Zyxin Regulates Cell Migration and Differentiation in EMT during Chicken AV Valve Morphogenesis

2013 ◽  
Vol 19 (4) ◽  
pp. 842-854 ◽  
Author(s):  
Na Li ◽  
Richard L. Goodwin ◽  
Jay D. Potts
Keyword(s):  
Cell Migration ◽  
Collagen Gel ◽  
Mesenchymal Cells ◽  
Valve Development ◽  
Focal Adhesion Protein ◽  
Endocardial Cell ◽  
Mesenchymal Transformation ◽  
And Function ◽  
Valve Formation

AbstractDuring heart valve development, epithelial–mesenchymal transformation (EMT) is a key process for valve formation. EMT leads to the generation of mesenchymal cells that will eventually become the interstitial cells (fibroblasts) of the mature valve. During EMT, cell architecture and motility change markedly; significant changes are also observed in various signaling pathways. Here we systematically examined the expression, localization, and function of zyxin, a focal adhesion protein, in EMT during atrioventricular (AV) valve morphogenesis. Expression and localization studies showed that zyxin was expressed in the AV canal region during crucial stages of valve development. An in vitro 3D collagen gel culture system was used to determine zyxin function either after siRNA gene knockdown or after overexpression. Our studies revealed that zyxin overexpression inhibited endocardial cell migration and cell differentiation and also led to a decrease in the number of migrating mesenchymal cells. Moreover, correlative cytoskeletal changes were apparent in response to both overexpression and knockdown treatments. Thus, zyxin appears to play a role as a regulator of cell migration and differentiation during EMT in chicken AV valve formation.

scholarly journals PDGFRα: Expression and Function during Mitral Valve Morphogenesis

2021 ◽  
Vol 8 (3) ◽  
pp. 28
Author(s):  
Kelsey Moore ◽  
Diana Fulmer ◽  
Lilong Guo ◽  
Natalie Koren ◽  
Janiece Glover ◽  
...  
Keyword(s):  
Mitral Valve ◽  
Primary Cilia ◽  
Growth Factor Receptor ◽  
Akt Phosphorylation ◽  
Valve Development ◽  
Biochemical Assays ◽  
Factor Receptor Alpha ◽  
Mesenchymal Transformation ◽  
And Function

Mitral valve prolapse (MVP) is a common form of valve disease and can lead to serious secondary complications. The recent identification of MVP causal mutations in primary cilia-related genes has prompted the investigation of cilia-mediated mechanisms of disease inception. Here, we investigate the role of platelet-derived growth factor receptor-alpha (PDGFRα), a receptor known to be present on the primary cilium, during valve development using genetically modified mice, biochemical assays, and high-resolution microscopy. While PDGFRα is expressed throughout the ciliated valve interstitium early in development, its expression becomes restricted on the valve endocardium by birth and through adulthood. Conditional ablation of Pdgfra with Nfatc1-enhancer Cre led to significantly enlarged and hypercellular anterior leaflets with disrupted endothelial adhesions, activated ERK1/2, and a dysregulated extracellular matrix. In vitro culture experiments confirmed a role in suppressing ERK1/2 activation while promoting AKT phosphorylation. These data suggest that PDGFRα functions to suppress mesenchymal transformation and disease phenotypes by stabilizing the valve endocardium through an AKT/ERK pathway.

scholarly journals Epithelia suspended in collagen gels can lose polarity and express characteristics of migrating mesenchymal cells.

1982 ◽  
Vol 95 (1) ