scholarly journals Adhesive multiplicity in the interaction of embryonic fibroblasts and myoblasts with extracellular matrices.

1984 ◽  
Vol 99 (4) ◽  
pp. 1398-1404 ◽  
Author(s):  
C Decker ◽  
R Greggs ◽  
K Duggan ◽  
J Stubbs ◽  
A Horwitz
Keyword(s):  
Skeletal Muscle ◽  
Cardiac Muscle ◽  
Cardiac Fibroblasts ◽  
Cell Types ◽  
Extracellular Matrices ◽  
Common Denominator ◽  
Cell Matrix ◽  
Skeletal Myoblasts ◽  
A Cell ◽  
Cell Matrix Adhesion

Neff et al. (1982, J. Cell Biol., 95:654-666) have described a monoclonal antibody, CSAT, directed against a cell surface antigen that participates in the adhesion of skeletal muscle to extracellular matrices. We used the same antibody to compare and parse the determinants of adhesion and morphology on myogenic and fibrogenic cells. We report here that the antigen is present on skeletal and cardiac muscle and on tendon, skeletal, dermal, and cardiac fibroblasts; however, its contribution to their morphology and adhesion is different. The antibody produces large alterations in the morphology and adhesion of skeletal myoblasts and tendon fibroblasts; in contrast, its effects on the cardiac fibroblasts are not readily detected. The effects of CSAT on the other cell types, i.e., dermal and skeletal fibroblasts, cardiac muscle, 5-bromodeoxyuridine-treated skeletal muscle, lie between these extremes. The effects of CSAT on the skeletal myoblasts depends on the calcium concentration in the growth medium and on the culture age. We interpret these differential responses to CSAT as revealing differences in the adhesion of the various cells to extracellular matrices. This interpretation is supported by parallel studies using quantitative assays of cell-matrix adhesion. The likely origin of these adhesive differences is the progressive display of different kinds of adhesion-related molecules and their organizational complexes on increasingly adhesive cells. The antigen to which CSAT is directed is present on all of the above cells and thus appears to be a lowest common denominator of their adhesion to extracellular matrices.

scholarly journals Occurrence and immunolocalization of plectin in tissues.

1983 ◽  
Vol 97 (3) ◽  
pp. 887-901 ◽  
Author(s):  
G Wiche ◽  
R Krepler ◽  
U Artlieb ◽  
R Pytela ◽  
H Denk
Keyword(s):  
Skeletal Muscle ◽  
Smooth Muscle ◽  
Urinary Bladder ◽  
Cardiac Muscle ◽  
Muscle Cells ◽  
Cell Types ◽  
Cell Junctions ◽  
Cell Periphery ◽  
Attachment Sites ◽  
Cytoplasmic Filaments

Various tissues from rat were examined for the occurrence and cellular localization of plectin, a 300,000-dalton polypeptide component present in intermediate filament-enriched cytoskeletons prepared from cultured cells by treatment with nonionic detergent and high salt solution. The extraction of liver, heart, skeletal muscle, tongue, and urinary bladder with 1% Triton/0.6 M KCl yielded insoluble cell residues that contained polypeptides of Mr 300,000 in variable amounts. These high Mr polypeptide species and a few bands of slightly lower Mr (most likely proteolytic breakdown products) were shown to react with antibodies to rat glioma C6 cell plectin using immunoautoradiography and/or immunoprecipitation. By indirect immunofluorescence microscopy using frozen sections (4 micron) of stomach, kidney, small intestine, liver, uterus, urinary bladder, and heart, antigens reacting with antibodies to plectin were found in fibroblast, endothelial, smooth, skeletal, and cardiac muscle, nerve, and epithelial cells of various types. Depending on the cell type, staining was observed either throughout the cytoplasm, or primarily at the periphery of cells, or in both locations. In hepatocytes, besides granular staining at the cell periphery, conspicuous staining of junctions sealing bile canaliculi was seen. In cardiac muscle strong staining was seen at intercalated disks and, as in skeletal muscle, at Z-lines. In cross sections through smooth muscle, most strikingly of urinary bladder, antibodies to plectin specifically decorated regularly spaced, spot-like structures at the cell periphery. By immunoelectron microscopy using the peroxidase technique, antiplectin-reactive material was found along cell junctions of hepatocytes and was particularly enriched at desmosomal plaques and structures associated with their cytoplasmic surfaces. A specific immunoreaction with desmosomes was also evident in sections through tongue. In cardiac muscle, besides Z-lines, intercalated disks were reactive along almost their entire surface, suggesting that plectin was associated with the fascia adherens, desmosomes, and probably gap junctions. In smooth muscle cells, regularly spaced lateral densities probably representing myofilament attachment sites were immunoreactive with plectin antibodies. The results show that plectin is of widespread occurrence with regard to tissues and cell types. Furthermore, immunolocalization by light and electron microscopy at junctional sites of various cell types and at attachment sites of cytoplasmic filaments in epithelial and muscle cells suggests that plectin possibly plays a universal role in the formation of cell junctions and the anchorage of cytoplasmic filaments.

scholarly journals Perturbing integrin function inhibits microtubule growth from centrosomes, spindle assembly, and cytokinesis

2006 ◽  
Vol 174 (4) ◽  
pp. 491-497 ◽  
Author(s):  
Carlos G. Reverte ◽  
Angela Benware ◽  
Christopher W. Jones ◽  
Susan E. LaFlamme
Keyword(s):  
Mitotic Spindle ◽  
Cell Types ◽  
Cellular Systems ◽  
Integrin Β1 ◽  
Dynamic Changes ◽  
Matrix Adhesion ◽  
Cell Matrix ◽  
Microtubule Growth ◽  
Cell Matrix Adhesion

In many mammalian cell types, integrin-mediated cell-matrix adhesion is required for the G1–S transition of the cell cycle. As cells approach mitosis, a dramatic remodeling of their cytoskeleton accompanies dynamic changes in matrix adhesion, suggesting a mechanistic link. However, the role of integrins in cell division remains mostly unexplored. Using two cellular systems, we demonstrate that a point mutation in the β1 cytoplasmic domain (β1 tail) known to decrease integrin activity supports entry into mitosis but inhibits the assembly of a radial microtubule array focused at the centrosome during interphase, the formation of a bipolar spindle at mitosis and cytokinesis. These events are restored by externally activating the mutant integrin with specific antibodies. This is the first demonstration that the integrin β1 tail can regulate centrosome function, the assembly of the mitotic spindle, and cytokinesis.

scholarly journals Mouse placental scaffolds: a three-dimensional environment model for recellularization

2019 ◽  
Vol 10 ◽  
pp. 204173141986796 ◽  
Author(s):  
Rodrigo SN Barreto ◽  
Patricia Romagnolli ◽  
Paula Fratini ◽  
Andrea Maria Mess ◽  
Maria Angelica Miglino
Keyword(s):  
Three Dimensional ◽  
Cell Nuclei ◽  
Embryonic Fibroblasts ◽  
Cell Matrix ◽  
Environment Model ◽  
Collagen Iii ◽  
Culture Substrate ◽  
A Cell ◽  
The Rich ◽  
Cell Matrix Adhesion

The rich extracellular matrix (ECM) and availability make placenta eligible as alternative biomaterial source. Herein we produced placental mouse scaffolds by decellularization, and structure, composition, and cytocompatibility were evaluated to be considered as a biomaterial. We obtained a cell-free scaffold containing 9.42 ± 5.2 ng dsDNA per mg of ECM, presenting well-preserved structure and composition. Proteoglycans were widespread throughout ECM without cell nuclei and cell remnants. Collagen I, weak in native placenta, clearly appears in the scaffold after recellularization, opposite distribution was observed for collagen III. Fibronectin was well-observed in placental scaffolds whereas laminin and collagen IV were strong expressed. Placental scaffolds recellularization potential was confirmed after mouse embryonic fibroblasts 3D dynamic culture, resulting in massive scaffold repopulation with cell–cell interactions, cell-matrix adhesion, and maintenance of natural morphology. Our small size scaffolds provide a useful tool for tissue engineering to produce grafts and organ fragments, as well as for cellular biology purposes for tridimensional culture substrate.

Embryonic Stem Cells Differentiated in Vitro as a Novel Source of Cells for Transplantation

1996 ◽  
Vol 5 (2) ◽  
pp. 131-143 ◽  
Author(s):  
Jonathan Dinsmore ◽  
Judson Ratliff ◽  
Terry Deacon ◽  
Peyman Pakzaba ◽  
Douglas Jacoby ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Es Cells ◽  
Embryonic Stem ◽  
Muscle Cells ◽  
Cell Types ◽  
Cell Type ◽  
Skeletal Myoblasts ◽  
Muscle Cell Type

The controlled differentiation of mouse embryonic stem (ES) cells into near homogeneous populations of both neurons and skeletal muscle cells that can survive and function in vivo after transplantation is reported. We show that treatment of pluripotent ES cells with retinoic acid (RA) and dimethylsulfoxide (DMSO) induce differentiation of these cells into highly enriched populations of γ-aminobutyric acid (GABA) expressing neurons and skeletal myoblasts, respectively. For neuronal differentiation, RA alone is sufficient to induce ES cells to differentiate into neuronal cells that show properties of postmitotic neurons both in vitro and in vivo. In vivo function of RA-induced neuronal cells was demonstrated by transplantation into the quinolinic acid lesioned striatum of rats (a rat model for Huntington's disease), where cells integrated and survived for up to 6 wk. The response of embryonic stem cells to DMSO to form muscle was less dramatic than that observed for RA. DMSO-induced ES cells formed mixed populations of muscle cells composed of cardiac, smooth, and skeletal muscle instead of homogeneous populations of a single muscle cell type. To determine whether the response of ES cells to DMSO induction could be further controlled, ES cells were stably transfected with a gene coding for the muscle-specific regulatory factor, MyoD. When induced with DMSO, ES cells constitutively expressing high levels of MyoD differentiated exclusively into skeletal myoblasts (no cardiac or smooth muscle cells) that fused to form myotubes capable of spontaneous contraction. Thus, the specific muscle cell type formed was controlled by the expression of MyoD. These results provided evidence that the specific cell type formed (whether it be muscle, neuronal, or other cell types) can be controlled in vitro. Further, these results demonstrated that ES cells can provide a source of multiple differentiated cell types that can be used for transplantation.

scholarly journals Global phosphotyrosinylated protein profile of cell-matrix adhesion complexes of trabecular meshwork cells

2020 ◽  
Vol 319 (2) ◽  
pp. C288-C299
Author(s):  
Rupalatha Maddala ◽  
Ponugoti Vasantha Rao
Keyword(s):  
Trabecular Meshwork ◽  
Protein Profile ◽  
Cell Types ◽  
Regulatory Mechanisms ◽  
Trabecular Meshwork Cells ◽  
Phosphorylated Proteins ◽  
Cell Matrix ◽  
Intracellular Activities ◽  
Adhesive Interactions ◽  
Cell Matrix Adhesion

Dysregulation of the mechanical properties and cell adhesive interactions of trabecular meshwork (TM) are known to impair aqueous humor drainage and elevate intraocular pressure in glaucoma patients. The identity of regulatory mechanisms underlying TM mechanotransduction, however, remains elusive. Here we analyzed the phosphotyrosine proteome of human TM cell-extracellular matrix (ECM) adhesion complexes, which play a key role in sensing and transducing extracellular chemical and mechanical cues into intracellular activities, using a two-level affinity pull-down (phosphotyrosine antibody and titanium dioxide beads) method and mass spectrometry. This analysis identified ~1,000 tyrosine-phosphorylated proteins of TM cell-ECM adhesion complexes. Many consensus adhesome proteins were found to be tyrosine phosphorylated. Interestingly, several of the phosphotyrosinylated proteins found in TM cell-ECM adhesion complexes are known to be required for podocyte glomerular filtration, indicating the existence of molecular parallels that are likely relevant to the shared fluid barrier and filtration functions of the two mechanosensitive cell types.

scholarly journals Cilia, Centrosomes and Skeletal Muscle

2021 ◽  
Vol 22 (17) ◽  
pp. 9605
Author(s):  
Dominic C. H. Ng ◽  
Uda Y. Ho ◽  
Miranda D. Grounds
Keyword(s):  
Cell Cycle ◽  
Skeletal Muscle ◽  
Cell Fate ◽  
Developmental Disorders ◽  
Cell Cycle Progression ◽  
Primary Cilia ◽  
Striated Muscle ◽  
Cell Types ◽  
Extracellular Milieu ◽  
A Cell

Primary cilia are non-motile, cell cycle-associated organelles that can be found on most vertebrate cell types. Comprised of microtubule bundles organised into an axoneme and anchored by a mature centriole or basal body, primary cilia are dynamic signalling platforms that are intimately involved in cellular responses to their extracellular milieu. Defects in ciliogenesis or dysfunction in cilia signalling underlie a host of developmental disorders collectively referred to as ciliopathies, reinforcing important roles for cilia in human health. Whilst primary cilia have long been recognised to be present in striated muscle, their role in muscle is not well understood. However, recent studies indicate important contributions, particularly in skeletal muscle, that have to date remained underappreciated. Here, we explore recent revelations that the sensory and signalling functions of cilia on muscle progenitors regulate cell cycle progression, trigger differentiation and maintain a commitment to myogenesis. Cilia disassembly is initiated during myoblast fusion. However, the remnants of primary cilia persist in multi-nucleated myotubes, and we discuss their potential role in late-stage differentiation and myofiber formation. Reciprocal interactions between cilia and the extracellular matrix (ECM) microenvironment described for other tissues may also inform on parallel interactions in skeletal muscle. We also discuss emerging evidence that cilia on fibroblasts/fibro–adipogenic progenitors and myofibroblasts may influence cell fate in both a cell autonomous and non-autonomous manner with critical consequences for skeletal muscle ageing and repair in response to injury and disease. This review addresses the enigmatic but emerging role of primary cilia in satellite cells in myoblasts and myofibers during myogenesis, as well as the wider tissue microenvironment required for skeletal muscle formation and homeostasis.

