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The plasma membrane is a physical boundary made of amphiphilic lipid molecules, proteins and carbohydrates extensions. Its role in mechanotransduction generates increasing attention in animal systems, where membrane tension is mainly induced by cortical actomyosin. In plant cells, cortical tension is of osmotic origin. Yet, because the plasma membrane in plant cells has comparable physical properties, findings fromanimal systems likely apply to plant cells too. Recent results suggest that this is indeed the case, with a role of membrane tension in vesicle trafficking, mechanosensitive channel opening or cytoskeleton organization in plant cells. Prospects for the plant science community are at least three fold: (i) to develop and use probes to monitor membrane tension in tissues, in parallel with other biochemical probes, with implications for protein activity and nanodomain clustering, (ii) to develop single cell approaches to decipher the mechanisms operating at the plant cell cortex at high spatio-temporal resolution, and (iii) to revisit the role of membrane composition at cell and tissue scale, by considering the physical implications of phospholipid properties and interactions in mechanotransduction. [ABSTRACT FROM AUTHOR]

Microtubules are dynamic polymers, permanently assembling and disassembling, that serve as tracks for intra-cellular transport by molecular motors. We recently found that the low energy of tubulin dimer interactions allows for spontaneous loss of tubulin dimers from the microtubule lattice [1]. This raised the possibility that the mechanical work produced by molecular motors as they move on microtubules can break dimer interactions and trigger microtubule disassembly. In a recent study, we tested this hypothesis by studying the interplay between microtubules and moving molecular motors in vitro [2]. Our results show that molecular motors can remove tubulin dimers from the lattice and rapidly destroy microtubules. We also found that dimer removal by motors was compensated by the insertion of free tubulin dimers into the microtubule lattice. This self-repair mechanism allows microtubules to survive the damage induced by molecular motors as they move along their tracks. Our study reveals the existence of coupling between the motion ofmolecular motors and the renewal of the microtubule lattice. [ABSTRACT FROM AUTHOR]

Asymmetrically differentiating cells are formed with the aid of RNA-binding proteins (RBPs), which can bind, stabilize, regulate, and transport target mRNAs. The loss of RBPs in neurons may lead to severe neurodevelopmental diseases such as the Fragile X Syndrome with the absence of the Fragile X Mental Retardation Protein (FMRP). Because the latter is ubiquitous and shares many similarities with other RBPs involved in the development of peripheral cells, we suggest that FMRP would have a role in the differentiation of all tissues where it is expressed. A MEG-01 differentiation model was, therefore, established to study the global developmental functions of FMRP. PMA induction of MEG-01 cells causes important morphological changes driven by cytoskeletal dynamics. Cytoskeleton change and colocalization analyses were performed by confocal microscopy and sucrose gradient fractionation. Total cellular protein content and de novo synthesis were also analyzed. Microtubular transport mediates the displacement of FMRP and other RBP-containing mRNP complexes towards regions of the cell in development. De novo protein synthesis decreases significantly upon differentiation and total protein content composition is altered. Because those results are comparable with those obtained in neurons, the absence of FMRP would have significant consequences in cells everywhere in the body. The latter should be further investigated to give a better understanding of the systemic implications of imbalances of FMRP and other functionally similar RBPs. [ABSTRACT FROM AUTHOR]

Airway smooth muscle (ASM) plays a central role in the excessive narrowing of the airway that characterizes the primary functional impairment in asthma. This phenomenon is known as airway hyper-responsiveness (AHR). Emerging evidence suggests that the development and maintenance of ASM force involves dynamic reorganization of the subcellular filament network in both the cytoskeleton and the contractile apparatus. In this review, evidence is presented to support the view that regulation of ASM contraction extends beyond the classical actomyosin interaction and involves processes within the cytoskeleton and at the interfaces between the cytoskeleton, the contractile apparatus, and the extracellular matrix. These processes are initiated when the muscle is activated, and collectively they cause the cytoskeleton and the contractile apparatus to undergo structural transformation, resulting in a more connected and solid state that allows force generated by the contractile apparatus to be transmitted to the extracellular domain. Solidification of the cytoskeleton also serves to stiffen the muscle and hence the airway. Oscillatory strain from tidal breathing and deep inspiration is believed to be the counter balance that prevents hypercontraction and stiffening of ASM in vivo. Dysregulation of this balance could lead to AHR seen in asthma. [ABSTRACT FROM AUTHOR]

