exploit the fact that osmotic volume flow alters the concentration of aqueous reporter dyes adjacent to the epithelia. We measure these changes by fluorescence correlation spectroscopy, which allows the calculation of both P(f,c) and osmolyte dilution within the unstirred layer. Shifting the focus of the laser from the aqueous solution to the apical and basolateral membranes allowed the FCS-based determination of n. Here we validate the new technique by determining the p(f) of aquaporin 5 in Madin-Darby canine kidney cell monolayers. Because inhibition and subsequent activity rescue are monitored on the same sample, drug effects on exocytosis or endocytosis can be dissected from those on p(f).

  7. Isobaric tags for relative and absolute quantification- based comparative proteomics reveals the features of plasma membrane-associated proteomes of pollen grains and pollen tubes from Lilium davidii.

    Science.gov (United States)

    Han, Bing; Chen, Sixue; Dai, Shaojun; Yang, Ning; Wang, Tai

    2010-12-01

    Mature pollen grains (PGs) from most plant species are metabolically quiescent. However, once pollinated onto stigma, they quickly hydrate and germinate. A PG can give rise to a vegetative cell-derived polarized pollen tube (PT), which represents a specialized polar cell. The polarized PT grows by the tip and requires interaction of different signaling molecules localized in the apical plasma membrane and active membrane trafficking. The mechanisms underlying the interaction and membrane trafficking are not well understood. In this work, we purified PG and PT plasma-membrane vesicles from Lilium davidii Duch. using the aqueous two-phase partition technique, then enriched plasma membrane proteins by using Brij58 and KCl to remove loosely bound contaminants. We identified 223 integral and membrane-associated proteins in the plasma membrane of PGs and PTs by using isobaric tags for relative and absolute quantification (iTRAQ) and 2-D high-performance liquid chromatography-tandem mass spectrometry. More than 68% of the proteins have putative transmembrane domains and/or lipid-modified motifs. Proteins involved in signal transduction, membrane trafficking and transport are predominant in the plasma-membrane proteome. We revealed most components of the clathrin-dependent endocytosis pathway. Statistical analysis revealed 14 proteins differentially expressed in the two development stages: in PTs, six upregulated and eight downregulated are mainly involved in signaling, transport and membrane trafficking. These results provide novel insights into polarized PT growth. © 2010 Institute of Botany, Chinese Academy of Sciences.

  8. Gene expression profile indicates involvement of NO in Camellia sinensis pollen tube growth at low temperature.

    Science.gov (United States)

    Pan, Junting; Wang, Weidong; Li, Dongqin; Shu, Zaifa; Ye, Xiaoli; Chang, Pinpin; Wang, Yuhua

    2016-10-18

    Nitric oxide (NO) functions as a critical signaling molecule in the low-temperature stress responses in plants, including polarized pollen tube growth in Camellia sinensis. Despite this, the potential mechanisms underlying the participation of NO in pollen tube responses to low temperature remain unclear. Here, we investigate alterations to gene expression in C. sinensis pollen tubes exposed to low-temperature stress and NO using RNA-Seq technology, in order to find the potential candidate genes related to the regulation of pollen tube elongation by NO under low-temperature stress. Three libraries were generated from C. sinensis cv. 'Longjingchangye' pollen tubes cultured at 25 °C (CsPT-CK) and 4 °C (CsPT-LT) or with 25 μM DEA NONOate (CsPT-NO). The number of unigenes found for the three biological replications were 39,726, 40,440 and 41,626 for CsPT-CK; 36,993, 39,070 and 39,439 for CsPT-LT; and 39,514, 38,298 and 39,061 for CsPT-NO. A total of 36,097 unique assembled and annotated sequences from C. sinensis pollen tube reads were found in a BLAST search of the following databases: NCBI non-redundant nucleotide, Swiss-prot protein, Kyoto Encyclopedia of Genes and Genomes, Cluster of Orthologous Groups of proteins, and Gene Ontology. The absolute values of log 2 Ratio > 1 and probability > 0.7 were used as the thresholds for significantly differential gene expression, and 766, 497 and 929 differentially expressed genes (DEGs) were found from the comparison analyses of the CK-VS-LT, CK-VS-NO and LT-VS-NO libraries, respectively. Genes related to metabolism and signaling pathways of plant hormones, transcription factors (TFs), vesicle polarized trafficking, cell wall biosynthesis, the ubiquitination machinery of the ubiquitin system and species-specific secondary metabolite pathways were mainly observed in the CK-VS-LT and CK-VS-NO libraries. Differentially expressed unigenes related to the inhibition of C. sinensis pollen tube growth under low

  9. Tests of a polarized source of hydrogen and deuterium based on spin-exchange optical pumping and a storage cell for polarized deuterium

    International Nuclear Information System (INIS)

    Holt, R.J.; Gilman, R.; Kinney, E.R.

    1988-01-01

    A novel laser-driven polarized source of hydrogen and deuterium which is based on the principle of spin-exchange optical pumping has been developed at Argonne. The advantages of this method over conventional polarized sources for internal target experiments is discussed. At present, the laser-driven polarized source delivers hydrogen 8 x 10 16 atoms/s with a polarization of 24% and deuterium at 6 x 10 16 atoms/s with a polarization of 25%. A passive storage cell for polarized deuterium was tested in the VEPP-3 electron storage ring. The storage cell was found to increase the target thickness by approximately a factor of three and no loss in polarization was observed. 10 refs., 4 figs., 2 tabs

  10. A Localized Complex of Two Protein Oligomers Controls the Orientation of Cell Polarity

    Directory of Open Access Journals (Sweden)

    Adam M. Perez

    2017-02-01

    Full Text Available Signaling hubs at bacterial cell poles establish cell polarity in the absence of membrane-bound compartments. In the asymmetrically dividing bacterium Caulobacter crescentus, cell polarity stems from the cell cycle-regulated localization and turnover of signaling protein complexes in these hubs, and yet the mechanisms that establish the identity of the two cell poles have not been established. Here, we recapitulate the tripartite assembly of a cell fate signaling complex that forms during the G1-S transition. Using in vivo and in vitro analyses of dynamic polar protein complex formation, we show that a polymeric cell polarity protein, SpmX, serves as a direct bridge between the PopZ polymeric network and the cell fate-directing DivJ histidine kinase. We demonstrate the direct binding between these three proteins and show that a polar microdomain spontaneously assembles when the three proteins are coexpressed heterologously in an Escherichia coli test system. The relative copy numbers of these proteins are essential for complex formation, as overexpression of SpmX in Caulobacter reorganizes the polarity of the cell, generating ectopic cell poles containing PopZ and DivJ. Hierarchical formation of higher-order SpmX oligomers nucleates new PopZ microdomain assemblies at the incipient lateral cell poles, driving localized outgrowth. By comparison to self-assembling protein networks and polar cell growth mechanisms in other bacterial species, we suggest that the cooligomeric PopZ-SpmX protein complex in Caulobacter illustrates a paradigm for coupling cell cycle progression to the controlled geometry of cell pole establishment.

  11. Polarity governed selective amplification of through plane proton shuttling in proton exchange membrane fuel cells.

    Science.gov (United States)

    Gautam, Manu; Chattanahalli Devendrachari, Mruthyunjayachari; Thimmappa, Ravikumar; Raja Kottaichamy, Alagar; Pottachola Shafi, Shahid; Gaikwad, Pramod; Makri Nimbegondi Kotresh, Harish; Ottakam Thotiyl, Musthafa

    2017-03-15

    Graphene oxide (GO) anisotropically conducts protons with directional dominance of in plane ionic transport (σ IP) over the through plane (σ TP). In a typical H 2 -O 2 fuel cell, since the proton conduction occurs through the plane during its generation at the fuel electrode, it is indeed inevitable to selectively accelerate GO's σ TP for advancement towards a potential fuel cell membrane. We successfully achieved ∼7 times selective amplification of GO's σ TP by tuning the polarity of the dopant molecule in its nanoporous matrix. The coexistence of strongly non-polar and polar domains in the dopant demonstrated a synergistic effect towards σ TP with the former decreasing the number of water molecules coordinated to protons by ∼3 times, diminishing the effects of electroosmotic drag exerted on ionic movements, and the latter selectively accelerating σ TP across the catalytic layers by bridging the individual GO planes via extensive host guest H-bonding interactions. When they are decoupled, the dopant with mainly non-polar or polar features only marginally enhances the σ TP, revealing that polarity factors contribute to fuel cell relevant transport properties of GO membranes only when they coexist. Fuel cell polarization and kinetic analyses revealed that these multitask dopants increased the fuel cell performance metrics of the power and current densities by ∼3 times compared to the pure GO membranes, suggesting that the functional group factors of the dopants are of utmost importance in GO-based proton exchange membrane fuel cells.

  12. Hierarchy of mechanisms involved in generating Na/K-ATPase polarity in MDCK epithelial cells

    NARCIS (Netherlands)

    Mays, R.W.; Siemers, K.A.; Fritz, B.A.; Lowe, A.W.; van Meer, G.; Nelson, W.J.

    1995-01-01

    We have studied mechanisms involved in generating a polarized distribution of Na/K-ATPase in the basal-lateral membrane of two clones of MDCK II cells. Both clones exhibit polarized distributions of marker proteins of the apical and basal-lateral membranes, including Na/K-ATPase, at steady state.

  13. Evolutionarily conserved phenylpropanoid pattern on angiosperm pollen.

    Science.gov (United States)

    Fellenberg, Christin; Vogt, Thomas

    2015-04-01

    The male gametophyte of higher plants appears as a solid box containing the essentials to transmit genetic material to the next generation. These consist of haploid generative cells that are required for reproduction, and an invasive vegetative cell producing the pollen tube, both mechanically protected by a rigid polymer, the pollen wall, and surrounded by a hydrophobic pollen coat. This coat mediates the direct contact to the biotic and abiotic environments. It contains a mixture of compounds required not only for fertilization but also for protection against biotic and abiotic stressors. Among its metabolites, the structural characteristics of two types of phenylpropanoids, hydroxycinnamic acid amides and flavonol glycosides, are highly conserved in Angiosperm pollen. Structural and functional aspects of these compounds will be discussed. Copyright © 2015 Elsevier Ltd. All rights reserved.

  14. Mathematical analysis of steady-state solutions in compartment and continuum models of cell polarization.

    Science.gov (United States)

    Zheng, Zhenzhen; Chou, Ching-Shan; Yi, Tau-Mu; Nie, Qing

    2011-10-01

    Cell polarization, in which substances previously uniformly distributed become asymmetric due to external or/and internal stimulation, is a fundamental process underlying cell mobility, cell division, and other polarized functions. The yeast cell S. cerevisiae has been a model system to study cell polarization. During mating, yeast cells sense shallow external spatial gradients and respond by creating steeper internal gradients of protein aligned with the external cue. The complex spatial dynamics during yeast mating polarization consists of positive feedback, degradation, global negative feedback control, and cooperative effects in protein synthesis. Understanding such complex regulations and interactions is critical to studying many important characteristics in cell polarization including signal amplification, tracking dynamic signals, and potential trade-off between achieving both objectives in a robust fashion. In this paper, we study some of these questions by analyzing several models with different spatial complexity: two compartments, three compartments, and continuum in space. The step-wise approach allows detailed characterization of properties of the steady state of the system, providing more insights for biological regulations during cell polarization. For cases without membrane diffusion, our study reveals that increasing the number of spatial compartments results in an increase in the number of steady-state solutions, in particular, the number of stable steady-state solutions, with the continuum models possessing infinitely many steady-state solutions. Through both analysis and simulations, we find that stronger positive feedback, reduced diffusion, and a shallower ligand gradient all result in more steady-state solutions, although most of these are not optimally aligned with the gradient. We explore in the different settings the relationship between the number of steady-state solutions and the extent and accuracy of the polarization. Taken together

  15. Networking for proteins : A yeast two-hybrid and RNAi profiling approach to uncover C. elegans cell polarity regulators

    NARCIS (Netherlands)

    Koorman, T.

    2016-01-01

    Cell polarity is a near universal trait of life and guides many aspects of animal development. Although a number of key polarity proteins have been identified, many interactions with proteins acting downstream likely remain to be elucidated. Mutations in polarity proteins or deregulation of polarity

  16. Airway epithelial cell-derived insulin-like growth factor-1 triggers skewed CD8(+) T cell polarization.

    Science.gov (United States)

    Zou, Jian-Yong; Huang, Shao-hong; Li, Yun; Chen, Hui-guo; Rong, Jian; Ye, Sheng

    2014-10-01

    Skewed CD8(+) T cell responses are important in airway inflammation. This study investigates the role of the airway epithelial cell-derived insulin-like growth factor 1 (IGF1) in contributing to CD8(+) T cell polarization. Expression of IGF1 in the airway epithelial cell line, RPMI2650 cells, was assessed by quantitative real time RT-PCR and Western blotting. The role of IGF1 in regulating CD8(+) T cell activation was observed by coculture of mite allergen-primed RPMI2650 cells and naïve CD8(+) T cells. CD8(+) T cell polarization was assessed by the carboxyfluorescein succinimidyl ester-dilution assay and the determination of cytotoxic cytokine levels in the culture medium. Exposure to mite allergen, Der p1, increased the expression of IGF1 by RPMI2650 cells. The epithelial cell-derived IGF1 prevented the activation-induced cell death by inducing the p53 gene hypermethylation. Mite allergen-primed RPMI2650 cells induced an antigen-specific CD8(+) T cell polarization. We conclude that mite allergens induce airway epithelial cell line, RPMI2650 cells, to produce IGF1; the latter contributes to antigen-specific CD8(+) T cell polarization. © 2014 International Federation for Cell Biology.

  17. Cell shape, spreading symmetry, and the polarization of stress-fibers in cells

    Energy Technology Data Exchange (ETDEWEB)

    Zemel, A [Institute of Dental Sciences, Faculty of Dental Medicine, and the Fritz Haber Center for Molecular Dynamics, Hebrew University-Hadassah Medical Center, Jerusalem, 91120 (Israel); Rehfeldt, F [III. Physikalisches Institut, Georg-August-Universitaet, 37077 Goettingen (Germany); Brown, A E X [Department of Physics and Astronomy, University of Pennsylvania, Philadelphia, PA 19104 (United States); Discher, D E [Graduate Group of Physics and Astronomy, University of Pennsylvania, Philadelphia, PA 19104 (United States); Safran, S A [Department of Materials and Interfaces, Weizmann Institute of Science, Rehovot 76100 (Israel)

    2010-05-19

    The active regulation of cellular forces during cell adhesion plays an important role in the determination of cell size, shape, and internal structure. While on flat, homogeneous and isotropic substrates some cells spread isotropically, others spread anisotropically and assume elongated structures. In addition, in their native environment as well as in vitro experiments, the cell shape and spreading asymmetry can be modulated by the local distribution of adhesive molecules and topography of the environment. We present a simple elastic model and experiments on stem cells to explain the variation of cell size with the matrix rigidity. In addition, we predict the experimental consequences of two mechanisms of acto-myosin polarization and focus here on the effect of the cell spreading asymmetry on the regulation of the stress-fiber alignment in the cytoskeleton. We show that when cell spreading is sufficiently asymmetric the alignment of acto-myosin forces in the cell increases monotonically with the matrix rigidity; however, in general this alignment is non-monotonic, as shown previously. These results highlight the importance of the symmetry characteristics of cell spreading in the regulation of cytoskeleton structure and suggest a mechanism by which different cell types may acquire different morphologies and internal structures in different mechanical environments.

  18. Polarization splitting phenomenon of photonic crystals constructed by two-fold rotationally symmetric unit-cells

    Science.gov (United States)

    Yasa, U. G.; Giden, I. H.; Turduev, M.; Kurt, H.

    2017-09-01

    We present an intrinsic polarization splitting characteristic of low-symmetric photonic crystals (PCs) formed by unit-cells with C 2 rotational symmetry. This behavior emerges from the polarization sensitive self-collimation effect for both transverse-magnetic (TM) and transverse-electric (TE) modes depending on the rotational orientations of the unit-cell elements. Numerical analyzes are performed in both frequency and time domains for different types of square lattice two-fold rotational symmetric PC structures. At incident wavelength of λ = 1550 nm, high polarization extinction ratios with ˜26 dB (for TE polarization) and ˜22 dB (for TM polarization) are obtained with an operating bandwidth of 59 nm. Moreover, fabrication feasibilities of the designed structure are analyzed to evaluate their robustness in terms of the unit-cell orientation: for the selected PC unit-cell composition, corresponding extinction ratios for both polarizations still remain to be over 18 dB for the unit-cell rotation interval of θ = [40°-55°]. Taking all these advantages, two-fold rotationally symmetric PCs could be considered as an essential component in photonic integrated circuits for polarization control of light.

  19. Reactions of airway epithelial cells to birch pollen grains previously exposed to in situ atmospheric Pb concentrations: a preliminary assay of allergenicity.

    Science.gov (United States)

    Bellanger, Anne-Pauline; Bosch-Cano, Floriane; Millon, Laurence; Ruffaldi, Pascale; Franchi, Marielle; Bernard, Nadine

    2012-12-01

    A growing body of evidence suggests that interactions between pollen grains and environmental pollutants, especially air pollutants, could be of critical importance with regard to the increase in allergic responses observed in the past decades. Using birch pollen grains (BPG), a major allergy source in European countries, and lead (Pb), a highly toxic metal trace element (MTE) present in urban areas, the immune response of human epithelial cells exposed to BPG or to Pb-associated BPG was compared. The cellular response after exposure either to BPG, BPG exposed to 30 mg/L of Pb (BPG-30), or BPG exposed to 60 mg/L of Pb (BPG-60) was evaluated after two time lapses (2 and 6 h) by measuring mRNA levels of four mediators, including two inflammatory (interleukin-8 and interleukin-6) and two allergic (interleukin-5 [IL-5] and interleukin-13) cytokines. After 2 h of exposure, significant upregulation of the IL-5 gene was observed after exposure to BPG-60 in comparison with exposure to BPG and BPG-30 (N (IL-5) = 1.9, Mann-Whitney test, p = 0.003). After 6 h of exposure, significant upregulation of the IL-5 gene was observed after exposure to BPG-30 with N (IL-5) = 1.8 and to BPG-60 with N (IL-5) = 2.3 (Mann-Whitney test, p = 0.0029) in comparison with exposure to BPG. This first attempt to investigate the influence of pollution by MTE on pollen grain showed a dose-time-dependent increase in IL-5 gene expression after exposure to BPG combined to Pb.

  20. MCAM contributes to the establishment of cell autonomous polarity in myogenic and chondrogenic differentiation

    Directory of Open Access Journals (Sweden)

    Artal Moreno-Fortuny

    2017-11-01

    Full Text Available Cell polarity has a fundamental role in shaping the morphology of cells and growing tissues. Polarity is commonly thought to be established in response to extracellular signals. Here we used a minimal in vitro assay that enabled us to monitor the determination of cell polarity in myogenic and chondrogenic differentiation in the absence of external signalling gradients. We demonstrate that the initiation of cell polarity is regulated by melanoma cell adhesion molecule (MCAM. We found highly polarized localization of MCAM, Moesin (MSN, Scribble (SCRIB and Van-Gogh-like 2 (VANGL2 at the distal end of elongating myotubes. Knockout of MCAM or elimination of its endocytosis motif does not impair the initiation of myogenesis or myoblast fusion, but prevents myotube elongation. MSN, SCRIB and VANGL2 remain uniformly distributed in MCAM knockout cells. We show that MCAM is also required at early stages of chondrogenic differentiation. In both myogenic and chondrogenic differentiation MCAM knockout leads to transcriptional downregulation of Scrib and enhanced MAP kinase activity. Our data demonstrates the importance of cell autonomous polarity in differentiation.

  1. Mechanisms of Cell Polarity-Controlled Epithelial Homeostasis and Immunity in the Intestine

    NARCIS (Netherlands)

    Klunder, Leon J.; Faber, Klaas Nico; Dijkstra, Gerard; van IJzendoorn, Sven C. D.

    Intestinal epithelial cell polarity is instrumental to maintain epithelial homeostasis and balance communications between the gut lumen and bodily tissue, thereby controlling the defense against gastrointestinal pathogens and maintenance of immune tolerance to commensal bacteria. In this review, we

  2. Concomitant use of polarization and positive phase contrast microscopy for the study of microbial cells

    Czech Academy of Sciences Publication Activity Database

    Žižka, Zdeněk; Gabriel, Jiří

    2015-01-01

    Roč. 60, č. 6 (2015), s. 545-550 ISSN 0015-5632 Institutional support: RVO:61388971 Keywords : polarization microscopy * microbial cells * positive phase contrast Subject RIV: EE - Microbiology, Virology Impact factor: 1.335, year: 2015

  3. Gametophytic Pollen Tube Guidance: Attractant Peptides, Gametic Controls, and Receptors1[OPEN

    Science.gov (United States)

    2017-01-01

    Pollen tube guidance in flowering plants is a unique and critical process for successful sexual reproduction. The pollen tube that grows from pollen, which is the male gametophyte, precisely navigates to the embryo sac, which is the female gametophyte, within the pistil. Recent advances have clarified the molecular framework of gametophytic pollen tube guidance. Multiple species-specific attractant peptides are secreted from synergid cells, the proper development and function of which are regulated by female gametes. Multiple receptor-like kinases on the pollen tube tip are involved in sensing species-specific attractant peptides. In this Update article, recent progress in our understanding of the mechanism of gametophytic pollen tube guidance is reviewed, including attraction by synergid cells, control of pollen tube guidance by female gametes, and directional growth of the pollen tube by directional cue sensing. Future directions in the study of pollen tube guidance also are discussed. PMID:27920159

  4. Zebrafish models of non-canonical Wnt/planar cell polarity signalling: fishing for valuable insight into vertebrate polarized cell behavior.

    Science.gov (United States)

    Jussila, Maria; Ciruna, Brian

    2017-05-01

    Planar cell polarity (PCP) coordinates the uniform orientation, structure and movement of cells within the plane of a tissue or organ system. It is beautifully illustrated in the polarized arrangement of bristles and hairs that project from specialized cell surfaces of the insect abdomen and wings, and pioneering genetic studies using the fruit fly, Drosophila melanogaster, have defined a core signalling network underlying PCP. This core PCP/non-canonical Wnt signalling pathway is evolutionarily conserved, and studies in zebrafish have helped transform our understanding of PCP from a peculiarity of polarized epithelia to a more universal cellular property that orchestrates a diverse suite of polarized cell behaviors that are required for normal vertebrate development. Furthermore, application of powerful genetics, embryonic cell-transplantation, and live-imaging capabilities afforded by the zebrafish model have yielded novel insights into the establishment and maintenance of vertebrate PCP, over the course of complex and dynamic morphogenetic events like gastrulation and neural tube morphogenesis. Although key questions regarding vertebrate PCP remain, with the emergence of new genome-editing technologies and the promise of endogenous labeling and Cre/LoxP conditional targeting strategies, zebrafish remains poised to deliver fundamental new insights into the function and molecular dynamic regulation of PCP signalling from embryonic development through to late-onset phenotypes and adult disease states. WIREs Dev Biol 2017, 6:e267. doi: 10.1002/wdev.267 For further resources related to this article, please visit the WIREs website. © 2017 Wiley Periodicals, Inc.

  5. Mechanisms of Cell Polarity-Controlled Epithelial Homeostasis and Immunity in the Intestine.

    Science.gov (United States)

    Klunder, Leon J; Faber, Klaas Nico; Dijkstra, Gerard; van IJzendoorn, Sven C D

    2017-07-05

    Intestinal epithelial cell polarity is instrumental to maintain epithelial homeostasis and balance communications between the gut lumen and bodily tissue, thereby controlling the defense against gastrointestinal pathogens and maintenance of immune tolerance to commensal bacteria. In this review, we highlight recent advances with regard to the molecular mechanisms of cell polarity-controlled epithelial homeostasis and immunity in the human intestine. Copyright © 2017 Cold Spring Harbor Laboratory Press; all rights reserved.

  6. CONTRIBUTIONS TO THE MORPHOLOGY OF POLLEN FROM SOME VARIETIES OF MALUS DOMESTICA, PYRUS COMMUNIS, PRUNUS DOMESTICA, PRUNUS PERSICA AND PRUNUS ARMENIACA

    Directory of Open Access Journals (Sweden)

    Nicoleta IANOVICI

    2010-01-01

    Full Text Available The objective of this study was to compare Rosaceae genotypes from for their pollen viability and morphology. The pollen of some varieties of Malus domestica, Pyrus communis, Prunus domestica, Prunus persica and Prunus armeniaca was identified. Pollen from mature anthers was collected. Pollen grains of all genera of Rosaceae surveyed occur as radially symmetric isopolar monads. The main aspect studied were the shape of pollen in the polar view (polar perimeter and polar area of pollen. Shape of pollen is triangular-obtuse-convex in polar view. The pollen class is trizonocolpate-obtus-triangular while the sculpturing of exine surface is striate. The number of colpi is three while the spines are absent. Apart from fully developed pollen grains, also much smaller, not completely developed pollen were found in the samples of the examined species. The viability of the pollen was determined on 1% 2,3,5-triphenyl tetrazolium chloride (TTC. Viable, semi-viable and dead pollen numbers and their percentages were determined. This stain test may be used to determine pollen viability in these species to provide only a rough estimate of viability. The results presented here are important for improving our understanding of Rosaceae reproduction biology. Knowledge of reproduction biology, particularly pollen viability and quality, is critical for the newly-developed cultivars.

  7. Porcine aminopeptidase N mediated polarized infection by porcine epidemic diarrhea virus in target cells

    Energy Technology Data Exchange (ETDEWEB)

    Cong, Yingying; Li, Xiaoxue; Bai, Yunyun [College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030 (China); Lv, Xiaonan [College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030 (China); CAS Key Lab for Biomedical Effects of Nanomaterials and Nanosafety, National Center for Nanoscience & Technology of China, Beijing 100090 (China); Herrler, Georg [Institute for Virology, University of Veterinary Medicine, Hannover D-30559 (Germany); Enjuanes, Luis [Department of Molecular and Cell Biology, Centro Nacional de Biotecnología (CNB-CSIC), Campus Universidad Autónoma de Madrid, Cantoblanco, Madrid (Spain); Zhou, Xingdong [College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030 (China); Qu, Bo [Faculty of Life Sciences, Northeast Agricultural University, Harbin 150030 (China); Meng, Fandan [Institute for Virology, University of Veterinary Medicine, Hannover D-30559 (Germany); Cong, Chengcheng [College Animal Husbandry and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110161 (China); Ren, Xiaofeng; Li, Guangxing [College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030 (China)

    2015-04-15

    Infection of polarized intestinal epithelial cells by porcine epidemic diarrhea virus (PEDV) was characterized. Indirect immunofluorescence assay, real-time PCR, and transmission electron microscopy confirmed PEDV can be successfully propagated in immortalized swine small intestine epithelial cells (IECs). Infection involved porcine aminpeptidase N (pAPN), a reported cellular receptor for PEDV, transient expression of pAPN and siRNA targeted pAPN increased and decreased the infectivity of PEDV in IECs, respectively. Subsequently, polarized entry into and release from both Vero E6 and IECs was analyzed. PEDV entry into polarized cells and pAPN grown on membrane inserts occurs via apical membrane. The progeny virus released into the medium was also quantified which demonstrated that PEDV is preferentially released from the apical membrane. Collectively, our data demonstrate that pAPN, the cellular receptor for PEDV, mediates polarized PEDV infection. These results imply the possibility that PEDV infection may proceed by lateral spread of virus in intestinal epithelial cells. - Highlights: • PEDV infection of polarized intestinal epithelial cells (IECs) was characterized. • Porcine aminpeptidase N (pAPN) facilitated PEDV infection in IECs. • PEDV entry into and release from polarized cell via its apical membrane. • PEDV infection may proceed by lateral spread of virus in IECs.

  8. Pollen morphology of the Onobrychis sect. Heliobrychis in Iran

    Directory of Open Access Journals (Sweden)

    Roya Karamian

    2010-01-01

    Full Text Available Pollen morphology of 20 species belonging to Onobrychis sect. Heliobrychis was studied in Iran. The pollen grains from the herbarium specimens were prepared by acetolysis method and then 8 quantitative characters were studied by light microscopy. Data analysis was carried out by cluster analysis and principal component analysis methods. Pollen grains of the sections were 3-colpate with reticulate ornamentation. Results of the study showed that although the pollen grains in the section were comparatively homogenous, but had a considerable variation in the studied quantitative characters. However among the studied characters, equatorial and polar diameters of pollen grains were the most significant characters. The members of the section can be divided into 3 groups based on these characters.

  9. Cytological evidences of pollen abortion in Ornithogalum caudatum Ait

    African Journals Online (AJOL)

    Administrator

    2011-10-19

    Oct 19, 2011 ... Ornithogalum caudatum Ait exhibits strong physiological adaptability; however, it suffers a serious menace of pollen abortion, as almost no seed is formed in the plants. In the current study, chromosome karyotype of root-tip cells (RTC) during mitosis, and chromosome behavior of pollen mother cells.

  10. Cytological evidences of pollen abortion in Ornithogalum caudatum Ait

    African Journals Online (AJOL)

    Ornithogalum caudatum Ait exhibits strong physiological adaptability; however, it suffers a serious menace of pollen abortion, as almost no seed is formed in the plants. In the current study, chromosome karyotype of root-tip cells (RTC) during mitosis, and chromosome behavior of pollen mother cells (PMC) during meiosis in ...

  11. Comprehensive analysis of tobacco pollen transcriptome unveils common pathways in polar cell expansion and underlying heterochronic shift during spermatogenesis

    Czech Academy of Sciences Publication Activity Database

    Hafidh, Said; Breznenová, Katarína; Růžička, Petr; Feciková, Jana; Čapková, Věra; Honys, David

    2012-01-01

    Roč. 12, č. 24 (2012) ISSN 1471-2229 R&D Projects: GA ČR GPP501/11/P321; GA ČR(CZ) GAP501/11/1462; GA MŠk(CZ) OC10054 Institutional research plan: CEZ:AV0Z50380511 Keywords : MALE GAMETOPHYTE DEVELOPMENT * TUBE GROWTH * GENE-EXPRESSION Subject RIV: ED - Physiology Impact factor: 4.354, year: 2012

  12. The viral spike protein is not involved in the polarized sorting of coronaviruses in epithelial cells

    NARCIS (Netherlands)

    Horzinek, M.C.; Rossen, J.W.A.; Beer, R. de; Godeke, G.J.; Raamsman, M.J.; Vennema, H.; Rottier, P.J.M.

    1998-01-01

    Coronaviruses are assembled by budding into a pre-Golgi compartment from which they are transported along the secretory pathway to leave the cell. In cultured epithelial cells, they are released in a polarized fashion; depending on the virus and cell type, they are sorted preferentially either to

  13. Anti-inflammatory effect of stem cells against spinal cord injury via regulating macrophage polarization

    Directory of Open Access Journals (Sweden)

    Cheng ZJ

    2017-02-01

    Full Text Available Zhijian Cheng, Xijing He Department of Orthopedics, The Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an, Shaanxi, People’s Republic of China Abstract: Spinal cord injury (SCI is a traumatic event that involves not just an acute physical injury but also inflammation-driven secondary injury. Macrophages play a very important role in secondary injury. The effects of macrophages on tissue damage and repair after SCI are related to macrophage polarization. Stem cell transplantation has been studied as a promising treatment for SCI. Recently, increasing evidence shows that stem cells, including mesenchymal stem, neural stem/progenitor, and embryonic stem cells, have an anti-inflammatory capacity and promote functional recovery after SCI by inducing macrophages M1/M2 phenotype transformation. In this review, we will discuss the role of stem cells on macrophage polarization and its role in stem cell-based therapies for SCI. Keywords: stem cells, macrophages, spinal cord injury, polarization

  14. A Comparison of the Human Buccal Cell Assay and the Pollen Abortion Assay in Assessing Genotoxicity in an Urban-Rural Gradient

    Science.gov (United States)

    Fleck, Alan da Silveira; Vieira, Mariana; Amantéa, Sergio Luís; Rhoden, Claudia Ramos

    2014-01-01

    Air pollution is exacerbated near heavy traffic roads in cities. Air pollution concentration and composition vary by region and depend on urban-rural gradients. The aim of this study was to evaluate the distribution of air pollution in areas of varying population densities and to compare plant biomonitoring with an established biomarker of human exposure to traffic-related air pollution in children. The areas of study were selected near a major street in 3 different regions. Areas A, B and C represent high, intermediate and low population densities, respectively. Micronucleus assay, an established biomarker of human exposure, was performed in children from these areas. For a plant biomonitoring assay, the pollen abortion assay was performed on Bauhinia variegata in these areas. NO2 and O3 concentrations were determined by passive sampling. We report here that the pollen abortion frequency in Bauhinia variegata is correlated with NO2 concentration (P = 0.004) and is strongly associated with vehicular flow and population density in the studied areas. Micronuclei frequency in buccal cells of children was higher in the regions with more degree of urbanization (P air pollutants in Porto Alegre are related to both human and plant genotoxicity. Areas with different concentration of pollutants demonstrated to have an urbanization gradient dependent pattern which also reflected on genotoxic damage among these areas. PMID:25166920

  15. A Comparison of the Human Buccal Cell Assay and the Pollen Abortion Assay in Assessing Genotoxicity in an Urban-Rural Gradient

    Directory of Open Access Journals (Sweden)

    Alan da Silveira Fleck

    2014-08-01

    Full Text Available Air pollution is exacerbated near heavy traffic roads in cities. Air pollution concentration and composition vary by region and depend on urban-rural gradients. The aim of this study was to evaluate the distribution of air pollution in areas of varying population densities and to compare plant biomonitoring with an established biomarker of human exposure to traffic-related air pollution in children. The areas of study were selected near a major street in 3 different regions. Areas A, B and C represent high, intermediate and low population densities, respectively. Micronucleus assay, an established biomarker of human exposure, was performed in children from these areas. For a plant biomonitoring assay, the pollen abortion assay was performed on Bauhinia variegata in these areas. NO2 and O3 concentrations were determined by passive sampling. We report here that the pollen abortion frequency in Bauhinia variegata is correlated with NO2 concentration (P = 0.004 and is strongly associated with vehicular flow and population density in the studied areas. Micronuclei frequency in buccal cells of children was higher in the regions with more degree of urbanization (P < 0.001 following the same pattern of O3 concentrations (P = 0.030. In conclusion, our results demonstrate that high concentrations of air pollutants in Porto Alegre are related to both human and plant genotoxicity. Areas with different concentration of pollutants demonstrated to have an urbanization gradient dependent pattern which also reflected on genotoxic damage among these areas.

  16. Kif3a regulates planar polarization of auditory hair cells through both ciliary and non-ciliary mechanisms

    Science.gov (United States)

    Sipe, Conor W.; Lu, Xiaowei

    2011-01-01

    Auditory hair cells represent one of the most prominent examples of epithelial planar polarity. In the auditory sensory epithelium, planar polarity of individual hair cells is defined by their V-shaped hair bundle, the mechanotransduction organelle located on the apical surface. At the tissue level, all hair cells display uniform planar polarity across the epithelium. Although it is known that tissue planar polarity is controlled by non-canonical Wnt/planar cell polarity (PCP) signaling, the hair cell-intrinsic polarity machinery that establishes the V-shape of the hair bundle is poorly understood. Here, we show that the microtubule motor subunit Kif3a regulates hair cell polarization through both ciliary and non-ciliary mechanisms. Disruption of Kif3a in the inner ear led to absence of the kinocilium, a shortened cochlear duct and flattened hair bundle morphology. Moreover, basal bodies are mispositioned along both the apicobasal and planar polarity axes of mutant hair cells, and hair bundle orientation was uncoupled from the basal body position. We show that a non-ciliary function of Kif3a regulates localized cortical activity of p21-activated kinases (PAK), which in turn controls basal body positioning in hair cells. Our results demonstrate that Kif3a-PAK signaling coordinates planar polarization of the hair bundle and the basal body in hair cells, and establish Kif3a as a key component of the hair cell-intrinsic polarity machinery, which acts in concert with the tissue polarity pathway. PMID:21752934

  17. Plasma membrane protein polarity and trafficking in RPE cells: Past, present and future

    Science.gov (United States)

    Lehmann, Guillermo L.; Benedicto, Ignacio; Philp, Nancy J.; Rodriguez-Boulan, Enrique

    2015-01-01

    The retinal pigment epithelium (RPE) comprises a monolayer of polarized pigmented epithelial cells that is strategically interposed between the neural retina and the fenestrated choroid capillaries. The RPE performs a variety of vectorial transport functions (water, ions, metabolites, nutrients and waste products) that regulate the composition of the subretinal space and support the functions of photoreceptors (PRs) and other cells in the neural retina. To this end, RPE cells display a polarized distribution of channels, transporters and receptors in their plasma membrane (PM) that is remarkably different from that found in conventional extra-ocular epithelia, e.g. intestine, kidney, and gall bladder. This characteristic PM protein polarity of RPE cells depends on the interplay of sorting signals in the RPE PM proteins and sorting mechanisms and biosynthetic/recycling trafficking routes in the RPE cell. Although considerable progress has been made in our understanding of the RPE trafficking machinery, most available data have been obtained from immortalized RPE cell lines that only partially maintain the RPE phenotype and by extrapolation of data obtained in the prototype Madin–Darby Canine Kidney (MDCK) cell line. The increasing availability of RPE cell cultures that more closely resemble the RPE in vivo together with the advent of advanced live imaging microscopy techniques provides a platform and an opportunity to rapidly expand our understanding of how polarized protein trafficking contributes to RPE PM polarity. PMID:25152359

  18. n3 PUFAs reduce mouse CD4+ T-cell ex vivo polarization into Th17 cells.

    Science.gov (United States)

    Monk, Jennifer M; Hou, Tim Y; Turk, Harmony F; McMurray, David N; Chapkin, Robert S

    2013-09-01

    Little is known about the impact of n3 (ω3) PUFAs on polarization of CD4(+) T cells into effector subsets other than Th1 and Th2. We assessed the effects of dietary fat [corn oil (CO) vs. fish oil (FO)] and fermentable fiber [cellulose (C) vs. pectin (P)] (2 × 2 design) in male C57BL/6 mice fed CO-C, CO-P, FO-C, or FO-P diets for 3 wk on the ex vivo polarization of purified splenic CD4(+) T cells (using magnetic microbeads) into regulatory T cells [Tregs; forkhead box P3 (Foxp3(+)) cells] or Th17 cells [interleukin (IL)-17A(+) and retinoic acid receptor-related orphan receptor (ROR) γτ(+) cells] by flow cytometry. Treg polarization was unaffected by diet; however, FO independently reduced the percentage of both CD4(+) IL-17A(+) (P diets enriched in eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), or DHA + EPA similarly reduced Th17-cell polarization in comparison to CO by reducing expression of the Th17-cell signature cytokine (IL-17A; P = 0.0015) and transcription factor (RORγτ P = 0.02), whereas Treg polarization was unaffected. Collectively, these data show that n3 PUFAs exert a direct effect on the development of Th17 cells in healthy mice, implicating a novel n3 PUFA-dependent, anti-inflammatory mechanism of action via the suppression of the initial development of this inflammatory T-cell subset.

  19. Osmotically induced cell swelling versus cell shrinking elicits specific changes in phospholipid signals in tobacco pollen tubes

    Czech Academy of Sciences Publication Activity Database

    Zonia, Laura; Munnik, T.

    2004-01-01

    Roč. 134, č. 2 (2004), s. 813-823 ISSN 0032-0889 R&D Projects: GA MŠk LN00A081 Institutional research plan: CEZ:AV0Z5038910 Keywords : ARABIDOPSIS-THALIANA CELLS * DEPENDENT PROTEIN-KINASE * SPIRODELA-POLYRHIZA L. Subject RIV: EF - Botanics Impact factor: 5.881, year: 2004

  20. The morphology of pollen presenter and polymorphism of pollen grains Taraxacum officinale F. H. Wigg.

    Directory of Open Access Journals (Sweden)

    Elżbieta Weryszko-Chmielewska

    2012-12-01

    Full Text Available The study of the structure of the pollen presenter of Taraxacum officinale and morphology of pollen grains was conducted based on plant material sampled from three different sites. One of them was a forest meadow situated away from the city, and the other two were located in the city centre of Lublin. Light and electron scanning microscopy were used in the study. The pollen presenter in Taraxacum officinale occurs at the upper part of the style situated over the androecium and on the outer part of the stigma. Numerous unicellular trichomes are found on the entire surface of the epidermis of the presenter. The function of the presenter consists in transferring pollen grains above the androecium and corolla petals. Its activity does not stop after pollen release from anthers. Taraxacum pollen grains represent the Crepis - type. Most frequently, they are tricolporate, radially symmetric and isopolar. In terms of the size, they are included in medium-sized grains. In the material examined, many deformed and asymmetric grains were observed, though they were marked by high viability at the level of 96.5-99%. Grains with the largest average lengths of the equatorial and polar axes were found in plants sampled from the meadow situated out of town. In the plant material from all sites, grains with disturbances of the external structure occurred.

  1. Differential effects of Mycobacterium bovis - derived polar and apolar lipid fractions on bovine innate immune cells

    Directory of Open Access Journals (Sweden)

    Pirson Chris

    2012-06-01

    Full Text Available Abstract Mycobacterial lipids have long been known to modulate the function of a variety of cells of the innate immune system. Here, we report the extraction and characterisation of polar and apolar free lipids from Mycobacterium bovis AF 2122/97 and identify the major lipids present in these fractions. Lipids found included trehalose dimycolate (TDM and trehalose monomycolate (TMM, the apolar phthiocerol dimycocersates (PDIMs, triacyl glycerol (TAG, pentacyl trehalose (PAT, phenolic glycolipid (PGL, and mono-mycolyl glycerol (MMG. Polar lipids identified included glucose monomycolate (GMM, diphosphatidyl glycerol (DPG, phenylethanolamine (PE and a range of mono- and di-acylated phosphatidyl inositol mannosides (PIMs. These lipid fractions are capable of altering the cytokine profile produced by fresh and cultured bovine monocytes as well as monocyte derived dendritic cells. Significant increases in the production of IL-10, IL-12, MIP-1β, TNFα and IL-6 were seen after exposure of antigen presenting cells to the polar lipid fraction. Phenotypic characterisation of the cells was performed by flow cytometry and significant decreases in the expression of MHCII, CD86 and CD1b were found after exposure to the polar lipid fraction. Polar lipids also significantly increased the levels of CD40 expressed by monocytes and cultured monocytes but no effect was seen on the constitutively high expression of CD40 on MDDC or on the levels of CD80 expressed by any of the cells. Finally, the capacity of polar fraction treated cells to stimulate alloreactive lymphocytes was assessed. Significant reduction in proliferative activity was seen after stimulation of PBMC by polar fraction treated cultured monocytes whilst no effect was seen after lipid treatment of MDDC. These data demonstrate that pathogenic mycobacterial polar lipids may significantly hamper the ability of the host APCs to induce an appropriate immune response to an invading pathogen.

  2. The keratin-binding protein Albatross regulates polarization of epithelial cells.

    Science.gov (United States)

    Sugimoto, Masahiko; Inoko, Akihito; Shiromizu, Takashi; Nakayama, Masanori; Zou, Peng; Yonemura, Shigenobu; Hayashi, Yuko; Izawa, Ichiro; Sasoh, Mikio; Uji, Yukitaka; Kaibuchi, Kozo; Kiyono, Tohru; Inagaki, Masaki

    2008-10-06

    The keratin intermediate filament network is abundant in epithelial cells, but its function in the establishment and maintenance of cell polarity is unclear. Here, we show that Albatross complexes with Par3 to regulate formation of the apical junctional complex (AJC) and maintain lateral membrane identity. In nonpolarized epithelial cells, Albatross localizes with keratin filaments, whereas in polarized epithelial cells, Albatross is primarily localized in the vicinity of the AJC. Knockdown of Albatross in polarized cells causes a disappearance of key components of the AJC at cell-cell borders and keratin filament reorganization. Lateral proteins E-cadherin and desmoglein 2 were mislocalized even on the apical side. Although Albatross promotes localization of Par3 to the AJC, Par3 and ezrin are still retained at the apical surface in Albatross knockdown cells, which retain intact microvilli. Analysis of keratin-deficient epithelial cells revealed that keratins are required to stabilize the Albatross protein, thus promoting the formation of AJC. We propose that keratins and the keratin-binding protein Albatross are important for epithelial cell polarization.

  3. Mammalian aPKC/Par polarity complex mediated regulation of epithelial division orientation and cell fate

    Energy Technology Data Exchange (ETDEWEB)

    Vorhagen, Susanne; Niessen, Carien M., E-mail: carien.niessen@uni-koeln.de

    2014-11-01

    Oriented cell division is a key regulator of tissue architecture and crucial for morphogenesis and homeostasis. Balanced regulation of proliferation and differentiation is an essential property of tissues not only to drive morphogenesis but also to maintain and restore homeostasis. In many tissues orientation of cell division is coupled to the regulation of differentiation producing daughters with similar (symmetric cell division, SCD) or differential fate (asymmetric cell division, ACD). This allows the organism to generate cell lineage diversity from a small pool of stem and progenitor cells. Division orientation and/or the ratio of ACD/SCD need to be tightly controlled. Loss of orientation or an altered ratio can promote overgrowth, alter tissue architecture and induce aberrant differentiation, and have been linked to morphogenetic diseases, cancer and aging. A key requirement for oriented division is the presence of a polarity axis, which can be established through cell intrinsic and/or extrinsic signals. Polarity proteins translate such internal and external cues to drive polarization. In this review we will focus on the role of the polarity complex aPKC/Par3/Par6 in the regulation of division orientation and cell fate in different mammalian epithelia. We will compare the conserved function of this complex in mitotic spindle orientation and distribution of cell fate determinants and highlight common and differential mechanisms in which this complex is used by tissues to adapt division orientation and cell fate to the specific properties of the epithelium.

  4. Cell polarity and neurogenesis in embryonic stem cell-derived neural rosettes.

    Science.gov (United States)

    Banda, Erin; McKinsey, Anna; Germain, Noelle; Carter, James; Anderson, Nickesha Camille; Grabel, Laura

    2015-04-15

    Embryonic stem cells (ESCs) undergoing neural differentiation form radial arrays of neural stem cells, termed neural rosettes. These structures manifest many of the properties associated with embryonic and adult neurogenesis, including cell polarization, interkinetic nuclear migration (INM), and a gradient of neuronal differentiation. We now identify novel rosette structural features that serve to localize key regulators of neurogenesis. Cells within neural rosettes have specialized basal as well as apical surfaces, based on localization of the extracellular matrix receptor β1 integrin. Apical processes of cells in mature rosettes terminate at the lumen, where adherens junctions are apparent. Primary cilia are randomly distributed in immature rosettes and tightly associated with the neural stem cell's apical domain as rosettes mature. Components of two signaling pathways known to regulate neurogenesis in vivo and in rosettes, Hedgehog and Notch, are apically localized, with the Hedgehog effector Smoothened (Smo) associated with primary cilia and the Notch pathway γ-secretase subunit Presenilin 2 associated with the adherens junction. Increased neuron production upon treatment with the Notch inhibitor DAPT suggests a major role for Notch signaling in maintaining the neural stem cell state, as previously described. A less robust outcome was observed with manipulation of Hedgehog levels, though consistent with a role in neural stem cell survival or proliferation. Inhibition of both pathways resulted in an additive effect. These data support a model by which cells extending a process to the rosette lumen maintain neural stem cell identity whereas release from this association, either through asymmetric cell division or apical abscission, promotes neuronal differentiation.

  5. Calcium gradients in conifer pollen tubes; dynamic properties differ from those seen in angiosperms.

    Science.gov (United States)

    Lazzaro, Mark D; Cardenas, Luis; Bhatt, Aadra P; Justus, Charles D; Phillips, Monique S; Holdaway-Clarke, Terena L; Hepler, Peter K

    2005-10-01

    Pollen tubes are an established model system for examining polarized cell growth. The focus here is on pollen tubes of the conifer Norway spruce (Picea abies, Pinaceae); examining the relationship between cytosolic free Ca2+, tip elongation, and intracellular motility. Conifer pollen tubes show important differences from their angiosperm counterparts; they grow more slowly and their organelles move in an unusual fountain pattern, as opposed to reverse fountain, in the tip. Ratiometric ion imaging of growing pollen tubes, microinjected with fura-2-dextran, reveals a tip-focused [Ca2+]i gradient extending from 450 nM at the extreme apex to 225 nM at the base of the tip clear zone. Injection of 5,5' dibromo-BAPTA does not dissipate the apical gradient, but stops cell elongation and uniquely causes rapid, transient increases of apical free Ca2+. The [Ca2+]i gradient is, however, dissipated by reversible perfusion of extracellular caffeine. When the basal cytosolic free Ca2+ concentration falls below 150 nM, again a large increase in apical [Ca2+]i occurs. An external source of calcium is not required for germination but significantly enhances elongation. However, both germination and elongation are significantly inhibited by the inclusion of calcium channels blockers, including lanthanum, gadolinium, or verapamil. Modulation of intracellular calcium also affects organelle position and motility. Extracellular perfusion of lanthanides reversibly depletes the apical [Ca2+]i gradient, altering organelle positioning in the tip. Later, during recovery from lanthanide perfusion, organelle motility switches direction to a reverse fountain. When taken together these data show a unique interplay in Picea abies pollen tubes between intracellular calcium and the motile processes controlling cellular organization.