Mini Review: Recent Advances in the Cell-Based Therapies for Cardiac Regeneration

2020 ◽  
Vol 15 (8) ◽  
pp. 649-660 ◽  
Author(s):  
Lan Luo ◽  
Tao-Sheng Li
Keyword(s):  
Cardiac Fibroblasts ◽  
Cardiac Regeneration ◽  
Cell Types ◽  
Cardiac Stem Cells ◽  
Direct Reprogramming ◽  
Promising Candidate ◽  
Surgical Interventions ◽  
Skeletal Myoblasts ◽  
Novel Approach ◽  
Induced Pluripotent

Cardiovascular diseases (CVDs) are the leading cause of morbidity and mortality globally, and traditional pharmaceutical and surgical interventions delay the progression of CVDs. Recently, stem cell therapy has emerged as a promising candidate for treating and preventing heart failure. Increasing efforts have been devoted towards the exploration and identification of potential cell types to repair the injured heart, such as skeletal myoblasts, embryonic, induced pluripotent, bone marrowderived, mesenchymal, and resident cardiac stem cells. In addition, direct reprogramming of cardiac fibroblasts into cardiomyocytes represents a novel approach to cardiac regeneration. Herein, we summarize the recent progress in the use of various cell types for cardiac regeneration and discuss major challenges and future perspectives of cell-based therapies for CVDs.

Anisotropic Strain Effects on Vascular Smooth Muscle Cell Physiology

2007 ◽  
Author(s):  
Vadim Tsvankin ◽  
Dmitry Belchenko ◽  
Devon Scott ◽  
Wei Tan
Keyword(s):  
Critical Role ◽  
Stem Cell Differentiation ◽  
Cell Physiology ◽  
Cell Matrix ◽  
Pathological Conditions ◽  
Wide Range ◽  
A Cell ◽  
Biological Development ◽  
Sense And Respond ◽  
Cell Matrix Adhesion

Biological development is a complex and highly-regulated process, a significant part of which is controlled by mechanostimulus, or the strain imparted on a cell by its environment. Mechanostimulus is important for stem cell differentiation, from cytoskeletal assembly to cell-cell and cell-matrix adhesion [1]. The mechanics of cells and tissues play a critical role in organisms, under both physiological and pathological conditions; abnormal mechanotransduction — the mechanism by which cells sense and respond to strain — has been implicated in a wide range of clinical pathologies [2,3].

scholarly journals Electromechanical Coupling between Skeletal and Cardiac Muscle

2000 ◽  
Vol 149 (3) ◽  
pp. 731-740 ◽  
Author(s):  
Hans Reinecke ◽  
Glen H. MacDonald ◽  
Stephen D. Hauschka ◽  
Charles E. Murry
Keyword(s):  
Skeletal Muscle ◽  
Gap Junctions ◽  
Cardiac Muscle ◽  
Gap Junction ◽  
Electromechanical Coupling ◽  
Skeletal Myoblasts ◽  
Adult Cardiomyocytes ◽  
Synchronous Contraction ◽  
Skeletal Myotubes

Skeletal myoblasts form grafts of mature muscle in injured hearts, and these grafts contract when exogenously stimulated. It is not known, however, whether cardiac muscle can form electromechanical junctions with skeletal muscle and induce its synchronous contraction. Here, we report that undifferentiated rat skeletal myoblasts expressed N-cadherin and connexin43, major adhesion and gap junction proteins of the intercalated disk, yet both proteins were markedly downregulated after differentiation into myo-tubes. Similarly, differentiated skeletal muscle grafts in injured hearts had no detectable N-cadherin or connexin43; hence, electromechanical coupling did not occur after in vivo grafting. In contrast, when neonatal or adult cardiomyocytes were cocultured with skeletal muscle, ∼10% of the skeletal myotubes contracted in synchrony with adjacent cardiomyocytes. Isoproterenol increased myotube contraction rates by 25% in coculture without affecting myotubes in monoculture, indicating the cardiomyocytes were the pacemakers. The gap junction inhibitor heptanol aborted myotube contractions but left spontaneous contractions of individual cardiomyocytes intact, suggesting myotubes were activated via gap junctions. Confocal microscopy revealed the expression of cadherin and connexin43 at junctions between myotubes and neonatal or adult cardiomyocytes in vitro. After microinjection, myotubes transferred dye to neonatal cardiomyocytes via gap junctions. Calcium imaging revealed synchronous calcium transients in cardiomyocytes and myotubes. Thus, cardiomyocytes can form electromechanical junctions with some skeletal myotubes in coculture and induce their synchronous contraction via gap junctions. Although the mechanism remains to be determined, if similar junctions could be induced in vivo, they might be sufficient to make skeletal muscle grafts beat synchronously with host myocardium.

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