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Cancer cells often exhibit characteristic aberrations in their nuclear architecture, which are indicative of their malignant potential. In this study, we have examined the nuclear and cytoskeletal composition, attachment configuration dynamics, and osmotic or drug treatment response of invasive (Hs578T and MDA-MB-231) and non-invasive (MCF-10A and MCF-7) breast cancer cell lines. Unlike MCF-10A and MCF-7, Hs578T and MDA-MB-231 cells showed extensive nuclear elasticity and deformability and displayed distinct kinetic profiles during substrate attachment. The nuclear shape of MCF-10A and MCF-7 cells remained almost unaffected upon detachment, hyperosmotic shock, or cytoskeleton depolymerization, while Hs578T and MDA-MB-231 revealed dramatic nuclear contour malformations following actin reorganization. [ABSTRACT FROM AUTHOR]

En 2011, l'Université Joseph Fourier (UJF), s'appuyant sur la force de la recherche interdisciplinaire des laboratoires grenoblois dans les Nanosciences et Nanotechnologies, a ouvert deux majeures : un M1 « Physique fondamentale et Nanosciences » et un M1 « Chimie et Nanosciences ». Dans ce cadre, des Travaux Pratiques interdisciplinaires et interplateformes ont été mis en place afin d'illustrer de manière concrète la convergence de la physique, de la chimie et de la biologie à l'échelle micro/nanométrique qui s'appuie sur les micro et nanotechnologies. Cet article décrit en détail ces TP et expose leur intérêt pédagogique pour la formation des étudiants ainsi que pour des enseignants-chercheurs. [ABSTRACT FROM AUTHOR]

Podocytes play a critical role in maintaining glomerular permselectivity. It has been long recognized that their intricate actin-based structures are tightly associated with their normal function; however, the precise mechanisms by which podocytes form and maintain their complex structure had been poorly understood until the intensive investigations on podocyte biology began in 1998, triggered by the breakthrough discovery of nephrin. This review summarizes the recent discoveries of the molecular mechanisms by which the actin cytoskeleton is regulated in podocytes. A particular focus will be on the role of the Rho-family of small GTPases, represented by RhoA, Rac1, and Cdc42. Rho-GTPases are known for their versatile cellular functions, most importantly for the actin regulatory roles. We will also discuss the potential roles of the 3 groups of proteins known to regulate Rho-GTPases, namely GTPase-activating proteins, guanine nucleotide exchange factors, and guanine nucleotide dissociation inhibitors. [ABSTRACT FROM AUTHOR]

We provide an overview of our studies on cadmium and the actin cytoskeleton in mesangial cells, from earlier work on the effects of Cd2+ on actin polymerization in vivo and in vitro, to a role of disruption or stabilization of the cytoskeleton in apoptosis and apoptosis-like death. More recent studies implicate cadmium-dependent association of gelsolin and the Ca2+/calmodulin-dependent protein kinase II (CaMK-II) with actin filaments in cytoskeletal effects. We also present previously unpublished data concerning cadmium and the disruption of focal adhesions. The work encompasses studies on rat, mouse, and human mesangial cells. The major conclusions are that Cd2+ acts independently of direct effects on cellular Ca2+ levels to nevertheless activate Ca2+-dependent proteins that shift the actin polymerization-depolymerization in favour of depolymerization. Cadmium-dependent translocation of CaMK-IIδ, gelsolin, and a 50 kDa gelsolin cleavage fragment to the filamentous (F-)actin cytoskeleton appear to be involved. An intact filamentous actin cytoskeleton is required to initiate apoptotic and apoptotic-like death, but F-actin depolymerization is an eventual result. [ABSTRACT FROM AUTHOR]

The present work aimed to investigate whether exposure to static magnetic field (SMF) and extremely low frequency magnetic field (ELF-MF) can induce biomolecular changes on Tuber borchii hyphal growth. Tuber borchii mycelium was exposed for 1 h for 3 consecutive days to a SMF of 300 mT or an ELF-MF of 0.1 mT 50 Hz. Gene expression and biochemical analyses were performed. In mycelia exposed to ELF-MF, some genes involved in hyphal growth, investigated using quantitative real-time polymerase chain reaction, were upregulated, and the activity of many glycolytic enzymes was increased. On the contrary, no differences were observed in gene expression after exposure to SMF treatment, and only the activities of glucose 6-phosphate dehydrogenase and hexokinase increased. The data herein presented suggest that the electromagnetic field can act as an environmental factor in promoting hyphal growth and can be used for applicative purposes, such as the set up of new in vitro cultivation techniques. [ABSTRACT FROM AUTHOR]

Summary: This brief text aims at illustrating the interactions between connective tissue fibers and cell cytoskeleton fibers. These two networks are connected by molecular bridges at the level of the cell membrane of the cells of the connective and vascular tissues, allowing functional adjustments across the two domains, but also the transduction of forces and tensions into a biochemical alphabet. The signaling between the cell kern and its environment, but equally the other way round, from the environment to the core of the cell, depends on it. [Copyright &y& Elsevier]