  6. Origin and Functional Prediction of Pollen Allergens in Plants.

    Science.gov (United States)

    Chen, Miaolin; Xu, Jie; Devis, Deborah; Shi, Jianxin; Ren, Kang; Searle, Iain; Zhang, Dabing

    2016-09-01

    Pollen allergies have long been a major pandemic health problem for human. However, the evolutionary events and biological function of pollen allergens in plants remain largely unknown. Here, we report the genome-wide prediction of pollen allergens and their biological function in the dicotyledonous model plant Arabidopsis (Arabidopsis thaliana) and the monocotyledonous model plant rice (Oryza sativa). In total, 145 and 107 pollen allergens were predicted from rice and Arabidopsis, respectively. These pollen allergens are putatively involved in stress responses and metabolic processes such as cell wall metabolism during pollen development. Interestingly, these putative pollen allergen genes were derived from large gene families and became diversified during evolution. Sequence analysis across 25 plant species from green alga to angiosperms suggest that about 40% of putative pollen allergenic proteins existed in both lower and higher plants, while other allergens emerged during evolution. Although a high proportion of gene duplication has been observed among allergen-coding genes, our data show that these genes might have undergone purifying selection during evolution. We also observed that epitopes of an allergen might have a biological function, as revealed by comprehensive analysis of two known allergens, expansin and profilin. This implies a crucial role of conserved amino acid residues in both in planta biological function and allergenicity. Finally, a model explaining how pollen allergens were generated and maintained in plants is proposed. Prediction and systematic analysis of pollen allergens in model plants suggest that pollen allergens were evolved by gene duplication and then functional specification. This study provides insight into the phylogenetic and evolutionary scenario of pollen allergens that will be helpful to future characterization and epitope screening of pollen allergens. © 2016 American Society of Plant Biologists. All rights reserved.

  7. Expanding signaling-molecule wavefront model of cell polarization in the Drosophila wing primordium.

    Science.gov (United States)

    Wortman, Juliana C; Nahmad, Marcos; Zhang, Peng Cheng; Lander, Arthur D; Yu, Clare C

    2017-07-01

    In developing tissues, cell polarization and proliferation are regulated by morphogens and signaling pathways. Cells throughout the Drosophila wing primordium typically show subcellular localization of the unconventional myosin Dachs on the distal side of cells (nearest the center of the disc). Dachs localization depends on the spatial distribution of bonds between the protocadherins Fat (Ft) and Dachsous (Ds), which form heterodimers between adjacent cells; and the Golgi kinase Four-jointed (Fj), which affects the binding affinities of Ft and Ds. The Fj concentration forms a linear gradient while the Ds concentration is roughly uniform throughout most of the wing pouch with a steep transition region that propagates from the center to the edge of the pouch during the third larval instar. Although the Fj gradient is an important cue for polarization, it is unclear how the polarization is affected by cell division and the expanding Ds transition region, both of which can alter the distribution of Ft-Ds heterodimers around the cell periphery. We have developed a computational model to address these questions. In our model, the binding affinity of Ft and Ds depends on phosphorylation by Fj. We assume that the asymmetry of the Ft-Ds bond distribution around the cell periphery defines the polarization, with greater asymmetry promoting cell proliferation. Our model predicts that this asymmetry is greatest in the radially-expanding transition region that leaves polarized cells in its wake. These cells naturally retain their bond distribution asymmetry after division by rapidly replenishing Ft-Ds bonds at new cell-cell interfaces. Thus we predict that the distal localization of Dachs in cells throughout the pouch requires the movement of the Ds transition region and the simple presence, rather than any specific spatial pattern, of Fj.

  8. Dystrophin expression in muscle stem cells regulates their polarity and asymmetric division.

    Science.gov (United States)

    Dumont, Nicolas A; Wang, Yu Xin; von Maltzahn, Julia; Pasut, Alessandra; Bentzinger, C Florian; Brun, Caroline E; Rudnicki, Michael A

    2015-12-01

    Dystrophin is expressed in differentiated myofibers, in which it is required for sarcolemmal integrity, and loss-of-function mutations in the gene that encodes it result in Duchenne muscular dystrophy (DMD), a disease characterized by progressive and severe skeletal muscle degeneration. Here we found that dystrophin is also highly expressed in activated muscle stem cells (also known as satellite cells), in which it associates with the serine-threonine kinase Mark2 (also known as Par1b), an important regulator of cell polarity. In the absence of dystrophin, expression of Mark2 protein is downregulated, resulting in the inability to localize the cell polarity regulator Pard3 to the opposite side of the cell. Consequently, the number of asymmetric divisions is strikingly reduced in dystrophin-deficient satellite cells, which also display a loss of polarity, abnormal division patterns (including centrosome amplification), impaired mitotic spindle orientation and prolonged cell divisions. Altogether, these intrinsic defects strongly reduce the generation of myogenic progenitors that are needed for proper muscle regeneration. Therefore, we conclude that dystrophin has an essential role in the regulation of satellite cell polarity and asymmetric division. Our findings indicate that muscle wasting in DMD not only is caused by myofiber fragility, but also is exacerbated by impaired regeneration owing to intrinsic satellite cell dysfunction.

  9. Regulation of vascular endothelial cell polarization and migration by Hsp70/Hsp90-organizing protein.

    Science.gov (United States)

    Li, Jingyu; Sun, Xiaodong; Wang, Zaizhu; Chen, Li; Li, Dengwen; Zhou, Jun; Liu, Min

    2012-01-01

    Hsp70/Hsp90-organizing protein (HOP) is a member of the co-chaperone family, which directly binds to chaperones to regulate their activities. The participation of HOP in cell motility and endothelial cell functions remains largely unknown. In this study, we demonstrate that HOP is critically involved in endothelial cell migration and angiogenesis. Tube formation and capillary sprouting experiments reveal that depletion of HOP expression significantly inhibits vessel formation from endothelial cells. Wound healing and transwell migration assays show that HOP is important for endothelial cell migration. By examination of centrosome reorientation and membrane ruffle dynamics, we find that HOP plays a crucial role in the establishment of cell polarity in response to migratory stimulus. Furthermore, our data show that HOP interacts with tubulin and colocalizes with microtubules in endothelial cells. These findings indicate HOP as a novel regulator of angiogenesis that functions through promoting vascular endothelial cell polarization and migration.

  10. Intracellular trafficking and PIN-mediated cell polarity during tropic responses in plants.

    Science.gov (United States)

    Rakusová, Hana; Fendrych, Matyáš; Friml, Jiří

    2015-02-01

    Subcellular trafficking and cell polarity are basic cellular processes crucial for plant development including tropisms - directional growth responses to environmental stimuli such as light or gravity. Tropisms involve auxin gradient across the stimulated organ that underlies the differential cell elongation and bending. The perception of light or gravity is followed by changes in the polar, cellular distribution of the PIN auxin transporters. Such re-specification of polar trafficking pathways is a part of the mechanism, by which plants adjust their phenotype to environmental changes. Recent genetic and biochemical studies provided the important insights into mechanisms of PIN polarization during tropisms. In this review, we summarize the present state of knowledge on dynamic PIN repolarization and its specific regulations during hypocotyl tropisms. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. Tracking maize pollen development by the Leaf Collar Method.

    Science.gov (United States)

    Begcy, Kevin; Dresselhaus, Thomas

    2017-12-01

    An easy and highly reproducible nondestructive method named the Leaf Collar Method is described to identify and characterize the different stages of pollen development in maize. In plants, many cellular events such as meiosis, asymmetric cell division, cell cycle regulation, cell fate determination, nucleus movement, vacuole formation, chromatin condensation and epigenetic modifications take place during pollen development. In maize, pollen development occurs in tassels that are confined within the internal stalk of the plant. Hence, identification of the different pollen developmental stages as a tool to investigate above biological processes is impossible without dissecting the entire plant. Therefore, an efficient and reproducible method is necessary to isolate homogeneous cell populations at individual stages throughout pollen development without destroying the plant. Here, we describe a method to identify the various stages of pollen development in maize. Using the Leaf Collar Method in the maize inbreed line B73, we have determined the duration of each stage from pollen mother cells before meiosis to mature tricellular pollen. Anther and tassel size as well as percentage of pollen stages were correlated with vegetative stages, which are easily recognized. The identification of stage-specific genes indicates the reproducibility of the method. In summary, we present an easy and highly reproducible nondestructive method to identify and characterize the different stages of pollen development in maize. This method now opens the way for many subsequent physiological, morphological and molecular analyses to study, for instance, transcriptomics, metabolomics, DNA methylation and chromatin patterns during normal and stressful conditions throughout pollen development in one of the economically most important grass species.

  12. Effect of III-nitride polarization on V{sub OC} in p-i-n and MQW solar cells

    Energy Technology Data Exchange (ETDEWEB)

    Namkoong, Gon; Boland, Patrick; Foe, Kurniawan; Latimer, Kevin [Department of Electrical and Computer Engineering, Old Dominion University, Applied Research Center, 12050 Jefferson Avenue, Newport News, VA 23606 (United States); Bae, Si-Young; Shim, Jae-Phil; Lee, Dong-Seon [School of Information and Communications, Gwangju Institute of Science and Technology, 261 Cheomdan-gwagiro (Oryong-dong), Buk-gu, Gwangju 500-712 (Korea, Republic of); Jeon, Seong-Ran [Korea Photonics Technology Institute, 971-35, Wolchul-dong, Buk-gu, Gwangju, 500-779 (Korea, Republic of); Doolittle, W. Alan [School of Electrical and Computer Engineering, Georgia Institute of Technology, Atlanta, GA 30332 (United States)

    2011-02-15

    We performed detailed studies of the effect of polarization on III-nitride solar cells. Spontaneous and piezoelectric polarizations were assessed to determine their impacts upon the open circuit voltages (V{sub OC}) in p-i(InGaN)-n and multi-quantum well (MQW) solar cells. We found that the spontaneous polarization in Ga-polar p-i-n solar cells strongly modifies energy band structures and corresponding electric fields in a way that degrades V{sub OC} compared to non-polar p-i-n structures. In contrast, we found that piezoelectric polarization in Ga-polar MQW structures does not have a large influence on V{sub OC} compared to non-polar MQW structures. (copyright 2011 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

  13. The polarity protein Par1b/EMK/MARK2 regulates T cell receptor-induced microtubule-organizing center polarization.

    Science.gov (United States)

    Lin, Joseph; Hou, Kirk K; Piwnica-Worms, Helen; Shaw, Andrey S

    2009-07-15

    Engagement of a T cell to an APC induces the formation of an immunological synapse as well as reorientation of the microtubule-organizing center (MTOC) toward the APC. How signals emanating from the TCR induce MTOC polarization is not known. One group of proteins known to play a critical role in asymmetric cell division and cell polarization is the partitioning defective (Par) family of proteins. In this study we found that Par1b, a member of the Par family of proteins, was inducibly phosphorylated following TCR stimulation. This phosphorylation resulted in 14-3-3 protein binding and caused the relocalization of Par1b from the membrane into the cytoplasm. Because a dominant-negative form of Par1b blocked TCR-induced MTOC polarization, our data suggest that Par1b functions in the establishment of T cell polarity following engagement to an APC.

  14. How honey bees carry pollen

    Science.gov (United States)

    Matherne, Marguerite E.; Anyanwu, Gabriel; Leavey, Jennifer K.; Hu, David L.

    2017-11-01

    Honey bees are the tanker of the skies, carrying thirty percent of their weight in pollen per foraging trip using specialized orifices on their body. How do they manage to hang onto those pesky pollen grains? In this experimental study, we investigate the adhesion force of pollen to the honeybee. To affix pollen to themselves, honey bees form a suspension of pollen in nectar, creating a putty-like pollen basket that is skewered by leg hairs. We use tensile tests to show that the viscous force of the pollen basket is more than ten times the honeybee's flight force. This work may provide inspiration for the design of robotic flying pollinators.

  15. Effects of Yamase climatic condition during the pollen mother cell developmental stage on concentrations of Cs and Sr in brown rice

    International Nuclear Information System (INIS)

    Yanai, Masumi; Kobayashi, Daisuke; Yamagami, Mutsumi; Takaku, Yuichi; Hisamatsu, Shun'ichi

    2007-01-01

    Effects of the Yamase climatic condition on the concentration of alkali metals and alkaline earth metals in brown rice were examined in relation to the soil-to-plant transfer factors of 137 Cs and 90 Sr. Rice plants (Oryza sativa L cv. Yumeakari) were cultivated in an artificial climate chamber, and exposed to a simulated Yamase condition for 3 or 7 d during the pollen mother cell developmental stage. In these simulated treatments, temperature and light intensity were set to 5degC lower and 50% lower than the respective control values. Fog was generated with visibility of 70±30 m. Concentrations of alkali metals, alkaline earth metals and the other minor elements in brown rice samples were analyzed. Measured Cs and Sr concentrations were found to be unaffected by the Yamase treatments, while concentrations of Fe, Cu, Zn and Mo were increased. (author)

  16. Evidence for Nuclear Tensor Polarization of Deuterium Molecules in Storage Cells

    International Nuclear Information System (INIS)

    van den Brand, J.; Bulten, H.; Zhou, Z.; Unal, O.; van den Brand, J.; Ferro-Luzzi, M.; Botto, T.; Bouwhuis, M.; Heimberg, P.; de Jager, C.; de Lange, D.; Nooren, G.; Papadakis, N.; Passchier, I.; Poolman, H.; Steijger, J.; Vodinas, N.; de Vries, H.; van den Brand, J.; Ferro-Luzzi, M.; Lang, J.; Alarcon, R.; Dolfini, S.; Ent, R.; Higinbotham, D.

    1997-01-01

    Deuterium molecules were obtained by recombination, on a copper surface, of deuterium atoms prepared in specific hyperfine states. The molecules were stored for about 5ms in an open-ended cylindrical cell, placed in a 23mT magnetic field, and their tensor polarization was measured by elastic scattering of 704MeV electrons. The results of the measurements are consistent with the deuterium molecules retaining the tensor polarization of the initial atoms. copyright 1997 The American Physical Society

  17. Tropospheric entrainment as a source of ground level aerosols within the polar Antarctic cell

    Science.gov (United States)

    Humphries, R. S.; Schofield, R.; Keywood, M.; Wilson, S. R.; Klekociuk, A. R.; Paton-Walsh, C.

    2013-12-01

    The Antarctic region is a pristine environment without any significant anthropogenic influence. Measurements of aerosols in this environment therefore allow the study of natural aerosol properties and formation mechanisms in polar conditions, and also allow insight into polar atmospheric dynamics. Measurements in this region have been limited primarily to continental and coastal locations where permanent stations exist, with only one other measurement campaign passing through the sea ice region. The MAPS campaign (Measurements of Aerosols and Precursors during SIPEXII) occurred as part of SIPEX II (Sea Ice Physics and Ecosystems eXperiment II) voyage in Spring, 2012, and produced the first sea-ice focused aerosol dataset aimed at characterizing new particle formation processes in the pack ice off the coast of East Antarctica (~65°S, 120°E). Numerous atmospheric parameters and species were measured, including the number of aerosol particles in the 3-10 nm size range, the range associated with new particle formation. During the latitudinal transect through the sea ice, these measurements were used to identify the polar front - the boundary between the Polar cell and the Ferrel cell. Nuclei concentrations showed a clear and sudden change with latitude, averaging 51cm-3 north of the front in the Ferrel cell, and 766 cm-3 south of the front, in the Polar cell region. The latitudinal location of the polar front was also confirmed by wind directions which reflected global circulation patterns (Ferrel cell westerlies and Polar cell easterlies). Background aerosol populations in the Polar cell fluctuated significantly (3-10 nm particle concentrations ranged between 153 cm-3 to 2312 cm-3) but displayed no growth indicators, suggesting transport. Back-trajectories revealed that air parcels often descended from the free-troposphere within the previous 24-48 hrs. It is proposed that particle formation occurs in the free troposphere from precursors uplifted at the polar front

  18. Optically-driven red blood cell rotor in linearly polarized laser tweezers

    Indian Academy of Sciences (India)

    We have constructed a dual trap optical tweezers set-up around an inverted microscope where both the traps can be independently controlled and manipulated in all the three dimensions. Here we report our observations on rotation of red blood cells (RBCs) in a linearly polarized optical trap. Red blood cells deform and ...

  19. Barley disease susceptibility factor RACB acts in epidermal cell polarity and positioning of the nucleus.

    Science.gov (United States)

    Scheler, Björn; Schnepf, Vera; Galgenmüller, Carolina; Ranf, Stefanie; Hückelhoven, Ralph

    2016-05-01

    RHO GTPases are regulators of cell polarity and immunity in eukaryotes. In plants, RHO-like RAC/ROP GTPases are regulators of cell shaping, hormone responses, and responses to microbial pathogens. The barley (Hordeum vulgare L.) RAC/ROP protein RACB is required for full susceptibility to penetration by Blumeria graminis f.sp. hordei (Bgh), the barley powdery mildew fungus. Disease susceptibility factors often control host immune responses. Here we show that RACB does not interfere with early microbe-associated molecular pattern-triggered immune responses such as the oxidative burst or activation of mitogen-activated protein kinases. RACB also supports rather than restricts expression of defence-related genes in barley. Instead, silencing of RACB expression by RNAi leads to defects in cell polarity. In particular, initiation and maintenance of root hair growth and development of stomatal subsidiary cells by asymmetric cell division is affected by silencing expression of RACB. Nucleus migration is a common factor of developmental cell polarity and cell-autonomous interaction with Bgh RACB is required for positioning of the nucleus near the site of attack from Bgh We therefore suggest that Bgh profits from RACB's function in cell polarity rather than from immunity-regulating functions of RACB. © The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  20. Hugl1 and Hugl2 in mammary epithelial cells: polarity, proliferation, and differentiation.

    Directory of Open Access Journals (Sweden)

    Atlantis Russ

    Full Text Available Loss of epithelial polarity is described as a hallmark of epithelial cancer. To determine the role of Hugl1 and Hugl2 expression in the breast, we investigated their localization in human mammary duct tissue and the effects of expression modulation in normal and cancer cell lines on polarity, proliferation and differentiation. Expression of Hugl1 and Hugl2 was silenced in both MCF10A cells and Human Mammary Epithelial Cells and cell lines were grown in 2-D on plastic and in 3-D in Matrigel to form acini. Cells in monolayer were compared for proliferative and phenotypic changes while acini were examined for differences in size, ability to form a hollow lumen, nuclear size and shape, and localization of key domain-specific proteins as a measure of polarity. We detected overlapping but distinct localization of Hugl1 and Hugl2 in the human mammary gland, with Hugl1 expressed in both luminal and myoepithelium and Hugl2 largely restricted to myoepithelium. On a plastic surface, loss of Hugl1 or Hugl2 in normal epithelium induced a mesenchymal phenotype, and these cells formed large cellular masses when grown in Matrigel. In addition, loss of Hugl1 or Hugl2 expression in MCF10A cells resulted in increased proliferation on Matrigel, while gain of Hugl1 expression in tumor cells suppressed proliferation. Loss of polarity was also observed with knockdown of either Hugl1 or Hugl2, with cells growing in Matrigel appearing as a multilayered epithelium, with randomly oriented Golgi and multiple enlarged nuclei. Furthermore, Hugl1 knock down resulted in a loss of membrane identity and the development of cellular asymmetries in Human Mammary Epithelial Cells. Overall, these data demonstrate an essential role for both Hugl1 and Hugl2 in the maintenance of breast epithelial polarity and differentiated cell morphology, as well as growth control.

  1. Iron repletion relocalizes hephaestin to a proximal basolateral compartment in polarized MDCK and Caco2 cells

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Seung-Min [Department of Biological Sciences, University of Columbia, NY (United States); Department of Nutritional Science and Toxicology, University of California, Berkeley, CA (United States); Attieh, Zouhair K. [Department of Laboratory Science and Technology, American University of Science and Technology, Ashrafieh (Lebanon); Department of Nutritional Science and Toxicology, University of California, Berkeley, CA (United States); Son, Hee Sook [Department of Food Science and Human Nutrition, College of Human Ecology, Chonbuk National University (Korea, Republic of); Department of Nutritional Science and Toxicology, University of California, Berkeley, CA (United States); Chen, Huijun [Medical School, Nanjing University, Nanjing 210008, Jiangsu Province (China); Department of Nutritional Science and Toxicology, University of California, Berkeley, CA (United States); Bacouri-Haidar, Mhenia [Department of Biology, Faculty of Sciences (I), Lebanese University, Hadath (Lebanon); Department of Nutritional Science and Toxicology, University of California, Berkeley, CA (United States); Vulpe, Chris D., E-mail: vulpe@berkeley.edu [Department of Nutritional Science and Toxicology, University of California, Berkeley, CA (United States)

    2012-05-11

    Highlights: Black-Right-Pointing-Pointer Hephaestin localizes in the perinuclear space in non-polarized cells. Black-Right-Pointing-Pointer Hephaestin localizes in the perinuclear space in iron deficient and polarized cells. Black-Right-Pointing-Pointer Hephaestin with apical iron moves near to basolateral membrane of polarized cells. Black-Right-Pointing-Pointer Peri-basolateral location of hephaestin is accessible to the extracellular space. Black-Right-Pointing-Pointer Hephaestin is involved in iron mobilization from the intestine to circulation. -- Abstract: While intestinal cellular iron entry in vertebrates employs multiple routes including heme and non-heme routes, iron egress from these cells is exclusively channeled through the only known transporter, ferroportin. Reduced intestinal iron export in sex-linked anemia mice implicates hephaestin, a ferroxidase, in this process. Polarized cells are exposed to two distinct environments. Enterocytes contact the gut lumen via the apical surface of the cell, and through the basolateral surface, to the body. Previous studies indicate both local and systemic control of iron uptake. We hypothesized that differences in iron availability at the apical and/or basolateral surface may modulate iron uptake via cellular localization of hephaestin. We therefore characterized the localization of hephaestin in two models of polarized epithelial cell lines, MDCK and Caco2, with varying iron availability at the apical and basolateral surfaces. Our results indicate that hephaestin is expressed in a supra-nuclear compartment in non-polarized cells regardless of the iron status of the cells and in iron deficient and polarized cells. In polarized cells, we found that both apical (as FeSO{sub 4}) and basolateral iron (as the ratio of apo-transferrin to holo-transferrin) affect mobilization of hephaestin from the supra-nuclear compartment. We find that the presence of apical iron is essential for relocalization of hephaestin to a

  2. Chromatin dynamics in Pollen Mother Cells underpin a common scenario at the somatic-to-reproductive fate transition of both the male and female lineages in Arabidopsis

    Directory of Open Access Journals (Sweden)

    Wenjing eShe

    2015-04-01

    Full Text Available Unlike animals, where the germline is established early during embryogenesis, plants set aside their reproductive lineage late in development in dedicated floral organs. The specification of pollen mother cells (PMCs committed to meiosis takes place in the sporogenous tissue in anther locules and marks the somatic-to-reproductive cell fate transition towards the male reproductive lineage. Here we show that Arabidopsis PMCs differentiation is accompanied by large-scale changes in chromatin organization. This is characterized by significant increase in nuclear volume, chromatin decondensation, reduction in heterochromatin, eviction of linker histones and the H2AZ histone variant. These structural alterations are accompanied by dramatic, quantitative changes in histone modifications levels compared to that of surrounding somatic cells that do not share a sporogenic fate. All these changes are highly reminiscent of those we have formerly described in female megaspore mother cells (MMCs. This indicates that chromatin reprogramming is a common underlying scenario in the somatic-to-reproductive cell fate transition in both male and female lineages.

  3. Monitoring the initiation and kinetics of human dendritic cell-induced polarization of autologous naive CD4+ T cells.

    Directory of Open Access Journals (Sweden)

    Tammy Oth

    Full Text Available A crucial step in generating de novo immune responses is the polarization of naive cognate CD4+ T cells by pathogen-triggered dendritic cells (DC. In the human setting, standardized DC-dependent systems are lacking to study molecular events during the initiation of a naive CD4+ T cell response. We developed a TCR-restricted assay to compare different pathogen-triggered human DC for their capacities to instruct functional differentiation of autologous, naive CD4+ T cells. We demonstrated that this methodology can be applied to compare differently matured DC in terms of kinetics, direction, and magnitude of the naive CD4+ T cell response. Furthermore, we showed the applicability of this assay to study the T cell polarizing capacity of low-frequency blood-derived DC populations directly isolated ex vivo. This methodology for addressing APC-dependent instruction of naive CD4+ T cells in a human autologous setting will provide researchers with a valuable tool to gain more insight into molecular mechanisms occurring in the early phase of T cell polarization. In addition, it may also allow the study of pharmacological agents on DC-dependent T cell polarization in the human system.

  4. Label-free investigation of the effects of lithium niobate polarization on cell adhesion

    Science.gov (United States)

    Mandracchia, B.; Gennari, O.; Paturzo, M.; Grilli, S.; Ferraro, P.

    2017-06-01

    The determination of contact area is pivotal to understand how biomaterials properties influence cell adhesion. In particular, the influence of surface charges is well-known but still controversial, especially when new functional materials and methods are introduced. Here, we use for the first time Holographic Total Internal Reflection Microscopy (HoloTIRM) to study the influence of the spontaneous polarization of ferroelectric lithium niobate (LN) on the adhesion properties of fibroblast cells. The selective illumination of a very thin region directly above the substrate, achieved by Total Internal Reflection, provides high-contrast images of the contact regions. Holographic recording, on the other hand, allows for label-free quantitative phase imaging of the contact areas between cells and LN. Phase signal is more sensitive in the first 100nm and, thus more reliable in order to locate focal contacts. This work shows that cells adhering on negatively polarized LN present a significant increase of the contact area in comparison with cells adhering on the positively polarized LN substrate, as well as an intensification of contact vicinity. This confirms the potential of LN as a platform for investigating the role of charges on cellular processes. The similarity of cell adhesion behavior on negatively polarized LN and glass control also confirms the possibility to use LN as an active substrate without impairing cell behavior.

  5. Krüppel-like factor 4 regulates intestinal epithelial cell morphology and polarity.

    Directory of Open Access Journals (Sweden)

    Tianxin Yu

    Full Text Available Krüppel-like factor 4 (KLF4 is a zinc finger transcription factor that plays a vital role in regulating cell lineage differentiation during development and maintaining epithelial homeostasis in the intestine. In normal intestine, KLF4 is predominantly expressed in the differentiated epithelial cells. It has been identified as a tumor suppressor in colorectal cancer. KLF4 knockout mice demonstrated a decrease in number of goblet cells in the colon, and conditional ablation of KLF4 from the intestinal epithelium led to altered epithelial homeostasis. However, the role of KLF4 in differentiated intestinal cells and colon cancer cells, as well as the mechanism by which it regulates homeostasis and represses tumorigenesis in the intestine is not well understood. In our study, KLF4 was partially depleted in the differentiated intestinal epithelial cells by a tamoxifen-inducible Cre recombinase. We found a significant increase in the number of goblet cells in the KLF4-deleted small intestine, suggesting that KLF4 is not only required for goblet cell differentiation, but also required for maintaining goblet cell numbers through its function in inhibiting cell proliferation. The number and position of Paneth cells also changed. This is consistent with the KLF4 knockout study using villin-Cre [1]. Through immunohistochemistry (IHC staining and statistical analysis, we found that a stem cell and/or tuft cell marker, DCAMKL1, and a proliferation marker, Ki67, are affected by KLF4 depletion, while an enteroendocrine cell marker, neurotensin (NT, was not affected. In addition, we found KLF4 depletion altered the morphology and polarity of the intestinal epithelial cells. Using a three-dimensional (3D intestinal epithelial cyst formation assay, we found that KLF4 is essential for cell polarity and crypt-cyst formation in human colon cancer cells. These findings suggest that, as a tumor suppressor in colorectal cancer, KLF4 affects intestinal epithelial cell

  6. Effect of atomic noise on optical squeezing via polarization self-rotation in a thermal vapor cell

    DEFF Research Database (Denmark)

    Hsu, M.T.L.; Hetet, G.; Peng, A.

    2006-01-01

    The traversal of an elliptically polarized optical field through a thermal vapor cell can give rise to a rotation of its polarization axis. This process, known as polarization self-rotation (PSR), has been suggested as a mechanism for producing squeezed light at atomic transition wavelengths. We...

  7. Allergies, asthma, and pollen

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/patientinstructions/000489.htm Allergies, asthma, and pollen To use the sharing features ... this page, please enable JavaScript. Things that make allergies or asthma worse are called triggers. It is ...

  8. PrPC Undergoes Basal to Apical Transcytosis in Polarized Epithelial MDCK Cells.

    Directory of Open Access Journals (Sweden)

    Alexander Arkhipenko

    Full Text Available The Prion Protein (PrP is an ubiquitously expressed glycosylated membrane protein attached to the external leaflet of the plasma membrane via a glycosylphosphatidylinositol anchor (GPI. While the misfolded PrPSc scrapie isoform is the infectious agent of prion disease, the cellular isoform (PrPC is an enigmatic protein with unclear function. Of interest, PrP localization in polarized MDCK cells is controversial and its mechanism of trafficking is not clear. Here we investigated PrP traffic in MDCK cells polarized on filters and in three-dimensional MDCK cysts, a more physiological model of polarized epithelia. We found that, unlike other GPI-anchored proteins (GPI-APs, PrP undergoes basolateral-to-apical transcytosis in fully polarized MDCK cells. Following this event full-length PrP and its cleavage fragments are segregated in different domains of the plasma membrane in polarized cells in both 2D and 3D cultures.

  9. Lactobacilli activate human dendritic cells that skew T cells toward T helper 1 polarization.

    Science.gov (United States)

    Mohamadzadeh, Mansour; Olson, Scott; Kalina, Warren V; Ruthel, Gordon; Demmin, Gretchen L; Warfield, Kelly L; Bavari, Sina; Klaenhammer, Todd R

    2005-02-22

    Professional antigen-presenting dendritic cells (DCs) are critical in regulating T cell immune responses at both systemic and mucosal sites. Many Lactobacillus species are normal members of the human gut microflora and most are regarded as safe when administered as probiotics. Because DCs can naturally or therapeutically encounter lactobacilli, we investigated the effects of several well defined strains, representing three species of Lactobacillus on human myeloid DCs (MDCs) and found that they modulated the phenotype and functions of human MDCs. Lactobacillus-exposed MDCs up-regulated HLA-DR, CD83, CD40, CD80, and CD86 and secreted high levels of IL-12 and IL-18, but not IL-10. IL-12 was sustained in MDCs exposed to all three Lactobacillus species in the presence of LPS from Escherichia coli, whereas LPS-induced IL-10 was greatly inhibited. MDCs activated with lactobacilli clearly skewed CD4(+) and CD8(+) T cells to T helper 1 and Tc1 polarization, as evidenced by secretion of IFN-gamma, but not IL-4 or IL-13. These results emphasize a potentially important role for lactobacilli in modulating immunological functions of DCs and suggest that certain strains could be particularly advantageous as vaccine adjuvants, by promoting DCs to regulate T cell responses toward T helper 1 and Tc1 pathways.

  10. Overexpression of the tomato pollen receptor kinase LePRK1 rewires pollen tube growth to a blebbling mode

    Science.gov (United States)

    The tubular growth of a pollen tube cell is crucial for the sexual reproduction of flowering plants. LePRK1 is a pollen-specific and plasma membrane–localized receptor-like kinase from tomato (Solanum lycopersicum). LePRK1 interacts with another receptor, LePRK2, and with KINASE PARTNER PROTEIN (KPP...

  11. Cooking birch pollen-related food: divergent consequences for IgE- and T cell-mediated reactivity in vitro and in vivo

    NARCIS (Netherlands)

    Bohle, Barbara; Zwölfer, Bettina; Heratizadeh, Annice; Jahn-Schmid, Beatrice; Antonia, Yuliya Dall; Alter, Mareike; Keller, Walter; Zuidmeer, Laurian; van Ree, Ronald; Werfel, Thomas; Ebner, Christof

    2006-01-01

    BACKGROUND: The major birch pollen allergen Bet v 1 cross-reacts with homologous food allergens, resulting in IgE-mediated oral allergy syndromes (OASs). To avoid this food, allergy allergologists and guidebooks advise patients to consume birch pollen-related foods after heating. OBJECTIVE: We

  12. Vimentin intermediate filaments template microtubule networks to enhance persistence in cell polarity and directed migration

    OpenAIRE

    Gan, Zhuo; Ding, Liya; Burckhardt, Christoph J.; Lowery, Jason; Zaritsky, Assaf; Sitterley, Karlyndsay; Mota, Andressa; Costigliola, Nancy; Starker, Colby G.; Voytas, Daniel F.; Tytell, Jessica; Goldman, Robert D.; Danuser, Gaudenz

    2016-01-01

    Increased expression of vimentin intermediate filaments (VIF) enhances directed cell migration, but the mechanism behind VIF’s effect on motility is not understood. VIF interact with microtubules, whose organization contributes to polarity maintenance in migrating cells. Here we characterize the dynamic coordination of VIF and microtubule networks in wounded monolayers of Retinal Pigment Epithelial cells. By genome editing we fluorescently labelled endogenous vimentin and α-...

  13. Polarization Curve of a Non-Uniformly Aged PEM Fuel Cell

    Directory of Open Access Journals (Sweden)

    Andrei Kulikovsky

    2014-01-01

    Full Text Available We develop a semi-analytical model for polarization curve of a polymer electrolyte membrane (PEM fuel cell with distributed (aged along the oxygen channel MEA transport and kinetic parameters of the membrane–electrode assembly (MEA. We show that the curve corresponding to varying along the channel parameter, in general, does not reduce to the curve for a certain constant value of this parameter. A possibility to determine the shape of the deteriorated MEA parameter along the oxygen channel by fitting the model equation to the cell polarization data is demonstrated.

  14. Cdc42 is not essential for filopodium formation, directed migration, cell polarization, and mitosis in fibroblastoid cells

    DEFF Research Database (Denmark)

    Czuchra, Aleksandra; Wu, Xunwei; Meyer, Hannelore

    2005-01-01

    Cdc42 is a small GTPase involved in the regulation of the cytoskeleton and cell polarity. To test whether Cdc42 has an essential role in the formation of filopodia or directed cell migration, we generated Cdc42-deficient fibroblastoid cells by conditional gene inactivation. We report here that loss...... of Cdc42 did not affect filopodium or lamellipodium formation and had no significant influence on the speed of directed migration nor on mitosis. Cdc42-deficient cells displayed a more elongated cell shape and had a reduced area. Furthermore, directionality during migration and reorientation of the Golgi...... apparatus into the direction of migration was decreased. However, expression of dominant negative Cdc42 in Cdc42-null cells resulted in strongly reduced directed migration, severely reduced single cell directionality, and complete loss of Golgi polarization and of directionality of protrusion formation...

  15. Rap1 integrates tissue polarity, lumen formation, and tumorigenicpotential in human breast epithelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Itoh, Masahiko; Nelson, Celeste M.; Myers, Connie A.; Bissell,Mina J.

    2006-09-29

    Maintenance of apico-basal polarity in normal breast epithelial acini requires a balance between cell proliferation, cell death, and proper cell-cell and cell-extracellular matrix signaling. Aberrations in any of these processes can disrupt tissue architecture and initiate tumor formation. Here we show that the small GTPase Rap1 is a crucial element in organizing acinar structure and inducing lumen formation. Rap1 activity in malignant HMT-3522 T4-2 cells is appreciably higher than in S1 cells, their non-malignant counterparts. Expression of dominant-negative Rap1 resulted in phenotypic reversion of T4-2 cells, led to formation of acinar structures with correct apico-basal polarity, and dramatically reduced tumor incidence despite the persistence of genomic abnormalities. The resulting acini contained prominent central lumina not observed when other reverting agents were used. Conversely, expression of dominant-active Rap1 in T4-2 cells inhibited phenotypic reversion and led to increased invasiveness and tumorigenicity. Thus, Rap1 acts as a central regulator of breast architecture, with normal levels of activation instructing apical polarity during acinar morphogenesis, and increased activation inducing tumor formation and progression to malignancy.

  16. Evolution and development of hair cell polarity and efferent function in the inner ear.

    Science.gov (United States)

    Sienknecht, Ulrike J; Köppl, Christine; Fritzsch, Bernd

    2014-01-01

    The function of the inner ear critically depends on mechanoelectrically transducing hair cells and their afferent and efferent innervation. The first part of this review presents data on the evolution and development of polarized vertebrate hair cells that generate a sensitive axis for mechanical stimulation, an essential part of the function of hair cells. Beyond the cellular level, a coordinated alignment of polarized hair cells across a sensory epithelium, a phenomenon called planar cell polarity (PCP), is essential for the organ's function. The coordinated alignment of hair cells leads to hair cell orientation patterns that are characteristic of the different sensory epithelia of the vertebrate inner ear. Here, we review the developmental mechanisms that potentially generate molecular and morphological asymmetries necessary for the control of PCP. In the second part, this review concentrates on the evolution, development and function of the enigmatic efferent neurons terminating on hair cells. We present evidence suggestive of efferents being derived from motoneurons and synapsing predominantly onto a unique but ancient cholinergic receptor. A review of functional data shows that the plesiomorphic role of the efferent system likely was to globally shut down and protect the peripheral sensors, be they vestibular, lateral line or auditory hair cells, from desensitization and damage during situations of self-induced sensory overload. The addition of a dedicated auditory papilla in land vertebrates appears to have favored the separation of vestibular and auditory efferents and specializations for more sophisticated and more diverse functions. © 2014 S. Karger AG, Basel.

  17. n3 PUFAs Reduce Mouse CD4+ T-Cell Ex Vivo Polarization into Th17 Cells123

    Science.gov (United States)

    Monk, Jennifer M.; Hou, Tim Y.; Turk, Harmony F.; McMurray, David N.; Chapkin, Robert S.

    2013-01-01

    Little is known about the impact of n3 (ω3) PUFAs on polarization of CD4+ T cells into effector subsets other than Th1 and Th2. We assessed the effects of dietary fat [corn oil (CO) vs. fish oil (FO)] and fermentable fiber [cellulose (C) vs. pectin (P)] (2 × 2 design) in male C57BL/6 mice fed CO-C, CO-P, FO-C, or FO-P diets for 3 wk on the ex vivo polarization of purified splenic CD4+ T cells (using magnetic microbeads) into regulatory T cells [Tregs; forkhead box P3 (Foxp3+) cells] or Th17 cells [interleukin (IL)-17A+ and retinoic acid receptor-related orphan receptor (ROR) γτ+ cells] by flow cytometry. Treg polarization was unaffected by diet; however, FO independently reduced the percentage of both CD4+ IL-17A+ (P diets enriched in eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), or DHA + EPA similarly reduced Th17-cell polarization in comparison to CO by reducing expression of the Th17-cell signature cytokine (IL-17A; P = 0.0015) and transcription factor (RORγτ P = 0.02), whereas Treg polarization was unaffected. Collectively, these data show that n3 PUFAs exert a direct effect on the development of Th17 cells in healthy mice, implicating a novel n3 PUFA–dependent, anti-inflammatory mechanism of action via the suppression of the initial development of this inflammatory T-cell subset. PMID:23864512

  18. [High temperature stres on crop pollen: a review].

    Science.gov (United States)

    Lu, Ming-Hui; Gong, Zhen-Hui; Chen, Ru-Gang; Huang, Wei; Li, Da-Wei

    2009-06-01

    High temperature has already become a noticeable environmental factor for crop production, while plant pollen was the most sensitive organ to high temperature stress. In this paper, the cytological, physiological, and molecular biological studies on the high temperature stress on crop pollen were reviewed, aimed to provide ideas for maintaining high productive ability of crops under high temperature stress. The cytological effects of high temperature on crop pollen included the changes of arrangement patterns of rough endoplasmic reticulum in tapetum cells, the irregularity of vascular bundle sheath cells in connective tissue, and the reduction of vesicle production by dictyosomes of pollen tube, etc.; physiological effects involved the incapacity of timely recovery of Ca2+ homeostasis, the changes of growth regulators contents, and the slowing down of carbohydrate metabolism, etc.; and molecular biological effects manifested in the insufficient induction of heat shock proteins and the inhibition of other functional genes for pollen development, etc.

  19. Proliferative effects of apical, but not basal, matrix metalloproteinase-7 activity in polarized MDCK cells

    International Nuclear Information System (INIS)

    Harrell, Permila C.; McCawley, Lisa J.; Fingleton, Barbara; McIntyre, J. Oliver; Matrisian, Lynn M.

    2005-01-01

    Matrix metalloproteinase-7 (MMP-7) is primarily expressed in glandular epithelium. Therefore, its mechanism of action may be influenced by its regulated vectorial release to either the apical and/or basolateral compartments, where it would act on its various substrates. To gain a better understanding of where MMP-7 is released in polarized epithelium, we have analyzed its pattern of secretion in polarized MDCK cells expressing stably transfected human MMP-7 (MDCK-MMP-7), and HCA-7 and Caco2 human colon cancer cell lines. In all cell lines, latent MMP-7 was secreted to both cellular compartments, but was 1.5- to 3-fold more abundant in the basolateral compartment as compared to the apical. However, studies in the MDCK system demonstrated that MMP-7 activity was 2-fold greater in the apical compartment of MDCK-MMP-7 HIGH -polarized monolayers, which suggests the apical co-release of an MMP-7 activator. In functional assays, MMP-7 over-expression increased cell saturation density as a result of increased cell proliferation with no effect on apoptosis. Apical MMP-7 activity was shown to be responsible for the proliferative effect, which occurred, as demonstrated by media transfer experiments, through cleavage of an apical substrate and not through the generation of a soluble factor. Taken together, our findings demonstrate the importance of MMP-7 secretion in relation to its mechanism of action when expressed in a polarized epithelium

  20. IKKα Promotes Intestinal Tumorigenesis by Limiting Recruitment of M1-like Polarized Myeloid Cells

    Directory of Open Access Journals (Sweden)

    Serkan I. Göktuna

    2014-06-01

    Full Text Available The recruitment of immune cells into solid tumors is an essential prerequisite of tumor development. Depending on the prevailing polarization profile of these infiltrating leucocytes, tumorigenesis is either promoted or blocked. Here, we identify IκB kinase α (IKKα as a central regulator of a tumoricidal microenvironment during intestinal carcinogenesis. Mice deficient in IKKα kinase activity are largely protected from intestinal tumor development that is dependent on the enhanced recruitment of interferon γ (IFNγ-expressing M1-like myeloid cells. In IKKα mutant mice, M1-like polarization is not controlled in a cell-autonomous manner but, rather, depends on the interplay of both IKKα mutant tumor epithelia and immune cells. Because therapies aiming at the tumor microenvironment rather than directly at the mutated cancer cell may circumvent resistance development, we suggest IKKα as a promising target for colorectal cancer (CRC therapy.

  1. The role of secretory and endocytic pathways in the maintenance of cell polarity.

    Science.gov (United States)

    Ang, Su Fen; Fölsch, Heike

    2012-01-01

    Epithelial cells line virtually every organ cavity in the body and are important for vectorial transport through epithelial monolayers such as nutrient uptake or waste product excretion. Central to these tasks is the establishment of epithelial cell polarity. During organ development, epithelial cells set up two biochemically distinct plasma membrane domains, the apical and the basolateral domain. Targeting of correct constituents to each of these regions is essential for maintaining epithelial cell polarity. Newly synthesized transmembrane proteins destined for the basolateral or apical membrane domain are sorted into separate transport carriers either at the TGN (trans-Golgi network) or in perinuclear REs (recycling endosomes). After initial delivery, transmembrane proteins, such as nutrient receptors, frequently undergo multiple rounds of endocytosis followed by re-sorting in REs. Recent work in epithelial cells highlights the REs as a potent sorting station with different subdomains representing individual targeting zones that facilitate the correct surface delivery of transmembrane proteins.

  2. Lol p I-induced IL-4 and IFN-gamma production by peripheral blood mononuclear cells of atopic and nonatopic subjects during and out of the pollen season.

    Science.gov (United States)

    Gagnon, R; Akoum, A; Hébert, J

    1993-04-01

    The reciprocal effects of IL-4 and IFN-gamma on IgE synthesis have been well established. It has also been shown that these two lymphokines are secreted by different subsets of CD4+ T cells (TH1 and TH2), and that TH2 helper T lymphocytes could be involved in the pathophysiology of allergic diseases. But little is known about the effects of an allergen on the profile of lymphokine synthesis by human peripheral blood mononuclear cells (PBMCs) of allergic and nonallergic subjects. We studied the production of IL-4 and IFN-gamma by PBMCs of atopic and nonatopic donors after in vitro stimulation by the group 1 allergen from Lolium perenne pollen (Lol p I), during and out of the grass pollen season. On natural exposure to pollen, Lol p I-induced IL-4 production was observed only with atopic donors (6 of 8), whereas the synthesis of IFN-gamma was observed for all nonatopic donors (7 of 7) and most allergic patients (5 of 7). At the time of the study, higher amounts of IFN-gamma were produced by PBMCs of nonatopic donors than by PBMCs of atopic patients. Out of the pollen season the production of IL-4 was not observed either by atopic (n = 11) or by nonatopic subjects (n = 5). On the other hand, IFN-gamma was produced by PBMCs of most subjects (atopic, 10 of 11; nonatopic, 5 of 5), but at the time of the study no difference was observed between the two groups. These results show that Lol p I induces different profiles of IL-4 and IFN-gamma production by PBMCs of atopic and nonatopic subjects. In atopic subjects this profile of lymphokine synthesis is influenced by the natural exposure to pollen, which is in keeping with the seasonal rise of IgE antibodies.

  3. Requirement for Dlgh-1 in planar cell polarity and skeletogenesis during vertebrate development.

    Directory of Open Access Journals (Sweden)

    Charlene Rivera

    Full Text Available The development of specialized organs is tightly linked to the regulation of cell growth, orientation, migration and adhesion during embryogenesis. In addition, the directed movements of cells and their orientation within the plane of a tissue, termed planar cell polarity (PCP, appear to be crucial for the proper formation of the body plan. In Drosophila embryogenesis, Discs large (dlg plays a critical role in apical-basal cell polarity, cell adhesion and cell proliferation. Craniofacial defects in mice carrying an insertional mutation in Dlgh-1 suggest that Dlgh-1 is required for vertebrate development. To determine what roles Dlgh-1 plays in vertebrate development, we generated mice carrying a null mutation in Dlgh-1. We found that deletion of Dlgh-1 caused open eyelids, open neural tube, and misorientation of cochlear hair cell stereociliary bundles, indicative of defects in planar cell polarity (PCP. Deletion of Dlgh-1 also caused skeletal defects throughout the embryo. These findings identify novel roles for Dlgh-1 in vertebrates that differ from its well-characterized roles in invertebrates and suggest that the Dlgh-1 null mouse may be a useful animal model to study certain human congenital birth defects.

  4. Effect of Polarization on Airway Epithelial Conditioning of Monocyte-Derived Dendritic Cells

    DEFF Research Database (Denmark)

    Papazian, Dick; Chhoden, Tashi; Arge, Maria

    2015-01-01

    ' immunoregulatory properties; thus, previous observations obtained using traditional setups should be considered with caution. Using the optimized setup, AEC conditioning of MDDCs led to increased expression of programmed death 1 ligand 1 (PD-L1), Immunoglobulin-Like Transcript 3, CD40, CD80 and CD23...... were allowed to polarize on filter inserts, and MDDCs were allowed to adhere to the epithelial basal side. In an optimized setup, the cell application was reversed, and the culture conditions were modified to preserve cellular polarization and integrity. These two parameters were crucial for the MDDCs...... to sample allergens administered to the apical side. Allergen uptake depended on both polarization and the nature of the allergen. AEC conditioning led to decreased birch allergen-specific proliferation of autologous T cells and a trend toward decreased secretion of the Th2-specific cytokines IL-5 and IL-13...

  5. Planar Cell Polarity Breaks the Symmetry of PAR Protein Distribution prior to Mitosis in Drosophila Sensory Organ Precursor Cells.

    Science.gov (United States)

    Besson, Charlotte; Bernard, Fred; Corson, Francis; Rouault, Hervé; Reynaud, Elodie; Keder, Alyona; Mazouni, Khalil; Schweisguth, François

    2015-04-20

    During development, cell-fate diversity can result from the unequal segregation of fate determinants at mitosis. Polarization of the mother cell is essential for asymmetric cell division (ACD). It often involves the formation of a cortical domain containing the PAR complex proteins Par3, Par6, and atypical protein kinase C (aPKC). In the fly notum, sensory organ precursor cells (SOPs) divide asymmetrically within the plane of the epithelium and along the body axis to generate two distinct cells. Fate asymmetry depends on the asymmetric localization of the PAR complex. In the absence of planar cell polarity (PCP), SOPs divide with a random planar orientation but still asymmetrically, showing that PCP is dispensable for PAR asymmetry at mitosis. To study when and how the PAR complex localizes asymmetrically, we have used a quantitative imaging approach to measure the planar polarization of the proteins Bazooka (Baz, fly Par3), Par6, and aPKC in living pupae. By using imaging of functional GFP-tagged proteins with image processing and computational modeling, we find that Baz, Par6, and aPKC become planar polarized prior to mitosis in a manner independent of the AuroraA kinase and that PCP is required for the planar polarization of Baz, Par6, and aPKC during interphase. This indicates that a "mitosis rescue" mechanism establishes asymmetry at mitosis in PCP mutants. This study therefore identifies PCP as the initial symmetry-breaking signal for the planar polarization of PAR proteins in asymmetrically dividing SOPs. Copyright © 2015 Elsevier Ltd. All rights reserved.