Abstract: Aim: The goal of this study was to understand whether dense fibrillar collagen matrices, with a hierarchical structure resembling native collagen matrices, could be useful to study collagen receptor function, in a more physiological context. The receptor analysed here was integrin α11β1, already shown to be involved in cell attachment and migration on collagen-coated plastic, and also in contraction of loose fibrillar collagen hydrogels. Materials and methods: Collagen matrices prepared here corresponded to dense fibrillar hydrogels concentrated at 5mg/ml. The behaviour of α11β1 deficient fibroblasts seeded on these concentrated matrices was assessed in terms of adhesion, morphology and migration, then compared to that observed on classical hydrogels at 1mg/ml, corresponding to loose collagen matrices. Results: Short-term attachment assays showed disturbed interactions between α11β1 deficient cells and collagen matrices in a concentration-dependent manner. Long-term assays revealed reduced cell spreading of alpha 11−/− cells on the dense collagen matrices, associated with a disturbed cytoskeleton network. Moreover, anoikis was observed when alpha 11−/− cells were seeded on 5mg/ml matrices, and not on looser 1mg/ml matrices. In scratch wound in vitro assays, carried out with cells on 5mg/ml fibrillar collagen matrices, alpha 11−/− cells migrated much better than their wild-type counterparts. In contrast, no significant difference was observed between wild and knock-out cells seeded on plastic. Conclusions: The present study demonstrates the validity of in vivo-like dense fibrillar collagen matrices to evaluate cell receptor functions more significantly than with 2D cell cultures or loose hydrogels. [Copyright &y& Elsevier]

Abstract: A group of four proteins – spectrin, ankyrin, 4.1 and adducin – evolved with the metazoa. These membrane-cytoskeletal proteins cross-link actin on the cytoplasmic face of plasma membranes and link a variety of transmembrane proteins to the cytoskeleton. In this paper, the evolution of these proteins is analysed. Genomics indicate that spectrin was the first to appear, since the genome of the choanoflagellate Monosiga brevicolis contains genes for α, β and βH spectrin. This organism represents a lineage of free-living and colonial protists from which the metazoa are considered to have diverged. This indicates that spectrin emerged in evolution before the animals. Simple animals such as the placozoan Trichoplax adherens also contain recognizable precursors of 4.1, ankyrin and adducin, but these could probably not bind spectrin. Ankyrin and adducin seem to have acquired spectrin-binding activity with the appearance of tissues since they appear to have largely the same domain structure in all eumetazoa. 4.1 was adapted more recently, with the emergence of the vertebrates, to bind spectrin and promote its interaction with actin. A simple hypothesis is that spectrin was prerequisite (but not sufficient) for animal life; that spectrin interaction with ankyrin and adducin was required for evolution of major tissues; and that 4.1 acquired a spectrin-actin binding activity as animal size increased with the appearance of vertebrates. The spectrin/ankyrin/adducin/4.1 complex represents a remarkable system that underpins animal life; it has been adapted to many different functions at different times during animal evolution. [ABSTRACT FROM AUTHOR]

Cadmium induces apoptotic cell death in mouse mesangial cells that is in part dependent on reactive oxygen species (ROS). Cadmium also activates multiple kinases in these cells, including the Ca2+/calmodulin-dependent protein kinase II (CaMK-II) and p38 kinase, and also leads to disruption of the filamentous actin cytoskeleton. We investigated the role of the cytoskeleton in Cd2+-induced cell death. Cell viability was decreased by Cd2+ and two types of apoptotic death, defined by flow cytometry, were increased. Disruption of actin filaments with cytochalasin D was partially protective, whereas stabilization of the cytoskeleton with jasplakinolide was without effect, indicating that cytoskeletal disruption contributes to, but is not necessary for, induction of apoptosis. Inhibition of CaMK-II and p38 kinase, mitochondrial stabilization with cyclosporine A, and the antioxidant N-acetyl cysteine all protected against apoptosis and prevented disruption of the cytoskeleton. Cytochalasin D decreased Cd2+-dependent ROS production, reduced the decline in mitochondrial membrane potential, and decreased phosphorylation of p38 kinase. We conclude that Cd2+-dependent actin disruption is a downstream event facilitating apoptotic death. Cadmium-dependent cell death involves actin-dependent mitochondrial changes, ROS production, and p38 activation. Le cadmium (Cd2+) induit la mort par apoptose dans les cellules mésangiales des souris, laquelle dépend en partie des espèces réactives de l’oxygène (ROS). Le Cd2+ active aussi de nombreuses kinases, y compris la Ca2+/protéine kinase II dépendante de la calmoduline (CaMK-II) et la p38 kinase, et provoque la rupture du cytosquelette d’actine. Nous avons examiné le rôle du cytosquelette dans l’apoptose induite par le Cd2+. Le Cd2+ a diminué la viabilité cellulaire, et deux types d’apoptose, identifiés par cytométrie de flux, ont augmenté. La rupture des filaments d’actine au moyen de cytochalasine D a eu un effet protecteur partiel, alors que la stabilisation du cytosquelette au moyen de jasplakinolide n’a pas eu d’effet, ce qui indique que la rupture du cytosquelette contribue, mais n’est pas essentielle, à l’induction de l’apoptose. L’inhibition de la CaMKII et de la p38 kinase, la stabilisation mitochondriale par la cyclosporine A et l’utilisation de l’antioxydant N-acétyl cystéine ont eu des effets protecteurs contre l’apoptose et prévenu la rupture du cytosquelette. La cytochalasine D a diminué la production de ROS dépendante du Cd2+, ralenti la diminution du potentiel de membrane mitochondrial et atténué la prohosphorylation de la p38 kinase. Nous concluons que la rupture des filaments d’actine dépendante du Cd2+ est un événement en aval favorisant l’apoptose. L’apoptose induite par le cadmium implique des modifications mitochondriales dépendantes de l’actine, la production de ROS et l’activation de la kinase p38. [ABSTRACT FROM AUTHOR]

Release of angiotensin II (Ang II) after vascular injury promotes tissue repair by stimulating phenotypic modulation of smooth muscle cells, which enables cell proliferation and migration. This process requires cytoskeleton remodeling, which involves cortactin, a scaffold protein that is phosphorylated by Src kinase in response to Ang II. Since insulin-like growth factor (IGF)-1 receptor transactivation mediates intracellular signals originating from the Ang II type 1 (AT1) receptor in a Src kinase-dependent manner, we examined whether IGF-1 receptor transactivation was also required for cortactin phosphorylation. Treatment of quiescent smooth muscle cells with Ang II resulted in both cortactin phosphorylation and its translocation to the plasma membrane. Both events were prevented by 1-(1,1-dimethylethyl)-1-(4-methylphenyl)-1H-pyrazolo(3,4-d)pyrimidin-4-amine (PP1), a Src kinase inhibitor, and by AG1024, an inhibitor of the IGF-1 receptor tyrosine kinase. Additionally, PP1 and AG1024 blocked the association of cortactin with actin-related protein (Arp) 3, an actin nucleation factor. These results indicate that Src kinase and the IGF-1 receptor kinase are necessary for activating cortactin. Phosphorylation of Src kinase in Ang II-treated cells was subsequently examined and was shown to be prevented by AG1024. Furthermore, Src kinase phosphorylation was blocked by inhibitors of protein kinase C (PKC), but not by inhibitors of phosphatidylinositol (PI) 3-kinase. These data establish that IGF-1 receptor transactivation is required for Src kinase-mediated cortactin phosphorylation and cytoskeletal reorganization in response to Ang II. [ABSTRACT FROM AUTHOR]

We examined the interaction of smooth muscle myosin with α-actin and β-actin isoforms during the contraction of A7r5 smooth muscle cells and rat aortic smooth muscle. The techniques of confocal microscopy and fluorescence resonance energy transfer (FRET) analysis were utilized in examining A7r5 cells and rat aortic rings contracted with phorbol 12,13-dibutyrate. Visual evaluation of confocal images of A7r5 smooth muscle cells contracted by phorbol 12,13-dibutyrate indicated significant disassociation of myosin from α-actin but not β-actin. Whole-cell FRET analysis confirmed these observations (α-actin-myosin -67%, β-actin-myosin -2%). Time course studies further showed that α-actin-myosin complex increased significantly (40%) within 1.5 min after the addition of phorbol 12,13-dibutyrate and then declined as contraction progressed. FRET analysis of rat aortic rings at different intervals of contraction indicated significant increases in α-actin-myosin at the initiation (79%) and plateau (67%) in force development, but not during the intermediate period of slowly developing tension (-4%). By comparison, β-actin-myosin complex was unchanged except during slow force development, in which the association was significantly decreased (-30%). Similar to that of α-actin-myosin, Alexa 488 - phalloidin staining fluorescence indicated increased tissue F-actin content at the initiation (21%) and plateau (62%) in force. FRET images indicated the development of thickened cables and patches of α-actin-myosin in tissue throughout the interval of contraction. The results provide direct evidence of dynamic remodeling of the contractile protein during vascular smooth muscle contraction and suggest that FRET analysis may be a powerful tool for assessment of tissue protein-protein associations. [ABSTRACT FROM AUTHOR]

Prestressed system mechanics applied to the cytoskeleton structure. These works show the pertinence and usefulness in modelling the cytoskeleton by mechanical prestressed systems. They are in the field of cellular and tissular biomechanics and two applications are proposed. We present first the form-finding of an isolated cell by non regular tensegrity systems, and then we propose to combine granular models and tensegrity analogy. A prospective extension to multicellular assemblies considering the concept of shared self-stresses, by the way of tensegrity grids, is subsequently presented. [ABSTRACT FROM AUTHOR]

Physical exercise was used as a model of the physiological modulator of free radical production to examine the effects of exercise-induced oxidative modifications on the physico-biochemical properties of erythrocyte membrane. The aim of our work was to investigate conformational changes of erythrocyte membrane proteins, membrane fluidity, and membrane susceptibility to disintegration. Venous blood was taken before, immediately after, and 1 h after an exhaustive incremental cycling test (30 W·min-1 ramp), performed by 11 healthy untrained males on balanced diets (mean age, 22 ± 2 years; mean body mass index, 25 ± 4.5 kg·m-2). In response to this exercise, individual maximum heart rate was 195 ± 12 beats·min-1 and maximum wattage was 292 ± 27 W. Electron paramagnetic resonance spectroscopy was used to investigate alterations in membrane proteins and membrane dynamics, and to measure production of radical species. The reducing potential of plasma (RPP) was measured using the reduction of 1,1-diphenyl-2-picrylhydrazyl (DPPH) and the ferric-reducing ability of plasma. Exercise induced decreases in erythrocyte membrane fluidity in the polar region (p < 0.0001) and alterations in the conformational state of membrane proteins (p < 0.05). An increase in RPP was observed immediately after exercise (p < 0.001), with a further increase 1 h postexercise (p < 0.0001). Supporting measurements of lipid peroxidation showed an increase in thiobarbituric acid reactive substances immediately after exercise (p < 0.05) and at 1 h of recovery (p < 0.001); however, free radicals were not detected. Results indicate the existence of early postexercise mild oxidative stress after single-exercise performance, which induced structural changes in erythrocyte membrane components (protein aggregation) and in the membrane organization (lipids rigidization) that followed lipid peroxidation but did not lead to cellular hemolysis. [ABSTRACT FROM AUTHOR]

Enteropathogenic Escherichia coli (EPEC) colonizes the intestinal mucosa and causes a cell lesion known as attachment and effacement (A/E) lesion. The molecular mechanisms for A/E lesions include injection of Tir, which is a receptor for an adhesin named intimin. The Tir-intimin interaction causes rearrangement of the cytoskeleton forming actin-rich structures called pedestals. Unfortunately, the formation of the A/E lesions and the dynamics of the actin cytoskeleton during this rearrangement induced by EPEC cannot be studied in the natural host. However, there are EPEC strains that infect rabbit (REPEC) that are genetically and pathologically similar to EPEC. Here, we used REPEC for the infection of rabbit kidney epithelial cells, line RK13, as a model to understand the actin cytoskeleton dynamics during pedestal formation. Actin-rich pedestal formation during the infection of RK13 cells by REPEC was analyzed by electron and confocal microscopy. The kinetics of infection along with the use of antibiotics for eliminating the bacteria, as well as reinfection, evidenced the plasticity of the actin cytoskeleton during pedestal formation. Thus, this model is a helpful tool for studying the dynamics of actin cytoskeleton and for correlating the data with those observed in in vivo models in rabbits experimentally infected with REPEC. Escherichia coli entérophathogène (EPEC) colonise la muqueuse intestinale et cause des lésions cellulaires appelées lésions d’attachement et d’effacement (A/E). Les mécanismes moléculaires associés aux lésions incluent l’implication de Tir, le récepteur d’une adhésine, l’intimine. L’interaction Tir-intimine cause un réarrangement du cytosquelette formant une structure riche en actine, le piédestal. Malheureusement, la formation de lésions A/E et la dynamique du cytosquelette d’actine lors du réarrangement induit par les EPEC ne peuvent être étudiées chez l’hôte naturel. Cependant, il existe des souches d’EPEC qui peuvent infecter le lapin (REPEC) et qui sont génétiquement et pathologiquement similaires aux EPEC. Nous avons utilisé ici des REPEC pour infecter des cellules épithéliales rénales de lapin, les cellules de la lignée RK13, un modèle permettant de comprendre la dynamique du cytosquelette d’actine lors de la formation de piédestaux. La formation d’un piédestal riche en actine lors de l’infection des cellules RK13 par les REPEC a été analysée par microscopie électronique et par microscopie confocale. La cinétique d’infection et l’utilisation d’antibiotiques pour éliminer les bactéries, ainsi que la réinfection ont mis en évidence la plasticité du cytosquelette d’actine lors de la formation du piédestal. Ainsi, ce modèle constitue un outil pratique pour étudier la dynamique du cytosquelette d’actine et pour corréler les résultats avec les observations faites dans des modèles in vivo chez le lapin infecté par les REPEC. [ABSTRACT FROM AUTHOR]

Hydrostatic pressure (HP) is a mechanical stimulus that has received relatively little attention in the field of the cell biology of mechanotransduction. Generalized models, such as the tensegrity model, do not provide a detailed explanation of how HP might be detected. This is significant, because HP is an important mechanical stimulus, directing cell behaviour in a variety of tissues, including cartilage, bone, airways, and the vasculature. HP sensitivity may also be an important factor in certain clinical situations, as well as under unique environmental conditions such as microgravity. While downstream cellular effects have been well characterized, the initial HP sensation mechanism remains unclear. In vitro evidence shows that HP affects cytoskeletal polymerization, an effect that may be crucial in triggering the cellular response. The balance between free monomers and cytoskeletal polymers is shifted by alterations in HP, which could initiate a cellular response by releasing and (or) activating cytoskeleton-associated proteins. This new model fits well with the basic tenets of the existing tensegrity model, including mechanisms in which cellular HP sensitivity could be tuned to accommodate variable levels of stress. [ABSTRACT FROM AUTHOR]

Airway hyperresponsiveness is the excessive narrowing of the airway lumen caused by stimuli that would cause little or no narrowing in the normal individual. It is one of the cardinal features of asthma, but its mechanisms remain unexplained. In asthma, the key end-effector of acute airway narrowing is contraction of the airway smooth muscle cell that is driven by myosin motors exerting their mechanical effects within an integrated cytoskeletal scaffolding. In just the past few years, however, our understanding of the rules that govern muscle biophysics has dramatically changed, as has their classical relationship to airway mechanics. It has become well established, for example, that muscle length is equilibrated dynamically rather than statically, and that in a dynamic setting nonclassical features of muscle biophysics come to the forefront, including unanticipated interactions between the muscle and its time-varying load, as well as the ability of the muscle cell to adapt (remodel) its internal microstructure rapidly in response to its ever-changing mechanical environment. Here, we consider some of these emerging concepts and, in particular, focus on structural remodeling of the airway smooth muscle cell as it relates to excessive airway narrowing in asthma. [ABSTRACT FROM AUTHOR]

The sliding filament theory of contraction that was developed for striated muscle is generally believed to be also applicable to smooth muscle. However, the well-organized myofilament lattice (i.e., the sarcomeric structure) found in striated muscle has never been clearly delineated in smooth muscle. There is evidence that the myofilament lattice in some smooth muscles, such as airway smooth muscle, is malleable; it can be reshaped to fit a large range of cell dimensions while the maximal overlap between the contractile filaments is maintained. In this review, some early models of the structurally static contractile apparatus of smooth muscle are described. The focus of the review, however, is on the recent findings supporting a model of structurally dynamic contractile apparatus and cytoskeleton for airway smooth muscle. A list of unanswered questions regarding smooth muscle ultrastructure is also proposed in this review, in the hope that it will provide some guidance for future research. [ABSTRACT FROM AUTHOR]

Exercise-induced mechanical loading can have potent effects on skeletal form and health. Both intrinsic and extrinsic factors contribute to bone structure and function. Mechanical simuli (e.g., strain magnitude, frequency, rate, and gradients, as well as fluid flow and shear stress) have potent influences on bone-cell cytoskeleton and associated signalling pathways. Although the immature skeleton may be more able to benefit from exercise, a skeletally mature population can also benefit from exercise programs aimed at increasing the functional loads to which the skeleton is exposed. The definitive explanation of mechanical-loading and (or) bone-cell mechanotransductive phenomena, however, remains elusive. Here, we briefly review the structural and anatomical foundation for bone adaptation, focusing on mechanical loading effects on bone, linked to the roles of integrins, cytoskeleton, membrane channels, and auto- and paracrine factors in bone modeling and remodeling. [ABSTRACT FROM AUTHOR]

Abstract: The ability to model the mechanical behaviour of the cell cytoskeleton as realistically as possible is a key point in understanding numerous biological mechanisms. Tensegrity systems have already demonstrated their pertinence for this purpose. However, the structures considered until now are based only on models with simplified geometry and topology compared to the true complexity of cytoskeleton architecture. The aim of this Note is to propose a form-finding method for generating nonregular tensegrity shapes of higher diversity and complexity. The process relies on the use of the dynamic relaxation method. Further improvements have made it possible to control the computed morphologies and to modify them to approach experimentally observed configurations. Various examples illustrate the use of the method and the results obtained for different cell typologies. To cite this article: H. Baudriller et al., C. R. Mecanique 334 (2006). [Copyright &y& Elsevier]