  6. Chemical modification of coating of Pinus halepensis pollen by ozone exposure.

    Science.gov (United States)

    Naas, Oumsaad; Mendez, Maxence; Quijada, Melesio; Gosselin, Sylvie; Farah, Jinane; Choukri, Ali; Visez, Nicolas

    2016-07-01

    Pollen coating, located on the exine, includes an extractible lipid fraction. The modification of the pollen coating by air pollutants may have implications on the interactions of pollen with plant stigmas and human cells. Pinus halepensis pollen was exposed to ozone in vitro and the pollen coating was extracted with organic solvent and analyzed by GC-MS. Ozone has induced chemical changes in the coating as observed with an increase in dicarboxylic acids, short-chain fatty acids and aldehydes. 4-Hydroxybenzaldehyde was identified as the main reaction product and its formation was shown to occur both on native pollen and on defatted pollen. 4-Hydroxybenzaldehyde is very likely formed via the ozonolysis of acid coumaric-like monomers constitutive of the sporopollenin. Modification of pollen coating by air pollutants should be accounted for in further studies on effect of pollution on germination and on allergenicity. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. The subapical compartment and its role in intracellular trafficking and cell polarity

    NARCIS (Netherlands)

    Van Ijzendoorn, Sven C. D.; Maier, Olaf; Van Der Wouden, Johanna M.; Hoekstra, Dick

    In polarized epithelial cells and hepatocytes, apical and basolateral plasma membrane surfaces are maintained, each displaying a distinct molecular composition. In recent years, it has become apparent that a subapical compartment, referred to as SAC, plays a prominent if not crucial role in the

  8. LKB1 and AMPK Family Signaling: The Intimate Link Between Cell Polarity and Energy Metabolism

    NARCIS (Netherlands)

    Jansen, Marnix; ten Klooster, Jean Paul; Offerhaus, G. Johan; Clevers, Hans

    2009-01-01

    Jansen M, ten Klooster JP, Offerhaus GJ, Clevers H. LKB1 and AMPK Family Signaling: The Intimate Link Between Cell Polarity and Energy Metabolism. Physiol Rev 89: 777-798, 2009; doi:10.1152/physrev.00026.2008. Research on the LKB1 tumor suppressor protein mutated in cancer-prone Peutz-Jeghers

  9. TH1 and TH2 cell polarization increases with aging and is modulated by zinc supplementation

    OpenAIRE

    2008-01-01

    TH1 and TH2 cell polarization increases with aging and is modulated by zinc supplementation correspondence: Corresponding author. Tel.: +49 241 8080208; fax: +49 241 8082613. (Rink, Lothar) (Rink, Lothar) Institute of Immunology, University Hospital, RWTH Aachen University - Aachen--> - GERMANY (Uciechowski, Peter) Institute of Immunology, University Hospital, RWTH Aachen University - Aachen--> - GERMAN...

  10. Apical–basal polarity: why plant cells don't stand on their heads

    Czech Academy of Sciences Publication Activity Database

    Friml, J.; Benfey, P.; Benková, E.; Bennett, M. D.; Berleth, T.; Geldner, N.; Grebe, M.; Heisler, M.; Hejátko, J.; Jürgens, G.; Laux, T.; Lindsey, K.; Lukowitz, W.; Luschnig, Ch.; Offringa, R.; Scheres, B.; Swarup, R.; Torres-Ruiz, R.; Weijers, D.; Zažímalová, Eva

    2006-01-01

    Roč. 11, č. 1 (2006), s. 12-14 ISSN 1360-1385 R&D Projects: GA AV ČR IAA6038303 Institutional research plan: CEZ:AV0Z50380511 Keywords : Apical * Basal * Polarity of plant cell Subject RIV: EF - Botanics Impact factor: 8.000, year: 2006

  11. Mutation of the planar cell polarity gene VANGL1 in adolescent idiopathic scoliosis

    DEFF Research Database (Denmark)

    Andersen, Malene Rask; Farooq, Muhammad; Rasmussen, Karen Koefoed

    2017-01-01

    STUDY DESIGN: Mutation analysis of a candidate disease gene in a cohort of patients with moderate to severe Adolescent idiopathic scoliosis (AIS). OBJECTIVE: To investigate if damaging mutations in the planar cell polarity gene VANGL1 could be identified in AIS patients. SUMMARY OF BACKGROUND DATA...

  12. Measurement of cell wall depolarization of polarized hydrogen gas targets in a weak magnetic field

    International Nuclear Information System (INIS)

    Price, J.S.; Haeberli, W.

    1994-01-01

    Polarized gas targets using windowless storage cells are being developed for use as internal targets in medium and high energy particle storage rings. Tests were conducted to evaluate wall depolarization for different cell wall materials. Measurements of the target polarization were made on polarized vector H 0 gas targets in a weak magnetic field. Fifteen materials were tested in geometries corresponding to different average number of wall collisions, N 0 , from 40 to 380 collisions, for wall temperatures, T, from 20 K to 300 K. A method was developed to measure the polarization of a vector H 0 target in a 0.5 mT field: a beam of 50 keV D + picks up electrons from the target gas and the vector D 0 acquires a tensor polarization, p zz , which is measured by means of the 3 H( vector d, n) 4 He reaction. A simple model for depolarization at surfaces is proposed. Comparison to the data shows fair agreement, but the model is unrealistic in that it does not include the effects of the recombination of atoms on the surface to form molecules. ((orig.))

  13. Mathematical modeling of planar cell polarity signaling in the Drosophila melanogaster wing

    Science.gov (United States)

    Amonlirdviman, Keith

    Planar cell polarity (PCP) signaling refers to the coordinated polarization of cells within the plane of various epithelial tissues to generate sub-cellular asymmetry along an axis orthogonal to their apical-basal axes. For example, in the Drosophila wing, PCP is seen in the parallel orientation of hairs that protrude from each of the approximately 30,000 epithelial cells to robustly point toward the wing tip. Through a poorly understood mechanism, cell clones mutant for some PCP signaling components, including some, but not all alleles of the receptor frizzled, cause polarity disruptions of neighboring, wild-type cells, a phenomenon referred to as domineering nonautonomy. Previous models have proposed diffusible factors to explain nonautonomy, but no such factors have yet been found. This dissertation describes the mathematical modeling of PCP in the Drosophila wing, based on a contact dependent signaling hypothesis derived from experimental results. Intuition alone is insufficient to deduce that this hypothesis, which relies on a local feedback loop acting at the cell membrane, underlies the complex patterns observed in large fields of cells containing mutant clones, and others have argued that it cannot account for observed phenotypes. Through reaction-diffusion, partial differential equation modeling and simulation, the feedback loop is shown to fully reproduce PCP phenotypes, including domineering nonautonomy. The sufficiency of this model and the experimental validation of model predictions argue that previously proposed diffusible factors need not be invoked to explain PCP signaling and reveal how specific protein-protein interactions lead to autonomy or domineering nonautonomy. Based on these results, an ordinary differential equation model is derived to study the relationship of the feedback loop with upstream signaling components. The cadherin Fat transduces a cue to the local feedback loop, biasing the polarity direction of each cell toward the wing tip

  14. RELATIONSHIP BETWEEN RECENT POLLEN DEPOSITION AND AIRBORNE POLLEN CONCENTRATION

    NARCIS (Netherlands)

    SPIEKSMA, FTM; NIKKELS, BH; BOTTEMA, S

    In the reconstruction of past or recent vegetation the study of deposited pollen plays an important role. As reference value, very often the pollen content of moss polsters (''moss cushions'') is assessed to estimate the pollen deposition (''influx'') from the air. Recently, the data from

  15. Bipolar Plasma Membrane Distribution of Phosphoinositides and Their Requirement for Auxin-Mediated Cell Polarity and Patterning in Arabidopsis

    NARCIS (Netherlands)

    Tejos, R.; Sauer, M.; Vanneste, S.; Palacios-Gomez, M.; Li, H.; Heilmann, M.; van Wijk, R.; Vermeer, J.E.M.; Heilmann, I.; Munnik, T.; Friml, J.

    2014-01-01

    Cell polarity manifested by asymmetric distribution of cargoes, such as receptors and transporters, within the plasma membrane (PM) is crucial for essential functions in multicellular organisms. In plants, cell polarity (re)establishment is intimately linked to patterning processes. Despite the

  16. Expression of the major mugwort pollen allergen Art v 1 in tobacco plants and cell cultures: problems and perspectives for allergen production in plants.

    Science.gov (United States)

    Siegert, Marc; Pertl-Obermeyer, Heidi; Gadermaier, Gabriele; Ferreira, Fatima; Obermeyer, Gerhard

    2012-03-01

    An economic and cheap production of large amounts of recombinant allergenic proteins might become a prerequisite for the common use of microarray-based diagnostic allergy assays which allow a component-specific diagnosis. A molecular pharming strategy was applied to express the major allergen of Artemisia vulgaris pollen, Art v 1, in tobacco plants and tobacco cell cultures. The original Art v 1 with its endogenous signal peptide which directs Art v 1 to the secretory pathway, was expressed in transiently transformed tobacco leaves but was lost in stable transformed tobacco plants during the alternation of generations. Using a light-regulated promoter and "hiding" the recombinant Art v 1 in the ER succeeded in expression of Art v 1 over three generations of tobacco plants and in cell cultures generated from stable transformed plants. However, the amounts of the recombinant allergen were sufficient for analysis but not high enough to allow an economic production. Although molecular pharming has been shown to work well for the production of non-plant therapeutic proteins, it might be less efficient for closely related plant proteins.

  17. Polarity establishment, morphogenesis, and cultured plant cells in space

    Science.gov (United States)

    Krikorian, Abraham D.

    1989-01-01

    Plant development entails an orderly progression of cellular events both in terms of time and geometry. There is only circumstantial evidence that, in the controlled environment of the higher plant embryo sac, gravity may play a role in embryo development. It is still not known whether or not normal embryo development and differentiation in higher plants can be expected to take place reliably and efficiently in the micro g space environment. It seems essential that more attention be given to studying aspects of reproductive biology in order to be confident that plants will survive seed to seed to seed in a space environment. Until the time arrives when successive generations of plants can be grown, the best that can be done is utilize the most appropriate systems and begin, piece meal, to accumulate information on important aspects of plant reproduction. Cultured plant cells can play an important role in these activities since they can be grown so as to be morphogenetically competent, and thus can simulate those embryogenic events more usually identified with fertilized eggs in the embryo sac of the ovule in the ovary. Also, they can be manipulated with relative ease. The extreme plasticity of such demonstrably totipotent cell systems provides a means to test environmental effects such as micro g on a potentially free-running entity. The successful manipulation and management of plant cells and propagules in space also has significance for exploitation of biotechnologies in space since such systems, perforce, are an important vehicle whereby many genetic engineering manipulations are achieved.

  18. Basolateral invasion and trafficking of Campylobacter jejuni in polarized epithelial cells.

    Directory of Open Access Journals (Sweden)

    Lieneke I Bouwman

    Full Text Available Campylobacter jejuni is a major cause of bacterial diarrheal disease. Most enteropathogenic bacteria including C. jejuni can invade cultured eukaryotic cells via an actin- and/or microtubule-dependent and an energy-consuming uptake process. Recently, we identified a novel highly efficient C. jejuni invasion pathway that involves bacterial migration into the subcellular space of non-polarized epithelial cells (termed subvasion followed by invasion from the cell basis. Here we report cellular requirements of this entry mechanism and the subsequent intracellular trafficking route of C. jejuni in polarized islands of Caco-2 intestinal epithelial cells. Advanced microscopy on infected cells revealed that C. jejuni invades the polarized intestinal cells via the subcellular invasion pathway. Remarkably, invasion was not blocked by the inhibitors of microtubule dynamics colchicine or paclitaxel, and was even enhanced after disruption of host cell actin filaments by cytochalasin D. Invasion also continued after dinitrophenol-induced cellular depletion of ATP, whereas this compound effectively inhibited the uptake of invasive Escherichia coli. Confocal microscopy demonstrated that intracellular C. jejuni resided in membrane-bound CD63-positive cellular compartments for up to 24 h. Establishment of a novel luciferase reporter-based bacterial viability assay, developed to overcome the limitations of the classical bacterial recovery assay, demonstrated that a subset of C. jejuni survived intracellularly for up to 48 h. Taken together, our results indicate that C. jejuni is able to actively invade polarized intestinal epithelial cells via a novel actin- and microtubule-independent mechanism and remains metabolically active in the intracellular niche for up to 48 hours.

  19. Influenza H5N1 virus infection of polarized human alveolar epithelial cells and lung microvascular endothelial cells

    Directory of Open Access Journals (Sweden)

    Yuen Kit M

    2009-10-01

    Full Text Available Abstract Background Highly pathogenic avian influenza (HPAI H5N1 virus is entrenched in poultry in Asia and Africa and continues to infect humans zoonotically causing acute respiratory disease syndrome and death. There is evidence that the virus may sometimes spread beyond respiratory tract to cause disseminated infection. The primary target cell for HPAI H5N1 virus in human lung is the alveolar epithelial cell. Alveolar epithelium and its adjacent lung microvascular endothelium form host barriers to the initiation of infection and dissemination of influenza H5N1 infection in humans. These are polarized cells and the polarity of influenza virus entry and egress as well as the secretion of cytokines and chemokines from the virus infected cells are likely to be central to the pathogenesis of human H5N1 disease. Aim To study influenza A (H5N1 virus replication and host innate immune responses in polarized primary human alveolar epithelial cells and lung microvascular endothelial cells and its relevance to the pathogenesis of human H5N1 disease. Methods We use an in vitro model of polarized primary human alveolar epithelial cells and lung microvascular endothelial cells grown in transwell culture inserts to compare infection with influenza A subtype H1N1 and H5N1 viruses via the apical or basolateral surfaces. Results We demonstrate that both influenza H1N1 and H5N1 viruses efficiently infect alveolar epithelial cells from both apical and basolateral surface of the epithelium but release of newly formed virus is mainly from the apical side of the epithelium. In contrast, influenza H5N1 virus, but not H1N1 virus, efficiently infected polarized microvascular endothelial cells from both apical and basolateral aspects. This provides a mechanistic explanation for how H5N1 virus may infect the lung from systemic circulation. Epidemiological evidence has implicated ingestion of virus-contaminated foods as the source of infection in some instances and our

  20. Molecular evidence that most RNAs required for germination and pollen tube growth are stored in the mature pollen grain in petunia.

    Science.gov (United States)

    Ishimizu, Takeshi; Kodama, Hiroaki; Ando, Toshio; Watanabe, Masao

    2010-01-01

    After landing on the stigma, the pollen grain germinates and elongates a tube to deliver its generative nuclei to the egg cell of the ovule. The molecular mechanisms involved in the drastic morphological changes in the pollen grain during this fertilization process remain largely unknown. In this study, the expression of 732 randomly selected genes in petunia pollen and pollen tubes was analyzed by microarray and quantitative PCR analyses. We found no evidence for up-regulation of any of these genes in the pollen tube. Our findings provide support at the gene level for the longstanding hypothesis that pollen germination and tube growth are not dependent on new RNA synthesis and that the large number of RNAs required for germination and tube growth are stored in mature pollen grains.

  1. Apolar and polar transitions drive the conversion between amoeboid and mesenchymal shapes in melanoma cells.

    Science.gov (United States)

    Cooper, Sam; Sadok, Amine; Bousgouni, Vicky; Bakal, Chris

    2015-11-05

    Melanoma cells can adopt two functionally distinct forms, amoeboid and mesenchymal, which facilitates their ability to invade and colonize diverse environments during the metastatic process. Using quantitative imaging of single living tumor cells invading three-dimensional collagen matrices, in tandem with unsupervised computational analysis, we found that melanoma cells can switch between amoeboid and mesenchymal forms via two different routes in shape space--an apolar and polar route. We show that whereas particular Rho-family GTPases are required for the morphogenesis of amoeboid and mesenchymal forms, others are required for transitions via the apolar or polar route and not amoeboid or mesenchymal morphogenesis per se. Altering the transition rates between particular routes by depleting Rho-family GTPases can change the morphological heterogeneity of cell populations. The apolar and polar routes may have evolved in order to facilitate conversion between amoeboid and mesenchymal forms, as cells are either searching for, or attracted to, particular migratory cues, respectively. © 2015 Cooper et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

  2. Effects of low-energy argon ion implantation on the dynamic organization of the actin cytoskeleton during maize pollen germination.

    Science.gov (United States)

    Deng, F; Zhu, S W; Wu, L J; Cheng, B J

    2010-04-27

    The relationship between pollen germination and the dynamic organization of the actin cytoskeleton during pollen germination is a central theme in plant reproductive biology research. Maize (Zea mays) pollen grains were implanted with 30 keV argon ion (Ar(+)) beams at doses ranging from 0.78 x 10(15) to 13 x 10(15) ions/cm(2). The effects of low-energy ion implantation on pollen germination viability and the dynamic organization of the actin cytoskeleton during pollen germination were studied using confocal laser scanning microscopy. Maize pollen germination rate increased remarkably with Ar(+) dose, in the range from 3.9 x 10(15) to 6.5 x 10(15) ions/cm(2); the germination rate peaked at an Ar(+) dose of 5.2 x 10(15) ions/cm(2). When the implantation dose exceeded 7.8 x 10(15) ions/cm(2), the rate of pollen germination decreased sharply. The actin filaments assembled in pollen grains implanted with 5.2 x 10(15) ions/cm(2) Ar(+) much earlier than in controls. The actin filaments organized as longer parallel bundles and extended into the emerging pollen tube in treated pollen grains, while they formed random and loose fine bundles and were gathered at the pollen aperture in the control. The reorganization of actin cytoskeleton in the pollen implanted with 9.1 x 10(15) ions/cm(2) Ar(+) was slower than in controls. There was a positive correlation between pollen germination and the dynamic organization of the actin cytoskeleton during pollen germination. Ion implantation into pollen did not cause changes in the polarization of actin filaments and organelle dynamics in the pollen tubes. The effects of Ar(+) implantation on pollen germination could be mediated by changes in the polymerization and rearrangement of actin polymers.

  3. New, simple theory-based, accurate polarization microscope for birefringence imaging of biological cells

    Science.gov (United States)

    Shin, In Hee; Shin, Sang-Mo; Kim, Dug Young

    2010-01-01

    We propose a new, simple theory-based, accurate polarization microscope for birefringence imaging of cytoskeletal structures of biological cells. The new theory lets us calculate very easily the phase retardation and the orientation of the principal axis of a particular area of a biological living cell in media by simply measuring the intensity variation of a pixel of a CCD camera while rotating a single polarizer. Just from the measured intensity maxima and minima, the amount of phase retardation δ between the fast and the slow axis of the sample area is obtained with an accuracy of 5.010+/-0.798×10-3 rad. The orientation of the principal axis is calculated from the angle of the polarizer for the intensity maximum. We have compared our microscopes with two previously reported polarization microscopes for birefringence imaging of cytoskeletal structures and demonstrated the utility of our microscope with the phase retardation and orientation images of weakly invasive MCF7 and highly invasive MDA MB 231 human breast cancer cells as an example.

  4. Hybridizing pines with diluted pollen

    Science.gov (United States)

    Robert Z. Callaham

    1967-01-01

    Diluted pollens would have many uses by the tree breeder. Dilutions would be particularly advantageous in making many controlled pollinations with a limited amount of pollen. They also would be useful in artificial mass pollinations of orchards or single trees. Diluted pollens might help overcome troublesome genetic barriers to crossing. Feasibility o,f using diluted...

  5. Germination and storage of pollen

    NARCIS (Netherlands)

    Visser, T.

    1955-01-01

    Germination of pear pollen markedly improved when boric acid was added to the medium. The pollen was more sensitive to boron in water than in 10 % sugar solution. Supplying weak solutions of boron to pear branches before flowering resulted in a good germination of the pollen in sugar solution

  6. Concomitant use of polarization and positive phase contrast microscopy for the study of microbial cells.

    Science.gov (United States)

    Žižka, Zdeněk; Gabriel, Jiří

    2015-11-01

    Polarization and positive phase contrast microscope were concomitantly used in the study of the internal structure of microbial cells. Positive phase contrast allowed us to view even the fine cell structure with a refractive index approaching that of the surrounding environment, e.g., the cytoplasm, and transferred the invisible phase image to a visible amplitude image. With polarization microscopy, crossed polarizing filters together with compensators and a rotary stage showed the birefringence of different cell structures. Material containing algae was collected in ponds in Sýkořice and Zbečno villages (Křivoklát region). The objects were studied in laboratory microscopes LOMO MIN-8 Sankt Petersburg and Polmi A Carl Zeiss Jena fitted with special optics for positive phase contrast, polarizers, analyzers, compensators, rotary stages, and digital SLR camera Nikon D 70 for image capture. Anisotropic granules were found in the cells of flagellates of the order Euglenales, in green algae of the orders Chlorococcales and Chlorellales, and in desmid algae of the order Desmidiales. The cell walls of filamentous algae of the orders Zygnematales and Ulotrichales were found to exhibit significant birefringence; in addition, relatively small amounts of small granules were found in the cytoplasm. A typical shape-related birefringence of the cylindrical walls and the septa between the cells differed in intensity, which was especially apparent when using a Zeiss compensator RI-c during its successive double setting. In conclusion, the anisotropic granules found in the investigated algae mostly showed strong birefringence and varied in number, size, and location of the cells. Representatives of the order Chlorococcales contained the highest number of granules per cell, and the size of these granules was almost double than that of the other monitored microorganisms. Very strong birefringence was exhibited by cell walls of filamentous algae; it differed in the intensity

  7. Participation of the cell polarity protein PALS1 to T-cell receptor-mediated NF-κB activation.

    Directory of Open Access Journals (Sweden)

    Gabrielle Carvalho

    Full Text Available BACKGROUND: Beside their established function in shaping cell architecture, some cell polarity proteins were proposed to participate to lymphocyte migration, homing, scanning, as well as activation following antigen receptor stimulation. Although PALS1 is a central component of the cell polarity network, its expression and function in lymphocytes remains unknown. Here we investigated whether PALS1 is present in T cells and whether it contributes to T Cell-Receptor (TCR-mediated activation. METHODOLOGY/PRINCIPAL FINDINGS: By combining RT-PCR and immunoblot assays, we found that PALS1 is constitutively expressed in human T lymphocytes as well as in Jurkat T cells. siRNA-based knockdown of PALS1 hampered TCR-induced activation and optimal proliferation of lymphocyte. We further provide evidence that PALS1 depletion selectively hindered TCR-driven activation of the transcription factor NF-κB. CONCLUSIONS: The cell polarity protein PALS1 is expressed in T lymphocytes and participates to the optimal activation of NF-κB following TCR stimulation.

  8. Pollen of Syzygium (Myrtaceae) from SE Asia, especially Thailand

    NARCIS (Netherlands)

    Parnell, J.

    2003-01-01

    Pollen of 57 species of Syzygium from SE Asia were surveyed by SEM. The grains are all ± triangular in polar view, uniformly small (mean diameter 10.4 μm) with few surface sculpturing features. All species examined exhibit a distinct apocolpium and, usually, apocolpial field. The presence of an

  9. Polarization of migrating monocytic cells is independent of PI 3-kinase activity.

    Directory of Open Access Journals (Sweden)

    Silvia Volpe

    Full Text Available BACKGROUND: Migration of mammalian cells is a complex cell type and environment specific process. Migrating hematopoietic cells assume a rapid amoeboid like movement when exposed to gradients of chemoattractants. The underlying signaling mechanisms remain controversial with respect to localization and distribution of chemotactic receptors within the plasma membrane and the role of PI 3-kinase activity in cell polarization. METHODOLOGY/PRINCIPAL FINDINGS: We present a novel model for the investigation of human leukocyte migration. Monocytic THP-1 cells transfected with the alpha(2A-adrenoceptor (alpha(2AAR display comparable signal transduction responses, such as calcium mobilization, MAP-kinase activation and chemotaxis, to the noradrenaline homologue UK 14'304 as when stimulated with CCL2, which binds to the endogenous chemokine receptor CCR2. Time-lapse video microscopy reveals that chemotactic receptors remain evenly distributed over the plasma membrane and that their internalization is not required for migration. Measurements of intramolecular fluorescence resonance energy transfer (FRET of alpha(2AAR-YFP/CFP suggest a uniform activation of the receptors over the entire plasma membrane. Nevertheless, PI 3-kinase activation is confined to the leading edge. When reverting the gradient of chemoattractant by moving the dispensing micropipette, polarized monocytes--in contrast to neutrophils--rapidly flip their polarization axis by developing a new leading edge at the previous posterior side. Flipping of the polarization axis is accompanied by re-localization of PI-3-kinase activity to the new leading edge. However, reversal of the polarization axis occurs in the absence of PI 3-kinase activation. CONCLUSIONS/SIGNIFICANCE: Accumulation and internalization of chemotactic receptors at the leading edge is dispensable for cell migration. Furthermore, uniformly distributed receptors allow the cells to rapidly reorient and adapt to changes in the

  10. Nuclear fallout provides a new link between aPKC and polarized cell trafficking.

    Science.gov (United States)

    Calero-Cuenca, Francisco J; Espinosa-Vázquez, José Manuel; Reina-Campos, Miguel; Díaz-Meco, María T; Moscat, Jorge; Sotillos, Sol

    2016-04-18

    Cell polarity, essential for cell physiology and tissue coherence, emerges as a consequence of asymmetric localization of protein complexes and directional trafficking of cellular components. Although molecules required in both processes are well known their relationship is still poorly understood. Here we show a molecular link between Nuclear Fallout (Nuf), an adaptor of Rab11-GTPase to the microtubule motor proteins during Recycling Endosome (RE) trafficking, and aPKC, a pivotal kinase in the regulation of cell polarity. We demonstrate that aPKC phosphorylates Nuf modifying its subcellular distribution. Accordingly, in aPKC mutants Nuf and Rab11 accumulate apically indicating altered RE delivery. We show that aPKC localization in the apico-lateral cortex is dynamic. When we block exocytosis, by means of exocyst-sec mutants, aPKC accumulates inside the cells. Moreover, apical aPKC concentration is reduced in nuf mutants, suggesting aPKC levels are maintained by recycling. We demonstrate that active aPKC interacts with Nuf, phosphorylating it and, as a result, modifying its subcellular distribution. We propose a regulatory loop by which Nuf promotes aPKC apical recycling until sufficient levels of active aPKC are reached. Thus, we provide a novel link between cell polarity regulation and traffic control in epithelia.

  11. Trafficking through COPII stabilises cell polarity and drives secretion during Drosophila epidermal differentiation.

    Directory of Open Access Journals (Sweden)

    Michaela Norum

    2010-05-01

    Full Text Available The differentiation of an extracellular matrix (ECM at the apical side of epithelial cells implies massive polarised secretion and membrane trafficking. An epithelial cell is hence engaged in coordinating secretion and cell polarity for a correct and efficient ECM formation.We are studying the molecular mechanisms that Drosophila tracheal and epidermal cells deploy to form their specific apical ECM during differentiation. In this work we demonstrate that the two genetically identified factors haunted and ghost are essential for polarity maintenance, membrane topology as well as for secretion of the tracheal luminal matrix and the cuticle. We show that they code for the Drosophila COPII vesicle-coating components Sec23 and Sec24, respectively, that organise vesicle transport from the ER to the Golgi apparatus.Taken together, epithelial differentiation during Drosophila embryogenesis is a concerted action of ECM formation, plasma membrane remodelling and maintenance of cell polarity that all three rely mainly, if not absolutely, on the canonical secretory pathway from the ER over the Golgi apparatus to the plasma membrane. Our results indicate that COPII vesicles constitute a central hub for these processes.

  12. ErbB receptors and cell polarity: New pathways and paradigms for understanding cell migration and invasion

    International Nuclear Information System (INIS)

    Feigin, Michael E.; Muthuswamy, Senthil K.

    2009-01-01

    The ErbB family of receptor tyrosine kinases is involved in initiation and progression of a number of human cancers, and receptor activation or overexpression correlates with poor patient survival. Research over the past two decades has elucidated the molecular mechanisms underlying ErbB-induced tumorigenesis, which has resulted in the development of effective targeted therapies. ErbB-induced signal transduction cascades regulate a wide variety of cell processes, including cell proliferation, apoptosis, cell polarity, migration and invasion. Within tumors, disruption of these core processes, through cooperative oncogenic lesions, results in aggressive, metastatic disease. This review will focus on the ErbB signaling networks that regulate migration and invasion and identify a potential role for cell polarity pathways during cancer progression

  13. Identification and characterization of TcCRP1, a pollen tube attractant from Torenia concolor.

    Science.gov (United States)

    Kanaoka, Masahiro M; Kawano, Nao; Matsubara, Yoshiyuki; Susaki, Daichi; Okuda, Satohiro; Sasaki, Narie; Higashiyama, Tetsuya

    2011-09-01

    During sexual reproduction in higher angiosperms, the pollen tubes are directed to the ovules in the pistil to deliver sperm cells. This pollen tube attraction is highly species specific, and a group of small secreted proteins, TfCRPs, are necessary for this process in Torenia fournieri. A candidate pollen tube attractant protein in Torenia concolor, a related species of T. fournieri, was isolated and the attractant abilities between them were compared. TcCRP1, an orthologous gene of TfCRP1 from T. concolor, is expressed predominantly in the synergid cell. The gene product attracted pollen tubes in a concentration-dependent manner, but attracted fewer pollen tubes from the other species. The results indicated that this class of CRP proteins is a common pollen tube attractant in Torenia species. The sequence diversity of these proteins is important for species-specific pollen tube attraction.

  14. Ion Implantation Hampers Pollen Tube Growth and Disrupts Actin Cytoskeleton Organization in Pollen Tubes of Pinus thunbergii

    International Nuclear Information System (INIS)

    Li Guoping; Yang Lusheng; Huang Qunce; Qin Guangyong

    2008-01-01

    Pollen grains of Pinus thunbergii Parl. (Japanese black pine) were implanted with 30 keV nitrogen ion beams and the effects of nitrogen ion implantation on pollen tube growth in vitro and the organization of actin cytoskeleton in the pollen tube cell were investigated using a confocal laser scanning microscope after fluorescence labeling. Treatment with ion implantation significantly blocked pollen tube growth. Confocal microscopy showed that ion implantation disrupted actin filament cytoskeleton organization in the pollen tube. It was found that there was a distinct correlation between the inhibition of pollen tube growth and the disruption of actin cytoskeleton organization, indicating that an intact actin cytoskeleton is essential for continuous pollen tube elongation in Pinus thunbergii. Although the detailed mechanism for the ion-implantation-induced bioeffect still remains to be elucidated, the present study assumes that the cytoskeleton system in pollen grains may provide a key target in response to ion beam implantation and is involved in mediating certain subsequent cytological changes.

  15. Ultrastructural analyses of somatic embryo initiation, development and polarity establishment from mesophyll cells of Dactylis glomerata

    Science.gov (United States)

    Vasilenko, A.; McDaniel, J. K.; Conger, B. V.

    2000-01-01

    Somatic embryos initiate and develop directly from single mesophyll cells in in vitro-cultured leaf segments of orchardgrass (Dactylis glomerata L.). Embryogenic cells establish themselves in the predivision stage by formation of thicker cell walls and dense cytoplasm. Electron microscopy observations for embryos ranging from the pre-cell-division stage to 20-cell proembryos confirm previous light microscopy studies showing a single cell origin. They also confirm that the first division is predominantly periclinal and that this division plane is important in establishing embryo polarity and in determining the embryo axis. If the first division is anticlinal or if divisions are in random planes after the first division, divisions may not continue to produce an embryo. This result may produce an embryogenic cell mass, callus formation, or no structure at all. Grant numbers: NAGW-3141, NAG10-0221.

  16. Diacylglycerol Kinases: Shaping Diacylglycerol and Phosphatidic Acid Gradients to Control Cell Polarity

    Directory of Open Access Journals (Sweden)

    Gianluca Baldanzi

    2016-11-01

    Full Text Available Diacylglycerol kinases (DGKs terminate diacylglycerol (DAG signaling and promote phosphatidic acid (PA production. Isoform specific regulation of DGKs activity and localization allows DGKs to shape the DAG and PA gradients. The capacity of DGKs to constrain the areas of DAG signaling is exemplified by their role in defining the contact interface between T cells and antigen presenting cells: the immune synapse. Upon T cell receptor engagement, both DGK α and ζ metabolize DAG at the immune synapse thus constraining DAG signaling. Interestingly, their activity and localization are not fully redundant because DGKζ activity metabolizes the bulk of DAG in the cell, whereas DGKα limits the DAG signaling area localizing specifically at the periphery of the immune synapse.When DGKs terminate DAG signaling, the local PA production defines a new signaling domain, where PA recruits and activates a second wave of effector proteins. The best-characterized example is the role of DGKs in protrusion elongation and cell migration. Indeed, upon growth factor stimulation, several DGK isoforms, such as α, ζ, and γ, are recruited and activated at the plasma membrane. Here, local PA production controls cell migration by finely modulating cytoskeletal remodeling and integrin recycling. Interestingly, DGK-produced PA also controls the localization and activity of key players in cell polarity such as aPKC, Par3, and integrin β1. Thus, T cell polarization and directional migration may be just two instances of the general contribution of DGKs to the definition of cell polarity by local specification of membrane identity signaling.

  17. The damaging effects of nitrogen ion beam implantation on upland cotton (Gossypium hirsutum L.) pollen grains

    Energy Technology Data Exchange (ETDEWEB)

    Yu Yanjie [College of Agronomy, Nanjing Agricultural University, Nanjing Jiangsu 210095 (China); Wu Lijun; Wu Yuejin [Key Laboratory of Ion Beam Bioengineering, Institute of Plasma Physics, Chinese Academy of Sciences, Hefei, Anhui 230031 (China); Wang Qingya [College of Life Science, Nanjing Agricultural University, Nanjing, Jiangsu 210095 (China); Tang Canming [College of Agronomy, Nanjing Agricultural University, Nanjing Jiangsu 210095 (China)], E-mail: tang20@jlonline.com

    2008-09-15

    With the aim to study the effects of an ion beam on plant cells, upland cotton (Gossypium hirsutum L.) cultivar 'Sumian 22' pollen grains were irradiated in vacuum (7.8 x 10{sup -3} Pa) by low-energy nitrogen ions with an energy of 20 keV at various fluences ranging from 0.26 x 10{sup 16} to 0.78 x 10{sup 16} N{sup +}/cm{sup 2}. The irradiation effects on pollen grains were tested, considering the ultrastructural changes in the exine and interior walls of pollen grains, their germination rate, the growth speed of the pollen tubes in the style, fertilization and boll development after the pistils were pollinated by the pollen grains which had been implanted with nitrogen ions. Nitrogen ions entered the pollen grains by etching and penetrating the exine and interior walls and destroying cell structures. A greater percentage of the pollen grains were destroyed as the fluence of N{sup +} ions increased. Obviously, the nitrogen ion beam penetrated the exine and interior walls of the pollen grains and produced holes of different sizes. As the ion fluence increased, the amount and the density of pollen grain inclusions decreased and the size of the lacuna and starch granules increased. Pollen grain germination rates decreased with increasing ion fluence. The number of pollen tubes in the style declined with increased ion implantation into pollen grains, but the growth speed of the tubes did not change. All of the pollen tubes reached the end of the style at 13 h after pollination. This result was consistent with that of the control. Also, the weight and the diameter of the ovary decreased and shortened with increased ion beam implantation fluence. No evident change in the fecundation time of the ovule was observed. These results indicate that nitrogen ions can enter pollen grains and cause a series of biological changes in pollen grains of upland cotton.

  18. The damaging effects of nitrogen ion beam implantation on upland cotton ( Gossypium hirsutum L.) pollen grains

    Science.gov (United States)

    Yu, Yanjie; Wu, Lijun; Wu, Yuejin; Wang, Qingya; Tang, Canming

    2008-09-01

    With the aim to study the effects of an ion beam on plant cells, upland cotton (Gossypium hirsutum L.) cultivar "Sumian 22" pollen grains were irradiated in vacuum (7.8 × 10-3 Pa) by low-energy nitrogen ions with an energy of 20 keV at various fluences ranging from 0.26 × 1016 to 0.78 × 1016 N+/cm2. The irradiation effects on pollen grains were tested, considering the ultrastructural changes in the exine and interior walls of pollen grains, their germination rate, the growth speed of the pollen tubes in the style, fertilization and boll development after the pistils were pollinated by the pollen grains which had been implanted with nitrogen ions. Nitrogen ions entered the pollen grains by etching and penetrating the exine and interior walls and destroying cell structures. A greater percentage of the pollen grains were destroyed as the fluence of N+ ions increased. Obviously, the nitrogen ion beam penetrated the exine and interior walls of the pollen grains and produced holes of different sizes. As the ion fluence increased, the amount and the density of pollen grain inclusions decreased and the size of the lacuna and starch granules increased. Pollen grain germination rates decreased with increasing ion fluence. The number of pollen tubes in the style declined with increased ion implantation into pollen grains, but the growth speed of the tubes did not change. All of the pollen tubes reached the end of the style at 13 h after pollination. This result was consistent with that of the control. Also, the weight and the diameter of the ovary decreased and shortened with increased ion beam implantation fluence. No evident change in the fecundation time of the ovule was observed. These results indicate that nitrogen ions can enter pollen grains and cause a series of biological changes in pollen grains of upland cotton.

  19. A Kinome RNAi Screen in Drosophila Identifies Novel Genes Interacting with Lgl, aPKC, and Crb Cell Polarity Genes in Epithelial Tissues

    NARCIS (Netherlands)

    Parsons, Linda M.; Grzeschik, Nicola A; Amaratunga, Kasun; Burke, Peter; Quinn, Leonie M; Richardson, Helena E

    2017-01-01

    In both Drosophila melanogaster and mammalian systems, epithelial structure and underlying cell polarity are essential for proper tissue morphogenesis and organ growth. Cell polarity interfaces with multiple cellular processes that are regulated by the phosphorylation status of large protein

  20. Fluorescence of Bacteria, Pollens, and Naturally Occurring Airborne Particles: Excitation/Emission Spectra

    National Research Council Canada - National Science Library

    Hill, Steven C; Mayo, Michael W; Chang, Richard K

    2009-01-01

    The fluorescence intensity as a function of excitation and emission wavelengths (EEM spectra) was measured for different species of bacteria, biochemical constituents of cells, pollens, and vegetation...

  1. Comparison of clinical grade type 1 polarized and standard matured dendritic cells for cancer immunotherapy

    DEFF Research Database (Denmark)

    Hansen, Morten; Hjortø, Gertrud Malene; Donia, Marco

    2013-01-01

    induction of type 1 effector T cells. Standard matured clinical grade DCs “sDCs” were compared with DCs matured with either of two type 1 polarizing maturation cocktails; the alpha-type-1 DCs “αDC1s” (TNF-α, IL-1β, IFN-γ, IFN-α, Poly(I:C)) and “mDCs” (monophosphoryl lipid A (MPL), IFN-γ) or a mixed cocktail....... αDC1s and mDCs were functionally superior to sDCs as they polarized naïve CD4+ T cells most efficiently into T helper type 1 effector cells and primed more functional MART-1 specific CD8+ T cells although with variation between donors. αDC1s and mDCs were transiently less capable of CCL21-directed......DCs and strikingly had the highest expression of the inhibitory molecules PD-L1 and CD25. Thus, further studies with type 1 polarized DCs are warranted for use in immunotherapy, but when combined with PGE2 as in mpDCs, they seems to be less optimal for maturation of DCs....

  2. Neuropeptide Y induces potent migration of human immature dendritic cells and promotes a Th2 polarization.

    Science.gov (United States)

    Buttari, Brigitta; Profumo, Elisabetta; Domenici, Giacomo; Tagliani, Angela; Ippoliti, Flora; Bonini, Sergio; Businaro, Rita; Elenkov, Ilia; Riganò, Rachele

    2014-07-01

    Neuropeptide Y (NPY), a major autonomic nervous system and stress mediator, is emerging as an important regulator of inflammation, implicated in autoimmunity, asthma, atherosclerosis, and cancer. Yet the role of NPY in regulating phenotype and functions of dendritic cells (DCs), the professional antigen-presenting cells, remains undefined. Here we investigated whether NPY could induce DCs to migrate, mature, and polarize naive T lymphocytes. We found that NPY induced a dose-dependent migration of human monocyte-derived immature DCs through the engagement of NPY Y1 receptor and the activation of ERK and p38 mitogen-activated protein kinases. NPY promoted DC adhesion to endothelial cells and transendothelial migration. It failed to induce phenotypic DC maturation, whereas it conferred a T helper 2 (Th2) polarizing profile to DCs through the up-regulation of interleukin (IL)-6 and IL-10 production. Thus, during an immune/inflammatory response NPY may exert proinflammatory effects through the recruitment of immature DCs, but it may exert antiinflammatory effects by promoting a Th2 polarization. Locally, at inflammatory sites, cell recruitment could be amplified in conditions of intense acute, chronic, or cold stress. Thus, altered or amplified signaling through the NPY-NPY-Y1 receptor-DC axis may have implications for the development of inflammatory conditions.-Buttari, B., Profumo, E., Domenici, G., Tagliani, A., Ippoliti, F., Bonini, S., Businaro, R., Elenkov, I., Riganò, R. Neuropeptide Y induces potent migration of human immature dendritic cells and promotes a Th2 polarization. © FASEB.

  3. B Cells Producing Type I IFN Modulate Macrophage Polarization in Tuberculosis.

    Science.gov (United States)

    Bénard, Alan; Sakwa, Imme; Schierloh, Pablo; Colom, André; Mercier, Ingrid; Tailleux, Ludovic; Jouneau, Luc; Boudinot, Pierre; Al-Saati, Talal; Lang, Roland; Rehwinkel, Jan; Loxton, Andre G; Kaufmann, Stefan H E; Anton-Leberre, Véronique; O'Garra, Anne; Sasiain, Maria Del Carmen; Gicquel, Brigitte; Fillatreau, Simon; Neyrolles, Olivier; Hudrisier, Denis

    2018-03-15

    In addition to their well-known function as antibody-producing cells, B lymphocytes can markedly influence the course of infectious or noninfectious diseases via antibody-independent mechanisms. In tuberculosis (TB), B cells accumulate in lungs, yet their functional contribution to the host response remains poorly understood. To document the role of B cells in TB in an unbiased manner. We generated the transcriptome of B cells isolated from Mycobacterium tuberculosis (Mtb)-infected mice and validated the identified key pathways using in vitro and in vivo assays. The obtained data were substantiated using B cells from pleural effusion of patients with TB. B cells isolated from Mtb-infected mice displayed a STAT1 (signal transducer and activator of transcription 1)-centered signature, suggesting a role for IFNs in B-cell response to infection. B cells stimulated in vitro with Mtb produced type I IFN, via a mechanism involving the innate sensor STING (stimulator of interferon genes), and antagonized by MyD88 (myeloid differentiation primary response 88) signaling. In vivo, B cells expressed type I IFN in the lungs of Mtb-infected mice and, of clinical relevance, in pleural fluid from patients with TB. Type I IFN expression by B cells induced an altered polarization of macrophages toward a regulatory/antiinflammatory profile in vitro. In vivo, increased provision of type I IFN by B cells in a murine model of B cell-restricted Myd88 deficiency correlated with an enhanced accumulation of regulatory/antiinflammatory macrophages in Mtb-infected lungs. Type I IFN produced by Mtb-stimulated B cells favors macrophage polarization toward a regulatory/antiinflammatory phenotype during Mtb infection.

  4. Coupling Mechanical Deformations and Planar Cell Polarity to Create Regular Patterns in the Zebrafish Retina

    Science.gov (United States)

    Salbreux, Guillaume; Barthel, Linda K.; Raymond, Pamela A.; Lubensky, David K.

    2012-01-01

    The orderly packing and precise arrangement of epithelial cells is essential to the functioning of many tissues, and refinement of this packing during development is a central theme in animal morphogenesis. The mechanisms that determine epithelial cell shape and position, however, remain incompletely understood. Here, we investigate these mechanisms in a striking example of planar order in a vertebrate epithelium: The periodic, almost crystalline distribution of cone photoreceptors in the adult teleost fish retina. Based on observations of the emergence of photoreceptor packing near the retinal margin, we propose a mathematical model in which ordered columns of cells form as a result of coupling between planar cell polarity (PCP) and anisotropic tissue-scale mechanical stresses. This model recapitulates many observed features of cone photoreceptor organization during retinal growth and regeneration. Consistent with the model's predictions, we report a planar-polarized distribution of Crumbs2a protein in cone photoreceptors in both unperturbed and regenerated tissue. We further show that the pattern perturbations predicted by the model to occur if the imposed stresses become isotropic closely resemble defects in the cone pattern in zebrafish lrp2 mutants, in which intraocular pressure is increased, resulting in altered mechanical stress and ocular enlargement. Evidence of interactions linking PCP, cell shape, and mechanical stresses has recently emerged in a number of systems, several of which show signs of columnar cell packing akin to that described here. Our results may hence have broader relevance for the organization of cells in epithelia. Whereas earlier models have allowed only for unidirectional influences between PCP and cell mechanics, the simple, phenomenological framework that we introduce here can encompass a broad range of bidirectional feedback interactions among planar polarity, shape, and stresses; our model thus represents a conceptual framework

  5. Co-occurrence of Artemisia and Ambrosia pollen seasons against the background of the synoptic situations in Poland

    Science.gov (United States)

    Stępalska, Danuta; Myszkowska, Dorota; Katarzyna, Leśkiewicz; Katarzyna, Piotrowicz; Katarzyna, Borycka; Kazimiera, Chłopek; Łukasz, Grewling; Idalia, Kasprzyk; Barbara, Majkowska-Wojciechowska; Małgorzata, Malkiewicz; Małgorzata, Nowak; Krystyna, Piotrowska-Weryszko; Małgorzata, Puc; Elżbieta, Weryszko-Chmielewska

    2017-04-01

    The Asteraceae family is one of the largest families, comprising 67 genera and 264 species in Poland. However, only a few genera, including Artemisia and Ambrosia are potential allergenic sources. The aim of the study was to estimate how often and to what degree Artemisia and Ambrosia pollen seasons co-occur intensifying human health risk, and how synoptic situations influence frequency of days with high pollen concentrations of both taxa. Artemisia and Ambrosia pollen data were collected, using the volumetric method, at 8 sites in Poland. Daily concentrations of Artemisia pollen equal to 30 grains or more and Ambrosia pollen equal to 10 grains or more were accepted as high values. Concentrations of more than 10 pollen grains were defined as high in the case of Ambrosia because its allergenicity is considered higher. High concentrations were confronted with synoptic situations. Analysis was performed on the basis of two calendars on circulation types of atmosphere in Poland (Niedźwiedź, 2006, 2015). Co-occurrence of Artemisia and Ambrosia pollen seasons is being found most often, when Ambrosia pollen season starts in the first half of August. If it happens in the last 10 days of August high pollen concentrations of Artemisia and Ambrosia do not occur at the same days. At three sites (Sosnowiec, Rzeszów, Lublin) high Ambrosia pollen concentrations during the Artemisia pollen season appear more often than in other sites under question. The high Artemisia pollen concentrations occur, when continental or polar maritime old air masses inflow into Poland. The impact of air masses on high Ambrosia pollen concentrations depends on site localizations. It is likely, that in the south-eastern part of Poland high Ambrosia pollen concentrations result from the pollen transport from east-south-south-westerly directions and the local sources. Co-occurrence of both taxa pollen seasons depends on the air masses inflow and appears more often in a south-eastern part of Poland.

  6. RCAN1.4 regulates VEGFR-2 internalisation, cell polarity and migration in human microvascular endothelial cells.

    Science.gov (United States)

    Alghanem, Ahmad F; Wilkinson, Emma L; Emmett, Maxine S; Aljasir, Mohammad A; Holmes, Katherine; Rothermel, Beverley A; Simms, Victoria A; Heath, Victoria L; Cross, Michael J

    2017-08-01

    Regulator of calcineurin 1 (RCAN1) is an endogenous inhibitor of the calcineurin pathway in cells. It is expressed as two isoforms in vertebrates: RCAN1.1 is constitutively expressed in most tissues, whereas transcription of RCAN1.4 is induced by several stimuli that activate the calcineurin-NFAT pathway. RCAN1.4 is highly upregulated in response to VEGF in human endothelial cells in contrast to RCAN1.1 and is essential for efficient endothelial cell migration and tubular morphogenesis. Here, we show that RCAN1.4 has a role in the regulation of agonist-stimulated VEGFR-2 internalisation and establishment of endothelial cell polarity. siRNA-mediated gene silencing revealed that RCAN1 plays a vital role in regulating VEGF-mediated cytoskeletal reorganisation and directed cell migration and sprouting angiogenesis. Adenoviral-mediated overexpression of RCAN1.4 resulted in increased endothelial cell migration. Antisense-mediated morpholino silencing of the zebrafish RCAN1.4 orthologue revealed a disrupted vascular development further confirming a role for the RCAN1.4 isoform in regulating vascular endothelial cell physiology. Our data suggest that RCAN1.4 plays a novel role in regulating endothelial cell migration by establishing endothelial cell polarity in response to VEGF.

  7. On the Relationship between Pollen Size and Genome Size

    Directory of Open Access Journals (Sweden)

    Charles A. Knight

    2010-01-01

    Full Text Available Here we test whether genome size is a predictor of pollen size. If it were, inferences of ancient genome size would be possible using the abundant paleo-palynolgical record. We performed regression analyses across 464 species of pollen width and genome size. We found a significant positive trend. However, regression analysis using phylogentically independent contrasts did not support the correlated evolution of these traits. Instead, a large split between angiosperms and gymnosperms for both pollen width and genome size was revealed. Sister taxa were not more likely to show a positive contrast when compared to deeper nodes. However, significantly more congeneric species had a positive trend than expected by chance. These results may reflect the strong selection pressure for pollen to be small. Also, because pollen grains are not metabolically active when measured, their biology is different than other cells which have been shown to be strongly related to genome size, such as guard cells. Our findings contrast with previously published research. It was our hope that pollen size could be used as a proxy for inferring the genome size of ancient species. However, our results suggest pollen is not a good candidate for such endeavors.

  8. Kv7.1 surface expression is regulated by epithelial cell polarization

    DEFF Research Database (Denmark)

    Andersen, Martin N; Olesen, Søren-Peter; Rasmussen, Hanne Borger

    2011-01-01

    The potassium channel K(V)7.1 is expressed in the heart where it contributes to the repolarization of the cardiac action potential. In addition, K(V)7.1 is expressed in epithelial tissues where it plays a role in salt and water transport. Mutations in the kcnq1 gene can lead to long QT syndrome...... and deafness, and several mutations have been described as trafficking mutations. To learn more about the basic mechanisms that regulate K(V)7.1 surface expression, we have investigated the trafficking of K(V)7.1 during the polarization process of the epithelial cell line Madin-Darby Canine Kidney (MDCK) using...... is regulated by signaling mechanisms involved in epithelial cell polarization in particular signaling cascades involving protein kinase C and PI3K....

  9. Localization of actin in pollen tubes of Ornithogalum virens L.

    Directory of Open Access Journals (Sweden)

    Małgorzata Stępka

    2014-01-01

    Full Text Available The germinating pollen grain (in vivo on the stigma or in vitro in germination medium forms a pollen tube which transports the vegetative nucleus and generative cell/two sperm cells participating in the process of double fertilization. The growth of the tube and the transport of organelles and the cells occur due to two major motor systems existing in the pollen tubes of higher plants: the tubuline-dynein/kinesin and the actin-myosin system. In pollen tubes of Ornithogalum virens the actin filaments were labelled with TRITC-phalloidin (2 µg/ml in the PIPES buffer and the 10% sucrose, without the fixative and DMSO. Omission of the fixative and permeabilizing agent (DMSO allowed better preservation of the structure, and the "fluorescence" of actin was observed in living pollen tubes. Observations in CLSM (confocal laser scanning microscope showed that actin is distributed in the vicinity of the cell membrane. This could support the view that actin filaments and the plasmalemma form the pollen tube cortex along which the cytoplasmic movement of organelles, and cell transport occurs.

  10. Characterization of a pancreatic islet cell tumor in a polar bear (Ursus maritimus).

    Science.gov (United States)

    Fortin, Jessica S; Benoit-Biancamano, Marie-Odile

    2014-01-01

    Herein, we report a 25-year-old male polar bear suffering from a pancreatic islet cell tumor. The aim of this report is to present a case of this rare tumor in a captive polar bear. The implication of potential risk factors such as high carbohydrate diet or the presence of amyloid fibril deposits was assessed. Necropsy examination revealed several other changes, including nodules observed in the liver, spleen, pancreas, intestine, and thyroid glands that were submitted for histopathologic analysis. Interestingly, the multiple neoplastic nodules were unrelated and included a pancreatic islet cell tumor. Immunohistochemistry of the pancreas confirmed the presence of insulin and islet amyloid polypeptide (IAPP) within the pancreatic islet cells. The IAPP gene was extracted from the paraffin-embedded liver tissue and sequenced. IAPP cDNA from the polar bear exhibits some differences as compared to the sequence published for several other species. Different factors responsible for neoplasms in bears such as diet, infectious agents, and industrial chemical exposure are reviewed. This case report raised several issues that further studies may address by evaluating the prevalence of cancers in captive or wild animals. © 2014 Wiley Periodicals, Inc.

  11. Self-organized spatiotemporal patterns of PIP3 and PTEN during spontaneous cell polarization

    Science.gov (United States)

    Knoch, Fabian; Tarantola, Marco; Rappel, Wouter-Jan; Bodenschatz, Eberhard

    2014-03-01

    During spontaneous cell polarization of Dictyostelium discoideum cells, PIP3 (phosphatidylinositol (3,4,5)-triphoshpate) and PTEN (phosphatase tensin homolog) have been identified as key signaling molecules, which govern the process of polarization in a self-organized manner. Gerisch et al. have shown that randomly triggered excitable PIP3 waves regulate the anti-correlated PTEN concentration. Here we show that this requires a switch-like dynamics of the overall membrane bound PTEN concentration in combination with two species of PTEN differing in their dephosphorylation rates. A quantitative modeling with a coupled reaction-diffusion system shows excellent agreement with experimental results and predicts a ratio σ of dephosphorylation rates acting on PIP3 of σ ~ 80 - 100. Our quantitative analysis suggests that surface-attached cell membrane spanning PIP3 waves are necessary for resetting the global actin network. This is evidenced by the experimentally observed delay between polarization-cycles also quantitatively captured by our analysis. Max Planck Society and Center for Theoretical Biological Physics.

  12. Ubisch bodies and pollen ontogeny in Oxalis articulata Savigny.

    Science.gov (United States)

    Rosenfeldt, Sonia; Galati, Beatriz G

    2005-12-01

    The correlation between the ontogeny of Ubisch bodies and pollen development in Oxalis articulata was studied with Transmission Electron Microscopy (TEM). The ultrastructural changes observed during the different stages of development in the tapetal cells are related to Ubisch bodies, sporopollenin and pollen-kitt formation. The pro-orbicules have the appearance of lipid globuli and their formation is related to the endoplasmic reticulum of rough type (ERr). The lipid globules or pro-orbicules disappear in the mature Ubisch bodies, and the places that they occupied remain free of contents or with pollen-kitt.

  13. Effect of Toxic Components on Microbial Fuel Cell-Polarization Curves and Estimation of the Type of Toxic Inhibition

    NARCIS (Netherlands)

    Stein, N.E.; Hamelers, H.V.M.; Straten, van G.; Keesman, K.J.

    2012-01-01

    Polarization curves are of paramount importance for the detection of toxic components in microbial fuel cell (MFC) based biosensors. In this study, polarization curves were made under non-toxic conditions and under toxic conditions after the addition of various concentrations of nickel, bentazon,

  14. Effect of toxic components on microbial fuel cell-polarization curves and estimation of the type of toxic inhibition

    NARCIS (Netherlands)

    Stein, N.E.; Hamelers, H.V.M.; Straten, G. van; Keesman, K.J.

    2012-01-01

    Polarization curves are of paramount importance for the detection of toxic components in microbial fuel cell (MFC) based biosensors. In this study, polarization curves were made under non-toxic conditions and under toxic conditions after the addition of various concentrations of nickel, bentazon,

  15. Polarity of fatty acid uptake and metabolism in a human intestinal cell line (CACO-2)

    Energy Technology Data Exchange (ETDEWEB)

    Trotter, P.J.; Storch, J. (Harvard School of Public Health, Boston, MA (United States))

    1990-02-26

    Free fatty acids (ffa) can enter the intestinal cell via the apical (AP) or basolateral (BL) membrane. The authors are using the Caco-2 intestinal cell line to examine the polarity of ffa uptake and metabolism in the enterocyte. Cells are grown on permeable polycarbonate Transwell filters in order to obtain access to both AP and BL compartments. Differentiated Caco-2 cells form tight polarized monolayers which express small intestine-specific enzymes and are impermeable to the fluid phase marker Lucifer Yellow. Submicellar concentrations of {sup 3}H-palmitic acid (2uM) were added to AP or BL sides of Caco-2 monolayers at 37{degrees}C and cells were incubated for various times between 2 and 120 minutes. Total AP and BL uptake is similar; however, when relative membrane surface areas are accounted for, AP uptake is about 2-fold higher. The metabolism of AP and BL ffa is not significantly different: triacylglycerol and phosphatidylcholine account for most of the metabolites (32{plus minus}4 and 24{plus minus}2% respectively at 5 minutes). Little ffa oxidation is observed. Preincubation with albumin-bound 2-monoolein (100uM) and palmitate (50uM) increases the level of TG metabolites. The results suggest that in this cell line the uptake of AP ffa may be greater than BL ffa, but that AP (dietary) ffa and BL (plasma) ffa are metabolized similarly.

  16. Polarity of fatty acid uptake and metabolism in a human intestinal cell line (CACO-2)

    International Nuclear Information System (INIS)

    Trotter, P.J.; Storch, J.

    1990-01-01

    Free fatty acids (ffa) can enter the intestinal cell via the apical (AP) or basolateral (BL) membrane. The authors are using the Caco-2 intestinal cell line to examine the polarity of ffa uptake and metabolism in the enterocyte. Cells are grown on permeable polycarbonate Transwell filters in order to obtain access to both AP and BL compartments. Differentiated Caco-2 cells form tight polarized monolayers which express small intestine-specific enzymes and are impermeable to the fluid phase marker Lucifer Yellow. Submicellar concentrations of 3 H-palmitic acid (2uM) were added to AP or BL sides of Caco-2 monolayers at 37 degrees C and cells were incubated for various times between 2 and 120 minutes. Total AP and BL uptake is similar; however, when relative membrane surface areas are accounted for, AP uptake is about 2-fold higher. The metabolism of AP and BL ffa is not significantly different: triacylglycerol and phosphatidylcholine account for most of the metabolites (32±4 and 24±2% respectively at 5 minutes). Little ffa oxidation is observed. Preincubation with albumin-bound 2-monoolein (100uM) and palmitate (50uM) increases the level of TG metabolites. The results suggest that in this cell line the uptake of AP ffa may be greater than BL ffa, but that AP (dietary) ffa and BL (plasma) ffa are metabolized similarly

  17. Polarity, cell division, and out-of-equilibrium dynamics control the growth of epithelial structures

    Science.gov (United States)

    Cerruti, Benedetta; Puliafito, Alberto; Shewan, Annette M.; Yu, Wei; Combes, Alexander N.; Little, Melissa H.; Chianale, Federica; Primo, Luca; Serini, Guido; Mostov, Keith E.; Celani, Antonio

    2013-01-01

    The growth of a well-formed epithelial structure is governed by mechanical constraints, cellular apico-basal polarity, and spatially controlled cell division. Here we compared the predictions of a mathematical model of epithelial growth with the morphological analysis of 3D epithelial structures. In both in vitro cyst models and in developing epithelial structures in vivo, epithelial growth could take place close to or far from mechanical equilibrium, and was determined by the hierarchy of time-scales of cell division, cell–cell rearrangements, and lumen dynamics. Equilibrium properties could be inferred by the analysis of cell–cell contact topologies, and the nonequilibrium phenotype was altered by inhibiting ROCK activity. The occurrence of an aberrant multilumen phenotype was linked to fast nonequilibrium growth, even when geometric control of cell division was correctly enforced. We predicted and verified experimentally that slowing down cell division partially rescued a multilumen phenotype induced by altered polarity. These results improve our understanding of the development of epithelial organs and, ultimately, of carcinogenesis. PMID:24145168

  18. Co-regulation of cell polarization and migration by caveolar proteins PTRF/Cavin-1 and caveolin-1.

    Directory of Open Access Journals (Sweden)

    Michelle M Hill

    Full Text Available Caveolin-1 and caveolae are differentially polarized in migrating cells in various models, and caveolin-1 expression has been shown to quantitatively modulate cell migration. PTRF/cavin-1 is a cytoplasmic protein now established to be also necessary for caveola formation. Here we tested the effect of PTRF expression on cell migration. Using fluorescence imaging, quantitative proteomics, and cell migration assays we show that PTRF/cavin-1 modulates cellular polarization, and the subcellular localization of Rac1 and caveolin-1 in migrating cells as well as PKCα caveola recruitment. PTRF/cavin-1 quantitatively reduced cell migration, and induced mesenchymal epithelial reversion. Similar to caveolin-1, the polarization of PTRF/cavin-1 was dependent on the migration mode. By selectively manipulating PTRF/cavin-1 and caveolin-1 expression (and therefore caveola formation in multiple cell systems, we unveil caveola-independent functions for both proteins in cell migration.

  19. Characteristics of anodic polarization of solid oxide fuel cells under pressurized conditions

    Energy Technology Data Exchange (ETDEWEB)

    Kikuchi, Ryuji; Yano, Tatsuya; Eguchi, Koichi [Department of Energy and Hydrocarbon Chemistry, Graduate School of Engineering, Kyoto University, Nishikyo-ku, Kyoto 615-8510 (Japan); Takeguchi, Tatsuya [Catalysis Research Center, Hokkaido University, Kita-ku, Sapporo 001-0021 (Japan)

    2004-10-29

    AC impedance measurements were carried out under pressurized conditions by using a Ni-Y{sub 2}O{sub 3}-stabilized zirconia (YSZ)/YSZ half cell in order to investigate anodic polarization at high-pressure conditions. AC impedance spectra were measured at 900 and 1000C in H{sub 2}-H{sub 2}O system with a constant H{sub 2}/H{sub 2}O ratio, or a constant partial pressure of H{sub 2} or H{sub 2}O for different total pressures of 1 to 10 atm. At high pressures, the resistance characterized by the semicircle at high frequency was lowered, whereas that at low frequency was raised. A model based on one-dimensional diffusion was developed to estimate concentration polarization based on the impedance measurements, and activation polarization was evaluated using a linear current-potential relation derived from the Butler-Volmer equation. The activation overvoltage was at most 40 mV at 10 mA/cm{sup 2}, irrespective of the total pressure. Concentration polarization was computed to increase as the total pressure was raised, whereas it was almost constant for temperature change. Large voltage drop at small current densities was calculated for the system with low partial pressure of oxygen.

  20. The C-terminal hypervariable domain targets Aradopsis ROP9 to the invaginated pollen tube plasma membrane

    Science.gov (United States)

    Rop9 is a small GTPase of the Type II class, whereas the often studied type I Rops play roles during pollen tube growth. In pollen, Rop9 is located at the invaginated plasma membrane that surrounds the sperm cells, whereas type I Rops are located at the apical membrane of the pollen tube. The C-ter...

  1. Human B cells induce dendritic cell maturation and favour Th2 polarization by inducing OX-40 ligand

    Science.gov (United States)

    Maddur, Mohan S.; Sharma, Meenu; Hegde, Pushpa; Stephen-Victor, Emmanuel; Pulendran, Bali; Kaveri, Srini V.; Bayry, Jagadeesh

    2015-01-01

    Dendritic cells (DCs) play a critical role in immune homeostasis by regulating the functions of various immune cells, including T and B cells. Notably, DCs also undergo education on reciprocal signalling by these immune cells and environmental factors. Various reports demonstrated that B cells have profound regulatory functions, although only few reports have explored the regulation of human DCs by B cells. Here we demonstrate that activated but not resting B cells induce maturation of DCs with distinct features to polarize Th2 cells that secrete interleukin (IL)-5, IL-4 and IL-13. B-cell-induced maturation of DCs is contact dependent and implicates signalling of B-cell activation molecules CD69, B-cell-activating factor receptor, and transmembrane activator and calcium-modulating cyclophilin ligand interactor. Mechanistically, differentiation of Th2 cells by B-cell-matured DCs is dependent on OX-40 ligand. Collectively, our results suggest that B cells have the ability to control their own effector functions by enhancing the ability of human DCs to mediate Th2 differentiation. PMID:24910129

  2. NKp46 clusters at the immune synapse and regulates NK cell polarization

    Directory of Open Access Journals (Sweden)

    Uzi eHadad

    2015-09-01

    Full Text Available Natural killer cells play an important role in first-line defense against tumor and virus-infected cells. The activity of NK cells is tightly regulated by a repertoire of cell-surface expressed inhibitory and activating receptors. NKp46 is a major NK cell activating receptor that is involved in the elimination of target cells. NK cells form different types of synapses that result in distinct functional outcomes: cytotoxic, inhibitory, and regulatory. Recent studies revealed that complex integration of NK receptor signaling controls cytoskeletal rearrangement and other immune synapse-related events. However the distinct nature by which NKp46 participates in NK immunological synapse formation and function remains unknown. In this study we determined that NKp46 forms microclusters structures at the immune synapse between NK cells and target cells. Over-expression of human NKp46 is correlated with increased accumulation of F-actin mesh at the immune synapse. Concordantly, knock-down of NKp46 in primary human NK cells decreased recruitment of F-actin to the synapse. Live cell imaging experiments showed a linear correlation between NKp46 expression and lytic granules polarization to the immune synapse. Taken together, our data suggest that NKp46 signaling directly regulates the NK lytic immune synapse from early formation to late function.

  3. All-optical clocked flip-flops and random access memory cells using the nonlinear polarization rotation effect of low-polarization-dependent semiconductor optical amplifiers

    Science.gov (United States)

    Wang, Yongjun; Liu, Xinyu; Tian, Qinghua; Wang, Lina; Xin, Xiangjun

    2018-03-01

    Basic configurations of various all-optical clocked flip-flops (FFs) and optical random access memory (RAM) based on the nonlinear polarization rotation (NPR) effect of low-polarization-dependent semiconductor optical amplifiers (SOA) are proposed. As the constituent elements, all-optical logic gates and all-optical SR latches are constructed by taking advantage of the SOA's NPR switch. Different all-optical FFs (AOFFs), including SR-, D-, T-, and JK-types as well as an optical RAM cell were obtained by the combination of the proposed all-optical SR latches and logic gates. The effectiveness of the proposed schemes were verified by simulation results and demonstrated by a D-FF and 1-bit RAM cell experimental system. The proposed all-optical clocked FFs and RAM cell are significant to all-optical signal processing.

  4. Evolutionarily conserved sites in yeast tropomyosin function in cell polarity, transport and contractile ring formation

    Directory of Open Access Journals (Sweden)

    Susanne Cranz-Mileva

    2015-08-01

    Full Text Available Tropomyosin is a coiled-coil protein that binds and regulates actin filaments. The tropomyosin gene in Schizosaccharomyces pombe, cdc8, is required for formation of actin cables, contractile rings, and polar localization of actin patches. The roles of conserved residues were investigated in gene replacement mutants. The work validates an evolution-based approach to identify tropomyosin functions in living cells and sites of potential interactions with other proteins. A cdc8 mutant with near-normal actin affinity affects patch polarization and vacuole fusion, possibly by affecting Myo52p, a class V myosin, function. The presence of labile residual cell attachments suggests a delay in completion of cell division and redistribution of cell patches following cytokinesis. Another mutant with a mild phenotype is synthetic negative with GFP-fimbrin, inferring involvement of the mutated tropomyosin sites in interaction between the two proteins. Proteins that assemble in the contractile ring region before actin do so in a mutant cdc8 strain that cannot assemble condensed actin rings, yet some cells can divide. Of general significance, LifeAct-GFP negatively affects the actin cytoskeleton, indicating caution in its use as a biomarker for actin filaments.

  5. Mycobacterium tuberculosislpdC, Rv0462, induces dendritic cell maturation and Th1 polarization

    Energy Technology Data Exchange (ETDEWEB)

    Heo, Deok Rim [Department of Microbiology and Immunology, School of Medicine, Pusan National University, Beom-eo Ri, Mulgum Eop, Yangsan, Gyeongsangnam-do 626-770 (Korea, Republic of); Shin, Sung Jae; Kim, Woo Sik [Department of Microbiology, College of Medicine, Chungnam National University, Munwha-Dong, Jung-Ku, Daejeon 301-747 (Korea, Republic of); Noh, Kyung Tae; Park, Jin Wook; Son, Kwang Hee [Department of Microbiology and Immunology, School of Medicine, Pusan National University, Beom-eo Ri, Mulgum Eop, Yangsan, Gyeongsangnam-do 626-770 (Korea, Republic of); Park, Won Sun [Department of Physiology, Kangwon National University, School of Medicine, Chuncheon 200-701 (Korea, Republic of); Lee, Min-Goo [Department of Physiology, Korea University, College of Medicine, Anam-dong, Sungbuk-Gu, Seoul 136-705 (Korea, Republic of); Kim, Daejin [Department of Anatomy, Chung-Ang University, College of Medicine, 221 Heuksuk-Dong, Dongjak-Ku, Seoul 156-756 (Korea, Republic of); Shin, Yong Kyoo [Department of Pharmacology, Chung-Ang University, College of Medicine, 221 Heuksuk-Dong, Dongjak-Ku, Seoul 156-756 (Korea, Republic of); Jung, In Duk, E-mail: jungid@pusan.ac.kr [Department of Microbiology and Immunology, School of Medicine, Pusan National University, Beom-eo Ri, Mulgum Eop, Yangsan, Gyeongsangnam-do 626-770 (Korea, Republic of); Research Institute of Convergence of Biomedical Science and Technology, Pusan National University Yangsan Hospital, Beom-eo Ri, Mulgum Eop, Yangsan, Gyeongsangnam-do 626-770 (Korea, Republic of); Park, Yeong-Min, E-mail: immunpym@pusan.ac.kr [Department of Microbiology and Immunology, School of Medicine, Pusan National University, Beom-eo Ri, Mulgum Eop, Yangsan, Gyeongsangnam-do 626-770 (Korea, Republic of); Research Institute of Convergence of Biomedical Science and Technology, Pusan National University Yangsan Hospital, Beom-eo Ri, Mulgum Eop, Yangsan, Gyeongsangnam-do 626-770 (Korea, Republic of)

    2011-08-05

    Highlights: {yields} Treatment with Rv0462 induces the expression of surface molecules and the production of cytokines in DCs. {yields} Rv0462 induces the activation of MAPKs. {yields} Rv0462-treated DCs enhances the proliferation of CD4{sup +} T cells. -- Abstract: Mycobacterium tuberculosis, the etiological factor of pulmonary tuberculosis, causes significant morbidity and mortality worldwide. Activation of host immune responses for containment of mycobacterial infections involves participation of innate immune cells, such as dendritic cells (DCs). In this study, we demonstrated that the gene encoding lipoamide dehydrogenase C (lpdC) from M. tuberculosis, Rv0462, induce maturation and activation of DCs involved in the MAPKs signaling pathway. Moreover, Rv0462-treated DCs activated naive T cells, polarized CD4{sup +} and CD8{sup +} T cells to secrete IFN-{gamma} in syngeneic mixed lymphocyte reactions, which would be expected to contribute to Th1 polarization of the immune response. Our results suggest that Rv0462 can contribute to the innate and adaptive immune responses during tuberculosis infection, and thus modulate the clinical course of tuberculosis.

  6. Dystroglycan loss disrupts polarity and beta-casein induction inmammary epithelial cells by perturbing laminin anchoring

    Energy Technology Data Exchange (ETDEWEB)

    Weir, M. Lynn; Oppizzi, Maria Luisa; Henry, Michael D.; Onishi,Akiko; Campbell, Kevin P.; Bissell, Mina J.; Muschler, John L.

    2006-02-17

    Precise contact between epithelial cells and their underlying basement membrane is critical to the maintenance of tissue architecture and function. To understand the role that the laminin receptor dystroglycan (DG) plays in these processes, we assayed cell responses to laminin-111 following conditional ablation of DG expression in cultured mammary epithelial cells (MECs). Strikingly, DG loss disrupted laminin-111-induced polarity and {beta}-casein production, and abolished laminin assembly at the step of laminin binding to the cell surface. DG re-expression restored these deficiencies. Investigations of mechanism revealed that DG cytoplasmic sequences were not necessary for laminin assembly and signaling, and only when the entire mucin domain of extracellular DG was deleted did laminin assembly not occur. These results demonstrate that DG is essential as a laminin-111 co-receptor in MECs that functions by mediating laminin anchoring to the cell surface, a process that allows laminin polymerization, tissue polarity, and {beta}-casein induction. The observed loss of laminin-111 assembly and signaling in DG-/-MECs provides insights into the signaling changes occurring in breast carcinomas and other cancers, where DG's laminin-binding function is frequently defective.

  7. T cell receptor reversed polarity recognition of a self-antigen major histocompatibility complex.

    Science.gov (United States)

    Beringer, Dennis X; Kleijwegt, Fleur S; Wiede, Florian; van der Slik, Arno R; Loh, Khai Lee; Petersen, Jan; Dudek, Nadine L; Duinkerken, Gaby; Laban, Sandra; Joosten, Antoinette; Vivian, Julian P; Chen, Zhenjun; Uldrich, Adam P; Godfrey, Dale I; McCluskey, James; Price, David A; Radford, Kristen J; Purcell, Anthony W; Nikolic, Tatjana; Reid, Hugh H; Tiganis, Tony; Roep, Bart O; Rossjohn, Jamie

    2015-11-01

    Central to adaptive immunity is the interaction between the αβ T cell receptor (TCR) and peptide presented by the major histocompatibility complex (MHC) molecule. Presumably reflecting TCR-MHC bias and T cell signaling constraints, the TCR universally adopts a canonical polarity atop the MHC. We report the structures of two TCRs, derived from human induced T regulatory (iT(reg)) cells, complexed to an MHC class II molecule presenting a proinsulin-derived peptide. The ternary complexes revealed a 180° polarity reversal compared to all other TCR-peptide-MHC complex structures. Namely, the iT(reg) TCR α-chain and β-chain are overlaid with the α-chain and β-chain of MHC class II, respectively. Nevertheless, this TCR interaction elicited a peptide-reactive, MHC-restricted T cell signal. Thus TCRs are not 'hardwired' to interact with MHC molecules in a stereotypic manner to elicit a T cell signal, a finding that fundamentally challenges our understanding of TCR recognition.

  8. Impact of cytomixis on meiosis, pollen viability and pollen size in ...

    Indian Academy of Sciences (India)

    Srinivas

    29. Ashraf M and Gohil R N 1994 Cytology of legumes of Kashmir. Himalaya V. Cytomixis and chromosome migration in. Astragalus subuliformis DC; Nucleus 37 119–122. Bahl J R and Tyagi B R 1988 Cytomixis in pollen mother cells of.

  9. CLAMP/Spef1 regulates planar cell polarity signaling and asymmetric microtubule accumulation in the Xenopus ciliated epithelia.

    Science.gov (United States)

    Kim, Sun K; Zhang, Siwei; Werner, Michael E; Brotslaw, Eva J; Mitchell, Jennifer W; Altabbaa, Mohamed M; Mitchell, Brian J

    2018-03-07

    Most epithelial cells polarize along the axis of the tissue, a feature known as planar cell polarity (PCP). The initiation of PCP requires cell-cell signaling via the noncanonical Wnt/PCP pathway. Additionally, changes in the cytoskeleton both facilitate and reflect this polarity. We have identified CLAMP/Spef1 as a novel regulator of PCP signaling. In addition to decorating microtubules (MTs) and the ciliary rootlet, a pool of CLAMP localizes at the apical cell cortex. Depletion of CLAMP leads to the loss of PCP protein asymmetry, defects in cilia polarity, and defects in the angle of cell division. Additionally, depletion of CLAMP leads to a loss of the atypical cadherin-like molecule Celrs2, suggesting that CLAMP facilitates the stabilization of junctional interactions responsible for proper PCP protein localization. Depletion of CLAMP also affects the polarized organization of MTs. We hypothesize that CLAMP facilitates the establishment of cell polarity and promotes the asymmetric accumulation of MTs downstream of the establishment of proper PCP. © 2018 Kim et al.

  10. Therapeutic effect of bee pollens on acute radiation sickness

    International Nuclear Information System (INIS)

    Wang Mingsuo; Huang Chaoqun; Chen Zhen; Huang Meiying; Jiang Ying; Wang Tao

    1997-09-01

    The therapeutic effect of bee pollens on acute radiation sickness were evaluated by observing the changes in the peripheral white blood cell (PWBC) count, the total activity of superoxide dismutase (SOD) and the levels of lipid peroxides (LPO) in sera of the irradiated rats following P.O. administration of bee pollens. It was found that bee pollens could remarkably help irradiated rats recover from radiation-induced injury. The functions of bee pollens might be summarized as follows: (1) Stimulating Proliferation of PWBC. The PWBC count of the bee pollens group showed no significant difference as compared with the normal control group on the 30 th day postirradiation. (2) Enhancing antioxidative effect of clearing free radicals. The total activity of serum SOD in the bee pollens group increased by 6.48% as compared with the normal control group on the 30 th day after irradiation, and the LPO levels i.e. MDA and POV in sera of the irradiated rats decreased by 54.73% and 21.60% respectively. The result suggests that using bee pollens as antiradiation and health-promoting agents in clinical treatment of acute radiation sickness and during radiotherapy of patients with tumors may has certain practical value. (12 refs., 2 figs., 2 tabs.)

  11. Simultaneously improving optical absorption of both transverse-electric polarized and transverse-magnetic polarized light for organic solar cells with Ag grating used as transparent electrode

    Directory of Open Access Journals (Sweden)

    Yongbing Long

    2014-08-01

    Full Text Available Theoretical simulations are performed to investigate optical performance of organic solar cells with Ag grating electrode. It is demonstrated that optical absorption for both transverse-electric (TE polarized and transverse-magnetic(TM polarized light is simultaneously improved when compared with that for the device without the Ag grating. The improvement is respectively attributed to the resonance and the surface plasmon polaritons within the device. After an additional WO3 layer is capped on the Ag grating, absorption of TE-polarized light is further improved due to resonance of double microcavities within the device, and absorption of TM-polarized light is improved by the combined effects of the microcavity resonance and the surface plasmon polaritons. Correspondingly, the short current density for randomly polarized light is improved by 18.1% from that of the device without the Ag grating. Finally, it is demonstrated that high transmission may not be an essential prerequisite for metallic gratings when they are used as transparent electrode since absorption loss caused by low transmission can be compensated by using a capping layer to optimize optical resonance of the WMC structure within the device.

  12. Kermit interacts with Gαo, Vang, and motor proteins in Drosophila planar cell polarity.

    Directory of Open Access Journals (Sweden)

    Chen Lin

    Full Text Available In addition to the ubiquitous apical-basal polarity, epithelial cells are often polarized within the plane of the tissue--the phenomenon known as planar cell polarity (PCP. In Drosophila, manifestations of PCP are visible in the eye, wing, and cuticle. Several components of the PCP signaling have been characterized in flies and vertebrates, including the heterotrimeric Go protein. However, Go signaling partners in PCP remain largely unknown. Using a genetic screen we uncover Kermit, previously implicated in G protein and PCP signaling, as a novel binding partner of Go. Through pull-down and genetic interaction studies, we find that Kermit interacts with Go and another PCP component Vang, known to undergo intracellular relocalization during PCP establishment. We further demonstrate that the activity of Kermit in PCP differentially relies on the motor proteins: the microtubule-based dynein and kinesin motors and the actin-based myosin VI. Our results place Kermit as a potential transducer of Go, linking Vang with motor proteins for its delivery to dedicated cellular compartments during PCP establishment.

  13. Spontaneous cell polarization: Feedback control of Cdc42 GTPase breaks cellular symmetry.

    Science.gov (United States)

    Martin, Sophie G

    2015-11-01

    Spontaneous polarization without spatial cues, or symmetry breaking, is a fundamental problem of spatial organization in biological systems. This question has been extensively studied using yeast models, which revealed the central role of the small GTPase switch Cdc42. Active Cdc42-GTP forms a coherent patch at the cell cortex, thought to result from amplification of a small initial stochastic inhomogeneity through positive feedback mechanisms, which induces cell polarization. Here, I review and discuss the mechanisms of Cdc42 activity self-amplification and dynamic turnover. A robust Cdc42 patch is formed through the combined effects of Cdc42 activity promoting its own activation and active Cdc42-GTP displaying reduced membrane detachment and lateral diffusion compared to inactive Cdc42-GDP. I argue the role of the actin cytoskeleton in symmetry breaking is not primarily to transport Cdc42 to the active site. Finally, negative feedback and competition mechanisms serve to control the number of polarization sites. © 2015 WILEY Periodicals, Inc.

  14. Grass Pollen Allergens

    Science.gov (United States)

    Augustin, Rosa

    1959-01-01

    Heat and pH stability studies and experiments with organic solvents show that the A-antigens discussed in the preceding paper (Augustin, 1959c) are much more labile than the I- (`inner ring') antigens. Breakdown products and/or aggregates are produced which no longer precipitate with antisera to the original extracts, but act as inhibitors. Solutions of pollen allergens, on the other hand, are found to withstand even autoclaving for 15 min. at 20 atm. and vigorous boiling over the naked flame of a bunsen burner. None of the carbohydrates tested has a demonstrable effect on skin reactivity which is, however, destroyed by crystalline pepsin, crystalline trypsin, a crystalline mould protease and a tissue protease (a partially purified extract from rabbit spleen). It follows that the bulk of the allergens—if not all—are proteins. The relation of skin reactivity, immuno-electrophoretic patterns, carbohydrate and protein reactions to the selective destruction of the pollen antigens is investigated. Pollen components prove to have a somewhat wider range of electrophoretic mobilities than serum proteins and are probably as complicated a mixture. The most and least highly negatively charged components are without skin reactivity in allergic subjects. The skin reactive allergens appear to have the mobilities of α- and β-globulins. Not all the hay fever subjects react equally to all the components, and Cocksfoot and Timothy activity patterns vary in different subjects. ImagesFIG. 5 PMID:13795119

  15. Cancer-associated fibroblasts as another polarized cell type of the tumor microenvironment

    Directory of Open Access Journals (Sweden)

    Martin eAugsten

    2014-03-01

    Full Text Available Tumor- or cancer-associated fibroblasts (CAFs are one of the most abundant stromal cell types in different carcinomas and comprise a heterogeneous cell population. Classically, CAFs are assigned with pro-tumorigenic effects stimulating tumor growth and progression. More recent studies demonstrated also tumor-inhibitory effects of CAFs suggesting that tumor-residing fibroblasts exhibit a similar degree of plasticity as other stromal cell types. Reciprocal interactions with the tumor milieu and different sources of origin are emerging as two important factors underlying CAF heterogeneity. This review highlights recent advances in our understanding of CAF biology and proposes to expand the term of cellular ´polarization´, previously introduced to describe different activation states of various immune cells, onto CAFs to reflect their phenotypic diversity.

  16. Hydrodynamic instabilities and concentration polarization coupled by osmotic pressure in a Taylor-Couette cell

    Science.gov (United States)

    Martinand, Denis; Tilton, Nils

    2016-11-01

    This study addresses analytically and numerically the coupling between hydrodynamic instabilities and osmotic pressure driven by concentration polarization. The configuration consists of a Taylor-Couette cell filled with a Newtonian fluid carrying a passive scalar. Whereas the concentric inner and outer cylinders are membranes permeable to the solvent, they totally reject the scalar. As a radial in- or outflow of solvent is imposed through both cylinders, a concentration boundary layer develops on the cylinder where the solvent exits, until an equilibrium steady state is reached. In addition, the rotation of the inner cylinder is used to drive centrifugal instabilities in the form of toroidal vortices, which interact with the concentration boundary layer. By means of the osmotic pressure, concentration polarization is found to promote or hinder the hydrodynamic instabilities, depending on capacity of the vortices and diffusion to increase the concentration field at the membrane. The results obtained by analytical stability analysis agree with dedicated Direct Numerical Simulations.

  17. A Kinome RNAi Screen inDrosophilaIdentifies Novel Genes Interacting with Lgl, aPKC, and Crb Cell Polarity Genes in Epithelial Tissues.

    Science.gov (United States)

    Parsons, Linda M; Grzeschik, Nicola A; Amaratunga, Kasun; Burke, Peter; Quinn, Leonie M; Richardson, Helena E

    2017-08-07

    In both Drosophila melanogaster and mammalian systems, epithelial structure and underlying cell polarity are essential for proper tissue morphogenesis and organ growth. Cell polarity interfaces with multiple cellular processes that are regulated by the phosphorylation status of large protein networks. To gain insight into the molecular mechanisms that coordinate cell polarity with tissue growth, we screened a boutique collection of RNAi stocks targeting the kinome for their capacity to modify Drosophila "cell polarity" eye and wing phenotypes. Initially, we identified kinase or phosphatase genes whose depletion modified adult eye phenotypes associated with the manipulation of cell polarity complexes (via overexpression of Crb or aPKC). We next conducted a secondary screen to test whether these cell polarity modifiers altered tissue overgrowth associated with depletion of Lgl in the wing. These screens identified Hippo, Jun kinase (JNK), and Notch signaling pathways, previously linked to cell polarity regulation of tissue growth. Furthermore, novel pathways not previously connected to cell polarity regulation of tissue growth were identified, including Wingless (Wg/Wnt), Ras, and lipid/Phospho-inositol-3-kinase (PI3K) signaling pathways. Additionally, we demonstrated that the "nutrient sensing" kinases Salt Inducible Kinase 2 and 3 ( SIK2 and 3 ) are potent modifiers of cell polarity phenotypes and regulators of tissue growth. Overall, our screen has revealed novel cell polarity-interacting kinases and phosphatases that affect tissue growth, providing a platform for investigating molecular mechanisms coordinating cell polarity and tissue growth during development. Copyright © 2017 Parsons et al.

  18. Arabidopsis thaliana CML25 mediates the Ca(2+) regulation of K(+) transmembrane trafficking during pollen germination and tube elongation.

    Science.gov (United States)

    Wang, Shuang-Shuang; Diao, Wen-Zhu; Yang, Xue; Qiao, Zhu; Wang, Mei; Acharya, Biswa R; Zhang, Wei

    2015-11-01

    The concentration alteration of cytosolic-free calcium ([Ca(2+) ]cyt ) is a well-known secondary messenger in plants and plays important roles during pollen grain germination and tube elongation. Here we demonstrate that CML25, a member of calmodulin-like proteins, has Ca(2+) -binding activity and plays a role in pollen grain germination, tube elongation and seed setting. CML25 transcript was abundant in mature pollen grains and pollen tubes, and its product CML25 protein was primarily directed to the cytoplasm. Two independent CML25 loss-of-function T-DNA insertion mutants suffered a major reduction in both the rate of pollen germination and the elongation of the pollen tube. Also, pollen grains of cml25 mutants were less sensitive to the external K(+) and Ca(2+) concentration than wild-type pollen. The disruption of CML25 increased the [Ca(2+) ]cyt in both the pollen grain and the pollen tube, which in turn impaired the Ca(2+) -dependent inhibition of whole-cell inward K(+) currents in protoplasts prepared from these materials (pollen grain and pollen tube). Complementation of cml25-1 mutant resulted in the recovery of wild-type phenotype. Our findings indicate that CML25 is an important transducer in the Ca(2+) -mediated regulation of K(+) influx during pollen germination and tube elongation. © 2015 John Wiley & Sons Ltd.

  19. Cor a 1-reactive T cells and IgE are predominantly cross-reactive to Bet v 1 in patients with birch pollen-associated food allergy to hazelnut.

    Science.gov (United States)

    Hofmann, Claudia; Scheurer, Stephan; Rost, Kathrin; Graulich, Edith; Jamin, Annette; Foetisch, Kay; Saloga, Joachim; Vieths, Stefan; Steinbrink, Kerstin; Adler, Henric S

    2013-05-01

    IgE- and T-cell cross-reactivity contribute to the birch pollen-food syndrome. We performed a comprehensive analysis of T-cell cross-reactivity in primary cell cultures, facilitating the identification of allergen-specific T-cell subpopulations from individual patients. Patients with birch pollen allergy and associated food allergy to hazelnuts, carrots, or both were analyzed for IgE cross-reactivity, T-cell responses, and T-cell cross-reactivity to recombinant Bet v 1.0101 (Bet v 1; birch), Cor a 1.0401 (Cor a 1; hazelnut), and Dau c 1.0104 (Dau c 1; carrot). A novel flow cytometry-based method using a 2-step staining process with fluorescent dyes was established to identify subpopulations of cross-reactive T cells. IgE-binding inhibition tests of individual sera revealed that the vast majority of Cor a 1-reactive IgE was cross-reactive to Bet v 1, whereas Bet v 1-reactive IgE was only partially inhibited by preincubation with Cor a 1. Primary stimulation of T cells with Bet v 1 or Cor a 1 resulted in a significant increase in specific responses to Cor a 1 or Bet v 1 after secondary stimulation, respectively, indicating T-cell cross-reactivity between birch and hazelnut allergens in all patients of the study cohort. Preactivation with Dau c 1 induced less pronounced effects. A novel flow cytometry-based proliferation assay identified a predominant Cor a 1/Bet v 1-cross-reactive T-cell subpopulation within highly Bet v 1/Cor a 1-responsive T cells. Analysis of primary allergen-specific T cells combined with flow cytometry-based proliferation assays facilitates investigation of allergen-specific T-cell subpopulations in subjects and might be helpful to evaluate the effect of birch-specific immunotherapy on pollen-associated food allergies. Copyright © 2012 American Academy of Allergy, Asthma & Immunology. Published by Mosby, Inc. All rights reserved.

  20. Structural polarity and dynamics of male germline stem cells in the milkweed bug (Oncopeltus fasciatus).

    Science.gov (United States)

    Schmidt, Esther D; Dorn, August

    2004-11-01

    The male germline stem cells (GSCs) of the milkweed bug present an extraordinary structural polarity that is, to our knowledge, unequalled by any other type of stem cells. They consist of a perikaryon and numerous projections arising from the cell pole directed toward the apical cells, the proposed niche of the GSCs. The projections can traverse a considerable distance until their terminals touch the apical cells. From hatching until death, the GSC projections undergo conspicuous changes, the sequence of which has been deduced from observations of all developmental stages. Projection formation starts from lobular cell protrusions showing trabecular ingrowths of the cell membrane. Finger-like projections result from a process of growth and "carving out". The newly formed projections contain mostly only free ribosomes other than a few mitochondria. A stereotyped degradation process commences in the projection terminals: profiles of circular, often concentric, cisternae of rough endoplasmic reticulum appear and turn into myelin bodies, whereas mitochondria become more numerous. The cytoplasm vesiculates, lysosomal bodies appear, and mitochondria become swollen. At the same time, the projection terminals are segregated by transverse ingrowths of the cell membrane. Finally, autophagic vacuoles and myelin bodies fill the segregated terminals, which then rupture. Simultaneously, new projections seem to sprout from the perikaryon of the GSCs. These dynamics, which are not synchronized among the GSCs, indicate that a novel type of signal exchange and transduction between the stem cells and their niche is involved in the regulation of asymmetric versus symmetric division of GSCs.

  1. Domain-specific control of germ cell polarity and migration by multifunction Tre1 GPCR

    Science.gov (United States)

    2017-01-01

    The migration of primordial germ cells (PGCs) from their place of origin to the embryonic gonad is an essential reproductive feature in many animal species. In Drosophila melanogaster, a single G protein–coupled receptor, Trapped in endoderm 1 (Tre1), mediates germ cell polarization at the onset of active migration and directs subsequent migration of PGCs through the midgut primordium. How these different aspects of cell behavior are coordinated through a single receptor is not known. We demonstrate that two highly conserved domains, the E/N/DRY and NPxxY motifs, have overlapping and unique functions in Tre1. The Tre1-NRY domain via G protein signaling is required for reading and responding to guidance and survival cues controlled by the lipid phosphate phosphatases Wunen and Wunen2. In contrast, the Tre1-NPIIY domain has a separate role in Rho1- and E-cadherin–mediated polarization at the initiation stage independent of G protein signaling. We propose that this bifurcation of the Tre1 G protein–coupled receptor signaling response via G protein–dependent and independent branches enables distinct spatiotemporal regulation of germ cell migration. PMID:28687666

  2. A Molecular Probe for the Detection of Polar Lipids in Live Cells.

    Directory of Open Access Journals (Sweden)

    Christie A Bader

    Full Text Available Lipids have an important role in many aspects of cell biology, including membrane architecture/compartment formation, intracellular traffic, signalling, hormone regulation, inflammation, energy storage and metabolism. Lipid biology is therefore integrally involved in major human diseases, including metabolic disorders, neurodegenerative diseases, obesity, heart disease, immune disorders and cancers, which commonly display altered lipid transport and metabolism. However, the investigation of these important cellular processes has been limited by the availability of specific tools to visualise lipids in live cells. Here we describe the potential for ReZolve-L1™ to localise to intracellular compartments containing polar lipids, such as for example sphingomyelin and phosphatidylethanolamine. In live Drosophila fat body tissue from third instar larvae, ReZolve-L1™ interacted mainly with lipid droplets, including the core region of these organelles. The presence of polar lipids in the core of these lipid droplets was confirmed by Raman mapping and while this was consistent with the distribution of ReZolve-L1™ it did not exclude that the molecular probe might be detecting other lipid species. In response to complete starvation conditions, ReZolve-L1™ was detected mainly in Atg8-GFP autophagic compartments, and showed reduced staining in the lipid droplets of fat body cells. The induction of autophagy by Tor inhibition also increased ReZolve-L1™ detection in autophagic compartments, whereas Atg9 knock down impaired autophagosome formation and altered the distribution of ReZolve-L1™. Finally, during Drosophila metamorphosis fat body tissues showed increased ReZolve-L1™ staining in autophagic compartments at two hours post puparium formation, when compared to earlier developmental time points. We concluded that ReZolve-L1™ is a new live cell imaging tool, which can be used as an imaging reagent for the detection of polar lipids in

  3. A Molecular Probe for the Detection of Polar Lipids in Live Cells.

    Science.gov (United States)

    Bader, Christie A; Shandala, Tetyana; Carter, Elizabeth A; Ivask, Angela; Guinan, Taryn; Hickey, Shane M; Werrett, Melissa V; Wright, Phillip J; Simpson, Peter V; Stagni, Stefano; Voelcker, Nicolas H; Lay, Peter A; Massi, Massimiliano; Plush, Sally E; Brooks, Douglas A

    2016-01-01

    Lipids have an important role in many aspects of cell biology, including membrane architecture/compartment formation, intracellular traffic, signalling, hormone regulation, inflammation, energy storage and metabolism. Lipid biology is therefore integrally involved in major human diseases, including metabolic disorders, neurodegenerative diseases, obesity, heart disease, immune disorders and cancers, which commonly display altered lipid transport and metabolism. However, the investigation of these important cellular processes has been limited by the availability of specific tools to visualise lipids in live cells. Here we describe the potential for ReZolve-L1™ to localise to intracellular compartments containing polar lipids, such as for example sphingomyelin and phosphatidylethanolamine. In live Drosophila fat body tissue from third instar larvae, ReZolve-L1™ interacted mainly with lipid droplets, including the core region of these organelles. The presence of polar lipids in the core of these lipid droplets was confirmed by Raman mapping and while this was consistent with the distribution of ReZolve-L1™ it did not exclude that the molecular probe might be detecting other lipid species. In response to complete starvation conditions, ReZolve-L1™ was detected mainly in Atg8-GFP autophagic compartments, and showed reduced staining in the lipid droplets of fat body cells. The induction of autophagy by Tor inhibition also increased ReZolve-L1™ detection in autophagic compartments, whereas Atg9 knock down impaired autophagosome formation and altered the distribution of ReZolve-L1™. Finally, during Drosophila metamorphosis fat body tissues showed increased ReZolve-L1™ staining in autophagic compartments at two hours post puparium formation, when compared to earlier developmental time points. We concluded that ReZolve-L1™ is a new live cell imaging tool, which can be used as an imaging reagent for the detection of polar lipids in different intracellular

  4. A Molecular Probe for the Detection of Polar Lipids in Live Cells

    Science.gov (United States)

    Bader, Christie A.; Shandala, Tetyana; Carter, Elizabeth A.; Ivask, Angela; Guinan, Taryn; Hickey, Shane M.; Werrett, Melissa V.; Wright, Phillip J.; Simpson, Peter V.; Stagni, Stefano; Voelcker, Nicolas H.; Lay, Peter A.; Massi, Massimiliano; Brooks, Douglas A.

    2016-01-01

    Lipids have an important role in many aspects of cell biology, including membrane architecture/compartment formation, intracellular traffic, signalling, hormone regulation, inflammation, energy storage and metabolism. Lipid biology is therefore integrally involved in major human diseases, including metabolic disorders, neurodegenerative diseases, obesity, heart disease, immune disorders and cancers, which commonly display altered lipid transport and metabolism. However, the investigation of these important cellular processes has been limited by the availability of specific tools to visualise lipids in live cells. Here we describe the potential for ReZolve-L1™ to localise to intracellular compartments containing polar lipids, such as for example sphingomyelin and phosphatidylethanolamine. In live Drosophila fat body tissue from third instar larvae, ReZolve-L1™ interacted mainly with lipid droplets, including the core region of these organelles. The presence of polar lipids in the core of these lipid droplets was confirmed by Raman mapping and while this was consistent with the distribution of ReZolve-L1™ it did not exclude that the molecular probe might be detecting other lipid species. In response to complete starvation conditions, ReZolve-L1™ was detected mainly in Atg8-GFP autophagic compartments, and showed reduced staining in the lipid droplets of fat body cells. The induction of autophagy by Tor inhibition also increased ReZolve-L1™ detection in autophagic compartments, whereas Atg9 knock down impaired autophagosome formation and altered the distribution of ReZolve-L1™. Finally, during Drosophila metamorphosis fat body tissues showed increased ReZolve-L1™ staining in autophagic compartments at two hours post puparium formation, when compared to earlier developmental time points. We concluded that ReZolve-L1™ is a new live cell imaging tool, which can be used as an imaging reagent for the detection of polar lipids in different intracellular

  5. Repeated evolution of tricellular (and bicellular) pollen.

    Science.gov (United States)

    Williams, Joseph H; Taylor, Mackenzie L; O'Meara, Brian C

    2014-04-01

    Male gametophytes of seed plants are sexually immature at the time they are dispersed as pollen, but approximately 30% of flowering plants have tricellular pollen containing fully formed sperm at anthesis. The classic study of Brewbaker (1967: American Journal of Botany 54: 1069-1083) provided a powerful confirmation of the long-standing hypothesis that tricellular pollen had many parallel and irreversible origins within angiosperms. We readdressed the main questions of that study with modern comparative phylogenetic methods. We used our own and more recent reports to greatly expand the Brewbaker data set. We modeled trait evolution for 2511 species on a time-calibrated angiosperm phylogeny using (1) Binary State Speciation and Extinction (BiSSE), which accounts for the effect of species diversification rates on character transition rates and, (2) the hidden rates model (HRM), which incorporates variation in transition rates across a phylogeny. Seventy percent of species had bicellular pollen. BiSSE found a 1.9-fold higher bicellular to tricellular transition rate than in the reverse direction, and bicellular lineages had a 1.8-fold higher diversification rate than tricellular lineages. HRM found heterogeneity in evolutionary rates, with bidirectional transition rates in three of four rate classes. The tricellular condition is not irreversible. Pollen cell numbers are maintained at intermediate frequencies because lower net diversification rates of tricellular lineages are counterbalanced by slower state shifts to the bicellular condition. That tricellular lineages diversify slowly and give rise to bicellular lineages slowly reflects a linkage between the evolution of sporophyte lifestyles and the developmental lability of male gametophytes.

  6. Pollen extracts and constituent sugars increase growth of a trypanosomatid parasite of bumble bees

    Directory of Open Access Journals (Sweden)

    Evan C. Palmer-Young

    2017-05-01

    Full Text Available Phytochemicals produced by plants, including at flowers, function in protection against plant diseases, and have a long history of use against trypanosomatid infection. Floral nectar and pollen, the sole food sources for many species of insect pollinators, contain phytochemicals that have been shown to reduce trypanosomatid infection in bumble and honey bees when fed as isolated compounds. Nectar and pollen, however, consist of phytochemical mixtures, which can have greater antimicrobial activity than do single compounds. This study tested the hypothesis that pollen extracts would inhibit parasite growth. Extracts of six different pollens were tested for direct inhibitory activity against cell cultures of the bumble bee trypanosomatid gut parasite Crithidia bombi. Surprisingly, pollen extracts increased parasite growth rather than inhibiting it. Pollen extracts contained high concentrations of sugars, mainly the monosaccharides glucose and fructose. Experimental manipulations of growth media showed that supplemental monosaccharides (glucose and fructose increased maximum cell density, while a common floral phytochemical (caffeic acid with inhibitory activity against other trypanosomatids had only weak inhibitory effects on Crithidia bombi. These results indicate that, although pollen is essential for bees and other pollinators, pollen may promote growth of intestinal parasites that are uninhibited by pollen phytochemicals and, as a result, can benefit from the nutrients that pollen provides.

  7. Live-cell imaging of phosphatidic acid dynamics in pollen tubes visualized by Spo20p-derived biosensor

    Czech Academy of Sciences Publication Activity Database

    Potocký, Martin; Pleskot, Roman; Pejchar, Přemysl; Vitale, N.; Kost, B.; Žárský, Viktor

    2014-01-01

    Roč. 203, č. 2 (2014), s. 483-494 ISSN 0028-646X R&D Projects: GA ČR GA13-19073S Institutional support: RVO:61389030 Keywords : live-cell microscopy * Nicotiana tabacum ( tobacco ) * phosphatidic acid (PA) Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 7.672, year: 2014 http://gateway.isiknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=Alerting&SrcApp=Alerting&DestApp=CCC&DestLinkType=FullRecord&UT=000337639800015

  8. Secretion of endogenous and exogenous proteins from polarized MDCK cell monolayers.

    Science.gov (United States)

    Gottlieb, T A; Beaudry, G; Rizzolo, L; Colman, A; Rindler, M; Adesnik, M; Sabatini, D D

    1986-04-01

    Confluent monolayers of MDCK (Madin-Darby canine kidney) cells provide a widely used system to study the biogenesis of epithelial cell polarity. We now report that these cells are also capable of the vectorial constitutive secretion of a major endogenous product, a glycoprotein of 81 kDa, which is released into the medium from the apical surface within 30 min of its synthesis. This release represents a bona fide exocytotic secretory process and is not the result of proteolytic cleavage of a plasma membrane-associated precursor since, in cells treated with chloroquine, a protein indistinguishable from the mature secretory product accumulated intracellularly. In contrast to the vectorial secretion of the endogenous product, a variety of exogenous exocrine and endocrine proteins synthesized in MDCK cells transfected with the corresponding genes were secreted from both the apical and basolateral surfaces. These included proteins such as rat growth hormone, chicken oviduct lysozyme, bovine gastric prochymosin, and rat salivary gland alpha 2u-globulin, which in their cells of origin are secreted via a regulated pathway, as well as the liver form of the alpha 2u-globulin and the immunoglobulin kappa chain, which are normally released constitutively. These results demonstrate the existence of secretory pathways that lead to both surfaces of MDCK cells and are accessible to the foreign secretory products. They are consistent with the operation of a sorting mechanism in which the polarized secretion of the endogenous product is effected through the recognition of signals that prevent its random distribution within the fluid phase in the cellular endomembrane system.

  9. Absence of the Polar Organizing Protein PopZ Results in Reduced and Asymmetric Cell Division in Agrobacterium tumefaciens.

    Science.gov (United States)

    Howell, Matthew; Aliashkevich, Alena; Salisbury, Anne K; Cava, Felipe; Bowman, Grant R; Brown, Pamela J B

    2017-09-01

    Agrobacterium tumefaciens is a rod-shaped bacterium that grows by polar insertion of new peptidoglycan during cell elongation. As the cell cycle progresses, peptidoglycan synthesis at the pole ceases prior to insertion of new peptidoglycan at midcell to enable cell division. The A. tumefaciens homolog of the Caulobacter crescentus polar organelle development protein PopZ has been identified as a growth pole marker and a candidate polar growth-promoting factor. Here, we characterize the function of PopZ in cell growth and division of A. tumefaciens Consistent with previous observations, we observe that PopZ localizes specifically to the growth pole in wild-type cells. Despite the striking localization pattern of PopZ, we find the absence of the protein does not impair polar elongation or cause major changes in the peptidoglycan composition. Instead, we observe an atypical cell length distribution, including minicells, elongated cells, and cells with ectopic poles. Most minicells lack DNA, suggesting a defect in chromosome segregation. Furthermore, the canonical cell division proteins FtsZ and FtsA are misplaced, leading to asymmetric sites of cell constriction. Together, these data suggest that PopZ plays an important role in the regulation of chromosome segregation and cell division. IMPORTANCE A. tumefaciens is a bacterial plant pathogen and a natural genetic engineer. However, very little is known about the spatial and temporal regulation of cell wall biogenesis that leads to polar growth in this bacterium. Understanding the molecular basis of A. tumefaciens growth may allow for the development of innovations to prevent disease or to promote growth during biotechnology applications. Finally, since many closely related plant and animal pathogens exhibit polar growth, discoveries in A. tumefaciens may be broadly applicable for devising antimicrobial strategies. Copyright © 2017 American Society for Microbiology.

  10. Pollen characteristics and in vitro pollen germination of Cedrus ...

    African Journals Online (AJOL)

    PRECIOUS

    2009-11-02

    Nov 2, 2009 ... Sources1020: Ankara Üniversitesi Ziraat Fakultesi Yayinlari, Ankara p.155. Siregar IB, Sweet GB (2000).The Impact of extraction and storage conditions on the viability of radiata pine pollen. Silva Genetica, 49: 10-14. Stairs GR, Troendle V (1967). Male bud pollen radiosensitivity in selected conifer Species.

  11. Pollen characteristics and in vitro pollen germination of Cedrus ...

    African Journals Online (AJOL)

    This study aims to determine the germination characteristics, pollen tube developments, effects of germination media and temperature and incubation durations of the pollens obtained from the four clones (11342, 11344, 11345 and 11351) of Cedrus libani A. Rich. (Lebanon Cedrus) obtained from clonal seed orchard (with ...

  12. Periplasmic Acid Stress Increases Cell Division Asymmetry (Polar Aging of Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Michelle W Clark

    Full Text Available Under certain kinds of cytoplasmic stress, Escherichia coli selectively reproduce by distributing the newer cytoplasmic components to new-pole cells while sequestering older, damaged components in cells inheriting the old pole. This phenomenon is termed polar aging or cell division asymmetry. It is unknown whether cell division asymmetry can arise from a periplasmic stress, such as the stress of extracellular acid, which is mediated by the periplasm. We tested the effect of periplasmic acid stress on growth and division of adherent single cells. We tracked individual cell lineages over five or more generations, using fluorescence microscopy with ratiometric pHluorin to measure cytoplasmic pH. Adherent colonies were perfused continually with LBK medium buffered at pH 6.00 or at pH 7.50; the external pH determines periplasmic pH. In each experiment, cell lineages were mapped to correlate division time, pole age and cell generation number. In colonies perfused at pH 6.0, the cells inheriting the oldest pole divided significantly more slowly than the cells inheriting the newest pole. In colonies perfused at pH 7.50 (near or above cytoplasmic pH, no significant cell division asymmetry was observed. Under both conditions (periplasmic pH 6.0 or pH 7.5 the cells maintained cytoplasmic pH values at 7.2-7.3. No evidence of cytoplasmic protein aggregation was seen. Thus, periplasmic acid stress leads to cell division asymmetry with minimal cytoplasmic stress.

  13. Loss of PodJ in Agrobacterium tumefaciens Leads to Ectopic Polar Growth, Branching, and Reduced Cell Division.

    Science.gov (United States)

    Anderson-Furgeson, James C; Zupan, John R; Grangeon, Romain; Zambryski, Patricia C

    2016-07-01

    Agrobacterium tumefaciens is a rod-shaped Gram-negative bacterium that elongates by unipolar addition of new cell envelope material. Approaching cell division, the growth pole transitions to a nongrowing old pole, and the division site creates new growth poles in sibling cells. The A. tumefaciens homolog of the Caulobacter crescentus polar organizing protein PopZ localizes specifically to growth poles. In contrast, the A. tumefaciens homolog of the C. crescentus polar organelle development protein PodJ localizes to the old pole early in the cell cycle and accumulates at the growth pole as the cell cycle proceeds. FtsA and FtsZ also localize to the growth pole for most of the cell cycle prior to Z-ring formation. To further characterize the function of polar localizing proteins, we created a deletion of A. tumefaciens podJ (podJAt). ΔpodJAt cells display ectopic growth poles (branching), growth poles that fail to transition to an old pole, and elongated cells that fail to divide. In ΔpodJAt cells, A. tumefaciens PopZ-green fluorescent protein (PopZAt-GFP) persists at nontransitioning growth poles postdivision and also localizes to ectopic growth poles, as expected for a growth-pole-specific factor. Even though GFP-PodJAt does not localize to the midcell in the wild type, deletion of podJAt impacts localization, stability, and function of Z-rings as assayed by localization of FtsA-GFP and FtsZ-GFP. Z-ring defects are further evidenced by minicell production. Together, these data indicate that PodJAt is a critical factor for polar growth and that ΔpodJAt cells display a cell division phenotype, likely because the growth pole cannot transition to an old pole. How rod-shaped prokaryotes develop and maintain shape is complicated by the fact that at least two distinct species-specific growth modes exist: uniform sidewall insertion of cell envelope material, characterized in model organisms such as Escherichia coli, and unipolar growth, which occurs in several

  14. Regulatory networks in pollen development under cold stress

    Directory of Open Access Journals (Sweden)

    Kamal Dev Sharma

    2016-03-01

    Full Text Available Cold stress modifies anthers’ metabolic pathways to induce pollen sterility. Cold-tolerant plants, unlike the susceptible ones, produce high proportion of viable pollen. Anthers in susceptible plants, when exposed to cold stress, increase abscisic acid (ABA metabolism and reduce ABA catabolism. Increased ABA negatively regulates expression of tapetum cell wall bound invertase and monosaccharide transport genes resulting in distorted carbohydrate pool in anther. Cold-stress also reduces endogenous levels of the bioactive gibberellins (GAs, GA4 and GA7, in susceptible anthers by repression of the GA biosynthesis genes. Here we discuss recent findings on mechanisms of cold susceptibility in anthers which determine pollen sterility. We also discuss differences in regulatory pathways between cold-stressed anthers of susceptible and tolerant plants that decide pollen sterility or viability.

  15. Polarization Sensitive Measurements of Molecular Reorientation in a Glass Capacitor Cell

    Science.gov (United States)

    Cooper, Nathan; Lawhead, Carlos; Anderson, Josiah; Shiver, Tegan; Prayaga, Chandra; Ujj, Laszlo

    2014-03-01

    It is well known that molecules having a permanent dipole moment tend to orient in the direction of the electric field at room temperature. The reorientation can be probed with the help of linear spectroscopy methods such as fluorescence anisotropy measurements. We have used nonlinear polarization sensitive Raman scattering spectroscopy to quantify the orientation effect of the dipoles. Vibrational spectra of the molecules has been recorded as a function of the external electric field. The polarization changes observed during the measurement are directly linked to the molecular reorientation rearrangement. Spectra has been recorded with a laser spectrometer comprised of a Nd:YAG laser and an optical parametric oscillator and an imaging spectrometer with a CCD detector. In order to make this measurement we have constructed a glass capacitor cell coated in TiO and applied a significant electric field (0-3 kV/mm) to the sample. Our measurements showed that the orientation effect is most significant for liquid crystals as observed previously with non-polarization sensitive CARS spectroscopy.

  16. Bipolar Plasma Membrane Distribution of Phosphoinositides and Their Requirement for Auxin-Mediated Cell Polarity and Patterning in Arabidopsis.

    Science.gov (United States)

    Tejos, Ricardo; Sauer, Michael; Vanneste, Steffen; Palacios-Gomez, Miriam; Li, Hongjiang; Heilmann, Mareike; van Wijk, Ringo; Vermeer, Joop E M; Heilmann, Ingo; Munnik, Teun; Friml, Jiří

    2014-05-01

    Cell polarity manifested by asymmetric distribution of cargoes, such as receptors and transporters, within the plasma membrane (PM) is crucial for essential functions in multicellular organisms. In plants, cell polarity (re)establishment is intimately linked to patterning processes. Despite the importance of cell polarity, its underlying mechanisms are still largely unknown, including the definition and distinctiveness of the polar domains within the PM. Here, we show in Arabidopsis thaliana that the signaling membrane components, the phosphoinositides phosphatidylinositol 4-phosphate (PtdIns4P) and phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P 2 ] as well as PtdIns4P 5-kinases mediating their interconversion, are specifically enriched at apical and basal polar plasma membrane domains. The PtdIns4P 5-kinases PIP5K1 and PIP5K2 are redundantly required for polar localization of specifically apical and basal cargoes, such as PIN-FORMED transporters for the plant hormone auxin. As a consequence of the polarity defects, instructive auxin gradients as well as embryonic and postembryonic patterning are severely compromised. Furthermore, auxin itself regulates PIP5K transcription and PtdIns4P and PtdIns(4,5)P 2 levels, in particular their association with polar PM domains. Our results provide insight into the polar domain-delineating mechanisms in plant cells that depend on apical and basal distribution of membrane lipids and are essential for embryonic and postembryonic patterning. © 2014 American Society of Plant Biologists. All rights reserved.

  17. Dishevelled is essential for neural connectivity and planar cell polarity in planarians.

    Science.gov (United States)

    Almuedo-Castillo, Maria; Saló, Emili; Adell, Teresa

    2011-02-15

    The Wingless/Integrated (Wnt) signaling pathway controls multiple events during development and homeostasis. It comprises multiple branches, mainly classified according to their dependence on β-catenin activation. The Wnt/β-catenin branch is essential for the establishment of the embryonic anteroposterior (AP) body axis throughout the phylogenetic tree. It is also required for AP axis establishment during planarian regeneration. Wnt/β-catenin-independent signaling encompasses several different pathways, of which the most extensively studied is the planar cell polarity (PCP) pathway, which is responsible for planar polarization of cell structures within an epithelial sheet. Dishevelled (Dvl) is the hub of Wnt signaling because it regulates and channels the Wnt signal into every branch. Here, we analyze the role of Schmidtea mediterranea Dvl homologs (Smed-dvl-1 and Smed-dvl-2) using gene silencing. We demonstrate that in addition to a role in AP axis specification, planarian Dvls are involved in at least two different β-catenin-independent processes. First, they are essential for neural connectivity through Smed-wnt5 signaling. Second, Smed-dvl-2, together with the S. mediterranea homologs of Van-Gogh (Vang) and Diversin (Div), is required for apical positioning of the basal bodies of epithelial cells. These data represent evidence not only of the function of the PCP network in lophotrocozoans but of the involvement of the PCP core elements Vang and Div in apical positioning of the cilia.

  18. Protocadherin FAT1 binds Ena/VASP proteins and is necessary for actin dynamics and cell polarization.

    Science.gov (United States)

    Moeller, Marcus J; Soofi, Abdulsalam; Braun, Gerald S; Li, Xiaodong; Watzl, Carsten; Kriz, Wilhelm; Holzman, Lawrence B

    2004-10-01

    Cell migration requires integration of cellular processes resulting in cell polarization and actin dynamics. Previous work using tools of Drosophila genetics suggested that protocadherin fat serves in a pathway necessary for determining cell polarity in the plane of a tissue. Here we identify mammalian FAT1 as a proximal element of a signaling pathway that determines both cellular polarity in the plane of the monolayer and directed actin-dependent cell motility. FAT1 is localized to the leading edge of lamellipodia, filopodia, and microspike tips where FAT1 directly interacts with Ena/VASP proteins that regulate the actin polymerization complex. When targeted to mitochondrial outer leaflets, FAT1 cytoplasmic domain recruits components of the actin polymerization machinery sufficient to induce ectopic actin polymerization. In an epithelial cell wound model, FAT1 knockdown decreased recruitment of endogenous VASP to the leading edge and resulted in impairment of lamellipodial dynamics, failure of polarization, and an attenuation of cell migration. FAT1 may play an integrative role regulating cell migration by participating in Ena/VASP-dependent regulation of cytoskeletal dynamics at the leading edge and by transducing an Ena/VASP-independent polarity cue.

  19. ImaEdge - a platform for quantitative analysis of the spatiotemporal dynamics of cortical proteins during cell polarization.

    Science.gov (United States)

    Zhang, Zhen; Lim, Yen Wei; Zhao, Peng; Kanchanawong, Pakorn; Motegi, Fumio

    2017-12-15

    Cell polarity involves the compartmentalization of the cell cortex. The establishment of cortical compartments arises from the spatial bias in the activity and concentration of cortical proteins. The mechanistic dissection of cell polarity requires the accurate detection of dynamic changes in cortical proteins, but the fluctuations of cell shape and the inhomogeneous distributions of cortical proteins greatly complicate the quantitative extraction of their global and local changes during cell polarization. To address these problems, we introduce an open-source software package, ImaEdge, which automates the segmentation of the cortex from time-lapse movies, and enables quantitative extraction of cortical protein intensities. We demonstrate that ImaEdge enables efficient and rigorous analysis of the dynamic evolution of cortical PAR proteins during Caenorhabditis elegans embryogenesis. It is also capable of accurate tracking of varying levels of transgene expression and discontinuous signals of the actomyosin cytoskeleton during multiple rounds of cell division. ImaEdge provides a unique resource for quantitative studies of cortical polarization, with the potential for application to many types of polarized cells.This article has an associated First Person interview with the first authors of the paper. © 2017. Published by The Company of Biologists Ltd.

  20. An Instructive Role for C. elegans HMR-1/E-cadherin in Translating Cell Contact Cues into Cortical Polarity

    Science.gov (United States)

    Klompstra, Diana; Anderson, Dorian C.; Yeh, Justin Y.; Zilberman, Yuliya; Nance, Jeremy

    2015-01-01

    Cell contacts provide spatial cues that polarize early embryos and epithelial cells. The homophilic adhesion protein E-cadherin is required for contact-induced polarity in many cells. However, it is debated whether E-cadherin functions instructively as a spatial cue, or permissively by ensuring adequate adhesion so that cells can sense other contact signals. In C. elegans, contacts polarize early embryonic cells by recruiting the RhoGAP PAC-1 to the adjacent cortex, inducing PAR protein asymmetry. Here we show that HMR-1/E-cadherin, which is dispensable for adhesion, functions together with HMP-1/α-catenin, JAC-1/p120 catenin, and the previously uncharacterized linker PICC-1/CCDC85/DIPA to bind PAC-1 and recruit it to contacts. Mislocalizing the HMR-1 intracellular domain to contact-free surfaces draws PAC-1 to these sites and depolarizes cells, demonstrating an instructive role for HMR-1 in polarization. Our findings identify an E-cadherin-mediated pathway that translates cell contacts into cortical polarity by directly recruiting a symmetry-breaking factor to the adjacent cortex. PMID:25938815

  1. Functional assessment of sodium chloride cotransporter NCC mutants in polarized mammalian epithelial cells

    DEFF Research Database (Denmark)

    Rosenbaek, Lena L; Rizzo, Federica; MacAulay, Nanna

    2017-01-01

    -free solutions. NCC was detected in the plasma membrane, and both membrane abundance and phosphorylation of NCC were increased by incubation in chloride-free solutions. Furthermore, in cells exposed for 15 min to low or high extracellular K(+), the levels of phosphorylated NCC increased and decreased...... constitutively active, even without chloride-free stimulation. In conclusion, this system allows the activity, cellular localization, and abundance of wild-type or mutant NCC to be examined in the same polarized mammalian expression system in a rapid, easy, and low-cost fashion....

  2. Pollen morphology of Rhizophora L. in Peninsular Malaysia

    Energy Technology Data Exchange (ETDEWEB)

    Mohd-Arrabe' , A. B.; Noraini, Talip Noraini [School of Environmental and Natural Resource Sciences, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor (Malaysia)

    2013-11-27

    Rhizophora L. are common mangrove genus in Peninsular Malaysia, it contains 3 species and 1 hybrid (R. apiculata Blume, R. mucronata Lam., R. stylosa Griff., R. x lamarckii Montrouz). This genus has some unique adaptation towards extreme environment. Rhizophora has looping aerial stilt-root and uniformly viviparous. The aim of this study is to investigate the variation in the pollen morphology of Rhizophora that can be related to their habitat. Methods include in this study is pollen observation under light and acetolysis method under scanning electron microscope. Pollen type of Rhizophora species studied except hybrid species is classified tricolporate, shape spheroidal based on ratio of length polar axis/ length of equatorial axis (1.03 - 1.09). The exine ornamentation is perforate-reticulate for R. apiculata and R. mucronata, while R. stylosa is perforate. For the only hybrid in Peninsular Malaysia, R. x lamarckii (R. apiculata x R. stylosa) differs from others, tricolpate with the absence of porate, shape is subprolate and exine ornamentation is reticulate and striate in equatorial region. Pollenkitt is present due to the salty and extreme environment. This may enhance the volume of pollenkitt present surrounding the pollen grains in Rhizophora for protection and adaptation purposes. Based on these findings, it is evident that pollen morphology is somehow related to its natural habitat.

  3. Epicutaneous Immunotherapy Compared with Sublingual Immunotherapy in Mice Sensitized to Pollen (Phleum pratense)

    OpenAIRE

    Mondoulet, Lucie; Dioszeghy, Vincent; Ligouis, Mélanie; Dhelft, Véronique; Puteaux, Emilie; Dupont, Christophe; Benhamou, Pierre-Henri

    2012-01-01

    Background. The aim of this study was to compare the efficacy of epicutaneous immunotherapy (EPIT) to sublingual immunotherapy (SLIT) in a model of mice sensitized to Phleum pratense pollen. Methods. BALB/c mice were sensitized by sub-cutaneous route to pollen protein extract mixed treated for 8 weeks, using sham, EPIT, or SLIT. Measurements involved the serological response and cytokine profile from reactivated splenocytes, plethysmography after aerosol challenge to pollen, cell, and cytokin...

  4. In vitro biocompatibility and proliferative effects of polar and non-polar extracts of cucurbita ficifolia on human mesenchymal stem cells.

    Science.gov (United States)

    Aristatile, Balakrishnan; Alshammari, Ghedeir M

    2017-05-01

    Cucurbita ficifolia (C. ficifolia) has been traditionally known for its medicinal properties as an antioxidant, anti-diabetic and anti-inflammatory agent. However, there has been an enduring attention towards the identification of unique method, to isolate the natural components for therapeutic applications. Our study focuses on different polar and non-polar solvents (methanol, hexane and chloroform) to extract the bioactive components from C. ficifolia (pumpkin) and to study the biocompatibility and cytotoxicity effects on human bone marrow-mesenchymal stem cells (hBM-MSCs). The extracts were screened for their effects on cytotoxicity, cell proliferation and cell cycle on the hBM-MSCs cell line. The assays demonstrated that the chloroform extract was highly biocompatible, with less cytotoxic effect, and enhanced the cell proliferation. The methanol extract did not exhibit significant cytotoxicity when compare to the control. Concordantly, the cell cycle analysis confirmed that chloroform extract enhances the proliferation at lower concentrations. On the other hand, hexane extract showed high level of cytotoxicity with apoptotic and necrotic changes in hBM-MSCs. Collectively, our data revealed that chloroform is a good candidate to extract the bioactive components from C. ficifolia. Furthermore, our results suggest that specific gravity and density of the solvent might play a crucial role in the extraction process, which warrants further investigations. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  5. Response of mouse splenic lymphocytes to timothy pollen antigens in a microculture system.

    Science.gov (United States)

    Fairchild, S S; Malley, A

    1975-12-01

    Spleen cells from LAF1 mice were stimulated in a microculture system with T and B cell mitogens or antigens of timothy pollen. Only cells from mice immunized with crude timothy pollen extract (WST) or a major antigen of timothy pollen conjugated to Ascaris (antigen B-Ascaris) responded to timothy antigens in vitro. Optimum responses were obtained at 120 to 144 hr of culture with 5 to 10 mug WST per culture and ranged from three to 10 times greater than cell background. No correlations could be found between the optimum antigen concentration or the maximum response and the immune status of the spleen cell donor. Response could be inhibited by a dialyzable fraction of timothy pollen, antigen D, which is a monovalent form of a major antigen of timothy pollen.

  6. Cloning and characterisation of a putative pollen-specific polygalacturonase gene (CpPG1) differentially regulated during pollen development in zucchini (Cucurbita pepo L.).

    Science.gov (United States)

    Carvajal, F; Garrido, D; Jamilena, M; Rosales, R

    2014-03-01

    Studies in zucchini (Cucurbita pepo L. spp. pepo) pollen have been limited to the viability and morphology of the mature pollen grain. The enzyme polygalacturonase (PG) is involved in pollen development and pollination in many species. In this work, we study anther and pollen development of C. pepo and present the cloning and characterisation of a putative PG CpPG1 (Accession no. HQ232488) from pollen cDNA in C. pepo. The predicted protein for CpPG1 has 416 amino acids, with a high homology to other pollen PGs, such as P22 from Oenothera organensis (76%) and PGA3 from Arabidopsis thaliana (73%). CpPG1 belongs to clade C, which comprises PGs expressed in pollen, and presents a 34 amino acid signal peptide for secretion towards the cell wall. DNA-blot analysis revealed that there are at least another two genes that code for PGs in C. pepo. The spatial and temporal accumulation of CpPG1 was studied by semi-quantitative- and qRT-PCR. In addition, mRNA was detected only in anthers, pollen and the rudimentary anthers of bisexual flowers (only present in some zucchini cultivars under certain environmental conditions that trigger anther development in the third whorl of female flowers). However, no expression was detected in cotyledons, stem or fruit. Furthermore, CpPG1 mRNA was accumulated throughout anther development, with the highest expression found in mature pollen. Similarly, exo-PG activity increased from immature anther stages to mature anthers and mature pollen. Overall, these data support the pollen specificity of this gene and suggest an involvement of CpPG1 in pollen development in C. pepo. © 2013 German Botanical Society and The Royal Botanical Society of the Netherlands.

  7. Impact of cytomixis on meiosis, pollen viability and pollen size in ...

    Indian Academy of Sciences (India)

    Srinivas

    mother cells of Medicago sativa L.; J. Heredity 94 512–516. Bhat T A, Parveen S and Khan A H 2006 MMS-induced cytomixis in pollen mother cells of broad bean (Vicia faba L.); Turk. J. Bot. 30 273–279. Bione N C P, Pagliarini M S and de Toledo J F F 2000 Meiotic behavior of several Brazilian soybean varieties; Genet.

  8. Ultrastructural aspects of pollen ontogeny in an endangered plant species, Pancratium maritimum L. (Amaryllidaceae).

    Science.gov (United States)

    Tütüncü Konyar, Sevil

    2017-03-01

    Pollen ontogeny in Pancratium maritimum L. was studied from the sporogenous cell to mature pollen grain stages using transmission electron, scanning electron, and light microscopy to determine whether the pollen development in P. maritimum follows the basic scheme in angiosperms or not. In the course of microsporogenesis and microgametogenesis, special attention was given to the considerable ultrastructural changes that are observed in the cytoplasm of microsporocytes, microspores, and mature pollen grains throughout the successive stages of pollen development. Microsporocyte differentiation concerning number and ultrastructure of organelles facilitates the transition of microsporocytes from the sporophytic phase to the gametophytic phase. However, cytoplasmic differentiation of generative and vegetative cells supports their functional distinctness and pollen maturation. Although microsporogenesis and microgametogenesis in P. maritimum generally follow the usual angiosperm pattern, abnormalities such as formation of unreduced gametes were observed. During normal microsporogenesis, meiocytes undergo meiosis and successive cytokinesis, resulting in the formation of isobilateral, decussate, and linear tetrads. Subsequent to the development of free and vacuolated microspores, the first mitotic division occurs and bicellular monosulcate pollen grains are produced. Pollen grains are shed from the anther at binucleate stage. During pollen ontogeny, three periods of vacuolization were observed: in meiocytes, in mononucleate free microspores, and in the generative cell.

  9. Normal and tumor-derived myoepithelial cells differ in their ability to interact with luminal breast epithelial cells for polarity and basement membrane deposition

    Energy Technology Data Exchange (ETDEWEB)

    Gudjonsson, Thorarinn; Ronnov-Jessen, Lone; Villadsen, Rene; Rank, Fritz; Bissell, Mina J.; Petersen, Ole William

    2001-10-04

    The signals that determine the correct polarity of breast epithelial structures in vivo are not understood. We have shown previously that luminal epithelial cells can be polarized when cultured within a reconstituted basement membrane gel. We reasoned that such cues in vivo may be given by myoepithelial cells. Accordingly, we used an assay where luminal epithelial cells are incorrectly polarized to test this hypothesis. We show that culturing human primary luminal epithelial cells within collagen-I gels leads to formation of structures with no lumina and with reverse polarity as judged by dual stainings for sialomucin, epithelial specific antigen or occludin. No basement membrane is deposited, and {beta}4-integrin staining is negative. Addition of purified human myoepithelial cells isolated from normal glands corrects the inverse polarity, and leads to formation of double-layered acini with central lumina. Among the laminins present in the human breast basement membrane (laminin-1, -5 and -10/11), laminin-1 was unique in its ability to substitute for myoepithelial cells in polarity reversal. Myoepithelial cells were purified also from four different breast cancer sources including a biphasic cell line. Three out of four samples either totally lacked the ability to interact with luminal epithelial cells, or conveyed only correction of polarity in a fraction of acini. This behavior was directly related to the ability of the tumor myoepithelial cells to produce {alpha}-1 chain of laminin. In vivo, breast carcinomas were either negative for laminin-1 (7/12 biopsies) or showed a focal, fragmented deposition of a less intensely stained basement membrane (5/12 biopsies). Dual staining with myoepithelial markers revealed that tumorassociated myoepithelial cells were either negative or weakly positive for expression of laminin-1, establishing a strong correlation between loss of laminin-1 and breast cancer. We conclude that the double-layered breast acinus may be

  10. Ror2 Enhances Polarity and Directional Migration of Primordial Germ Cells

    Science.gov (United States)

    Kissner, Michael D.; Zhou, Xin; Anderson, Kathryn V.

    2011-01-01

    The trafficking of primordial germ cells (PGCs) across multiple embryonic structures to the nascent gonads ensures the transmission of genetic information to the next generation through the gametes, yet our understanding of the mechanisms underlying PGC migration remains incomplete. Here we identify a role for the receptor tyrosine kinase-like protein Ror2 in PGC development. In a Ror2 mouse mutant we isolated in a genetic screen, PGC migration and survival are dysregulated, resulting in a diminished number of PGCs in the embryonic gonad. A similar phenotype in Wnt5a mutants suggests that Wnt5a acts as a ligand to Ror2 in PGCs, although we do not find evidence that WNT5A functions as a PGC chemoattractant. We show that cultured PGCs undergo polarization, elongation, and reorientation in response to the chemotactic factor SCF (secreted KitL), whereas Ror2 PGCs are deficient in these SCF-induced responses. In the embryo, migratory PGCs exhibit a similar elongated geometry, whereas their counterparts in Ror2 mutants are round. The protein distribution of ROR2 within PGCs is asymmetric, both in vitro and in vivo; however, this asymmetry is lost in Ror2 mutants. Together these results indicate that Ror2 acts autonomously to permit the polarized response of PGCs to KitL. We propose a model by which Wnt5a potentiates PGC chemotaxis toward secreted KitL by redistribution of Ror2 within the cell. PMID:22216013

  11. Ror2 enhances polarity and directional migration of primordial germ cells.

    Directory of Open Access Journals (Sweden)

    Diana J Laird

    2011-12-01

    Full Text Available The trafficking of primordial germ cells (PGCs across multiple embryonic structures to the nascent gonads ensures the transmission of genetic information to the next generation through the gametes, yet our understanding of the mechanisms underlying PGC migration remains incomplete. Here we identify a role for the receptor tyrosine kinase-like protein Ror2 in PGC development. In a Ror2 mouse mutant we isolated in a genetic screen, PGC migration and survival are dysregulated, resulting in a diminished number of PGCs in the embryonic gonad. A similar phenotype in Wnt5a mutants suggests that Wnt5a acts as a ligand to Ror2 in PGCs, although we do not find evidence that WNT5A functions as a PGC chemoattractant. We show that cultured PGCs undergo polarization, elongation, and reorientation in response to the chemotactic factor SCF (secreted KitL, whereas Ror2 PGCs are deficient in these SCF-induced responses. In the embryo, migratory PGCs exhibit a similar elongated geometry, whereas their counterparts in Ror2 mutants are round. The protein distribution of ROR2 within PGCs is asymmetric, both in vitro and in vivo; however, this asymmetry is lost in Ror2 mutants. Together these results indicate that Ror2 acts autonomously to permit the polarized response of PGCs to KitL. We propose a model by which Wnt5a potentiates PGC chemotaxis toward secreted KitL by redistribution of Ror2 within the cell.

  12. Modeling self-organized spatio-temporal patterns of PIP₃ and PTEN during spontaneous cell polarization.

    Science.gov (United States)

    Knoch, Fabian; Tarantola, Marco; Bodenschatz, Eberhard; Rappel, Wouter-Jan

    2014-08-01

    During spontaneous cell polarization of Dictyostelium discoideum cells, phosphatidylinositol (3,4,5)-triphoshpate (PIP3) and PTEN (phosphatase tensin homolog) have been identified as key signaling molecules which govern the process of polarization in a self-organized manner. Recent experiments have quantified the spatio-temporal dynamics of these signaling components. Surprisingly, it was found that membrane-bound PTEN can be either in a high or low state, that PIP3 waves were initiated in areas lacking PTEN through an excitable mechanism, and that PIP3 was degraded even though the PTEN concentration remained low. Here we develop a reaction-diffusion model that aims to explain these experimental findings. Our model contains bistable dynamics for PTEN, excitable dynamics for PIP3, and postulates the existence of two species of PTEN with different dephosphorylation rates. We show that our model is able to produce results that are in good qualitative agreement with the experiments, suggesting that our reaction-diffusion model underlies the self-organized spatio-temporal patterns observed in experiments.

  13. TNF-α decreases VEGF secretion in highly polarized RPE cells but increases it in non-polarized RPE cells related to crosstalk between JNK and NF-κB pathways.

    Directory of Open Access Journals (Sweden)

    Hiroto Terasaki

    Full Text Available Asymmetrical secretion of vascular endothelial growth factor (VEGF by retinal pigment epithelial (RPE cells in situ is critical for maintaining the homeostasis of the retina and choroid. VEGF is also involved in the development and progression of age-related macular degeneration (AMD. We studied the effect of tumor necrosis factor-α (TNF-α on the secretion of VEGF in polarized and non-polarized RPE cells (P-RPE cells and N-RPE cells, respectively in culture and in situ in rats. A subretinal injection of TNF-α caused a decrease in VEGF expression and choroidal atrophy. Porcine RPE cells were seeded on Transwell™ filters, and their maturation and polarization were confirmed by the asymmetrical VEGF secretion and trans electrical resistance. Exposure to TNF-α decreased the VEGF secretion in P-RPE cells but increased it in N-RPE cells in culture. TNF-α inactivated JNK in P-RPE cells but activated it in N-RPE cells, and TNF-α activated NF-κB in P-RPE cells but not in N-RPE cells. Inhibition of NF-κB activated JNK in both types of RPE cells indicating crosstalk between JNK and NF-κB. TNF-α induced the inhibitory effects of NF-κB on JNK in P-RPE cells because NF-κB is continuously inactivated. In N-RPE cells, however, it was not evident because NF-κB was already activated. The basic activation pattern of JNK and NF-κB and their crosstalk led to opposing responses of RPE cells to TNF-α. These results suggest that VEGF secretion under inflammatory conditions depends on cellular polarization, and the TNF-α-induced VEGF down-regulation may result in choroidal atrophy in polarized physiological RPE cells. TNF-α-induced VEGF up-regulation may cause neovascularization by non-polarized or non-physiological RPE cells.

  14. Stingless bees (Melipona subnitida) adjust brood production rather than foraging activity in response to changes in pollen stores.

    Science.gov (United States)

    Maia-Silva, Camila; Hrncir, Michael; Imperatriz-Fonseca, Vera Lucia; Schorkopf, Dirk Louis P

    2016-10-01

    Highly eusocial bees (honey bees and stingless bees) sustain their colonies through periods of resource scarcity by food stored within the nest. The protein supply necessary for successful brood production is ensured through adjustments of the colonies' pollen foraging according to the availability of this resource in the environment. In honey bees Apis mellifera, in addition, pollen foraging is regulated through the broods' demand for this resource. Here, we investigated the influence of the colony's pollen store level on pollen foraging and brood production in stingless bees (Melipona subnitida). When pollen was added to the nests, colonies increased their brood production and reduced their pollen foraging within 24 h. On the other hand, when pollen reserves were removed, colonies significantly reduced their brood production. In strong contrast to A. mellifera; however, M. subnitida did not significantly increase its pollen foraging activity under poor pollen store conditions. This difference concerning the regulation of pollen foraging may be due to differences regarding the mechanism of brood provisioning. Honey bees progressively feed young larvae and, consequently, require a constant pollen supply. Stingless bees, by contrast, mass-provision their brood cells and temporary absence of pollen storage will not immediately result in substantial brood loss.

  15. Pollen morphology of some asiatic species of genus salsola (chenopodiaceae) and its taxonomic relationships

    International Nuclear Information System (INIS)

    Toderich, K.N.; Shuyskaya, E.V.; Ozturk, M

    2010-01-01

    Comparative studies on the pollen grain morphology of 27 Asiatic species of the genus Salsola were conducted by using scanning electron microscope (SEM analysis) in order to assess the taxonomic value of pollen traits. The pollen are radially symmetrical isopolar, pantopolyporate, spherical or subspheroid. The pollen characters like size, pore number, chord (C/D ratio), pore diameter, exine thickness, level of sinking of pore, convexness of mesoporial exine, spinule and minute-hole densities and number of spines on pore membrane appeared to be useful characters in distinguishing the species. Interesting intraspecific variations in pollen grain morphology were recorded for the C/D ratio. This parameter is highly specific, supporting the delimitation of Salsola species, and appears to be more conservative than some flower and fruit characters. The numerical value of form index comprising the ratio between the length of polar axis and diameter (P/E) also was an informative trait for delimitation of the species investigated here. Three pollen types were recognized. Euclidean distance was used to compute the dissimilarity matrix and a cladogram prepared. The quantitative characters of pollen morphology allowed clustering of groups and ordination analyses of species from different sections/subsections within genus Salsola. These features indicated that overall pollen traits reflect the current taxonomic boundaries, except for the Salsola species allocated to Climacoptera and Halothamnus, which should be accepted as separate genera. (author)

  16. Origin and Functional Prediction of Pollen Allergens in Plants1[OPEN

    Science.gov (United States)

    Chen, Miaolin; Xu, Jie; Ren, Kang; Searle, Iain

    2016-01-01

    Pollen allergies have long been a major pandemic health problem for human. However, the evolutionary events and biological function of pollen allergens in plants remain largely unknown. Here, we report the genome-wide prediction of pollen allergens and their biological function in the dicotyledonous model plant Arabidopsis (Arabidopsis thaliana) and the monocotyledonous model plant rice (Oryza sativa). In total, 145 and 107 pollen allergens were predicted from rice and Arabidopsis, respectively. These pollen allergens are putatively involved in stress responses and metabolic processes such as cell wall metabolism during pollen development. Interestingly, these putative pollen allergen genes were derived from large gene families and became diversified during evolution. Sequence analysis across 25 plant species from green alga to angiosperms suggest that about 40% of putative pollen allergenic proteins existed in both lower and higher plants, while other allergens emerged during evolution. Although a high proportion of gene duplication has been observed among allergen-coding genes, our data show that these genes might have undergone purifying selection during evolution. We also observed that epitopes of an allergen might have a biological function, as revealed by comprehensive analysis of two known allergens, expansin and profilin. This implies a crucial role of conserved amino acid residues in both in planta biological function and allergenicity. Finally, a model explaining how pollen allergens were generated and maintained in plants is proposed. Prediction and systematic analysis of pollen allergens in model plants suggest that pollen allergens were evolved by gene duplication and then functional specification. This study provides insight into the phylogenetic and evolutionary scenario of pollen allergens that will be helpful to future characterization and epitope screening of pollen allergens. PMID:27436829

  17. Connexin 43-mediated modulation of polarized cell movement and the directional migration of cardiac neural crest cells.

    Science.gov (United States)

    Xu, Xin; Francis, Richard; Wei, Chih Jen; Linask, Kaari L; Lo, Cecilia W

    2006-09-01

    Connexin 43 knockout (Cx43alpha1KO) mice have conotruncal heart defects that are associated with a reduction in the abundance of cardiac neural crest cells (CNCs) targeted to the heart. In this study, we show CNCs can respond to changing fibronectin matrix density by adjusting their migratory behavior, with directionality increasing and speed decreasing with increasing fibronectin density. However, compared with wild-type CNCs, Cx43alpha1KO CNCs show reduced directionality and speed, while CNCs overexpressing Cx43alpha1 from the CMV43 transgenic mice show increased directionality and speed. Altered integrin signaling was indicated by changes in the distribution of vinculin containing focal contacts, and altered temporal response of Cx43alpha1KO and CMV43 CNCs to beta1 integrin function blocking antibody treatment. High resolution motion analysis showed Cx43alpha1KO CNCs have increased cell protrusive activity accompanied by the loss of polarized cell movement. They exhibited an unusual polygonal arrangement of actin stress fibers that indicated a profound change in cytoskeletal organization. Semaphorin 3A, a chemorepellent known to inhibit integrin activation, was found to inhibit CNC motility, but in the Cx43alpha1KO and CMV43 CNCs, cell processes failed to retract with semaphorin 3A treatment. Immunohistochemical and biochemical analyses suggested close interactions between Cx43alpha1, vinculin and other actin-binding proteins. However, dye coupling analysis showed no correlation between gap junction communication level and fibronectin plating density. Overall, these findings indicate Cx43alpha1 may have a novel function in mediating crosstalk with cell signaling pathways that regulate polarized cell movement essential for the directional migration of CNCs.

  18. Pollen rain and pollen representation across a forest-páramo ecotone in northern Ecuador

    NARCIS (Netherlands)

    Moscol Olivera, M.; Duivenvoorden, J.F.; Hooghiemstra, H.

    2009-01-01

    Modern pollen spectra were studied in forest and páramo vegetation from the Guandera area, northern Ecuador. Pollen representation was estimated by comparing the presence of plant taxa from a recent vegetation survey with the pollen spectra in moss polsters and pollen traps. In total, 73 pollen taxa

  19. Modulation of cell polarization by the Na+-K+-ATPase-associated protein FXYD5 (dysadherin).

    Science.gov (United States)

    Lubarski, Irina; Asher, Carol; Garty, Haim

    2014-06-01

    FXYD5 (dysadherin or also called a related to ion channel, RIC) is a transmembrane auxiliary subunit of the Na(+)-K(+)-ATPase shown to increase its maximal velocity (Vmax). FXYD5 has also been identified as a cancer-associated protein whose expression in tumor-derived cell lines impairs cytoskeletal organization and increases cell motility. Previously, we have demonstrated that the expression of FXYD5 in M1 cells derived from mouse kidney collecting duct impairs the formation of tight and adherence junctions. The current study aimed to further explore effects of FXYD5 at a single cell level. It was found that in M1, as well as three other cell lines, FXYD5 inhibits transformation of adhered single cells from the initial radial shape to a flattened, elongated shape in the first stage of monolayer formation. This is also correlated to less ordered actin cables and fewer focal points. Structure-function analysis has demonstrated that the transmembrane domain of FXYD5, and not its unique extracellular segment, mediates the inhibition of change in cell shape. This domain has been shown before to be involved in the association of FXYD5 with the Na(+)-K(+)-ATPase, which leads to the increase in Vmax. Furthermore, specific transmembrane point mutations in FXYD5 that either increase or decrease its effect on cell elongation had a corresponding effect on the coimmunoprecipitation of FXYD5 with α Na(+)-K(+)-ATPase. These findings lend support to the possibility that FXYD5 affects cell polarization through its transmembrane domain interaction with the Na(+)-K(+)-ATPase. Yet interaction of FXYD5 with other proteins cannot be excluded. Copyright © 2014 the American Physiological Society.

  20. MHV-A59 enters polarized murine epithelial cells through the apical surface but is released basolaterally

    NARCIS (Netherlands)

    Rossen, J W; Voorhout, W F; Horzinek, M C; van der Ende, A; Strous, G J; Rottier, P J

    1995-01-01

    Coronaviruses have a marked tropism for epithelial cells. Entry and release of the porcine transmissible gastroenteritis virus (TGEV) is restricted to apical surfaces of polarized epithelial cells, as we have recently shown (J. W. A. Rossen, C. P. J. Bekker, W. F. Voorhout, G. J. A. M. Strous, A.

  1. Clonal expansion of genome-intact HIV-1 in functionally polarized Th1 CD4+ T cells.

    Science.gov (United States)

    Lee, Guinevere Q; Orlova-Fink, Nina; Einkauf, Kevin; Chowdhury, Fatema Z; Sun, Xiaoming; Harrington, Sean; Kuo, Hsiao-Hsuan; Hua, Stephane; Chen, Hsiao-Rong; Ouyang, Zhengyu; Reddy, Kavidha; Dong, Krista; Ndung'u, Thumbi; Walker, Bruce D; Rosenberg, Eric S; Yu, Xu G; Lichterfeld, Mathias

    2017-06-30

    HIV-1 causes a chronic, incurable disease due to its persistence in CD4+ T cells that contain replication-competent provirus, but exhibit little or no active viral gene expression and effectively resist combination antiretroviral therapy (cART). These latently infected T cells represent an extremely small proportion of all circulating CD4+ T cells but possess a remarkable long-term stability and typically persist throughout life, for reasons that are not fully understood. Here we performed massive single-genome, near-full-length next-generation sequencing of HIV-1 DNA derived from unfractionated peripheral blood mononuclear cells, ex vivo-isolated CD4+ T cells, and subsets of functionally polarized memory CD4+ T cells. This approach identified multiple sets of independent, near-full-length proviral sequences from cART-treated individuals that were completely identical, consistent with clonal expansion of CD4+ T cells harboring intact HIV-1. Intact, near-full-genome HIV-1 DNA sequences that were derived from such clonally expanded CD4+ T cells constituted 62% of all analyzed genome-intact sequences in memory CD4 T cells, were preferentially observed in Th1-polarized cells, were longitudinally detected over a duration of up to 5 years, and were fully replication- and infection-competent. Together, these data suggest that clonal proliferation of Th1-polarized CD4+ T cells encoding for intact HIV-1 represents a driving force for stabilizing the pool of latently infected CD4+ T cells.

  2. Determination of apical membrane polarity in mammary epithelial cell cultures: The role of cell-cell, cell-substratum, and membrane-cytoskeleton interactions

    Energy Technology Data Exchange (ETDEWEB)

    Parry, G.; Beck, J.C.; Moss, L.; Bartley, J. (Lawrence Berkeley Lab., CA (United States)); Ojakian, G.K. (State Univ. of New York, Brooklyn (United States))

    1990-06-01

    The membrane glycoprotein, PAS-O, is a major differentiation antigen on mammary epithelial cells and is located exclusively in the apical domain of the plasma membrane. The authors have used 734B cultured human mammary carcinoma cells as a model system to study the role of tight junctions, cell-substratum contacts, and submembranous cytoskeletal elements in restricting PAS-O to the apical membrane. Immunofluorescence and immunoelectronmicroscopy experiments demonstrated that while tight junctions demarcate PAS-O distribution in confluent cultures, apical polarity could be established at low culture densities when cells could not form tight junctions with neighboring cells. They suggest, then, that interactions between vitronectin and its receptor, are responsible for establishment of membrane domains in the absence of tight junctions. The role of cytoskeletal elements in restricting PAS-O distribution was examined by treating cultures with cytochalasin D, colchicine, or acrylamide. Cytochalasin D led to a redistribution of PAS0O while colchicine and acrylamide did not. They hypothesize that PAS-O is restricted to the apical membrane by interactions with a microfilament network and that the cytoskeletal organization is dependent upon cell-cell and cell-substratum interactions.

  3. CHARACTERIZATION OF THE MAIZE POLLEN TRANSCRIPTOME

    Science.gov (United States)

    Pollen is a primary vehicle for transgene flow from engineered plants to their non-transgenic, native or weedy relatives. Hence, gene flow will be affected by pollen fitness (e.g., how well a particular pollen grain can outcompete other pollen present on the stigma and complete ...