In the present study, we investigated the reorganization of α- and β-actin in the contracting A7r5 smooth muscle cell. The remodeling of these actin variants was markedly different in response to increasing concentrations of phorbol 12, 13-dibutyrate (PDBu). At the lowest concentrations (≤10-7 mol/L), cells showed an ~70% loss in α-actin stress fibers with robust transport of this isoform to podosomes. By comparison, β-actin remained in stress fibers in cells stimulated at low concentrations (≤10-7 mol/L) of PDBu. However, at high concentrations (≥10-6 mol/L) ~50% of cells showed transport of β-actin to podosomes. Consistent with these findings, staining with phalloidin indicated a significant decrease in the whole-cell content of F-actin with PDBu treatment. However, staining with DNase I indicated no change in the cellular content of G-actin, suggesting reduced access of phalloidin to tightly packed actin in the podosome core. Inhibition of protein kinase C (staurosporine, bisindolymaleimide) blocked PDBu-induced (5 × 10-8 mol/L) loss in α-actin stress fibers or reversed podosome formation with re-establishment of α-actin stress fibers. By comparison, these inhibitors caused partial loss of β-actin stress fibers. The results support our earlier conclusion of independent remodeling of α- and β-actin cytoskeletal structure and suggest that the regulation of these structures is different. [ABSTRACT FROM AUTHOR]

Abstract: The cultures of neonatal rat cardiomyocytes represent a very useful tool for the observation and the understanding of the cellular aspects of the electrophysiological, contractile, morphological, metabolic and molecular properties of the myocardium. This model is characterized by a homogeneous population of cardiac muscular cells and by vast possibilities of control of the chemical and physical environment of the cells, allowing the in vitro mimicry of a wide range of cardiac pathological situations. The cardiomyocyte cultures are thus suited to very varied experimental protocols, allowing multiparametric analysis of the cardiocellular effects of different stress such as hypoxia–reoxygenation, of ischemia–reperfusion, of the free radical attack and of thermal shock. These investigations can be combined with the study of the effects and of the cytotoxicity of pharmacological agents, not limited to the putatively cardioactive drugs. The present review proposes an outline of the procedures for the isolation, the culture and the use of neonatal cardiomyocytes. To illustrate the potentialities of this preparation, we describe more specifically the protocols and the various consequences at the cellular scale of an in vitro model of myocardial ischemia–reperfusion. [Copyright &y& Elsevier]

Biomechanical studies aim at understanding the relationship between the mechanical properties of biological structures and their function. In cytomechanical investigations, this approach is brought down to the scale of cells and subcellular structures. In plant cells and the hyphae of fungi and water molds, interactions between turgor pressure, the cell wall, and the cytoskeleton are considered of primary importance. This review is an overview of how the mechanical properties of these individual features and their interactions have been measured and how the experimental data are used to produce theoretical mechanical models of cellular architecture and dynamics. Several models are discussed, and focusing on the example of tip-growing cells, various approaches to understanding the mechanical aspects of cellular morphogenesis are analyzed. [ABSTRACT FROM AUTHOR]

Recent studies are revealing plausible mechanisms that help explain how the two major cytoskeletal systems of plant cells interact to co-ordinate morphogenesis in diffusely expanding cells. In this article, we focus on the development of pavement cells typically found in the leaf epidermis, and highlight work that provides insights into the mechanisms that generate their complex morphology. Pavement cells interdigitate with adjacent cells, forming narrow neck regions interspersed with lobe-like projections. Earlier analysis demonstrated that distinct banding of cortical microtubules and associated accumulation of cell wall material was responsible for maintaining the neck regions during expansion. More recently, it has been determined that patches of fine actin microfilaments regulate the formation of lobing regions. This zonation into microtubule-rich bands and actin patches is coordinated by the activity of Rops, small GTPases that control a wide range of signalling pathways including ones that remodel both actin microfilament and microtubule arrays. Moreover, the formation of microtubule bands and actin patches seems to be self-reinforcing. Loss of microtubule polymers by drug treatment or mutation broadens actin patch formation, apparently by enhancing Rop interactions with a positive regulator of actin polymerization. Thus, cross-talk between microtubule and actin microfilament networks is essential for coordinating and reinforcing pavement cell morphogenesis. [ABSTRACT FROM AUTHOR]

In keeping with the myriad functions carried out by plants, their component cells display an amazing diversity of shapes and sizes. How is a precise cell form achieved? In recent years, the single-celled, branched, aerial epidermal trichome of Arabidopsis thaliana L. (Heynh) has emerged as a model cell for understanding the cell biological and molecular basis underlying the development of cell shape in plants. Here, I critique the recent information gleaned from dissecting trichome cell morphogenesis in Arabidopsis and identify areas and questions that can be further addressed using this unique cell type. [ABSTRACT FROM AUTHOR]

The cytoskeleton plays an important role in numerous cellular processes, including subcellular mRNA localization and translation. Several examples of mRNA localization have emerged in plant cells, and these appear to function in protein targeting, the establishment of polarity, and cell-to-cell trafficking. The identification of several cytoskeleton-associated RNA-binding proteins in plant cells has made available candidate proteins that mediate the interaction between mRNA and the cytoskeleton, and possibly play a role in mRNA localization and translational control. We propose a model that links mRNA–microtubule interactions to translational autoregulation, a process that may assist in the efficient and regulated binding of proteins to microtubules. [ABSTRACT FROM AUTHOR]

We evaluated the hypothesis that mechanical deformation of airway smooth muscle induces structural remodeling of airway smooth muscle cells, thereby modulating mechanical performance in subsequent contractions. This hypothesis implied that past experience of mechanical deformation was retained (or "memorized") as structural changes in airway smooth muscle cells, which modulated the cell's subsequent contractile responses. We termed this phenomenon mechanical strain memory. Preshortening has been found to induce attenuation of both force and isotonic shortening velocity in cholinergic receptor-activated airway smooth muscle. Rapid stretching of cholinergic receptor-activated airway smooth muscle from an initial length to a final length resulted in post-stretch force and myosin light chain phosphorylation that correlated significantly with initial length. Thus post-stretch muscle strips appeared to retain memory of the initial length prior to rapid stretch (mechanical strain memory). Cytoskeletal recruitment of actin- and integrin- binding proteins and Erk 1/2 MAPK appeared to be important mechanisms of mechanical strain memory. Sinusoidal length oscillation led to force attenuation during oscillation and in subsequent contractions in intact airway smooth muscle, and p38 MAPK appeared to be an important mechanism. In contrast, application of local mechanical strain to cultured airway smooth muscle cells induced local actin polymerization and cytoskeletal stiffening. It is conceivable that deep inspiration-induced bronchoprotection may be a manifestation of mechanical strain memory such that mechan ical deformation from past breathing cycles modulated the mechanical performance of airway smooth muscle in subsequent cycles in a continuous and dynamic manner. [ABSTRACT FROM AUTHOR]

Microtubules are key cytoskeletal components in the cytoplasm of eukaryotic cells where they have pleiotropic and vital roles in functions such as cell division, trafficking or morphogenesis. Microtubules are especially abundant in neurons. Although microtubules are in many cells dynamic polymers, they exhibit an extreme state of stability in neurons. Previous work has indicated a central role of microtubule associated proteins called STOPs in neuronal microtubule stabilization. We have recently developed STOP null mice. These mice were devoid of stable brain microtubules but to our surprise had nevertheless an apparently normal brain anatomy. However the mice showed synaptic defects affecting different forms of long- and short-term synaptic plasticity. These synaptic defects were associated with severe behavioral defects that showed a remarkable sensitivity to long-term treatment with neuroleptics. We discuss the relationship of the phenotypes observed in STOP null mice with current models of schizophrenia in which the multiple, severe, and neuroleptic sensitive mental disorders caused by the disease are due to a “disease of the synapse”. [Copyright &y& Elsevier]

Deciphering how Listeria monocytogenes exploits the host cell machinery to invade mammalian cells during infection is a key issue for the understanding how this food-borne pathogen causes a pleiotropic disease ranging from gastro-enteritis to meningitis and abortions. Using multidisciplinary approaches, essentially combining bacterial genetics and cell biology, we have identified two bacterial proteins critical for entry into target cells, InlA and InlB. Their cellular ligands have been also identified: InlA interacts with the adhesion molecule E-cadherin, while InlB interacts with the receptor for the globular head of the complement factor C1q (gC1q-R), with the hepatocyte growth factor receptor (c-Met) and with glycosaminoglycans (including heparan sulphate). The dynamic interaction between these cellular receptors and the actin cytoskeleton is currently under investigation. Several intracellular molecules have been recognized as key effectors for Listeria entry into target cells, including catenins (implicated in the connection of E-cadherin to actin) and the actin depolymerising factor/cofilin (involved in the rearrangement of the cytoskeleton in the InlB-dependent internalisation pathway). At the organism level, species specificity has been discovered concerning the interaction between InlA and E-cadherin, leading to the generation of transgenic mice expressing the human E-cadherin, in which the critical role of InlA in the crossing of the intestinal barrier has been clearly determined. Listeria appears as an instrumental model for addressing critical questions concerning both the complex process of bacterial pathogenesis and also fundamental molecular processes, such as phagocytosis. To cite this article: J. Pizarro-Cerda´ et al., C. R. Biologies 327 (2004). [Copyright &y& Elsevier]