  4. YAP and TAZ in epithelial stem cells: A sensor for cell polarity, mechanical forces and tissue damage.

    Science.gov (United States)

    Elbediwy, Ahmed; Vincent-Mistiaen, Zoé I; Thompson, Barry J

    2016-07-01

    The YAP/TAZ family of transcriptional co-activators drives cell proliferation in epithelial tissues and cancers. Yet, how YAP and TAZ are physiologically regulated remains unclear. Here we review recent reports that YAP and TAZ act primarily as sensors of epithelial cell polarity, being inhibited when cells differentiate an apical membrane domain, and being activated when cells contact the extracellular matrix via their basal membrane domain. Apical signalling occurs via the canonical Crumbs/CRB-Hippo/MST-Warts/LATS kinase cascade to phosphorylate and inhibit YAP/TAZ. Basal signalling occurs via Integrins and Src family kinases to phosphorylate and activate YAP/TAZ. Thus, YAP/TAZ is localised to the nucleus in basal stem/progenitor cells and cytoplasm in differentiated squamous cells or columnar cells. In addition, other signals such as mechanical forces, tissue damage and possibly receptor tyrosine kinases (RTKs) can influence MST-LATS or Src family kinase activity to modulate YAP/TAZ activity. © 2016 The Authors BioEssays Published by WILEY Periodicals, Inc.

  5. Epicutaneous Immunotherapy Compared with Sublingual Immunotherapy in Mice Sensitized to Pollen (Phleum pratense).

    Science.gov (United States)

    Mondoulet, Lucie; Dioszeghy, Vincent; Ligouis, Mélanie; Dhelft, Véronique; Puteaux, Emilie; Dupont, Christophe; Benhamou, Pierre-Henri

    2012-01-01

    Background. The aim of this study was to compare the efficacy of epicutaneous immunotherapy (EPIT) to sublingual immunotherapy (SLIT) in a model of mice sensitized to Phleum pratense pollen. Methods. BALB/c mice were sensitized by sub-cutaneous route to pollen protein extract mixed treated for 8 weeks, using sham, EPIT, or SLIT. Measurements involved the serological response and cytokine profile from reactivated splenocytes, plethysmography after aerosol challenge to pollen, cell, and cytokine contents in the bronchoalveolar lavages (BALs). Results. After immunotherapy, sIgE was significantly decreased in the treated groups compared to sham (P mice sensitized to pollen, EPIT was at least as efficient as SLIT.

  6. Endocytic machinery protein SlaB is dispensable for polarity establishment but necessary for polarity maintenance in hyphal tip cells of Aspergillus nidulans.

    Science.gov (United States)

    Hervás-Aguilar, América; Peñalva, Miguel A

    2010-10-01

    The Aspergillus nidulans endocytic internalization protein SlaB is essential, in agreement with the key role in apical extension attributed to endocytosis. We constructed, by gene replacement, a nitrate-inducible, ammonium-repressible slaB1 allele for conditional SlaB expression. Video microscopy showed that repressed slaB1 cells are able to establish but unable to maintain a stable polarity axis, arresting growth with budding-yeast-like morphology shortly after initially normal germ tube emergence. Using green fluorescent protein (GFP)-tagged secretory v-SNARE SynA, which continuously recycles to the plasma membrane after being efficiently endocytosed, we establish that SlaB is crucial for endocytosis, although it is dispensable for the anterograde traffic of SynA and of the t-SNARE Pep12 to the plasma and vacuolar membrane, respectively. By confocal microscopy, repressed slaB1 germlings show deep plasma membrane invaginations. Ammonium-to-nitrate medium shift experiments demonstrated reversibility of the null polarity maintenance phenotype and correlation of normal apical extension with resumption of SynA endocytosis. In contrast, SlaB downregulation in hyphae that had progressed far beyond germ tube emergence led to marked polarity maintenance defects correlating with deficient SynA endocytosis. Thus, the strict correlation between abolishment of endocytosis and disability of polarity maintenance that we report here supports the view that hyphal growth requires coupling of secretion and endocytosis. However, downregulated slaB1 cells form F-actin clumps containing the actin-binding protein AbpA, and thus F-actin misregulation cannot be completely disregarded as a possible contributor to defective apical extension. Latrunculin B treatment of SlaB-downregulated tips reduced the formation of AbpA clumps without promoting growth and revealed the formation of cortical "comets" of AbpA.

  7. Polarization of macrophages in the tumor microenvironment is influenced by EGFR signaling within colon cancer cells

    Science.gov (United States)

    Zhang, Weina; Chen, Lechuang; Ma, Kai; Zhao, Yahui; Liu, Xianghe; Wang, Yu; Liu, Mei; Liang, Shufang; Zhu, Hongxia; Xu, Ningzhi

    2016-01-01

    Epidermal growth factor receptor (EGFR) is a target of colon cancer therapy, but the effects of this therapy on the tumor microenvironment remain poorly understood. Our in vivo studies showed that cetuximab, an anti-EGFR monoclonal antibody, effectively inhibited AOM/DSS-induced, colitis-associated tumorigenesis, downregulated M2-related markers, and decreased F4/80+/CD206+ macrophage populations. Treatment with conditioned medium of colon cancer cells increased macrophage expression of the M2-related markers arginase-1 (Arg1), CCL17, CCL22, IL-10 and IL-4. By contrast, conditioned medium of EGFR knockout colon cancer cells inhibited expression of these M2-related markers and induced macrophage expression of the M1-related markers inducible nitric oxide synthase (iNOS), IL-12, TNF-α and CCR7. EGFR knockout in colon cancer cells inhibited macrophage-induced promotion of xenograft tumor growth. Moreover, colon cancer-derived insulin-like growth factor-1 (IGF-1) increased Arg1 expression, and treatment with the IGF1R inhibitor AG1024 inhibited that increase. These results suggest that inhibition of EGFR signaling in colon cancer cells modulates cytokine secretion (e.g. IGF-1) and prevents M1-to-M2 macrophage polarization, thereby inhibiting cancer cell growth. PMID:27683110

  8. Distinct chromatin environment associated with phosphorylated H3S10 histone during pollen mitosis I in orchids.

    Science.gov (United States)

    Sharma, Santosh Kumar; Yamamoto, Maki; Mukai, Yasuhiko

    2017-01-01

    Pollen developmental pathway in plants involving synchronized transferal of cellular divisions from meiosis (microsporogenesis) to mitosis (pollen mitosis I/II) eventually offers a unique "meiosis-mitosis shift" at pollen mitosis I. Since the cell type (haploid microspore) and fate of pollen mitosis I differ from typical mitosis (in meristem cells), it is immensely important to analyze the chromosomal distribution of phosphorylated H3S10 histone during atypical pollen mitosis I to comprehend the role of histone phosphorylation in pollen development. We investigated the chromosomal phosphorylation of H3S10 histone during pollen mitosis I in orchids using immunostaining technique. The chromosomal distribution of H3S10ph during pollen mitosis I revealed differential pattern than that of typical mitosis in plants, however, eventually following the similar trends of mitosis in animals where H3S10 phosphorylation begins in the pericentromeric regions first, later extending to the whole chromosomes, and finally declining at anaphase/early cytokinesis (differentiation of vegetative and generative cells). The study suggests that the chromosomal distribution of H3S10ph during cell division is not universal and can be altered between different cell types encoded for diverse cellular processes. During pollen development, phosphorylation of histone might play a critical role in chromosome condensation events throughout pollen mitosis I in plants.

  9. Modulation of Dendritic Cell Activation and Subsequent Th1 Cell Polarization by Lidocaine

    Science.gov (United States)

    Chung, Yeonseok

    2015-01-01

    Dendritic cells play an essential role in bridging innate and adaptive immunity by recognizing cellular stress including pathogen- and damage-associated molecular patterns and by shaping the types of antigen-specific T cell immunity. Although lidocaine is widely used in clinical settings that trigger cellular stress, it remains unclear whether such treatment impacts the activation of innate immune cells and subsequent differentiation of T cells. Here we showed that lidocaine inhibited the production of IL–6, TNFα and IL–12 from dendritic cells in response to toll-like receptor ligands including lipopolysaccharide, poly(I:C) and R837 in a dose-dependent manner. Notably, the differentiation of Th1 cells was significantly suppressed by the addition of lidocaine while the same treatment had little effect on the differentiation of Th17, Th2 and regulatory T cells in vitro. Moreover, lidocaine suppressed the ovalbumin-specific Th1 cell responses in vivo induced by the adoptive transfer of ovalbumin-pulsed dendritic cells. These results demonstrate that lidocaine inhibits the activation of dendritic cells in response to toll-like receptor signals and subsequently suppresses the differentiation of Th1 cell responses. PMID:26445366

  10. Daple coordinates organ-wide and cell-intrinsic polarity to pattern inner-ear hair bundles.

    Science.gov (United States)

    Siletti, Kimberly; Tarchini, Basile; Hudspeth, A J

    2017-12-26

    The establishment of planar polarization by mammalian cells necessitates the integration of diverse signaling pathways. In the inner ear, at least two systems regulate the planar polarity of sensory hair bundles. The core planar cell polarity (PCP) proteins coordinate the orientations of hair cells across the epithelial plane. The cell-intrinsic patterning of hair bundles is implemented independently by the G protein complex classically known for orienting the mitotic spindle. Although the primary cilium also participates in each of these pathways, its role and the integration of the two systems are poorly understood. We show that Dishevelled-associating protein with a high frequency of leucine residues (Daple) interacts with PCP and cell-intrinsic signals. Regulated by the cell-intrinsic pathway, Daple is required to maintain the polarized distribution of the core PCP protein Dishevelled and to position the primary cilium at the abneural edge of the apical surface. Our results suggest that the primary cilium or an associated structure influences the domain of cell-intrinsic signals that shape the hair bundle. Daple is therefore essential to orient and pattern sensory hair bundles. Copyright © 2017 the Author(s). Published by PNAS.

  11. Structural polarity and dynamics of male germline stem cells in an insect (milkweed bug Oncopeltus fasciatus).

    Science.gov (United States)

    Dorn, David C; Dorn, August

    2008-01-01

    Knowing the structure opens a door for a better understanding of function because there is no function without structure. Male germline stem cells (GSCs) of the milkweed bug (Oncopeltus fasciatus) exhibit a very extraordinary structure and a very special relationship with their niche, the apical cells. This structural relationship is strikingly different from that known in the fruit fly (Drosophila melanogaster) -- the most successful model system, which allowed deep insights into the signaling interactions between GSCs and niche. The complex structural polarity of male GSCs in the milkweed bug combined with their astonishing dynamics suggest that cell morphology and dynamics are causally related with the most important regulatory processes that take place between GSCs and niche and ensure maintenance, proliferation, and differentiation of GSCs in accordance with the temporal need of mature sperm. The intricate structure of the GSCs of the milkweed bug (and probably of some other insects, i.e., moths) is only accessible by electron microscopy. But, studying singular sections through the apical complex (i.e., GSCs and apical cells) is not sufficient to obtain a full picture of the GSCs; especially, the segregation of projection terminals is not tangible. Only serial sections and their overlay can establish whether membrane ingrowths merely constrict projections or whether a projection terminal is completely cut off. To sequence the GSC dynamics, it is necessary to include juvenile stages, when the processes start and the GSCs occur in small numbers. The fine structural analysis of segregating projection terminals suggests that these terminals undergo autophagocytosis. Autophagosomes can be labeled by markers. We demonstrated acid phosphatase and thiamine pyrophosphatase (TPPase). Both together are thought to identify autophagosomes. Using the appropriate substrate of the enzymes and cerium chloride, the precipitation of electron-dense cerium phosphate granules

  12. High expression of Rac1 is correlated with partial reversed cell polarity and poor prognosis in invasive ductal carcinoma of the breast.

    Science.gov (United States)

    Liu, Bingbing; Xiong, Jianhua; Liu, Guiqiu; Wu, Jing; Wen, Likun; Zhang, Qin; Zhang, Chuanshan

    2017-07-01

    The change of cell polarity is usually associated with invasion and metastasis. Partial reverse cell polarity in IDC-NOS may play a role in lymphatic tumor spread. Rac1 is a kind of polarity related protein. It plays an important role in invasion and metastasis in tumors. We here investigated the expression of Rac1 and partial reverse cell polarity status in breast cancer and evaluated their value for prognosis in breast cancer. The association of the expression of Rac1 and MUC-1 with clinicopathological parameters and prognostic significance was evaluated in 162 cases of IDC-NOS paraffin-embedded tissues by immunohistochemical method. The Rac1 messenger RNA expression was measured by real-time polymerase chain reaction in 30 breast cancer patients, which was divided into two groups of partial reverse cell polarity and no partial reverse cell polarity. We found that lymph node metastasis of partial reverse cell polarity patients was higher than no partial reverse cell polarity patients (Z = -4.030, p = 0.000). Rac1 was upregulated in partial reverse cell polarity group than no partial reverse cell polarity group (Z = -3.164, p = 0.002), and there was correlationship between the expression of Rac1 and partial reverse cell polarity status (r s  = 0.249, p = 0.001). The level of Rac1 messenger RNA expression in partial reverse cell polarity group was significantly higher compared to no partial reverse cell polarity group (t = -2.527, p = 0.017). Overexpression of Rac1 and partial reverse cell polarity correlates with poor prognosis of IDC-NOS patients (p = 0.011). Partial reverse cell polarity and lymph node metastasis remained as independent predictors for poor disease-free survival of IDC-NOS (p = 0.023, p = 0.046). Our study suggests that partial reverse cell polarity may lead to poor prognosis of breast cancer. Overexpression of Rac1 may lead to polarity change in IDC-NOS of the breast. Therefore, Rac1 could be a

  13. Pollen germination and pollen tube growth in ZP maize lines

    Directory of Open Access Journals (Sweden)

    Cerović Radosav

    2014-01-01

    Full Text Available The study was conducted on the in vitro pollen germination at 26°, 28°, 32° and 35°C for 24h of male parental lines, pollen tube growth in vivo in cross pollination of female and male parental lines that make couples in four hybrids: ZP 504 su (♀ ZPPL 51 × ♂ ZPPL 67; ZP 677 (♀ ZPPL 17 × ♂ ZPPL 201; ZP 704 (♀ ZPPL 109 × ♂ ZPPL 79, ZP 611 k (♀ ZPPL 126 × ♂ ZPPL 105, and the open pollination of female parental lines of the above mentioned hybrids. Pollen germination in vitro and pollen tube growth dynamics in vivo showed different genotypic specificities with the tests applied. The obtained results were discussed in the context of reproductive biology of ZP maize lines and aimed to create the preconditions for successful management and direction of the process in practice - seed production in certain environmental conditions.

  14. THE CONTENTS OF NEUTRAL AND POLAR LIPIDS IN CLOSTRIDIA CELLS UNDER CULTIVATION IN THE PRESENCE OF BUTANOL

    Directory of Open Access Journals (Sweden)

    S. I. Voychuk

    2017-02-01

    Full Text Available The aim of the study was to evaluate changes in the portion of polar and neutral lipids in the cells of Clostridium during their cultivation in the presence of butanol. Four natural isolates of Clostridium genus were studied with flow cytometry approaches. Under the optimal culture conditions, the polar lipids prevailed over neutral ones in bacterial cells; the content of neutral lipids doubled in spores of these microorganisms, while the content of polar ones was reduced. Strains No 1 and No 2 were able to grow at 1% butanol in the medium, and the strain No 4 was at 1.5%. When cultivated in the presence of different concentrations of butanol, the bacterial strains did not differ in such cytomorphological features as granularity and cell size. The quantitative content of polar and neutral lipids in the presence of butanol varied depending on the content of butanol in the medium, however this effect had a strain-specific character and did not show a correlation with the resistance of these bacteria to butanol. So, the content of polar and neutral lipids varied depending on butanol content in the medium. However this effect was strain-specific independently of resistance of these bacteria to butanol. The use of bacterial biomass as a source of lipids for the production of biofuels requires further optimization of the process to increase the content of the neutral lipid fraction in bacterial cells.

  15. Shifted T Helper Cell Polarization in a Murine Staphylococcus aureus Mastitis Model.

    Science.gov (United States)

    Zhao, Yanqing; Zhou, Ming; Gao, Yang; Liu, Heyuan; Yang, Wenyu; Yue, Jinhua; Chen, Dekun

    2015-01-01

    Mastitis, one of the most costly diseases in dairy ruminants, is an inflammation of the mammary gland caused by pathogenic infection. The mechanisms of adaptive immunity against pathogens in mastitis have not been fully elucidated. To investigate T helper cell-mediated adaptive immune responses, we established a mastitis model by challenge with an inoculum of 4 × 106 colony-forming units of Staphylococcus aureus in the mammary gland of lactating mice, followed by quantification of bacterial burden and histological analysis. The development of mastitis was accompanied by a significant increase in both Th17 and Th1 cells in the mammary gland. Moreover, the relative expression of genes encoding cytokines and transcription factors involved in the differentiation and function of these T helper cells, including Il17, Rorc, Tgfb, Il1b, Il23, Ifng, Tbx21, and Il12, was greatly elevated in the infected mammary gland. IL-17 is essential for neutrophil recruitment to infected mammary gland via CXC chemokines, whereas the excessive IL-17 production contributes to tissue damage in mastitis. In addition, a shift in T helper cell polarization toward Th2 and Treg cells was observed 5 days post-infection, and the mRNA expression of the anti-inflammatory cytokine Il10 was markedly increased at day 7 post-infection. These results indicate that immune clearance of Staphylococcus aureus in mastitis is facilitated by the enrichment of Th17, Th1 and Th2 cells in the mammary gland mediated by pro-inflammatory cytokine production, which is tightly regulated by Treg cells and the anti-inflammatory cytokine IL-10.

  16. Priming dendritic cells for Th2 polarization: lessons learned from helminths and implications for metabolic disorders

    Directory of Open Access Journals (Sweden)

    Leonie eHussaarts

    2014-10-01

    Full Text Available Nearly one quarter of the world’s population is infected with helminth parasites. A common feature of helminth infections is the manifestation of a type 2 immune response, characterized by T helper 2 (Th2 cells that mediate anti-helminth immunity. In addition, recent literature described a close association between type 2 immune responses and wound repair, suggesting that a Th2 response may concurrently mediate repair of parasite-induced damage. The molecular mechanisms that govern Th2 responses are poorly understood, although it is clear that dendritic cells (DCs, which are the most efficient antigen-presenting cells in the immune system, play a central role. Here, we review the molecular mechanisms by which DCs polarize Th2 cells, examining both helminth antigens and helminth-mediated tissue damage as Th2-inducing triggers. Finally, we discuss the implication of these findings in the context of metabolic disorders, as recent literature indicates that various aspects of the Th2-associated inflammatory response contribute to metabolic homeostasis.

  17. Epidermal wound repair is regulated by the planar cell polarity signaling pathway

    Science.gov (United States)

    Caddy, Jacinta; Wilanowski, Tomasz; Darido, Charbel; Dworkin, Sebastian; Ting, Stephen B.; Zhao, Quan; Rank, Gerhard; Auden, Alana; Srivastava, Seema; Papenfuss, Tony A.; Murdoch, Jennifer N.; Humbert, Patrick O.; Boulos, Nidal; Weber, Thomas; Zuo, Jian; Cunningham, John M.; Jane, Stephen M.

    2010-01-01

    SUMMARY The mammalian PCP pathway regulates diverse developmental processes requiring coordinated cellular movement, including neural tube closure and cochlear stereociliary orientation. Here, we show that epidermal wound repair is regulated by PCP signaling. Mice carrying mutant alleles of PCP genes Vangl2, Celsr1, PTK7, and Scrb1, and the transcription factor Grhl3, interact genetically, exhibiting failed wound healing, neural tube defects and disordered cochlear polarity. Using phylogenetic analysis, ChIP, and gene expression in Grhl3−/− mice, we identified RhoGEF19, a homologue of a RhoA activator involved in PCP signaling in Xenopus, as a direct target of GRHL3. Knockdown of Grhl3 or RhoGEF19 in keratinocytes induced defects in actin polymerisation, cellular polarity and wound healing, and re-expression of RhoGEF19 rescued these defects in Grhl3-kd cells. These results define a role for Grhl3 in PCP signaling, and broadly implicate this pathway in epidermal repair. PMID:20643356

  18. Analysis of polarization methods for elimination of power overshoot in microbial fuel cells

    KAUST Repository

    Watson, Valerie J.

    2011-01-01

    Polarization curves from microbial fuel cells (MFCs) often show an unexpectedly large drop in voltage with increased current densities, leading to a phenomenon in the power density curve referred to as "power overshoot". Linear sweep voltammetry (LSV, 1 mV s- 1) and variable external resistances (at fixed intervals of 20 min) over a single fed-batch cycle in an MFC both resulted in power overshoot in power density curves due to anode potentials. Increasing the anode enrichment time from 30 days to 100 days did not eliminate overshoot, suggesting that insufficient enrichment of the anode biofilm was not the primary cause. Running the reactor at a fixed resistance for a full fed-batch cycle (~ 1 to 2 days), however, completely eliminated the overshoot in the power density curve. These results show that long times at a fixed resistance are needed to stabilize current generation by bacteria in MFCs, and that even relatively slow LSV scan rates and long times between switching circuit loads during a fed-batch cycle may produce inaccurate polarization and power density results for these biological systems. © 2010 Elsevier B.V. All rights reserved.

  19. External application of gametophyte-specific ZmPMEI1 induces pollen tube burst in maize.

    Science.gov (United States)

    Woriedh, Mayada; Wolf, Sebastian; Márton, Mihaela L; Hinze, Axel; Gahrtz, Manfred; Becker, Dirk; Dresselhaus, Thomas

    2013-09-01

    Regulated demethylesterification of homogalacturonan, a major component of plant cell walls, by the activity of pectin methylesterases (PMEs), plays a critical role for cell wall stability and integrity. Especially fast growing plant cells such as pollen tubes secrete large amounts of PMEs toward their apoplasmic space. PME activity itself is tightly regulated by its inhibitor named as PME inhibitor and is thought to be required especially at the very pollen tube tip. We report here the identification and functional characterization of PMEI1 from maize (ZmPMEI1). We could show that the protein acts as an inhibitor of PME but not of invertases and found that its gene is strongly expressed in both gametophytes (pollen grain and embryo sac). Promoter reporter studies showed gene activity also during pollen tube growth toward and inside the transmitting tract. All embryo sac cells except the central cell displayed strong expression. Weaker signals were visible at sporophytic cells of the micropylar region. ZmPMEI1-EGFP fusion protein is transported within granules inside the tube and accumulates at the pollen tube tip as well as at sites where pollen tubes bend and/or change growth directions. The female gametophyte putatively influences pollen tube growth behavior by exposing it to ZmPMEI1. We therefore simulated this effect by applying recombinant protein at different concentrations on growing pollen tubes. ZmPMEI1 did not arrest growth, but destabilized the cell wall inducing burst. Compared with female gametophyte secreted defensin-like ZmES4, which induces burst at the very pollen tube tip, ZmPMEI1-induced burst occurs at the subapical region. These findings indicate that ZmPMEI1 secreted by the embryo sac likely destabilizes the pollen tube wall during perception and together with other proteins such as ZmES4 leads to burst and thus sperm release.

  20. Lactobacillus strains differentially modulate cytokine production by hPBMC from pollen allergic patients

    NARCIS (Netherlands)

    Vissers, Y.M.; Snel, J.; Zuurendonk, P.F.; Kleerebezem, M.; Wichers, H.J.; Savelkoul, H.F.J.

    2011-01-01

    The objective of this study was to assess the potential immunomodulatory effect of six Lactobacillus strains on human peripheral blood mononuclear cells (hPBMC) isolated from allergic patients. hPBMC from patients allergic to birch pollen or grass pollen were cultured in vitro in the presence or

  1. Effect of Turkish pollen and propolis extracts on caspase-3 activity in ...

    African Journals Online (AJOL)

    Management, High School of Health, University of Gumushane, Gumushane, 4Karadeniz Advanced Technology Research and. Application Centre ... Conclusion: Turkish pollen and propolis individually increase apoptosis and the activity of caspase-3 in. HL-60 cells. ... essential and aromatic oils, 5 % pollen and 5 %.

  2. The dual effects of polar methanolic extract of Hypericum perforatum L. in bladder cancer cells

    Science.gov (United States)

    Nseyo, U. O.; Nseyo, O. U.; Shiverick, K. T.; Medrano, T.; Mejia, M.; Stavropoulos, N.; Tsimaris, I.; Skalkos, D.

    2007-02-01

    Introduction and background: We have reported on the polar methanolic fraction (PMF) of Hypericum Perforatum L as a novel photosensitizing agent for photodynamic therapy (PDT) and photodynamic diagnosis (PDD). PMF has been tested in human leukemic cells, HL-60 cells, cord blood hemopoietic progenitor cells, bladder cancers derived from metastatic lymph node (T-24) and primary papillary bladder lesion (RT-4). However, the mechanisms of the effects of PMF on these human cell lines have not been elucidated. We have investigated mechanisms of PMF + light versus PMF-alone (dark experiment) in T-24 human bladder cancer cells. Methods: PMF was prepared from an aerial herb of HPL which was brewed in methanol and extracted with ether and methanol. Stock solutions of PMF were made in DSMO and stored in dark conditions. PMF contains 0.57% hypericin and 2.52% hyperforin. The T24 cell line was obtained from American Type Culture Collection (ATCC). In PDT treatment, PMF (60μg/ml) was incubated with cells, which were excited with laser light (630nm) 24 hours later. Apoptosis was determined by DNA fragmentation/laddering assay. DNA isolation was performed according to the manufacture's instructions with the Kit (Oncogene Kit#AM41). Isolated DNA samples were separated by electrophoresis in 1.5% in agarose gels and bands were visualized by ethidium bromide labeling. The initial cell cycle analysis and phase distribution was by flow cytometry. DNA synthesis was measured by [3H] thymidine incorporation, and cell cycle regulatory proteins were assayed by Western immunoblot. Results: The results of the flow cytometry showed PMF +light induced significant (40%) apoptosis in T24 cells, whereas Light or PMF alone produced little apoptosis. The percentage of cells in G 0/G I phase was decreased by 25% and in G2/M phase by 38%. The main impact was observed on the S phase which was blocked by 78% from the specific photocytotoxic process. DNA laddering analysis showed that PMF (60

  3. Endothelial Cell Migration and Vascular Endothelial Growth Factor Expression Are the Result of Loss of Breast Tissue Polarity

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Amy; Cuevas, Ileana; Kenny, Paraic A; Miyake, Hiroshi; Mace, Kimberley; Ghajar, Cyrus; Boudreau, Aaron; Bissell, Mina; Boudreau, Nancy

    2009-05-26

    Recruiting a new blood supply is a rate-limiting step in tumor progression. In a three-dimensional model of breast carcinogenesis, disorganized, proliferative transformed breast epithelial cells express significantly higher expression of angiogenic genes compared with their polarized, growth-arrested nonmalignant counterparts. Elevated vascular endothelial growth factor (VEGF) secretion by malignant cells enhanced recruitment of endothelial cells (EC) in heterotypic cocultures. Significantly, phenotypic reversion of malignant cells via reexpression of HoxD10, which is lost in malignant progression, significantly attenuated VEGF expression in a hypoxia-inducible factor 1{alpha}-independent fashion and reduced EC migration. This was due primarily to restoring polarity: forced proliferation of polarized, nonmalignant cells did not induce VEGF expression and EC recruitment, whereas disrupting the architecture of growth-arrested, reverted cells did. These data show that disrupting cytostructure activates the angiogenic switch even in the absence of proliferation and/or hypoxia and restoring organization of malignant clusters reduces VEGF expression and EC activation to levels found in quiescent nonmalignant epithelium. These data confirm the importance of tissue architecture and polarity in malignant progression.

  4. Planar cell polarity enables posterior localization of nodal cilia and left-right axis determination during mouse and Xenopus embryogenesis.

    Directory of Open Access Journals (Sweden)

    Dragana Antic

    2010-02-01

    Full Text Available Left-right asymmetry in vertebrates is initiated in an early embryonic structure called the ventral node in human and mouse, and the gastrocoel roof plate (GRP in the frog. Within these structures, each epithelial cell bears a single motile cilium, and the concerted beating of these cilia produces a leftward fluid flow that is required to initiate left-right asymmetric gene expression. The leftward fluid flow is thought to result from the posterior tilt of the cilia, which protrude from near the posterior portion of each cell's apical surface. The cells, therefore, display a morphological planar polarization. Planar cell polarity (PCP is manifested as the coordinated, polarized orientation of cells within epithelial sheets, or as directional cell migration and intercalation during convergent extension. A set of evolutionarily conserved proteins regulates PCP. Here, we provide evidence that vertebrate PCP proteins regulate planar polarity in the mouse ventral node and in the Xenopus gastrocoel roof plate. Asymmetric anterior localization of VANGL1 and PRICKLE2 (PK2 in mouse ventral node cells indicates that these cells are planar polarized by a conserved molecular mechanism. A weakly penetrant Vangl1 mutant phenotype suggests that compromised Vangl1 function may be associated with left-right laterality defects. Stronger functional evidence comes from the Xenopus GRP, where we show that perturbation of VANGL2 protein function disrupts the posterior localization of motile cilia that is required for leftward fluid flow, and causes aberrant expression of the left side-specific gene Nodal. The observation of anterior-posterior PCP in the mouse and in Xenopus embryonic organizers reflects a strong evolutionary conservation of this mechanism that is important for body plan determination.

  5. Effect of gamma rays doses on pollen germination, polysiphony and pollen tube elongation in Pinus patula Schiede et Deppe

    International Nuclear Information System (INIS)

    Katiyar, S.R.; Chauhan, Y.S.

    1987-01-01

    The present study aimed to study the effects of gamma radiation ( 60 Co) on pollen germination and pollen tube elongation in Pinus patula. Pollen germination and pollen tube elongation are stimulated by low doses of radiation. Although higher doses of radiation inhibit the germination of pollen, pollen tube elongation remains unaffected. Thus in Pinus patula pollen tube elongation is less radiosensitive than pollen germination. Compared to control pollen, irradiated pollen produced more number of long pollen tubes. Therefore pollen tube size can be improved using low doses of radiation. (author). 15 refs., 5 figs., 2 tables

  6. Absence of transepithelial anion exchange by rabbit OMCD: Evidence against reversal of cell polarity

    Energy Technology Data Exchange (ETDEWEB)

    Hayashi, Matsuhiko; Schuster, V.L.; Stokes, J.B. (Univ. of Iowa College of Medicine, Iowa City (USA))

    1988-08-01

    In the rabbit cortical collecting duct (CCD), Cl tracer crosses the epithelium predominantly via an anion exchange system that operates in either a Cl-Cl or Cl-HCO{sub 3} exchange mode. In the present study, the authors used the {sup 36}Cl lumen-to-bath rate coefficient (K{sub Cl}, nm/s), a sensitive measurement of CCD transepithelial anion transport, to investigate the nature of Cl transport in the medullary collecting duct dissected from inner stripe, outer medulla (OMCD). The K{sub Cl} in OMCD perfused and bathed in HCO{sub 3}-Ringer solution was low and similar to that value observed in the CCD when anion exchange is inhibited and Cl permeates the epithelium by diffusion. To test the hypothesis that metabolic alkalosis could reverse the polarity of intercalated cells and thus induce an apical Cl-HCO{sub 3} exchanger in H{sup +}-secreting OMCD cells, they measured K{sub Cl} in OMCD from rabbits make alkalotic by deoxycorticosterone and furosemide. Although the base-line K{sub Cl} was slightly higher than in OMCD from control rabbits, the value was still far lower than the K{sub Cl} under comparable conditions in CCD. They conclude (1) Cl transport across the MCD by anion exchange is immeasurably low or nonexistent; (2) unlike the CCD, Cl transport in OMCD is not responsive to cAMP; and (3) metabolic alkalosis does not induce an apical anion exchanger in OMCD, i.e., does not cause epithelial polarity reversal.

  7. Molecular and cellular characterization of the tomato pollen profilin, LePro1.

    Directory of Open Access Journals (Sweden)

    Long-Xi Yu

    Full Text Available Profilin is an actin-binding protein involved in the dynamic turnover and restructuring of the actin cytoskeleton in all eukaryotic cells. We previously cloned a profilin gene, designated as LePro1 from tomato pollen. To understand its biological role, in the present study, we investigated the temporal and spatial expression of LePro1 during pollen development and found that the transcript was only detected at late stages during microsporogenesis and pollen maturation. Using antisense RNA, we successfully knocked down the expression of LePro1 in tomato plants using stable transformation, and obtained two antisense lines, A2 and A3 showing significant down-regulation of LePro1 in pollen resulting in poor pollen germination and abnormal pollen tube growth. A disorganized F-actin distribution was observed in the antisense pollen. Down-regulation of LePro1 also appeared to affect hydration of pollen deposited on the stigma and arrested pollen tube elongation in the style, thereby affecting fertilization. Our results suggest that LePro1 in conjunction with perhaps other cytoskeletal proteins, plays a regulatory role in the proper organization of F-actin in tomato pollen tubes through promoting actin assembly. Down-regulation of LePro1 leads to interruption of actin assembly and disorganization of the actin cytoskeleton thus arresting pollen tube growth. Based on the present and previous studies, it is likely that a single transcript of profilin gives rise to multiple forms displaying multifunctionality in tomato pollen.

  8. Polarizing T and B cell responses by APC-targeted subunit vaccines.

    Directory of Open Access Journals (Sweden)

    Gunnveig eGrødeland

    2015-07-01

    Full Text Available Current influenza vaccines mostly aim at the induction of specific neutralizing antibodies. While antibodies are important for protection against a particular virus strain, T cells can recognize epitopes that will offer broader protection against influenza. We have previously developed a DNA vaccine format by which protein antigens can be targeted specifically to receptors on antigen presenting cells (APCs. The DNA-encoded vaccine proteins are homodimers, each chain consisting of a targeting unit, a dimerization unit, and an antigen. The strategy of targeting antigen to APCs greatly enhances immune responses as compared to non-targeted controls. Furthermore, targeting of antigen to different receptors on APCs can polarize the immune response to different arms of immunity. Here, we discuss how targeting of hemagglutinin (HA to MHC class II molecules increases Th2 and IgG1 antibody responses, whereas targeting to chemokine receptors XCR1 or CCR1/3/5 increases Th1 and IgG2a responses, in addition to CD8+ T cell responses. We also discuss these results in relation to work published by others on APC-targeting. Differential targeting of APC surface molecules may allow the induction of tailor-made phenotypes of adaptive immune responses that are optimal for protection against various infectious agents, including influenza virus.

  9. Directional cell migration establishes the axes of planar polarity in the posterior lateral-line organ of the zebrafish.

    Science.gov (United States)

    López-Schier, Hernán; Starr, Catherine J; Kappler, James A; Kollmar, Richard; Hudspeth, A J

    2004-09-01

    The proper orientation of mechanosensory hair cells along the lateral-line organ of a fish or amphibian is essential for the animal's ability to sense directional water movements. Within the sensory epithelium, hair cells are polarized in a stereotyped manner, but the mechanisms that control their alignment relative to the body axes are unknown. We have found, however, that neuromasts can be oriented either parallel or perpendicular to the anteroposterior body axis. By characterizing the strauss mutant zebrafish line and by tracking labeled cells, we have demonstrated that neuromasts of these two orientations originate from, respectively, the first and second primordia. Furthermore, altering the migratory pathway of a primordium reorients a neuromast's axis of planar polarity. We propose that the global orientation of hair cells relative to the body axes is established through an interaction between directional movement by primordial cells and the timing of neuromast maturation.

  10. Pollen counts in the television media.

    Science.gov (United States)

    Giberson, J E; Murray, L W; Frenz, D A

    1997-01-01

    Allergists routinely supply pollen counts to television stations to assist allergy sufferers and to publicize their medical practices. This investigation examined how widely pollen counts are used in television news programming. The authors telephoned the news director or a meteorologist at 121 broadcast television stations in the nation's 30 largest Designated Market Areas (DMAs). They were asked if pollen counts were included or would be included in news programming during the 1995 pollen season. At least one television station in 28 DMAs included pollen counts in news programming. All five stations in the Washington, DC, DMA aired pollen counts; however, no stations in the Pittsburgh or Milwaukee DMAs carried pollen information. Approximately 49 million television households have access to pollen counts through a broadcast television station. Because these data appear important to television stations and to the public, the authors outline four measures to assure the reliability and meaningfulness of pollen information.

  11. Pollen analyses for pollination research, acetolysis

    Directory of Open Access Journals (Sweden)

    Gretchen D. Jones

    2014-08-01

    Full Text Available Analysis of the pollen associated with pollinators can reveal their role in pollination, the habitats and plants they visit, from where they migrated, what they eat, and dispersal patterns in and around cropping systems. However, finding pollen on and or in a pollinator depends on the technique used to recover the pollen. Acetolysis can be used to recover pollen from internal insect tissues (gut, alimentary canal, crop, etc., external tissues (proboscis, legs, eyes, etc. or both. Acetolysis is the best technique for recovering pollen because any tissue is dissolved and lipids and debris are removed from the sample and the pollen grains. This makes the diagnostic characteristics of the pollen grains more visible so that accurate pollen identification can be made. By using the proper technique, better pollen recovery can be made and thus better data can be obtained.

  12. Characterization of a caleosin expressed during olive (Olea europaea L. pollen ontogeny

    Directory of Open Access Journals (Sweden)

    Rodríguez-García María

    2011-08-01

    Full Text Available Abstract Background The olive tree is an oil-storing species, with pollen being the second most active site in storage lipid biosynthesis. Caleosins are proteins involved in storage lipid mobilization during seed germination. Despite the existence of different lipidic structures in the anther, there are no data regarding the presence of caleosins in this organ to date. The purpose of the present work was to characterize a caleosin expressed in the olive anther over different key stages of pollen ontogeny, as a first approach to unravel its biological function in reproduction. Results A 30 kDa caleosin was identified in the anther tissues by Western blot analysis. Using fluorescence and transmission electron microscopic immunolocalization methods, the protein was first localized in the tapetal cells at the free microspore stage. Caleosins were released to the anther locule and further deposited onto the sculptures of the pollen exine. As anthers developed, tapetal cells showed the presence of structures constituted by caleosin-containing lipid droplets closely packed and enclosed by ER-derived cisternae and vesicles. After tapetal cells lost their integrity, the caleosin-containing remnants of the tapetum filled the cavities of the mature pollen exine, forming the pollen coat. In developing microspores, this caleosin was initially detected on the exine sculptures. During pollen maturation, caleosin levels progressively increased in the vegetative cell, concurrently with the number of oil bodies. The olive pollen caleosin was able to bind calcium in vitro. Moreover, PEGylation experiments supported the structural conformation model suggested for caleosins from seed oil bodies. Conclusions In the olive anther, a caleosin is expressed in both the tapetal and germ line cells, with its synthesis independently regulated. The pollen oil body-associated caleosin is synthesized by the vegetative cell, whereas the protein located on the pollen exine and

  13. Pollen Forecast and Dispersion Modelling

    Science.gov (United States)

    Costantini, Monica; Di Giuseppe, Fabio; Medaglia, Carlo Maria; Travaglini, Alessandro; Tocci, Raffaella; Brighetti, M. Antonia; Petitta, Marcello

    2014-05-01

    The aim of this study is monitoring, mapping and forecast of pollen distribution for the city of Rome using in-situ measurements of 10 species of common allergenic pollens and measurements of PM10. The production of daily concentration maps, associated to a mobile phone app, are innovative compared to existing dedicated services to people who suffer from respiratory allergies. The dispersal pollen is one of the most well-known causes of allergic disease that is manifested by disorders of the respiratory functions. Allergies are the third leading cause of chronic disease and it is estimated that tens millions of people in Italy suffer from it. Recent works reveal that during the last few years there was a progressive increase of affected subjects, especially in urban areas. This situation may depend: on the ability to transport of pollutants, on the ability to react between pollutants and pollen and from a combination of other irritants, existing in densely populated and polluted urban areas. The methodology used to produce maps is based on in-situ measurements time series relative to 2012, obtained from networks of air quality and pollen stations in the metropolitan area of Rome. The monitoring station aerobiological of University of Rome "Tor Vergata" is located at the Department of Biology. The instrument used to pollen monitoring is a volumetric sampler type Hirst (Hirst 1952), Model 2000 VPPS Lanzoni; the data acquisition is carried out as reported in Standard UNI 11008:2004 - "Qualità dell'aria - Metodo di campionamento e conteggio dei granuli pollinici e delle spore fungine aerodisperse" - the protocol that describes the procedure for measuring of the concentration of pollen grains and fungal spores dispersed into the atmosphere, and reported in the "Manuale di gestione e qualità della R.I.M.A" (Travaglini et. al. 2009). All 10 allergenic pollen are monitored since 1996. At Tor Vergata university is also operating a meteorological station (SP2000, CAE

  14. A Recombinant Fragment of Human Surfactant Protein D Suppresses Basophil Activation and T-Helper Type 2 and B-Cell Responses in Grass Pollen-induced Allergic Inflammation.

    Science.gov (United States)

    Qaseem, Asif S; Singh, Iesha; Pathan, Ansar A; Layhadi, Janice A; Parkin, Rebecca; Alexandra, Fedina; Durham, Stephen R; Kishore, Uday; Shamji, Mohamed H

    2017-12-15

    Recombinant fragment of human surfactant protein D (rfhSP-D) has been shown to suppress house dust mite- and Aspergillus fumigatus-induced allergic inflammation in murine models. We sought to elucidate the effect of rfhSP-D on high-affinity IgE receptor- and CD23-mediated, grass pollen-induced allergic inflammatory responses. rfhSP-D, containing homotrimeric neck and lectin domains, was expressed in Escherichia coli BL21(λDE3)pLysS cells. Peripheral blood mononuclear cells and sera were obtained from individuals with grass pollen allergy (n = 27). The effect of rfhSP-D on basophil activation and histamine release was measured by flow cytometry. IgE-facilitated allergen binding and presentation were assessed by flow cytometry. T-helper cell type 2 (Th2) cytokines were measured in cell culture supernatants. The effect of rfhSP-D on IgE production by B cells when stimulated with CD40L, IL-4, and IL-21 was also determined. rfhSP-D bound to Phleum pratense in a dose- and calcium-dependent manner. Allergen-induced basophil responsiveness and histamine release were inhibited in the presence of rfhSP-D, as measured by CD63, CD203c (P = 0.0086, P = 0.04205), and intracellularly labeled diamine oxidase (P = 0.0003, P = 0.0148). The binding of allergen-IgE complexes to B cells was reduced by 51% (P = 0.002) in the presence of rfhSP-D. This decrease was concomitant with reduction in CD23 expression on B cells (P D suppressed allergen-driven CD27 - CD4 + CRTh2 + T-cell proliferation (P P D inhibited CD40L/IL-4- and IL-21-mediated IgE production (77.12%; P = 0.02) by B cells. For the first time, to our knowledge, we show that rfhSP-D inhibited allergen-induced basophil responses at a single-cell level and suppressed CD23-mediated facilitated allergen presentation and Th2 cytokine production. In addition, rfhSP-D inhibited IgE synthesis by B cells, which is also a novel observation.

  15. Pollen cone anatomy of Classostrobus crossii sp. nov. (Cheirolepidiaceae)

    Energy Technology Data Exchange (ETDEWEB)

    Rothwell, Gar W.; Mapes, Gene [Department of Environmental and Plant Biology, Ohio University, Athens OH 45701 (United States); Hilton, Jason [Department of Earth Sciences, School of Geography, Earth and Environmental Sciences, The University of Birmingham, Edgbaston, Birmingham, B15 2TT (United Kingdom); Hollingworth, Neville T. [Centre for Ecology and Hydrology, Winfrith Technology Centre, Winfrith Newburgh, Dorchester, Dorset, DT2 8ZD (United Kingdom)

    2007-01-02

    Discovery of a permineralized fossil cone in Mesozoic deposits of southern England provides an opportunity to document the first detailed evidence of internal pollen cone anatomy for the extinct conifer family Cheirolepidiaceae. The specimen, described here as Classostrobus crossii sp. nov., occurs in a calcareous nodule recovered from Middle Jurassic marine sediments of the Lower Callovian Sigaloceras calloviense biozone, Kellaways, near Cirencester, England. The cone is 2.0 cm long and 1.8 cm wide. Sporophylls diverge helically from the axis. Each sporophyll displays a narrow stalk and a distal lamina approx. 11 mm long that tapers to a pointed tip. There is also a basal keel that bends inward at the bottom and sides to form a shallow pocket. A single vascular bundle diverges from the cone axis, extends distally into the sporophyll stalk at the contact of two distinctly different histological zones, and further expands into the distal lamina as transfusion tracheids. Several pollen sacs are attached abaxially at the juncture of the sporophyll stalk and keel. Pollen is roughly spheroidal, 26-35 {mu}m in diameter, with unequal polar caps separated by a striated belt with a subequatorial furrow. This specimen helps clarify the range of variation in the morphology of Mesozoic conifer pollen cones. (author)

  16. Ca2+ influx and phosphoinositide signalling are essential for the establishment and maintenance of cell polarity in monospores from the red alga Porphyra yezoensis.

    Science.gov (United States)

    Li, Lin; Saga, Naotsune; Mikami, Koji

    2009-01-01

    The asymmetrical distribution of F-actin directed by cell polarity has been observed during the migration of monospores from the red alga Porphyra yezoensis. The significance of Ca2+ influx and phosphoinositide signalling during the formation of cell polarity in migrating monospores was analysed pharmacologically. The results indicate that the inhibition of the establishment of cell polarity, as judged by the ability of F-actin to localize asymmetrically, cell wall synthesis, and development into germlings, occurred when monospores were treated with inhibitors of the Ca2+ permeable channel, phospholipase C (PLC), diacylglycerol kinase, and inositol-1,4,5-trisphosphate receptor. Moreover, it was also found that light triggered the establishment of cell polarity via photosynthetic activity but not its direction, indicating that the Ca2+ influx and PLC activation required for the establishment of cell polarity are light dependent. By contrast, inhibition of phospholipase D (PLD) prevented the migration of monospores but not the asymmetrical localization of F-actin. Taken together, these findings suggest that there is functional diversity between the PLC and PLD signalling systems in terms of the formation of cell polarity; the former being critical for the light-dependent establishment of cell polarity and the latter playing a role in the maintenance of established cell polarity.

  17. Alpha-type-1 polarized dendritic cells: A novel immunization tool with optimized CTL-inducing activity

    NARCIS (Netherlands)

    Mailliard, Robbie B.; Wankowicz-Kalinska, Anna; Cai, Quan; Wesa, Amy; Hilkens, Catharien M.; Kapsenberg, Martien L.; Kirkwood, John M.; Storkus, Walter J.; Kalinski, Pawel

    2004-01-01

    Using the principle of functional polarization of dendritic cells (DCs), we have developed a novel protocol to generate human DCs combining the three features critical for the induction of type-1 immunity: (a) fully mature status; (b) responsiveness to secondary lymphoid organ chemokines; and (c)

  18. Regulatory Cells and Immunosuppressive Cytokines: Parasite-Derived Factors Induce Immune Polarization

    Directory of Open Access Journals (Sweden)

    Ali Ouaissi

    2007-01-01

    Full Text Available Parasitic infections are prevalent in both tropical and subtropical areas. Most of the affected and/or exposed populations are living in developing countries where control measures are lacking or inadequately applied. Although significant progress has been made in our understanding of the immune response to parasites, no definitive step has yet been successfully done in terms of operational vaccines against parasitic diseases. Evidence accumulated during the past few years suggests that the pathology observed during parasitic infections is in part due to deregulation of normal components of the immune system, mainly cytokines, antibodies, and immune effector cell populations. A large number of studies that illustrate how parasites can modify the host immune system for their own benefit have been reported in both metazoan and protozoan parasites. The first line of defense against foreign organisms is barrier tissue such as skin, humoral factors, for instance the complement system and pentraxin, which upon activation of the complement cascade facilitate pathogen recognition by cells of innate immunity such as macrophages and DC. However, all the major groups of parasites studied have been shown to contain and/or to release factors, which interfere with both arms of the host immune system. Even some astonishing observations relate to the production by some parasites of orthologues of mammalian cytokines. Furthermore, chronic parasitic infections have led to the immunosuppressive environment that correlates with increased levels of myeloid and T suppressor cells that may limit the success of immunotherapeutic strategies based on vaccination. This minireview briefly analyzes some of the current data related to the regulatory cells and molecules derived from parasites that affect cellular function and contribute to the polarization of the immune response of the host. Special attention is given to some of the data from our laboratory illustrating the

  19. Auxin efflux carrier activity and auxin accumulation regulate cell division and polarity in tobacco cells

    Czech Academy of Sciences Publication Activity Database

    Petrášek, Jan; Elčkner, Miroslav; Morris, David; Zažímalová, Eva

    2002-01-01

    Roč. 216, - (2002), s. 302-308 ISSN 0032-0935 R&D Projects: GA ČR GA206/98/1510 Grant - others:INCO Copernicus(BE) IC15-CT98-0118 Institutional research plan: CEZ:AV0Z5038910 Keywords : Auxin carrier * 1,N,Naphthylphthalamic acid * Nicotiana ( cell culture) Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.960, year: 2002

  20. Lactobacilli Activate Human Dendritic Cells that Skew T Cells Toward T Helper 1 Polarization

    Science.gov (United States)

    2005-01-06

    as Klebsiella pneumoniae (21–23). To determine whether this difference might vary among species, live L . gasseri, L . johnsonii, and L . reuteri cells...or L . reuteri plus E. coli LPS for 2 d. Under these conditions, LPS promoted the production of IL-10 only in 11% of MDCs, compared with the high (60...induction of IL-10 with E. coli LPS in the absence of L . reuteri (Fig. 2 B and C). Lethally-irradiated Lactobacillus species (10 gml) also

  1. Coconut (Cocos nucifera l.) pollen cryopreservation.

    Science.gov (United States)

    Karun, A; Sjini, K K; Niral, V; Amarnth, C H; Remya, P; Rajesh, M K; Samsudeen, K; Jerard, B A; Engelmann, F

    2014-01-01

    Coconut genetic resources are threatened by pests and pathogens, natural hazards and human activities. Cryopreservation is the only method allowing the safe and cost-effective long-term conservation of recalcitrant seed species such as coconut. The objective of this work was to test the effect of cryopreservation and of cryostorage duration on coconut pollen germination and fertility. Pollen of two coconut varieties (West Coast Tall WWCTW and Chowghat Orange Dwarf CODC) was collected in March-May over three successive years, desiccated to 7.5 % moisture content (FW) and cryopreserved by direct immersion in liquid nitrogen. Germination and pollen tube length (PTL) of desiccated and cryopreserved pollen were not significantly different for both WCT and COD over the three harvest months of the three consecutive years of study. Pollen germination ranged from 24 to 32 % in desiccated pollen whereas it was between 26 and 29 % in cryopreserved COD pollen. In the case of WCT, germination ranged from 30 to 31 % in desiccated pollen, while it was between 28 and 32 % in cryopreserved pollen. PTL of cryopreserved pollen ranged between 224-390 nm and 226-396 mm for COD and WCT, respectively. Germination of COD pollen varied between 29.0 and 44.1 % after 4 years and 1.0/1.5 years cryostorage, respectively. Germination of WCT pollen did not change significantly between 0 and 6 years cryostorage, being comprised between 32 (24 h) and 40 % (1.5 years). Germination and vigour of cryopreserved pollen were generally higher compared to that of pollen dried in oven and non-cryopreserved. Normal seed set was observed in COD and WCT palms using pollen cryostored for 6 months and 4 years. Cryopreserved pollen of five Tall and five Dwarf accessions displayed 24-31 % and 25-49 % germination, respectively. These results show that it is now possible to establish pollen cryobanks to contribute to coconut germplasm long-term conservation.

  2. Pollen morphology and viability in Bromeliaceae

    Directory of Open Access Journals (Sweden)

    EVERTON H. SOUZA

    2017-12-01

    Full Text Available ABSTRACT Pollen morphology characterization is important in taxonomy, conservation and plant breeding, and pollen viability studies can support breeding programs. This study investigated pollen morphology and male fertility in 18 species of Bromeliaceae with ornamental potential. For morphological characterization, pollen grains were acetolyzed and characterization of exine was done using scanning and transmission electron microscopy. Pollen viability was investigated by in vitro germination and histochemical tests. Species belonging to Aechmea and Ananas genera presented medium size pollen, except for Ae. fasciata, with large pollen. Al. nahoumii, P. sagenarius and the Vriesea species analyzed showed large pollen, except for V. carinata, with very large pollen. Pollen of Aechmea, Ananas and P. sagenarius presented bilateral symmetry, diporate, exine varying from tectate to semitectate. Al. nahoumii and Vriesea species presented pollen with bilateral symmetry, monocolpate; exine was semitectate, reticulate and heterobrochate. Germination percentage and tube growth were greater in SM and BKM media. Histochemical tests showed pollen viability above 70% for all species, except for Ananas sp. (40%. Pollen morphology is important for the identification of species, especially in this family, which contains a large number of species. High rates of viability favor fertilization and seed production, essential for efficient hybrid production and conservation.

  3. Pollen morphology and viability in Bromeliaceae.

    Science.gov (United States)

    Souza, Everton H; Souza, Fernanda V D; Rossi, Mônica L; Packer, Renan M; Cruz-Barros, Maria Amelia V; Martinelli, Adriana P

    2017-01-01

    Pollen morphology characterization is important in taxonomy, conservation and plant breeding, and pollen viability studies can support breeding programs. This study investigated pollen morphology and male fertility in 18 species of Bromeliaceae with ornamental potential. For morphological characterization, pollen grains were acetolyzed and characterization of exine was done using scanning and transmission electron microscopy. Pollen viability was investigated by in vitro germination and histochemical tests. Species belonging to Aechmea and Ananas genera presented medium size pollen, except for Ae. fasciata, with large pollen. Al. nahoumii, P. sagenarius and the Vriesea species analyzed showed large pollen, except for V. carinata, with very large pollen. Pollen of Aechmea, Ananas and P. sagenarius presented bilateral symmetry, diporate, exine varying from tectate to semitectate. Al. nahoumii and Vriesea species presented pollen with bilateral symmetry, monocolpate; exine was semitectate, reticulate and heterobrochate. Germination percentage and tube growth were greater in SM and BKM media. Histochemical tests showed pollen viability above 70% for all species, except for Ananas sp. (40%). Pollen morphology is important for the identification of species, especially in this family, which contains a large number of species. High rates of viability favor fertilization and seed production, essential for efficient hybrid production and conservation.

  4. Peptide signalling during the pollen tube journey and double fertilization.

    Science.gov (United States)

    Qu, Li-Jia; Li, Ling; Lan, Zijun; Dresselhaus, Thomas

    2015-08-01

    Flowering seed plants (angiosperms) have evolved unique ways to protect their gametes from pathogen attack and from drying out. The female gametes (egg and central cell) are deeply embedded in the maternal tissues of the ovule inside the ovary, while the male gametes (sperm cells) are enclosed in the vegetative pollen tube cell. After germination of the pollen tube at the surface of papilla cells of the stigma the two immobile sperm cells are transported deep inside the sporophytic maternal tissues to be released inside the ovule for double fertilization. Angiosperms have evolved a number of hurdles along the pollen tube journey to prevent inbreeding and fertilization by alien sperm cells, and to maximize reproductive success. These pre-zygotic hybridization barriers require intensive communication between the male and female reproductive cells and the necessity to distinguish self from non-self interaction partners. General molecules such as nitric oxide (NO) or gamma-aminobutyric acid (GABA) therefore appear to play only a minor role in these species-specific communication events. The past 20 years have shown that highly polymorphic peptides play a leading role in all communication steps along the pollen tube pathway and fertilization. Here we review our current understanding of the role of peptides during reproduction with a focus on peptide signalling during self-incompatibility, pollen tube growth and guidance as well as sperm reception and gamete activation. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  5. The influence of non polar and polar molecules in mouse motile cells membranes and pure lipid bilayers.

    Directory of Open Access Journals (Sweden)

    Francisco J Sierra-Valdez

    Full Text Available We report an experimental study of mouse sperm motility that shows chief aspects characteristic of neurons: the anesthetic (produced by tetracaine and excitatory (produced by either caffeine or calcium effects and their antagonic action. While tetracaine inhibits sperm motility and caffeine has an excitatory action, the combination of these two substances balance the effects, producing a motility quite similar to that of control cells. We also study the effects of these agents (anesthetic and excitatory on the melting points of pure lipid liposomes constituted by 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC and dipalmitoyl phosphatidic acid (DPPA. Tetracaine induces a large fluidization of the membrane, shifting the liposomes melting transition temperature to much lower values. The effect of caffeine is null, but its addition to tetracaine-doped liposomes greatly screen the fluidization effect. A high calcium concentration stiffens pure lipid membranes and strongly reduces the effect of tetracaine. Molecular Dynamics Simulations are performed to further understand our experimental findings at the molecular level. We find a strong correlation between the effect of antagonic molecules that could explain how the mechanical properties suitable for normal cell functioning are affected and recovered.

  6. Non-canonical Wnt signaling regulates cell polarity in female reproductive tract development via van gogh-like 2

    Science.gov (United States)

    vandenBerg, Alysia L.; Sassoon, David A.

    2009-01-01

    Summary Wnt signaling effectors direct the development and adult remodeling of the female reproductive tract (FRT); however, the role of non-canonical Wnt signaling has not been explored in this tissue. The non-canonical Wnt signaling protein van gogh-like 2 is mutated in loop-tail (Lp) mutant mice (Vangl2Lp), which display defects in multiple tissues. We find that Vangl2Lp mutant uterine epithelium displays altered cell polarity, concommitant with changes in cytoskeletal actin and scribble (scribbled, Scrb1) localization. The postnatal mutant phenotype is an exacerbation of that seen at birth, exhibiting more smooth muscle and reduced stromal mesenchyme. These data suggest that early changes in cell polarity have lasting consequences for FRT development. Furthermore, Vangl2 is required to restrict Scrb1 protein to the basolateral epithelial membrane in the neonatal uterus, and an accumulation of fibrillar-like structures observed by electron microscopy in Vangl2Lp mutant epithelium suggests that mislocalization of Scrb1 in mutants alters the composition of the apical face of the epithelium. Heterozygous and homozygous Vangl2Lp mutant postnatal tissues exhibit similar phenotypes and polarity defects and display a 50% reduction in Wnt7a levels, suggesting that the Vangl2Lp mutation acts dominantly in the FRT. These studies demonstrate that the establishment and maintenance of cell polarity through non-canonical Wnt signaling are required for FRT development. PMID:19363157

  7. National Allergy Bureau Pollen and Mold Report

    Science.gov (United States)

    ... Search AAAAI National Allergy Bureau Pollen and Mold Report Date: May 01, 2018 Location: San Antonio (2), ... 30/2018 ( click here to view ). Our Allergen Report Email Service can automatically email you daily pollen ...

  8. Differential effects of carbohydrates on arabidopsis pollen germination

    Czech Academy of Sciences Publication Activity Database

    Hirsche, J.; Fernández, J. M. G.; Stabentheiner, E.; Großkinsky, D.K.; Roitsch, Thomas

    2017-01-01

    Roč. 58, č. 4 (2017), s. 691-701 ISSN 0032-0781 R&D Projects: GA MŠk(CZ) LO1415 Institutional support: RVO:67179843 Keywords : Arabidopsis thaliana * Carbohydrates * Metabolic regulation * Pollen germination * Signaling * Structure-function relationship Subject RIV: EF - Botanics OBOR OECD: Cell biology Impact factor: 4.760, year: 2016

  9. Pollen loads of eucalypt and other pollen types in birds in NW Spain

    Directory of Open Access Journals (Sweden)

    María Calviño-Cancela

    2015-12-01

    Full Text Available Here we present the amount of pollen of eucalypt and pollen of other types for birds captured in two bird ringing stations for 14 months (March 2014 to April 2015 in NW Spain. Common and latin names of all birds species captured, together with the number of captured individuals (N, prevalence of eucalypt pollen (percentage of individuals with eucalypt pollen and of pollen of other types and average pollen loads per individual for eucalypt and other pollen types is presented. See [1] for further information and discussion.

  10. [The epidemiology of pollen allergy].

    Science.gov (United States)

    Charpin, D; Caillaud, D

    2014-04-01

    The prevalence of seasonal allergic rhinitis can be established through surveys performed in a sample of the general population. These surveys are based on a questionnaire, which could lead to an overestimate of prevalence rates, and on measurements of specific IgE, which need to be interpreted in the light of the responses to the questionnaire. Such surveys are few in France and need to be updated. Risk factors for seasonal allergic rhinitis are genetic, epigenetic and environmental. Relationships between exposure to pollen and health can be documented through ecological and panel surveys. Panel surveys may give information on threshold levels and dose-response relationships. In addition to pollen exposure, global warming and air pollutants act as cofactors. Monitoring of both pollen exposure and its health effects should be encouraged and strengthened. Copyright © 2014 SPLF. Published by Elsevier Masson SAS. All rights reserved.

  11. Pollination with gamma-irradiated pollen and seed development in kiwifruit (Actinidia deliciosa var. deliciosa)

    International Nuclear Information System (INIS)

    Musial, K.

    1997-01-01

    Full text. The effects of pollen irradiation at 70 and 90 kr on seed set were studied in Actinidia deliciosa var. deliciosa. Pollination with irradiated pollen affected seed development and contents. Rising irradiation doses increased the percentages of empty seeds and decreased the percentages of seeds containing embryos with endosperm. Moreover, pollination with heavily irradiated pollen led to the formation of seeds containing the endosperm only. Embryo and endosperm size was also strongly influenced by irradiated pollen. The length of endosperms was reduced at all levels of pollen irradiation compared to the non-irradiated controls; the embryo development was conspicuously retarded. Cells in endosperm resulting from the treatments differed in the presence and number of starch grains. (author)

  12. T cells' immunological synapses induce polarization of brain astrocytes in vivo and in vitro: a novel astrocyte response mechanism to cellular injury.

    Science.gov (United States)

    Barcia, Carlos; Sanderson, Nicholas S R; Barrett, Robert J; Wawrowsky, Kolja; Kroeger, Kurt M; Puntel, Mariana; Liu, Chunyan; Castro, Maria G; Lowenstein, Pedro R

    2008-08-20

    Astrocytes usually respond to trauma, stroke, or neurodegeneration by undergoing cellular hypertrophy, yet, their response to a specific immune attack by T cells is poorly understood. Effector T cells establish specific contacts with target cells, known as immunological synapses, during clearance of virally infected cells from the brain. Immunological synapses mediate intercellular communication between T cells and target cells, both in vitro and in vivo. How target virally infected astrocytes respond to the formation of immunological synapses established by effector T cells is unknown. Herein we demonstrate that, as a consequence of T cell attack, infected astrocytes undergo dramatic morphological changes. From normally multipolar cells, they become unipolar, extending a major protrusion towards the immunological synapse formed by the effector T cells, and withdrawing most of their finer processes. Thus, target astrocytes become polarized towards the contacting T cells. The MTOC, the organizer of cell polarity, is localized to the base of the protrusion, and Golgi stacks are distributed throughout the protrusion, reaching distally towards the immunological synapse. Thus, rather than causing astrocyte hypertrophy, antiviral T cells cause a major structural reorganization of target virally infected astrocytes. Astrocyte polarization, as opposed to hypertrophy, in response to T cell attack may be due to T cells providing a very focused attack, and thus, astrocytes responding in a polarized manner. A similar polarization of Golgi stacks towards contacting T cells was also detected using an in vitro allogeneic model. Thus, different T cells are able to induce polarization of target astrocytes. Polarization of target astrocytes in response to immunological synapses may play an important role in regulating the outcome of the response of astrocytes to attacking effector T cells, whether during antiviral (e.g. infected during HIV, HTLV-1, HSV-1 or LCMV infection), anti

  13. T cells' immunological synapses induce polarization of brain astrocytes in vivo and in vitro: a novel astrocyte response mechanism to cellular injury.

    Directory of Open Access Journals (Sweden)

    Carlos Barcia

    2008-08-01

    Full Text Available Astrocytes usually respond to trauma, stroke, or neurodegeneration by undergoing cellular hypertrophy, yet, their response to a specific immune attack by T cells is poorly understood. Effector T cells establish specific contacts with target cells, known as immunological synapses, during clearance of virally infected cells from the brain. Immunological synapses mediate intercellular communication between T cells and target cells, both in vitro and in vivo. How target virally infected astrocytes respond to the formation of immunological synapses established by effector T cells is unknown.Herein we demonstrate that, as a consequence of T cell attack, infected astrocytes undergo dramatic morphological changes. From normally multipolar cells, they become unipolar, extending a major protrusion towards the immunological synapse formed by the effector T cells, and withdrawing most of their finer processes. Thus, target astrocytes become polarized towards the contacting T cells. The MTOC, the organizer of cell polarity, is localized to the base of the protrusion, and Golgi stacks are distributed throughout the protrusion, reaching distally towards the immunological synapse. Thus, rather than causing astrocyte hypertrophy, antiviral T cells cause a major structural reorganization of target virally infected astrocytes.Astrocyte polarization, as opposed to hypertrophy, in response to T cell attack may be due to T cells providing a very focused attack, and thus, astrocytes responding in a polarized manner. A similar polarization of Golgi stacks towards contacting T cells was also detected using an in vitro allogeneic model. Thus, different T cells are able to induce polarization of target astrocytes. Polarization of target astrocytes in response to immunological synapses may play an important role in regulating the outcome of the response of astrocytes to attacking effector T cells, whether during antiviral (e.g. infected during HIV, HTLV-1, HSV-1 or LCMV

  14. Cigarette smoke extract-treated mast cells promote alveolar macrophage infiltration and polarization in experimental chronic obstructive pulmonary disease.

    Science.gov (United States)

    Li, Hong; Yang, Tian; Ning, Qian; Li, Feiyan; Chen, Tianjun; Yao, Yan; Sun, Zhongmin

    2015-01-01

    Cigarette smoking is the main cause of chronic obstructive pulmonary disease (COPD) and may modulate the immune response of exposed individuals. Mast cell function can be altered by cigarette smoking, but the role of smoking in COPD remains poorly understood. The current study aimed to explore the role of cigarette smoke extract (CSE)-treated mast cells in COPD pathogenesis. Cytokine and chemokine expression as well as degranulation of bone marrow-derived mast cells (BMMCs) were detected in cells exposed to immunoglobulin E (IgE) and various doses of CSE. Adoptive transfer of CSE-treated BMMCs into C57BL/6J mice was performed, and macrophage infiltration and polarization were evaluated by fluorescence-activated cell sorting (FACS). Furthermore, a coculture system of BMMCs and macrophages was established to examine macrophage phenotype transition. The role of protease serine member S31 (Prss31) was also investigated in the co-culture system and in COPD mice. CSE exposure suppressed cytokine expression and degranulation in BMMCs, but promoted the expressions of chemokines and Prss31. Adoptive transfer of CSE-treated BMMCs induced macrophage infiltration and M2 polarization in the mouse lung. Moreover, CSE-treated BMMCs triggered macrophage M2 polarization via Prss31 secretion. Recombinant Prss31 was shown to activate interleukin (IL)-13/IL-13Rα/Signal transducers and activators of transcription (Stat) 6 signaling in macrophages. Additionally, a positive correlation was found between Prss31 expression and the number of M2 macrophages in COPD mice. In conclusion, CSE-treated mast cells may induce macrophage infiltration and M2 polarization via Prss31 expression, and potentially contribute to COPD progression.

  15. Evidence of pollen tubes in paleozoic pteridosperms.

    Science.gov (United States)

    Rothwell, G W

    1972-02-18

    A saccate pollen grain with a branched pollen tube has been discovered within the pollen chamber of a fossil seed-fern ovule of Middle Pennsylvanian age. This suggests that microgametophytes comparable to those of living gymnosperms were produced by some Paleozoic pteridosperms.

  16. A link between planar polarity and staircase-like bundle architecture in hair cells.

    Science.gov (United States)

    Tarchini, Basile; Tadenev, Abigail L D; Devanney, Nicholas; Cayouette, Michel

    2016-11-01

    Sensory perception in the inner ear relies on the hair bundle, the highly polarized brush of movement detectors that crowns hair cells. We previously showed that, in the mouse cochlea, the edge of the forming bundle is defined by the 'bare zone', a microvilli-free sub-region of apical membrane specified by the Insc-LGN-Gαi protein complex. We now report that LGN and Gαi also occupy the very tip of stereocilia that directly abut the bare zone. We demonstrate that LGN and Gαi are both essential for promoting the elongation and differential identity of stereocilia across rows. Interestingly, we also reveal that total LGN-Gαi protein amounts are actively balanced between the bare zone and stereocilia tips, suggesting that early planar asymmetry of protein enrichment at the bare zone confers adjacent stereocilia their tallest identity. We propose that LGN and Gαi participate in a long-inferred signal that originates outside the bundle to model its staircase-like architecture, a property that is essential for direction sensitivity to mechanical deflection and hearing. © 2016. Published by The Company of Biologists Ltd.

  17. Bipolar Plasma Membrane Distribution of Phosphoinositides and Their Requirement for Auxin-Mediated Cell Polarity and Patterning in Arabidopsis[W

    Science.gov (United States)

    Tejos, Ricardo; Sauer, Michael; Vanneste, Steffen; Palacios-Gomez, Miriam; Li, Hongjiang; Heilmann, Mareike; van Wijk, Ringo; Vermeer, Joop E.M.; Heilmann, Ingo; Munnik, Teun; Friml, Jiří

    2014-01-01

    Cell polarity manifested by asymmetric distribution of cargoes, such as receptors and transporters, within the plasma membrane (PM) is crucial for essential functions in multicellular organisms. In plants, cell polarity (re)establishment is intimately linked to patterning processes. Despite the importance of cell polarity, its underlying mechanisms are still largely unknown, including the definition and distinctiveness of the polar domains within the PM. Here, we show in Arabidopsis thaliana that the signaling membrane components, the phosphoinositides phosphatidylinositol 4-phosphate (PtdIns4P) and phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] as well as PtdIns4P 5-kinases mediating their interconversion, are specifically enriched at apical and basal polar plasma membrane domains. The PtdIns4P 5-kinases PIP5K1 and PIP5K2 are redundantly required for polar localization of specifically apical and basal cargoes, such as PIN-FORMED transporters for the plant hormone auxin. As a consequence of the polarity defects, instructive auxin gradients as well as embryonic and postembryonic patterning are severely compromised. Furthermore, auxin itself regulates PIP5K transcription and PtdIns4P and PtdIns(4,5)P2 levels, in particular their association with polar PM domains. Our results provide insight into the polar domain–delineating mechanisms in plant cells that depend on apical and basal distribution of membrane lipids and are essential for embryonic and postembryonic patterning. PMID:24876254

  18. Intracellular photoreceptive site for blue light-induced cell division in protonemata of the fern Adiantum [Pteridophyta]: Further analyses by polarized light irradiation and cell centrifugation

    International Nuclear Information System (INIS)

    Kadota, A.; Fushimi, Y.; Wada, M.

    1986-01-01

    The intracellular localization of the photoreceptive site for blue light-induced cell division in single-celled protonemata of Adiantum capillus-veneris L. was investigated using polarized light irradiation and protonemal cell centrifugation. The response to irradiation with polarized blue light showed no dependence on the direction of light polarization. However, centrifugation of the protonemata followed by microbeam irradiation showed that the site of blue light perception could be displaced together with the nucleus. Centrifugal treatment changed the distribution of intracellular organelles at the time of light exposure and basipetally displaced the nucleus about 90μm. This treatment had no effect on the induction of cell division with blue light if the protonemata were centrifuged again acropetally after the light treatment. Microbeam (30×30 μm2) irradiation with blue light of the apical 45–75 βm region, the receptive site of blue light in non-centrifuged cell, did not induce cell division. However, cell division was induced by irradiation of the nucleus-containing region, indicating that the photoreceptive site was displaced together with the nucleus by the centrifugation. These results suggest that the blue light receptor regulating cell division in Adiantum protonemata is not likely to be located on the plasma membrane. (author)

  19. Pollen characteristics and in vitro pollen germination of Cedrus ...

    African Journals Online (AJOL)

    PRECIOUS

    2009-11-02

    Nov 2, 2009 ... 1Forest Tree Seeds and Tree Breeding Research Directory, Ankara, Turkey. 2Zonguldak Karaelmas ... germination rate in MS medium was achieved in clone no.11342 with 84.77% among the pollen samples of 2005. On the other hand ... Among these species, Cedrus libani A. Rich. (Lebanon. Cedar) is a ...

  20. Par1b links lumen polarity with LGN-NuMA positioning for distinct epithelial cell division phenotypes.

    Science.gov (United States)

    Lázaro-Diéguez, Francisco; Cohen, David; Fernandez, Dawn; Hodgson, Louis; van Ijzendoorn, Sven C D; Müsch, Anne

    2013-10-28

    Columnar epithelia establish their luminal domains and their mitotic spindles parallel to the basal surface and undergo symmetric cell divisions in which the cleavage furrow bisects the apical domain. Hepatocyte lumina interrupt the lateral domain of neighboring cells perpendicular to two basal domains and their cleavage furrow rarely bifurcates the luminal domains. We determine that the serine/threonine kinase Par1b defines lumen position in concert with the position of the astral microtubule anchoring complex LGN-NuMA to yield the distinct epithelial division phenotypes. Par1b signaling via the extracellular matrix (ECM) in polarizing cells determined RhoA/Rho-kinase activity at cell-cell contact sites. Columnar MDCK and Par1b-depleted hepatocytic HepG2 cells featured high RhoA activity that correlated with robust LGN-NuMA recruitment to the metaphase cortex, spindle alignment with the substratum, and columnar organization. Reduced RhoA activity at the metaphase cortex in HepG2 cells and Par1b-overexpressing MDCK cells correlated with a single or no LGN-NuMA crescent, tilted spindles, and the development of lateral lumen polarity.

  1. Glutathione-S-transferase: a minor allergen in birch pollen due to limited release from hydrated pollen.

    Directory of Open Access Journals (Sweden)

    Stephan Deifl

    Full Text Available Recently, a protein homologous to glutathione-S-transferases (GST was detected in prominent amounts in birch pollen by proteomic profiling. As members of the GST family are relevant allergens in mites, cockroach and fungi we investigated the allergenic relevance of GST from birch (bGST.bGST was expressed in Escherichia coli, purified and characterized by mass spectrometry. Sera from 217 birch pollen-allergic patients were tested for IgE-reactivity to bGST by ELISA. The mediator-releasing activity of bGST was analysed with IgE-loaded rat basophil leukaemia cells (RBL expressing human FcεRI. BALB/c mice were immunized with bGST or Bet v 1. Antibody and T cell responses to either protein were assessed. IgE-cross-reactivity between bGST with GST from house dust mite, Der p 8, was studied with murine and human sera in ELISA. The release kinetics of bGST and Bet v 1 from birch pollen were assessed in water, simulated lung fluid, 0.9% NaCl and PBS. Eluted proteins were quantified by ELISA and analysed by immunoblotting.Only 13% of 217 birch pollen-allergic patients showed IgE-reactivity to bGST. In RBL assays bGST induced mediator release. Immunization of mice with bGST induced specific IgE and a Th2-dominated cellular immune response comparably to immunization with Bet v 1. bGST did not cross-react with Der p 8. In contrast to Bet v 1, only low amounts of bGST were released from pollen grains upon incubation in water and the different physiological solutions.Although bGST is abundant in birch pollen, immunogenic in mice and able to induce mediator release from effector cells passively loaded with specific IgE, it is a minor allergen for birch pollen-allergic patients. We refer this discrepancy to its limited release from hydrated pollen. Hence, bGST is an example demonstrating that allergenicity depends mainly on rapid elution from airborne particles.

  2. Rab17 and rab18, small GTPases with specificity for polarized epithelial cells: genetic mapping in the mouse.

    Science.gov (United States)

    McMurtrie, E B; Barbosa, M D; Zerial, M; Kingsmore, S F

    1997-11-01

    The Rab subfamily of small GTPases plays an important role in the regulation of membrane traffic in eukaryotic cells. While most Rab proteins are equally expressed in polarized and nonpolarized cells, Rab17 and Rab18 show epithelial cell specificity. Here we report the genetic mapping of Rab17 and Rab18 on mouse chromosomes 1 and 18, respectively. We also discuss some implications of Rab17 and Rab18 mapping, including their candidacy for the mouse mutations ln (leaden), Tw (twirler), and ax (ataxia). Copyright 1997 Academic Press.

  3. ASPP2 links the apical lateral polarity complex to the regulation of YAP activity in epithelial cells.

    Directory of Open Access Journals (Sweden)

    Christophe Royer

    Full Text Available The Hippo pathway, by tightly controlling the phosphorylation state and activity of the transcription cofactors YAP and TAZ is essential during development and tissue homeostasis whereas its deregulation may lead to cancer. Recent studies have linked the apicobasal polarity machinery in epithelial cells to components of the Hippo pathway and YAP and TAZ themselves. However the molecular mechanism by which the junctional pool of YAP proteins is released and activated in epithelial cells remains unknown. Here we report that the tumour suppressor ASPP2 forms an apical-lateral polarity complex at the level of tight junctions in polarised epithelial cells, acting as a scaffold for protein phosphatase 1 (PP1 and junctional YAP via dedicated binding domains. ASPP2 thereby directly induces the dephosphorylation and activation of junctional YAP. Collectively, this study unearths a novel mechanistic paradigm revealing the critical role of the apical-lateral polarity complex in activating this localised pool of YAP in vitro, in epithelial cells, and in vivo, in the murine colonic epithelium. We propose that this mechanism may commonly control YAP functions in epithelial tissues.

  4. Dependence of InGaN solar cell performance on polarization-induced electric field and carrier lifetime

    International Nuclear Information System (INIS)

    Yang Jing; Zhao De-Gang; Jiang De-Sheng; Liu Zong-Shun; Chen Ping; Li Liang; Wu Liang-Liang; Le Ling-Cong; Li Xiao-Jing; He Xiao-Guang; Yang Hui; Wang Hui; Zhu Jian-Jun; Zhang Shu-Ming; Zhang Bao-Shun

    2013-01-01

    The effects of Mg-induced net acceptor doping concentration and carrier lifetime on the performance of a p—i—n InGaN solar cell are investigated. It is found that the electric field induced by spontaneous and piezoelectric polarization in the i-region could be totally shielded when the Mg-induced net acceptor doping concentration is sufficiently high. The polarization-induced potential barriers are reduced and the short circuit current density is remarkably increased from 0.21 mA/cm 2 to 0.95 mA/cm 2 by elevating the Mg doping concentration. The carrier lifetime determined by defect density of i-InGaN also plays an important role in determining the photovoltaic properties of solar cell. The short circuit current density severely degrades, and the performance of InGaN solar cell becomes more sensitive to the polarization when carrier lifetime is lower than the transit time. This study demonstrates that the crystal quality of InGaN absorption layer is one of the most important challenges in realizing high efficiency InGaN solar cells. (interdisciplinary physics and related areas of science and technology)

  5. Study of Collagen Birefringence in Different Grades of Oral Squamous Cell Carcinoma Using Picrosirius Red and Polarized Light Microscopy

    Directory of Open Access Journals (Sweden)

    Pillai Arun Gopinathan

    2015-01-01

    Full Text Available Objectives. The present study was done to evaluate birefringence pattern of collagen fibres in different grades of oral squamous cell carcinoma using Picrosirius red stain and polarization microscopy and to determine if there is a change in collagen fibres between different grades of oral squamous cell carcinoma. Materials and Methods. Picrosirius red stained 5 μm thick sections of previously diagnosed different grades of squamous cell carcinoma and normal oral mucosa were studied under polarization microscopy for arrangement as well as birefringence of collagen fibres around tumour islands. Results. It was found that thin collagen fibres increased and thick collagen fibres decreased with dedifferentiation of OSCC (P<0.0001 . It was observed that there was change in polarization colours of thick fibres from yellowish orange to greenish yellow with dedifferentiation of OSCC indicating loosely packed fibres (P<0.0001. Conclusion. There was a gradual change of birefringence of collagen from yellowish orange to greenish yellow from well to poorly differentiated squamous cell carcinoma, indicating that there is a change from mature form of collagen to immature form as tumour progresses. Studying collagen fibres with Picrosirius red for stromal changes around tumour islands along with routine staining may help in predicting the prognosis of tumour.

  6. A new source of Southern Ocean and Antarctic aerosol from tropospheric polar cell chemistry of sea ice emissions

    Science.gov (United States)

    Humphries, R. S.; Klekociuk, A. R.; Schofield, R.; Robinson, A. D.; Harris, N. R. P.; Keywood, M.; Ward, J.; Galbally, I.; Molloy, S.; Thomas, A.; Wilson, S. R.

    2014-12-01

    The Antarctic region is a pristine environment with minimal anthropogenic influence. Aerosol measurements in this environment allow the study of natural aerosols and polar atmospheric dynamics. Measurements in this region have been limited primarily to continental and coastal locations where permanent stations exist, with a handful of measurements in the sea ice region. The MAPS campaign (Measurements of Aerosols and Precursors during SIPEXII) occurred as part of SIPEX II (Sea Ice Physics and Ecosystems eXperiment II) voyage in Spring, 2012, and produced the first Antarctic pack-ice focused aerosol dataset aimed at characterizing new particle formation processes off the coast of East Antarctica (~65°S, 120°E). Numerous atmospheric parameters and species were measured, including the number of aerosol particles in the 3-10 nm size range, the range associated with nucleating particle formation. A latitudinal transect through the sea ice identified the Polar Front from sudden changes in nucleating particle concentrations, averaging 51cm-3 north of the front in the Ferrel cell, and 766 cm-3 south of the front, in the Polar cell region. The Polar Front location was also confirmed by meteorological and back-trajectory data. Background aerosol populations in the Polar cell fluctuated significantly but displayed no growth indicators, suggesting transport. Back-trajectories revealed that air parcels often descended from the free-troposphere within the previous 24-48 hrs. It is proposed that particle formation occurs in the free troposphere from precursors uplifted at the polar front region which, being a sea-ice/ocean region, is a significant precursor source. After tropospheric formation, populations descending at the poles are transported northward and reach the sea ice surface, missing continental stations. Current measurements of Antarctic aerosol suggest very low loading which may be explained by these circulation patterns and may underestimate total regional loading

  7. Photon harvesting, coloring and polarizing in photovoltaic cell integrated color filters: efficient energy routing strategies for power-saving displays.

    Science.gov (United States)

    Wen, Long; Chen, Qin; Song, Shichao; Yu, Yan; Jin, Lin; Hu, Xin

    2015-07-03

    We describe the integral electro-optical strategies that combine the functionalities of photovoltaic (PV) electricity generation and color filtering as well as polarizing to realize more efficient energy routing in display technology. Unlike the conventional pigment-based filters and polarizers, which absorb substantial amounts of unwanted spectral components and dissipate them in the form of heat, we propose converting the energy of those photons into electricity by constructing PV cell-integrated color filters based on a selectively transmitting aluminum (Al) rear electrode perforated with nanoholes (NHs). Combining with a dielectric-metal-dielectric (DMD) front electrode, the devices were optimized to enable efficient cavity-enhanced photon recycling in the PV functional layers. We perform a comprehensive theoretical and numerical analysis to explore the extraordinary optical transmission (EOT) through the Al NHs and identify basic design rules for achieving structural coloring or polarizing in our PV color filters. We show that the addition of thin photoactive polymer layers on the symmetrically configured Al NH electrode narrows the bandwidth of the EOT-assisted high-pass light filtering due to the strongly damped anti-symmetric coupling of the surface modes excited on the front and rear surface of the Al NHs, which facilitates the whole visible coloring with relatively high purity for the devices. By engineering the cut-off characteristics of the plasmonic waveguide mode supported by the circular or ellipsoidal Al NHs, beyond the photon recycling capacity, PV color filters and PV polarizing color filters that allow polarization-insensitive and strong polarization-anisotropic color filtering were demonstrated. The findings presented here may shed some light on expanding the utilization of PV electricity generation across new-generation energy-saving electrical display devices.

  8. Photon harvesting, coloring and polarizing in photovoltaic cell integrated color filters: efficient energy routing strategies for power-saving displays

    International Nuclear Information System (INIS)

    Wen, Long; Chen, Qin; Song, Shichao; Yu, Yan; Jin, Lin; Hu, Xin

    2015-01-01

    We describe the integral electro-optical strategies that combine the functionalities of photovoltaic (PV) electricity generation and color filtering as well as polarizing to realize more efficient energy routing in display technology. Unlike the conventional pigment-based filters and polarizers, which absorb substantial amounts of unwanted spectral components and dissipate them in the form of heat, we propose converting the energy of those photons into electricity by constructing PV cell-integrated color filters based on a selectively transmitting aluminum (Al) rear electrode perforated with nanoholes (NHs). Combining with a dielectric-metal-dielectric (DMD) front electrode, the devices were optimized to enable efficient cavity-enhanced photon recycling in the PV functional layers. We perform a comprehensive theoretical and numerical analysis to explore the extraordinary optical transmission (EOT) through the Al NHs and identify basic design rules for achieving structural coloring or polarizing in our PV color filters. We show that the addition of thin photoactive polymer layers on the symmetrically configured Al NH electrode narrows the bandwidth of the EOT-assisted high-pass light filtering due to the strongly damped anti-symmetric coupling of the surface modes excited on the front and rear surface of the Al NHs, which facilitates the whole visible coloring with relatively high purity for the devices. By engineering the cut-off characteristics of the plasmonic waveguide mode supported by the circular or ellipsoidal Al NHs, beyond the photon recycling capacity, PV color filters and PV polarizing color filters that allow polarization-insensitive and strong polarization-anisotropic color filtering were demonstrated. The findings presented here may shed some light on expanding the utilization of PV electricity generation across new-generation energy-saving electrical display devices. (paper)

  9. A Simple, Inexpensive Pollen Trap

    Science.gov (United States)

    P. E. Hoekstra

    1965-01-01

    Pollen plays a role of vital importance in the sexual reproduction of all plants but it is especially important in forestry. With few exceptions, sexual reproduction is the only link between succeeding generations in the forest. To be sure, vegetative reproduction is important for special purposes, but it will probably not be used on a mass scale in timber...

  10. Comparative Pollen Development in Dioscoreales

    NARCIS (Netherlands)

    Schols, P.; Furness, C.; Merckx, V.; Wilkin, P.; Smets, E.

    2005-01-01

    Pollen and orbicule ontogeny in representatives of three genera of Dioscoreales—Narthecium ossifragum Huds. (Nartheciaceae), Tacca artocarpifolia Seem. and Tacca chantrieri Andre´ (Dioscoreaceae), and Dioscorea communis (L.) Caddick & Wilkin (Dioscoreaceae)—is described and illustrated using LM,

  11. Pollen grains are efficient cloud condensation nuclei

    Energy Technology Data Exchange (ETDEWEB)

    Pope, F D, E-mail: fdp21@cam.ac.uk [Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW (United Kingdom)

    2010-10-15

    This letter presents a laboratory study investigating the ability of pollen grains to act as cloud condensation nuclei. The hygroscopicity of pollen is measured under subsaturated relative humidities using an electrodynamic balance. It is found, along with other results, that pollen exhibits bulk uptake of water under subsaturated conditions. Through the use of an environmental scanning electron microscope it was observed that the surface of pollen is wettable at high subsaturated humidities. The hygroscopic response of the pollen to subsaturated relative humidities is parametrized using {kappa}-Koehler theory and values of the parameter {kappa} for pollen are between 0.05 and 0.1. It is found that while pollen grains are only moderately hygroscopic, they can activate at critical supersaturations of 0.001% and lower, and thus pollen grains will readily act as cloud condensation nuclei. While the number density of pollen grains is too low for them to represent a significant global source of cloud condensation nuclei, the large sizes of pollen grains suggest that they will be an important source of giant cloud condensation nuclei. Low temperature work using the environmental scanning electron microscope indicated that pollen grains do not act as deposition ice nuclei at temperatures warmer than - 15 deg. C.

  12. Taraxacum officinale pollen depresses seed set of montane wildflowers through pollen allelopathy

    Directory of Open Access Journals (Sweden)

    Deirdre Loughnan

    2014-08-01

    Full Text Available Plant species that share pollinators can suffer from interspecific pollen deposition. Male reproductive success is inevitably reduced by the loss of pollen to flowers of another species. Female reproductive success can be affected by reduced stigmatic area or, more strongly, through allelopathic effects by which the admixture of some foreign pollen reduces seed or fruit set. We tested for allelopathic effects of Taraxacum officinale (Asteracaeae pollen on the seed set of montane wildflowers Erythronium grandiflorum (Liliaceae and Erysimum capitatum (Brassicaceae, by hand-pollinating plants with pollen mixtures. Taraxacum is a common invasive species, which produces allelopathic chemicals in its root and vegetative tissue, making it a likely candidate for pollen allelopathy. Flowers of both species produced fewer well-developed seeds when pollinated with pollen mixtures containing Taraxacum pollen. The pollen-allelopathic potential of weedy dandelion may add to its ability to disrupt communities that it invades.

  13. Polarization Imaging Apparatus for Cell and Tissue Imaging and Diagnostics, Phase II

    Data.gov (United States)

    National Aeronautics and Space Administration — This work proposes to capitalize on our Phase I success in a novel visible-near infrared Stokes polarization imaging technology based on high performance fast...

  14. Polarization Imaging Apparatus for Cell and Tissue Imaging and Diagnostics, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — In recent years there has been an increasing interest in the propagation of polarized light in randomly scattering media. The investigation of backscattered light is...

  15. Polarization Imaging Apparatus for Cell and Tissue Imaging and Diagnostics Project

    Data.gov (United States)

    National Aeronautics and Space Administration — This work proposes to capitalize on our Phase I success in a novel visible-near infrared Stokes polarization imaging technology based on high performance fast...

  16. Pollen selection under acid rain stress

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Y.

    1994-01-01

    To investigate whether acid rain stress induces pollen selection in nature, three different approaches were used, based on the assumption that the response of pollen grains to acid rain is controlled by an acid sensitive gene product. Germination of pollen from homozygous and heterozygous individuals under acid rain stress was examined to detect any differences in rate of germination between populations of homogeneous and heterogeneous pollen grains. In vitro and in vivo bulked segregant analysis using RAPDs was used to search for differences in DNA constitution between the survivors of acid rain stressed and non-acid rain stressed pollen populations in vitro and between the progenies of acid rain stressed and non-acid rain stressed populations during pollination, respectively. No evidence for the pollen selection under acid rain stress was obtained in any of the test systems. Inhibition of protein synthesis using cycloheximide led to significant reduction of tube elongation at 4 hr and had no effect on pollen germination at any time interval tested. Total proteins extracted from control and acid rain stressed pollen grain populations exhibited no differences. The reduction of corn pollen germination in vitro under acid rain stress was mainly due to pollen rupture. The present data indicates the reduction of pollen germination and tube growth under acid rain stress may be a physiological response rather than a genetic response. A simple, nontoxic, and effective method to separate germinated from ungerminated pollen grains has been developed using pollen from corn (Zea mays, L. cv. Pioneer 3747). The separated germinated pollen grains retained viability and continued tube growth when placed in culture medium.

  17. Grass pollen immunotherapy: where are we now.

    Science.gov (United States)

    Würtzen, Peter A; Gupta, Shashank; Brand, Stephanie; Andersen, Peter S

    2016-01-01

    During allergen immunotherapy (AIT), the allergic patient is exposed to the disease-inducing antigens (allergens) in order to induce clinical and immunological tolerance and obtain disease modification. Large trials of grass AIT with highly standardized subcutaneous and sublingual tablet vaccines have been conducted to document the clinical effect. Induction of blocking antibodies as well as changes in the balance between T-cell phenotypes, including induction of regulatory T-cell subtypes, have been demonstrated for both treatment types. These observations increase the understanding of the immunological mechanism behind the clinical effect and may make it possible to use the immunological changes as biomarkers of clinical effect. The current review describes the recent mechanistic findings for subcutaneous immunotherapy and sublingual immunotherapy/tablet treatment and discusses how the observed immunological changes translate into a scientific foundation for the observed clinical effects of grass pollen immunotherapy and lead to new treatment strategies for grass AIT.

  18. Mammalian diaphanous-related formin 1 regulates GSK3β-dependent microtubule dynamics required for T cell migratory polarization.

    Directory of Open Access Journals (Sweden)

    Baoxia Dong

    Full Text Available The mammalian diaphanous-related formin (mDia1, a Rho-regulated cytoskeletal modulator, has been shown to promote T lymphocyte chemotaxis and interaction with antigen presenting cells, but the mechanisms underpinning mDia1 roles in these processes have not been defined. Here we show that mDia1(-/- T cells exhibit impaired lymphocyte function-associated antigen 1 (LFA-1-mediated T cell adhesion, migration and in vivo trafficking. These defects are associated with impaired microtubule (MT polarization and stabilization, altered MT dynamics and reduced peripheral clustering of the MT plus-end-protein, adenomatous polyposis coli (APC in migrating T cells following LFA-1-engagement. Loss of mDia1 also leads to impaired inducible inactivation of the glycogen synthase kinase (GSK 3β as well as hyperphosphorylation and reduced levels of APC in migrating T cells. These findings identify essential roles for the mDia1 formin in modulating GSK3β-dependent MT contributions to induction of T-cell polarity, adhesion and motility.

  19. Polarization Optics

    OpenAIRE

    Fressengeas, Nicolas

    2010-01-01

    The physics of polarization optics *Polarized light propagation *Partially polarized light; DEA; After a brief introduction to polarization optics, this lecture reviews the basic formalisms for dealing with it: Jones Calculus for totally polarized light and Stokes parameters associated to Mueller Calculus for partially polarized light.

  20. Promoter activity of a putative pollen monosaccharide transporter in Petunia hybrida and characterisation of a transposon insertion mutant

    NARCIS (Netherlands)

    Garrido, D.; Busscher-Lange, J.; Tunen, van A.J.

    2006-01-01

    For the growth of the male reproductive cells of plants, the pollen, the presence of sufficient sucrose or monosaccharides is of vital importance. From Petunia hybrida a pollen-specific putative monosaccharide transporter designated PMT1 (for petunia monosaccharide transporter) has been identified

  1. Water-polysaccharide interactions in the primary cell wall of Arabidopsis thaliana from polarization transfer solid-state NMR.

    Science.gov (United States)

    White, Paul B; Wang, Tuo; Park, Yong Bum; Cosgrove, Daniel J; Hong, Mei

    2014-07-23

    Polysaccharide-rich plant cell walls are hydrated under functional conditions, but the molecular interactions between water and polysaccharides in the wall have not been investigated. In this work, we employ polarization transfer solid-state NMR techniques to study the hydration of primary-wall polysaccharides of the model plant, Arabidopsis thaliana. By transferring water (1)H polarization to polysaccharides through distance- and mobility-dependent (1)H-(1)H dipolar couplings and detecting it through polysaccharide (13)C signals, we obtain information about water proximity to cellulose, hemicellulose, and pectins as well as water mobility. Both intact and partially extracted cell wall samples are studied. Our results show that water-pectin polarization transfer is much faster than water-cellulose polarization transfer in all samples, but the extent of extraction has a profound impact on the water-polysaccharide spin diffusion. Removal of calcium ions and the consequent extraction of homogalacturonan (HG) significantly slowed down spin diffusion, while further extraction of matrix polysaccharides restored the spin diffusion rate. These trends are observed in cell walls with similar water content, thus they reflect inherent differences in the mobility and spatial distribution of water. Combined with quantitative analysis of the polysaccharide contents, our results indicate that calcium ions and HG gelation increase the amount of bound water, which facilitates spin diffusion, while calcium removal disrupts the gel and gives rise to highly dynamic water, which slows down spin diffusion. The recovery of spin diffusion rates after more extensive extraction is attributed to increased water-exposed surface areas of the polysaccharides. Water-pectin spin diffusion precedes water-cellulose spin diffusion, lending support to the single-network model of plant primary walls in which a substantial fraction of the cellulose surface is surrounded by pectins.

  2. Pollen, Tapetum, and Orbicule Development in Colletia paradoxa and Discaria americana (Rhamnaceae

    Directory of Open Access Journals (Sweden)

    M. Gotelli

    2012-01-01

    Full Text Available Tapetum, orbicule, and pollen grain ontogeny in Colletia paradoxa and Discaria americana were studied with transmission electron microscopy (TEM. The ultrastructural changes observed during the different stages of development in the tapetal cells and related to orbicule and pollen grain formation are described. The proorbicules have the appearance of lipid globule, and their formation is related to the endoplasmic reticulum of rough type (ERr. This is the first report on the presence of orbicules in the family Rhamnaceae. Pollen grains are shed at the bicellular stage.

  3. Differential expression of p-ERM, a marker of cell polarity, in benign and neoplastic oviductal epithelium.

    Science.gov (United States)

    Ning, Gang; Bijron, Jonathan G; Yuan, Ju; Hirsch, Michelle S; McKeon, Frank D; Nucci, Marisa R; Crum, Christopher P; Xian, Wa

    2013-07-01

    Serous tubal intraepithelial carcinoma (STIC) is a noninvasive phase of pelvic serous cancer at risk for metastasizing. Because of its biologic significance, its accurate distinction from nonmalignant mimics is important. Loss of cell orientation is an important feature of STIC. We sought to determine whether the immunohistochemical localization of cytoskeletal-organizing proteins phospho-ezrin-radaxin-moesin (p-ERM) would be useful in making this distinction. The benign oviductal entities (normal and p53 signatures), premalignant atypias (tubal intraepithelial lesions in transition), serous intraepithelial carcinomas (STICs), and carcinomas were analyzed for 5 staining patterns and compared. Linear or uniform luminal p-ERM staining was strongly associated with benign mucosa in contrast to STICs, in which it was lost and often replaced by nonlinear or nonuniform patterns highlighting individually cell groups or single cells. Premalignant atypias were similar to benign mucosa by p-ERM staining and retained the linear luminal pattern. This study shows, for the first time, that patterns of staining for an immunohistochemical correlate of cell polarity (p-ERM) differ between STICs, their benign counterparts and premalignant atypias that do not fulfill the criteria for STICs. If confirmed, these findings warrant further analysis of indices of cell polarity as objective markers for the diagnosis and mapping of the evolution of pelvic serous precursors.

  4. Effects of low molecular weight heparin on the polarization and cytokine profile of macrophages and T helper cells in vitro.

    Science.gov (United States)

    Bruno, Valentina; Svensson-Arvelund, Judit; Rubér, Marie; Berg, Göran; Piccione, Emilio; Jenmalm, Maria C; Ernerudh, Jan

    2018-03-08

    Low molecular weight heparin (LMWH) is widely used in recurrent miscarriage treatment. The anti-coagulant effects are established, while immunological effects are not fully known. Our aim was to assess LMWH effects on activation and polarization of central regulatory immune cells from healthy women, and on placenta tissues from women undergoing elective abortions. Isolated blood monocytes and T helper (Th) cells under different activation and polarizing conditions were cultured with or without LMWH. Flow cytometry showed that LMWH exposure induced increased expression of HLA-DR and CD206 in macrophages. This phenotype was associated with increased secretion of Th17-associated CCL20, and decreased secretion of CCL2 (M2-associated) and CCL22 (Th2), as measured by multiplex bead array. In accordance, LMWH exposure to Th cells reduced the proportion of CD25highFoxp3+ regulatory T-cells, intensified IFN-γ secretion and showed a tendency to increase the lymphoblast proportions. Collectively, a mainly pro-inflammatory effect was noted on two essential tolerance-promoting cells. Although the biological significancies of these in vitro findings are uncertain and need to be confirmed in vivo, they suggest the possibility that immunological effects of LMWH may be beneficial mainly at an earlier gestational age to provide an appropriate implantation process in women with recurrent miscarriage.

  5. The long pollen tube journey and in vitro pollen germination of Phalaenopsis orchids.

    Science.gov (United States)

    Chen, Jhun-Chen; Fang, Su-Chiung

    2016-06-01

    Pollen biology in P. aphrodite. Orchids have a distinct reproductive program. Pollination triggers ovule development and differentiation within flowers, and fertilization occurs days to months after pollination. It is unclear how pollen tubes travel through the developing ovaries during ovule development and when pollen tubes arrive at the mature embryo sac to achieve fertilization. Here, we report a robust staining protocol to image and record the timing of pollen germination, progressive growth of pollen tubes in ovaries, and arrival of pollen tubes at embryo sacs in Phalaenopsis aphrodite. The pollen germinated and pollen tubes entered the ovary 3 days after pollination. Pollen tubes continued to grow and filled the entire cavity of the ovary as the ovary elongated and ovules developed. Pollen tubes were found to enter the matured embryo sacs at approximately 60-65 days after pollination in an acropetal manner. Moreover, these temporal changes in developmental events such as growth of pollen tubes and fertilization were associated with expression of molecular markers. In addition, we developed an in vitro pollen germination protocol, which is valuable to enable studies on pollen tube guidance and tip growth regulation in Phalaenopsis orchids and possibly in other orchid species.

  6. Pollen parameters estimates of genetic variability among newly ...

    African Journals Online (AJOL)

    Estimates of some pollen parameters where used to assess the genetic diversity among some newly selected Nigerian Roselle (Hibiscus sabdariffa L.). Standard procedures were used to determine the pollen parameters such as: percentage pollen fertility, percentage pollen sterility, pollen diameters as well as anther ...

  7. Pollen tube energetics: respiration, fermentation and the race to the ovule.

    Science.gov (United States)

    Rounds, Caleb M; Winship, Lawrence J; Hepler, Peter K

    2011-01-01

    Pollen tubes grow by transferring chemical energy from stored cellular starch and newly assimilated sugars into ATP. This drives myriad processes essential for cell elongation, directly or through the creation of ion gradients. Respiration plays a central role in generating and regulating this energy flow and thus in the success of plant reproduction. Pollen tubes are easily grown in vitro and have become an excellent model for investigating the contributions of respiration to plant cellular growth and morphogenesis at the molecular, biochemical and physiological levels. In recent decades, pollen tube research has become increasingly focused on the molecular mechanisms involved in cellular processes. Yet, effective growth and development requires an intact, integrated set of cellular processes, all supplied with a constant flow of energy. Here we bring together information from the current and historical literature concerning respiration, fermentation and mitochondrial physiology in pollen tubes, and assess the significance of more recent molecular and genetic investigations in a physiological context. The rapid growth of the pollen tube down the style has led to the evolution of high rates of pollen tube respiration. Respiration rates in lily predict a total energy turnover of 40-50 fmol ATP s(-1) per pollen grain. Within this context we examine the energetic requirements of cell wall synthesis, osmoregulation, actin dynamics and cyclosis. At present, we can only estimate the amount of energy required, because data from growing pollen tubes are not available. In addition to respiration, we discuss fermentation and mitochondrial localization. We argue that the molecular pathways need to be examined within the physiological context to understand better the mechanisms that control tip growth in pollen tubes.

  8. Membrane potential of cells and its regulation during aging. 2. Report: the effect of hormones on the level of the cellular plasma membrane polarization.

    Science.gov (United States)

    Frolkis, V V; Tanin, S A; Gorban, E N; Bogatskaya, L N; Sabko, V E

    1987-01-01

    Age-dependent changes in the polarization of plasma membranes (PM) of various cell types and the mechanisms responsible for its regulation were studied in the experiments on the adult (6-8 and old (28-32 months) Wistar male rats. It was found that the effect of the hormones on the PM polarization level is altered during aging. This being related to shifts in the number and affinity of the hormonal receptors, energetic processes and protein synthesis in the cell.

  9. The Pollen Coat Proteome: At the Cutting Edge of Plant Reproduction

    Directory of Open Access Journals (Sweden)

    Juan David Rejón

    2016-01-01

    Full Text Available The tapetum is a single layer of secretory cells which encloses the anther locule and sustains pollen development and maturation. Upon apoptosis, the remnants of the tapetal cells, consisting mostly of lipids and proteins, fill the pits of the sculpted exine to form the bulk of the pollen coat. This extracellular matrix forms an impermeable barrier that protects the male gametophyte from water loss and UV light. It also aids pollen adhesion and hydration and retains small signaling compounds involved in pollen–stigma communication. In this study, we have updated the list of the pollen coat’s protein components and also discussed their functions in the context of sexual reproduction

  10. Identification and expression analysis of BoMF25, a novel polygalacturonase gene involved in pollen development of Brassica oleracea.

    Science.gov (United States)

    Lyu, Meiling; Liang, Ying; Yu, Youjian; Ma, Zhiming; Song, Limin; Yue, Xiaoyan; Cao, Jiashu

    2015-06-01

    BoMF25 acts on pollen wall. Polygalacturonase (PG) is a pectin-digesting enzyme involved in numerous plant developmental processes and is described to be of critical importance for pollen wall development. In the present study, a PG gene, BoMF25, was isolated from Brassica oleracea. BoMF25 is the homologous gene of At4g35670, a PG gene in Arabidopsis thaliana with a high expression level at the tricellular pollen stage. Collinear analysis revealed that the orthologous gene of BoMF25 in Brassica campestris (syn. B. rapa) genome was probably lost because of genome deletion and reshuffling. Sequence analysis indicated that BoMF25 contained four classical conserved domains (I, II, III, and IV) of PG protein. Homology and phylogenetic analyses showed that BoMF25 was clustered in Clade F. The putative promoter sequence, containing classical cis-acting elements and pollen-specific motifs, could drive green fluorescence protein expression in onion epidermal cells. Quantitative RT-PCR analysis suggested that BoMF25 was mainly expressed in the anther at the late stage of pollen development. In situ hybridization analysis also indicated that the strong and specific expression signal of BoMF25 existed in pollen grains at the mature pollen stage. Subcellular localization showed that the fluorescence signal was observed in the cell wall of onion epidermal cells, which suggested that BoMF25 may be a secreted protein localized in the pollen wall.

  11. Near-surface and columnar measurements with a micro pulse lidar of atmospheric pollen in Barcelona, Spain

    Directory of Open Access Journals (Sweden)

    M. Sicard

    2016-06-01

    Full Text Available We present for the first time continuous hourly measurements of pollen near-surface concentration and lidar-derived profiles of particle backscatter coefficients and of volume and particle depolarization ratios during a 5-day pollination event observed in Barcelona, Spain, between 27 and 31 March 2015. Daily average concentrations ranged from 1082 to 2830 pollen m−3. Platanus and Pinus pollen types represented together more than 80 % of the total pollen. Maximum hourly pollen concentrations of 4700 and 1200 m−3 were found for Platanus and Pinus, respectively. Every day a clear diurnal cycle caused by the vertical transport of the airborne pollen was visible on the lidar-derived profiles with maxima usually reached between 12:00 and 15:00 UT. A method based on the lidar polarization capabilities was used to retrieve the contribution of the pollen to the total aerosol optical depth (AOD. On average the diurnal (09:00–17:00 UT pollen AOD was 0.05, which represented 29 % of the total AOD. Maximum values of the pollen AOD and its contribution to the total AOD reached 0.12 and 78 %, respectively. The diurnal means of the volume and particle depolarization ratios in the pollen plume were 0.08 and 0.14, with hourly maxima of 0.18 and 0.33, respectively. The diurnal mean of the height of the pollen plume was found at 1.24 km with maxima varying in the range of 1.47–1.78 km. A correlation study is performed (1 between the depolarization ratios and the pollen near-surface concentration to evaluate the ability of the former parameter to monitor pollen release and (2 between the depolarization ratios as well as pollen AOD and surface downward solar fluxes, which cause the atmospheric turbulences responsible for the particle vertical motion, to examine the dependency of the depolarization ratios and the pollen AOD upon solar fluxes. For the volume depolarization ratio the first correlation study yields to correlation

  12. Multiple BiP genes of Arabidopsis thaliana are required for male gametogenesis and pollen competitiveness.

    Science.gov (United States)

    Maruyama, Daisuke; Sugiyama, Tomoyuki; Endo, Toshiya; Nishikawa, Shuh-Ichi

    2014-04-01

    Immunoglobulin-binding protein (BiP) is a molecular chaperone of the heat shock protein 70 (Hsp70) family. BiP is localized in the endoplasmic reticulum (ER) and plays key roles in protein translocation, protein folding and quality control in the ER. The genomes of flowering plants contain multiple BiP genes. Arabidopsis thaliana has three BiP genes. BIP1 and BIP2 are ubiquitously expressed. BIP3 encodes a less well conserved BiP paralog, and it is expressed only under ER stress conditions in the majority of organs. Here, we report that all BiP genes are expressed and functional in pollen and pollen tubes. Although the bip1 bip2 double mutation does not affect pollen viability, the bip1 bip2 bip3 triple mutation is lethal in pollen. This result indicates that lethality of the bip1 bip2 double mutation is rescued by BiP3 expression. A decrease in the copy number of the ubiquitously expressed BiP genes correlates well with a decrease in pollen tube growth, which leads to reduced fitness of mutant pollen during fertilization. Because an increased protein secretion activity is expected to increase the protein folding demand in the ER, the multiple BiP genes probably cooperate with each other to ensure ER homeostasis in cells with active secretion such as rapidly growing pollen tubes.

  13. The influence of brood on the pollen consumption of worker bees (Apis mellifera L.).

    Science.gov (United States)

    Hrassnigg, Norbert; Crailsheim, Karl

    1998-05-01

    (1) In midgut dry weight (tissue plus contents) of worker bees we found a representative parameter for pollen consumption. Midguts of bees of successive ages were analyzed and correlated with various parameters. The relative proportions of sugar, protein and water were either constant or negatively correlated with midgut weight. Only the relative pollen weight (percent of midgut dry weight) increased. (2) To investigate the influence of different levels of brood on pollen consumption of individual bees, midgut dry weights from 2 normally breeding control colonies and 2 brood-reduced experimental colonies were analyzed. In bees from control colonies the pollen consumption increased up to the nursing age (3-10d), remained on an elevated level in middle-aged-bees (10-18d) and decreased relatively sharply towards the foraging ages (>21d). When queens were caged in the experimental colonies, the following decline of brood cells affected the consumption of pollen differently. After 6 days of caging, with a reduction of open brood only, no effect was seen. After 15 days, and even more pronounced after 23 days when no brood was present, the pollen consumption in young and middle-aged (10, 14, 18d) worker bees was significantly reduced, while it was clearly elevated in older bees. We discuss pollen consumption as an adaptation to reduced necessity to nurse brood in young and middle-aged bees, and to enhance life span in older animals.

  14. Pollen tube and root-hair tip growth is disrupted in a mutant of Arabidopsis thaliana.

    Science.gov (United States)

    Schiefelbein, J; Galway, M; Masucci, J; Ford, S

    1993-11-01

    The expansion of both root hairs and pollen tubes occurs by a process known as tip growth. In this report, an Arabidopsis thaliana mutant (tip1) is described that displays defects in both root-hair and pollen-tube growth. The root hairs of the tip1 mutant plants are shorter than those of the wild-type plants and branched at their base. The tip1 pollen-tube growth defect was identified by the aberrant segregation ratio of phenotypically normal to mutant seeds in siliques from self-pollinated, heterozygous plants. Homozygous mutant seeds are not randomly distributed in the siliques, comprising only 14.4% of the total seeds, 5.3% of the seeds from the bottom half, and 2.2% of the seeds from the bottom quarter of the heterozygous siliques. Studies of pollen-tube growth in vivo showed that mutant pollen tubes grow more slowly than wild-type pollen through the transmitting tissue of wild-type flowers. Cosegregation studies indicate that the root-hair and pollen-tube defects are caused by the same genetic lesion. Based on these findings, the TIP1 gene is likely to encode a product involved in a fundamental aspect of tip growth in plant cells.

  15. Living Cells and Dynamic Molecules Observed with the Polarized Light Microscope: the Legacy of Shinya Inoué.

    Science.gov (United States)

    Tani, Tomomi; Shribak, Michael; Oldenbourg, Rudolf

    2016-08-01

    In 1948, Shinya Inoué arrived in the United States for graduate studies at Princeton. A year later he came to Woods Hole, starting a long tradition of summer research at the Marine Biological Laboratory (MBL), which quickly became Inoué's scientific home. Primed by his Japanese mentor, Katsuma Dan, Inoué followed Dan's mantra to work with healthy, living cells, on a fundamental problem (mitosis), with a unique tool set that he refined for precise and quantitative observations (polarized light microscopy), and a fresh and brilliant mind that was unafraid of challenging current dogma. Building on this potent combination, Inoué contributed landmark observations and concepts in cell biology, including the notion that there are dynamic, fine structures inside living cells, in which molecular assemblies such as mitotic spindle fibers exist in delicate equilibrium with their molecular building blocks suspended in the cytoplasm. In the late 1970s and 1980s, Inoué and others at the MBL were instrumental in conceiving video microscopy, a groundbreaking technique which married light microscopy and electronic imaging, ushering in a revolution in how we know and what we know about living cells and the molecular mechanisms of life. Here, we recount some of Inoué's accomplishments and describe how his legacy has shaped current activities in polarized light imaging at the MBL. © 2016 Marine Biological Laboratory.

  16. Type I collagen gel induces Madin-Darby canine kidney cells to become fusiform in shape and lose apical-basal polarity.

    Science.gov (United States)

    Zuk, A; Matlin, K S; Hay, E D

    1989-03-01

    In the embryo, epithelia give rise to mesenchyme at specific times and places. Recently, it has been reported (Greenburg, G., and E. D. Hay. 1986. Dev. Biol. 115:363-379; Greenberg, G., and E. D. Hay. 1988. Development (Camb.). 102:605-622) that definitive epithelia can give rise to fibroblast-like cells when suspended within type I collagen gels. We wanted to know whether Madin-Darby canine kidney (MDCK) cells, an epithelial line, can form mesenchyme under similar conditions. Small explants of MDCK cells on basement membrane were suspended within or placed on top of extracellular matrix gels. MDCK cells on basement membrane gel are tall, columnar in shape, and ultrastructurally resemble epithelia transporting fluid and ions. MDCK explants cultured on type I collagen gel give rise to isolated fusiform-shaped cells that migrate over the gel surface. The fusiform cells extend pseudopodia and filopodia, lose cell membrane specializations, and develop an actin cortex around the entire cell. Unlike true mesenchymal cells, which express vimentin and type I collagen, fusiform cells produce both keratin and vimentin, continue to express laminin, and do not turn on type I collagen. Fusiform cells are not apically-basally polarized, but show mesenchymal cell polarity. Influenza hemagglutinin and virus budding localize to the front end or entire cell surface. Na,K-ATPase occurs intracellularly and also symmetrically distributes on the cell surface. Fodrin becomes diffusely distributed along the plasma membrane, ZO-1 cannot be detected, and desmoplakins distribute randomly in the cytoplasm. The loss of epithelial polarity and acquisition of mesenchymal cell polarity and shape by fusiform MDCK cells on type I collagen gel was previously unsuspected. The phenomenon may offer new opportunities for studying cytoplasmic and nuclear mechanisms regulating cell shape and polarity.

  17. MicroRNA and tasiRNA diversity in mature pollen of Arabidopsis thaliana

    Directory of Open Access Journals (Sweden)

    Hafidh Said

    2009-12-01

    Full Text Available Abstract Background New generation sequencing technology has allowed investigation of the small RNA populations of flowering plants at great depth. However, little is known about small RNAs in their reproductive cells, especially in post-meiotic cells of the gametophyte generation. Pollen - the male gametophyte - is the specialised haploid structure that generates and delivers the sperm cells to the female gametes at fertilisation. Whether development and differentiation of the male gametophyte depends on the action of microRNAs and trans-acting siRNAs guiding changes in gene expression is largely unknown. Here we have used 454 sequencing to survey the various small RNA populations present in mature pollen of Arabidopsis thaliana. Results In this study we detected the presence of 33 different microRNA families in mature pollen and validated the expression levels of 17 selected miRNAs by Q-RT-PCR. The majority of the selected miRNAs showed pollen-enriched expression compared with leaves. Furthermore, we report for the first time the presence of trans-acting siRNAs in pollen. In addition to describing new patterns of expression for known small RNAs in each of these classes, we identified 7 putative novel microRNAs. One of these, ath-MIR2939, targets a pollen-specific F-box transcript and we demonstrate cleavage of its target mRNA in mature pollen. Conclusions Despite the apparent simplicity of the male gametophyte, comprising just two different cell types, pollen not only utilises many miRNAs and trans-acting siRNAs expressed in the somatic tissues but also expresses novel miRNAs.

  18. The coexistence of bicellular and tricellular pollen in Annona cherimola (Annonaceae): Implications for pollen evolution

    OpenAIRE

    Lora, Jorge; Herrero Romero, María; Hormaza Urroz, José Ignacio

    2009-01-01

    Most angiosperms release bicellular pollen. However, in about one-third of extant angiosperms, the second pollen mitosis occurs before anthesis such that pollen is tricellular upon release. The shift from bicellular to tricellular development has occurred several times independently, but its causes are largely unknown. In this work, we observed the coexistence of both kinds of pollen at anther dehiscence in Annona cherimola, a species that belongs to the basal angiosperm family Annonaceae. Ex...

  19. Exocytosis and polarity in plant cells: insights by studying cellulose synthase complexes and the exocyst

    NARCIS (Netherlands)

    Ying Zhang, Ying

    2012-01-01

    The work presented in this thesis covers aspects of exocytosis, plant cell growth and cell wall formation. These processes are strongly linked as cell growth and cell wall formation occur simultaneously and exocytosis is the process that delivers cell wall components to the existing cell wall

  20. Proinflammatory-Activated Glioma Cells Induce a Switch in Microglial Polarization and Activation Status, From a Predominant M2b Phenotype to a Mixture of M1 and M2a/B Polarized Cells

    Directory of Open Access Journals (Sweden)

    Lucia Lisi

    2014-04-01

    Full Text Available Malignant gliomas are primary brain tumors characterized by morphological and genetic complexities, as well as diffuse infiltration into normal brain parenchyma. Within gliomas, microglia/macrophages represent the largest tumor-infiltrating cell population, contributing by at least one-third to the total tumor mass. Bi-directional interactions between glioma cells and microglia may therefore play an important role on tumor growth and biology. In the present study, we have characterized the influence of glioma-soluble factors on microglial function, comparing the effects of media harvested under basal conditions with those of media obtained after inducing a pro-inflammatory activation state in glioma cells. We found that microglial cells undergo a different pattern of activation depending on the stimulus; in the presence of activated glioma-derived factors, i.e. a condition mimicking the late stage of pathology, microglia presents as a mixture of polarization phenotypes (M1 and M2a/b, with up-regulation of iNOS (inducible nitric oxide synthase, ARG (arginase and IL (interleukine-10. At variance, microglia exposed to basal glioma-derived factors, i.e. a condition resembling the early stage of pathology, shows a more specific pattern of activation, with increased M2b polarization status and up-regulation of IL-10 only. As far as viability and cell proliferation are concerned, both LI-CM [LPS (lipopolysaccharide—IFNγ (interferon γ conditioned media] and C-CM (control-conditioned media induce similar effects on microglial morphology. Finally, in human glioma tissue obtained from surgical resection of patients with IV grade glioblastoma, we detected a significant amount of CD68 positive cells, which is a marker of macrophage/microglial phagocytic activity, suggesting that in vitro findings presented here might have a relevance in the human pathology as well.

  1. Seed set, pollen morphology and pollen surface composition response to heat stress in field pea.

    Science.gov (United States)

    Jiang, Yunfei; Lahlali, Rachid; Karunakaran, Chithra; Kumar, Saroj; Davis, Arthur R; Bueckert, Rosalind A

    2015-11-01

    Pea (Pisum sativum L.) is a major legume crop grown in a semi-arid climate in Western Canada, where heat stress affects pollination, seed set and yield. Seed set and pod growth characteristics, along with in vitro percentage pollen germination, pollen tube growth and pollen surface composition, were measured in two pea cultivars (CDC Golden and CDC Sage) subjected to five maximum temperature regimes ranging from 24 to 36 °C. Heat stress reduced percentage pollen germination, pollen tube length, pod length, seed number per pod, and the seed-ovule ratio. Percentage pollen germination of CDC Sage was greater than CDC Golden at 36 °C. No visible morphological differences in pollen grains or the pollen surface were observed between the heat and control-treated pea. However, pollen wall (intine) thickness increased due to heat stress. Mid-infrared attenuated total reflectance (MIR-ATR) spectra revealed that the chemical composition (lipid, proteins and carbohydrates) of each cultivar's pollen grains responded differently to heat stress. The lipid region of the pollen coat and exine of CDC Sage was more stable compared with CDC Golden at 36 °C. Secondary derivatives of ATR spectra indicated the presence of two lipid types, with different amounts present in pollen grains from each cultivar. © 2015 John Wiley & Sons Ltd.

  2. In Vitro Pollen Viability and Pollen Germination in Cherry Laurel (Prunus laurocerasus L.

    Directory of Open Access Journals (Sweden)

    Melekber Sulusoglu

    2014-01-01

    Full Text Available Pollen quality is important for growers and breeders. This study was carried out to determine in vitro pollen viability and pollen germination in seven genotypes of cherry laurel (Prunus laurocerasus L.. Two pollen viability tests, TTC (2,3,5-triphenyl tetrazolium chloride and IKI (iodine potassium iodide, were used. Pollen traits of genotypes were studied using an in vitro medium containing 0%, 5%, 10%, 15%, and 20% sucrose to determine the best sucrose concentrations for germination. In the second step, the germinated pollen was counted 1, 4, 6, 10, 12, 24, and 48 hours later until there was no further germination. The viability rates were different according to genotypes and tests used. The IKI and TTC staining tests and pollen germination had low correlation (r2 = 0.0614 and r2 = 0.0015, resp.. Painted pollen rate was higher and pollen was well-stained with IKI test and pollen viability estimated with TTC staining test was better than that estimated with the IKI staining test. 15% sucrose gave the best germination rates in most of the genotypes. Pollen germination rates were recorded periodically from one hour to 48 hours in 15% sucrose and the results showed that pollen germination rates increased after 6 hours of being placed in culture media.

  3. In vitro pollen viability and pollen germination in cherry laurel (Prunus laurocerasus L.).

    Science.gov (United States)

    Sulusoglu, Melekber; Cavusoglu, Aysun

    2014-01-01

    Pollen quality is important for growers and breeders. This study was carried out to determine in vitro pollen viability and pollen germination in seven genotypes of cherry laurel (Prunus laurocerasus L.). Two pollen viability tests, TTC (2,3,5-triphenyl tetrazolium chloride) and IKI (iodine potassium iodide), were used. Pollen traits of genotypes were studied using an in vitro medium containing 0%, 5%, 10%, 15%, and 20% sucrose to determine the best sucrose concentrations for germination. In the second step, the germinated pollen was counted 1, 4, 6, 10, 12, 24, and 48 hours later until there was no further germination. The viability rates were different according to genotypes and tests used. The IKI and TTC staining tests and pollen germination had low correlation (r(2) = 0.0614 and r(2) = 0.0015, resp.). Painted pollen rate was higher and pollen was well-stained with IKI test and pollen viability estimated with TTC staining test was better than that estimated with the IKI staining test. 15% sucrose gave the best germination rates in most of the genotypes. Pollen germination rates were recorded periodically from one hour to 48 hours in 15% sucrose and the results showed that pollen germination rates increased after 6 hours of being placed in culture media.

  4. Controlling Hay Fever Symptoms with Accurate Pollen Counts

    Science.gov (United States)

    ... Hay fever and pollen counts Share | Controlling Hay Fever Symptoms with Accurate Pollen Counts This article has ... MD, FAAAAI Seasonal allergic rhinitis known as hay fever is caused by pollen carried in the air ...

  5. Experimental studies on mushroom and pollen drying

    Energy Technology Data Exchange (ETDEWEB)

    Midilli, Adnan; Olgun, Hayati; Ayhan, Teoman [Karadeniz Technical Univ., Mechanical Engineering Dept., Trabzon (Turkey)

    1999-07-01

    In this study, drying periods of mushrooms and pollens (for Anzer honey) were investigated. The experiments of mushrooms were carried out under both laboratory and shady- and sunny-atmospheric conditions and those of pollens were conducted in the drying cupboard heated by electric heater and under sunny atmospheric conditions. The temperature of the drying cupboard was regulated at 45degC during the drying experiment of pollen. The mass loss of pollens and the temperature of each shelf including pollens in the drying cupboard were measured. Drying curves of mushrooms and pollens for each experiment and the variation of the mass change ratio, the moisture ratio of pollens and the temperatures in the shelves with drying time were presented, respectively. Furthermore, their equations of drying curves and the average experimental uncertainty ratios were calculated based on the experimental results. It was concluded that mushrooms can be dried in the cupboards by using hot air at 50degC in a time period of 5-6 h, and the dried must be protected in vacuum, and the pollens also must be dried at temperatures between 40 and 45degC in a time period of 2.5-3 h without their losing the colour, flavour, smell and structure. The average experimental uncertainty ratio of mushrooms and pollens during the drying process were calculated to be 22 and 18% respectively. (Author)

  6. Monoclonal antibodies to the major Lolium perenne (rye grass) pollen allergen Lol p I (Rye I).

    Science.gov (United States)

    Kahn, C R; Marsh, D G

    1986-12-01

    Thirteen monoclonal antibodies (MAbs) were produced against Lol p I (Rye I), the major Lolium perenne (rye grass) pollen allergen. Spleen cells from A/J and SJL mice immunized with highly purified Lol p I (Lol I) were allowed to fuse with cells from the non-secreting Sp2/0-Ag14 myeloma cell line. Each MAb was analyzed for antigenic specificity by radioimmunoassay (RIA) using 125I-Lol I. The epitope specificities of seven of the MAbs were examined by competitive binding against a labelled standard MAb for the Lol I antigen (Ag). The dissociation constant, Kd, of one MAb (No. 3.2) that was studied most extensively was determined by double Ab RIA to be 3.5 X 10(-6) L/M. This MAb recognized the related 27,000-30,000 Group I glycoproteins found in the pollens of nine other species of grass pollens tested, including weak binding to Bermuda grass Group I (Cyn d I), which by conventional analysis using polyclonal anti-Lol I serum shows no detectable binding. Monoclonal antibody No. 3.2 was coupled covalently to Sepharose 4B and used to prepare highly purified Lol I from a partially purified rye pollen extract. Finally, an RIA was developed which permitted the analysis of the Group I components in rye grass and nine other grass pollen species. The latter assay is likely to prove useful in the standardization of grass pollen extracts according to their Group I contents.

  7. T helper type 1 polarizing γδ T cells and Scavenger receptors contribute to the pathogenesis of Pemphigus vulgaris.

    Science.gov (United States)

    Das, Dayasagar; Anand, Vivek; Khandpur, Sujay; Sharma, Vinod K; Sharma, Alpana

    2018-01-01

    γδ T cells and Scavenger receptors are key parts of the innate immune machinery, playing significant roles in regulating immune homeostasis at the epithelial surface. The roles of these immune components are not yet characterized for the autoimmune skin disorder Pemphigus vulgaris (PV). Phenotyping and frequency of γδ T cells estimated by flow cytometry have shown increased frequency of γδ T cells (6·7% versus 4·4%) producing interferon- γ (IFN-γ; 35·2% versus 26·68%) in the circulation of patients compared with controls. Dual cytokine-secreting (IFN-γ and interleukin-4) γδ T cells indicate the plasticity of these cells. The γδ T cells of patients with PV have shown higher cytotoxic potential and the higher frequency of γδ T cells producing IFN-γ shows T helper type 1 polarization. The increased expression of Scavenger receptors expression (CD36 and CD163) could be contributing to the elevated inflammatory environment and immune imbalance in this disease. Targeting the inflammatory γδ T cells and Scavenger receptors may pave the way for novel therapeutics. © 2017 John Wiley & Sons Ltd.

  8. Impedance of vapor feed direct methanol fuel cells--polarization dependence of elementary processes at the anode

    International Nuclear Information System (INIS)

    Fukunaga, Hiroshi; Ishida, Tomohiro; Teranishi, Nozomu; Arai, Chikao; Yamada, Koichi

    2004-01-01

    Membrane electrode assemblies of direct methanol fuel cells (DMFCs) with different catalyst and ionomer loading were prepared. Anode performance and impedance spectra were measured to clarify the characteristics of vapor feed DMFCs (VF-DMFCs). The impedance spectra were deconvolved into three semi-circles with different time constants, each showing a different dependence on the anodic polarization. The middle-frequency range arc decreased as the anodic polarization increased, indicating that this process represents the oxidation reaction of methanol. The high-frequency range arc showed little dependence on the anodic polarization, but increased with the thickness of the electrode, indicating that this process might be related to proton conduction through the electrode. The low-frequency range arc was observed only when the methanol concentration was low, in contrast to liquid feed DMFCs (LF-DMFCs), for which the removal of the product gas presents a large resistance. A simpler design can therefore be used for a VF-DMFC, giving it an advantage over an LF-DMFC. A decreasing ionomer to catalyst ratio (I/C) caused the interfacial conductivity (σ E ) to increase, but it intensively decreased when I/C was below 0.25. Thus, the connection of the catalysts is important for the anode's performance

  9. All in one - integrating cell polarity, meiosis, mitosis and mechanical forces in early oocyte differentiation in vertebrates.

    Science.gov (United States)

    Elkouby, Yaniv M

    2017-01-01

    While the differentiation of oocytes is key for embryonic development, and its investigation is crucial for advancing our understanding of human reproduction and fertility, many fundamental questions in oogenesis have been long standing. However, recent technical advances have led to several breakthroughs mainly in mice and zebrafish. Here I review these recent findings, including regulation and organization of the germline cyst, the mechanistics of chromosomal pairing, establishment of cell polarity, and formation of a universal mRNA-protein (mRNP) granule called the Balbiani body. I discuss common themes in oogenesis from frogs, fish and mouse and compare them to findings from C. elegans and Drosophila. The zebrafish juvenile ovary is an attractive model where these individual processes can be investigated, but also revealing how they are inter-coordinated in oocyte differentiation. A conserved cellular organizer was discovered in the zebrafish oocyte that seems to function at a nexus of oocyte differentiation. This organizer, termed the Meiotic Vegetal Center (MVC), is composed of the oocyte centrosome, and couples meiotic chromosomal pairing with oocyte polarization and Balbiani body formation. The MVC breaks the oocyte symmetry, is regulated by upstream mitotic division in the cyst and nucleates Balbiani body mRNPs prion-like aggregation downstream. These processes can shed new light on broad questions in biology, such as how mitosis contributes to cell polarity, and how prion aggregation which lead to neurodegenerative disease when awry, is regulated in a physiological context. Furthermore, novel cytoskeletal structures can unravel cytoplasmic mechanical functions in chromosomal pairing. Finally, together with recently developed tools, genome editing technology now enables a robust genetic analysis of these fundamental processes in the zebrafish, paving the way for a comprehensive cell and developmental view of vertebrate oogenesis.

  10. Studies on mutagenic activity of 60Co γ-ray irradiated rape pollen

    International Nuclear Information System (INIS)

    He Weishun; Liu Aihua; Lin Shiying; Xiong Xikun

    1989-01-01

    In the present study on disinfection, the rape pollen was irradiated with 2.5 kGy 60 Co γ-ray. Micronuclei, sister chromatid exchanges (SCE) of bone marrow cells and chromosomal aberrations of meiotic cells in mice were used as an indicater of chromosomal damage to study the mutagenicity of irradiated rape pollen. The results are as follows: (1) The frequency of micronuclei in polychromatic erythrocytes is 2.00 per mille; nucleated cells is 0.8 per mille in control group. In the numbers of polychromatic erythrocytes and nucleated cells with micronuclei, there is no obviously difference in irradiated and unirradiated groups. (2) SCE incidence of control group is 2.01 ± 0.12/cell. No significant difference in the frequency of SCE exists between non-irradiated rape pollen and the control groups. But the frequency of SCE in irradiated rape pollen group (3000 mg/kg/day x 7) is 2.36 ± 0.12/cell; high dose group (6000 mg/kg/day x 7) is 2.96 ± 0.14/cell. In comparison with control group, there is a significant difference. (3) The chromatid breaks, fragments, and univalents in primary spermatocytes have been obseved. The frequencies of chromosomal aberration showed no obviously difference among irradiated and non-irradiated rape pollen groups

  11. M2-polarized tumor-associated macrophages facilitated migration and epithelial-mesenchymal transition of HCC cells via the TLR4/STAT3 signaling pathway.

    Science.gov (United States)

    Yao, Rong-Rong; Li, Jing-Huan; Zhang, Rui; Chen, Rong-Xin; Wang, Yan-Hong

    2018-01-16

    M2-polarized macrophages are tumor-associated-macrophages (TAMs), which are important contents of tumor-infiltrating immune cells. Toll-like receptor 4 (TLR4) is a molecular biomarker of tumor aggressiveness and poor prognosis. Toll-like receptors (TLRs) have important roles in the immune system and M2-polarized macrophages. However, the effects of TLR4 on M2-polarized macrophages in hepatocellular carcinoma (HCC) are unknown. Here, TLR4 expressed on HCC cells mediates the pro-tumor effects and mechanisms of M2-polarized macrophages. THP-1 cells were induced to differentiate into M2-like macrophages through treatments with IL-4, IL-13, and phorbol myristate acetate (PMA). We used the HCC cell lines SMMC-7721 and MHCC97-H cultured in conditioned medium from M2-like macrophages (M2-CM) to investigate the migration potential of HCC cells and epithelial-mesenchymal transition (EMT)-associated molecular genetics. Signaling pathways that mediated M2-CM-promoted HCC migration were detected using western blotting. HCC cells cultured with M2-CM displayed a fibroblast-like morphology, an increased metastatic capability, and expression of EMT markers. TLR4 expression was markedly increased in M2-CM-treated HCC cells. TLR4 overexpression promoted HCC cell migration, and a TLR4-neutralizing antibody markedly inhibited HCC EMT in cells cultured with M2-CM. Furthermore, the TLR4/(signal transducer and activator of transcription 3 (STAT3) signaling pathway contributed to the effects of M2-CM on HCC cells. Taken together, M2-polarized macrophages facilitated the migration and EMT of HCC cells via the TLR4/STAT3 signaling pathway, suggesting that TLR4 may be a novel therapeutic target. These results improve our understanding of M2-polarized macrophages.

  12. [Fluorescence polarization used to investigate the cell membrane fluidity of Saccharomyces cerevisiae treated by pulsed electric field].

    Science.gov (United States)

    Zhang, Ying; Zeng, Xin-An; Wen, Qi-Biao; Li, Lin

    2008-01-01

    To know the lethal mechanism of microorganisms under pulsed electric field treatment, the relationship between the inactivation of Saccharomyces cerevisiae (CICC1308) cell and the permeability and fluidity changes of its cell membrane treated by pulsed electric field (0-25 kV x cm(-1), 0-266 ms) was investigated. With 1,6-diphenyl-1,3,5-hexatriene (DPH) used as a probe, the cell membrane fluidity of Saccharomyces cerevisiae treated by pulsed electric field was expressed by fluorescence polarization. Results showed that the cell membrane fluidity decreases when the electric flied strength is up to 5 kV x cm(-1), and decreases with the increase in electric field strength and treatment time. The plate counting method and ultraviolet spectrophotometer were used to determine the cell viability and to investigate the cell membrane permeability, respectively, treated by pulsed electric field. Results showed that the lethal ratio and the content of protein and nucleic acid leaked from intracellular plasma increased with the increase in the electric field strength and the extension of treatment time. Even in a quite lower electric field of 5 kV x cm(-1) with a tiny microorganism lethal level, the increase in UV absorption value and the decrease in fluidity were significant. It was demonstrated that the cell membrane fluidity decreases with the increase in lethal ratio and cell membrane permeability. The viscosity of cell membrane increases with the decrease in fluidity. These phenomena indicated that cell membrane is one of the most key sites during the pulsed electric field treatment, and the increased membrane permeability and the decreased cell membrane fluidity contribute to the cell death.

  13. Aerodynamics and pollen ultrastructure in Ephedra.

    Science.gov (United States)

    Bolinder, Kristina; Niklas, Karl J; Rydin, Catarina

    2015-03-01

    Pollen dispersal is affected by the terminal settling velocity (Ut) of the grains, which is determined by their size, bulk density, and by atmospheric conditions. The likelihood that wind-dispersed pollen is captured by ovulate organs is influenced by the aerodynamic environment created around and by ovulate organs. We investigated pollen ultrastructure and Ut of Ephedra foeminea (purported to be entomophilous), and simulated the capture efficiency of its ovules. Results were compared with those from previously studied anemophilous Ephedra species.• Ut was determined using stroboscopic photography of pollen in free fall. The acceleration field around an "average" ovule was calculated, and inflight behavior of pollen grains was predicted using computer simulations. Pollen morphology and ultrastructure were investigated using SEM and STEM.• Pollen wall ultrastructure was correlated with Ut in Ephedra. The relative proportion and amount of granules in the infratectum determine pollen bulk densities, and (together with overall size) determine Ut and thus dispersal capability. Computer simulations failed to reveal any functional traits favoring anemophilous pollen capture in E. foeminea.• The fast Ut and dense ultrastructure of E. foeminea pollen are consistent with functional traits that distinguish entomophilous species from anemophilous species. In anemophilous Ephedra species, ovulate organs create an aerodynamic microenvironment that directs airborne pollen to the pollination drops. In E. foeminea, no such microenvironment is created. Ephedroid palynomorphs from the Cretaceous share the ultrastructural characteristics of E. foeminea, and at least some may, therefore, have been produced by insect-pollinated plants. © 2015 Botanical Society of America, Inc.

  14. PINCH1 regulates cell-matrix and cell-cell adhesions, cell polarity and cell survival during the peri-implantation stage

    DEFF Research Database (Denmark)

    Li, Shaohua; Bordoy, Randi; Stanchi, Fabio

    2005-01-01

    PINCH1 is composed of 5 LIM domains, binds integrin-linked kinase (ILK) and locates to integrin-mediated adhesion sites. In order to investigate PINCH1 function we generated mice and embryonic stem (ES) cell-derived embryoid bodies (EBs) lacking the PINCH1 gene. Similar to mice lacking beta1...... integrin or Ilk, loss of PINCH1 arrested development at the peri-implantation stage. In contrast to beta1 integrin or Ilk mutants, however, disruption of the PINCH1 gene produced implantation chambers with visible cell clumps even at embryonic day 9.5. In order to define the phenotype leading to the peri...... with specific antibodies revealed no apparent alteration of PKB/Akt phosphorylation in PINCH1-deficient EBs. Altogether these data demonstrate an important role of PINCH1 for integrin function, actin organization, cell-cell adhesion and endodermal cell survival during the implanting of mouse embryos....

  15. Adipose-derived stem cells were impaired in restricting CD4+T cell proliferation and polarization in type 2 diabetic ApoE-/- mouse.

    Science.gov (United States)

    Liu, Ming-Hao; Li, Ya; Han, Lu; Zhang, Yao-Yuan; Wang, Di; Wang, Zhi-Hao; Zhou, Hui-Min; Song, Ming; Li, Yi-Hui; Tang, Meng-Xiong; Zhang, Wei; Zhong, Ming

    2017-07-01

    Atherosclerosis (AS) is the most common and serious complication of type 2 diabetes mellitus (T2DM) and is accelerated via chronic systemic inflammation rather than hyperglycemia. Adipose tissue is the major source of systemic inflammation in abnormal metabolic state. Pro-inflammatory CD4 + T cells play pivotal role in promoting adipose inflammation. Adipose-derived stem cells (ADSCs) for fat regeneration have potent ability of immunosuppression and restricting CD4 + T cells as well. Whether T2DM ADSCs are impaired in antagonizing CD4 + T cell proliferation and polarization remains unclear. We constructed type 2 diabetic ApoE -/- mouse models and tested infiltration and subgroups of CD4 + T cell in stromal-vascular fraction (SVF) in vivo. Normal/T2DM ADSCs and normal splenocytes with or without CD4 sorting were separated and co-cultured at different scales ex vivo. Immune phenotypes of pro- and anti-inflammation of ADSCs were also investigated. Flow cytometry (FCM) and ELISA were applied in the experiments above. CD4 + T cells performed a more pro-inflammatory phenotype in adipose tissue in T2DM ApoE -/- mice in vivo. Restriction to CD4 + T cell proliferation and polarization was manifested obviously weakened after co-cultured with T2DM ADSCs ex vivo. No obvious distinctions were found in morphology and growth type of both ADSCs. However, T2DM ADSCs acquired a pro-inflammatory immune phenotype, with secreting less PGE2 and expressing higher MHC-II and co-stimulatory molecules (CD40, CD80). Normal ADSCs could also obtain the phenotypic change after cultured with T2DM SVF supernatant. CD4 + T cell infiltration and pro-inflammatory polarization exist in adipose tissue in type 2 diabetic ApoE -/- mice. T2DM ADSCs had impaired function in restricting CD4 + T lymphocyte proliferation and pro-inflammatory polarization due to immune phenotypic changes. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Pollen productivity and morphology of pollen grains in two cultivars of honeyberry (Lonicera kamtschatica (Sevast. Pojark.

    Directory of Open Access Journals (Sweden)

    Małgorzata Bożek

    2012-12-01

    Full Text Available In the years 2004-2006, investigations on the abundance of pollen production in two cultivars of Lonicera kamtschatica (Sevast. Pojark. - Atut and Duet, were conducted at the Felin Experimental Farm of the Agricultural University in Lublin. Moreover, the viability of pollen grains was estimated and measurements of their size were taken. Ten flowers of both studied cultivars supplied 11.42 mg of pollen and the average pollen productivity per 1 ha of a several-years-old plantation was 30.04 kg. The pollen of the observed plants was eagerly collected by honey bees. Pollen grains of both cultivars are suboblate. Considering their size, they can be described as large ones (P=47.55 µm, E=60.37 µm. Pollen viability for both cultivars is high, about 95%.

  17. Apico-basal polarity complex and cancer

    Indian Academy of Sciences (India)

    Apico-basal polarity is a cardinal molecular feature of adult eukaryotic epithelial cells and appears to be involved in several key cellular processes including polarized cell migration and maintenance of tissue architecture. Epithelial cell polarity is maintained by three well-conserved polarity complexes, namely, PAR, Crumbs ...

  18. Transcriptional Evidence for Inferred Pattern of Pollen Tube-Stigma Metabolic Coupling during Pollination

    Science.gov (United States)

    Wang, Fang; Dong, YuXiu; Li, XingGuo; Zhang, Xian Sheng

    2014-01-01

    It is difficult to derive all qualitative proteomic and metabolomic experimental data in male (pollen tube) and female (pistil) reproductive tissues during pollination because of the limited sensitivity of current technology. In this study, genome-scale enzyme correlation network models for plants (Arabidopsis/maize) were constructed by analyzing the enzymes and metabolic routes from a global perspective. Then, we developed a data-driven computational pipeline using the “guilt by association” principle to analyze the transcriptional coexpression profiles of enzymatic genes in the consecutive steps for metabolic routes in the fast-growing pollen tube and stigma during pollination. The analysis identified an inferred pattern of pollen tube-stigma ethanol coupling. When the pollen tube elongates in the transmitting tissue (TT) of the pistil, this elongation triggers the mobilization of energy from glycolysis in the TT cells of the pistil. Energy-rich metabolites (ethanol) are secreted that can be taken up by the pollen tube, where these metabolites are incorporated into the pollen tube's tricarboxylic acid (TCA) cycle, which leads to enhanced ATP production for facilitating pollen tube growth. In addition, our analysis also provided evidence for the cooperation of kaempferol, dTDP-alpha-L-rhamnose and cell-wall-related proteins; phosphatidic-acid-mediated Ca2+ oscillations and cytoskeleton; and glutamate degradation IV for γ-aminobutyric acid (GABA) signaling activation in Arabidopsis and maize stigmas to provide the signals and materials required for pollen tube tip growth. In particular, the “guilt by association” computational pipeline and the genome-scale enzyme correlation network models (GECN) developed in this study was initiated with experimental “omics” data, followed by data analysis and data integration to determine correlations, and could provide a new platform to assist inachieving a deeper understanding of the co-regulation and inter

  19. The Fz-Dsh planar cell polarity pathway induces oriented cell division via Mud/NuMA in Drosophila and zebrafish.

    Science.gov (United States)

    Ségalen, Marion; Johnston, Christopher A; Martin, Charlotte A; Dumortier, Julien G; Prehoda, Kenneth E; David, Nicolas B; Doe, Chris Q; Bellaïche, Yohanns

    2010-11-16

    The Frizzled receptor and Dishevelled effector regulate mitotic spindle orientation in both vertebrates and invertebrates, but how Dishevelled orients the mitotic spindle is unknown. Using the Drosophila S2 cell "induced polarity" system, we find that Dishevelled cortical polarity is sufficient to orient the spindle and that Dishevelled's DEP domain mediates this function. This domain binds a C-terminal domain of Mud (the Drosophila NuMA ortholog), and Mud is required for Dishevelled-mediated spindle orientation. In Drosophila, Frizzled-Dishevelled planar cell polarity (PCP) orients the sensory organ precursor (pI) spindle along the anterior-posterior axis. We show that Dishevelled and Mud colocalize at the posterior cortex of pI, Mud localization at the posterior cortex requires Dsh, and Mud loss-of-function randomizes spindle orientation. During zebrafish gastrulation, the Wnt11-Frizzled-Dishevelled PCP pathway orients spindles along the animal-vegetal axis, and reducing NuMA levels disrupts spindle orientation. Overall, we describe a Frizzled-Dishevelled-NuMA pathway that orients division from Drosophila to vertebrates. Copyright © 2010 Elsevier Inc. All rights reserved.

  20. Pollen viability and membrane lipid composition

    NARCIS (Netherlands)

    Bilsen, van D.G.J.L.

    1993-01-01

    In this thesis membrane lipid composition is studied in relation to pollen viability during storage. Chapter 1 reviews pollen viability, membranes in the dry state and membrane changes associated with cellular aging. This chapter is followed by a study of age-related changes in phospholipid

  1. Pollen Evolution in Yams (Dioscorea: Dioscoreaceae)

    NARCIS (Netherlands)

    Schols, P.; Wilkin, P.; Furness, C.; Huysmans, S.; Smets, E.

    2005-01-01

    Pollen character evolution in yams (Dioscorea: Dioscoreaceae) was investigated in relation to the phylogeny obtained from a recent combined analysis of rbcL and matK gene sequences. The following characters were evaluated: pollen size, aperture number, sexine ornamentation, perforation density, and

  2. Practical aspects of daily pollen census

    NARCIS (Netherlands)

    Quarles van Ufford, W.J.

    1967-01-01

    It is important for diagnostic purposes to keep a daily pollen count. Correlating this with hayfever symptoms, it is then possible to gain insights into critical hours and circumstances which favor an outbreak. Thus we learn to predict the severity of a pollen season and to take preventive measures.

  3. A combinatorial approach to angiosperm pollen morphology.

    Science.gov (United States)

    Mander, Luke

    2016-11-30

    Angiosperms (flowering plants) are strikingly diverse. This is clearly expressed in the morphology of their pollen grains, which are characterized by enormous variety in their shape and patterning. In this paper, I approach angiosperm pollen morphology from the perspective of enumerative combinatorics. This involves generating angiosperm pollen morphotypes by algorithmically combining character states and enumerating the results of these combinations. I use this approach to generate 3 643 200 pollen morphotypes, which I visualize using a parallel-coordinates plot. This represents a raw morphospace. To compare real-world and theoretical morphologies, I map the pollen of 1008 species of Neotropical angiosperms growing on Barro Colorado Island (BCI), Panama, onto this raw morphospace. This highlights that, in addition to their well-documented taxonomic diversity, Neotropical rainforests also represent an enormous reservoir of morphological diversity. Angiosperm pollen morphospace at BCI has been filled mostly by pollen morphotypes that are unique to single plant species. Repetition of pollen morphotypes among higher taxa at BCI reflects both constraint and convergence. This combinatorial approach to morphology addresses the complexity that results from large numbers of discrete character combinations and could be employed in any situation where organismal form can be captured by discrete morphological characters. © 2016 The Author(s).

  4. Abnormal pollen mitoses (PM I and PM II) in an interspecific hybrid ...

    Indian Academy of Sciences (India)

    Unknown

    and the typical hemispherical cell plate was not detected as well. The usual asymmetry was not seen at first pollen mitosis (PM I), and microspores therefore lacked differ- entiation between the vegetative and the generative cell. After telophase, each cell entered PM II, but this was not always followed by cytokinesis. Tripolar ...

  5. ROCK and RHO Playlist for Preimplantation Development: Streaming to HIPPO Pathway and Apicobasal Polarity in the First Cell Differentiation.

    Science.gov (United States)

    Alarcon, Vernadeth B; Marikawa, Yusuke

    2018-01-01

    In placental mammalian development, the first cell differentiation produces two distinct lineages that emerge according to their position within the embryo: the trophectoderm (TE, placenta precursor) differentiates in the surface, while the inner cell mass (ICM, fetal body precursor) forms inside. Here, we discuss how such position-d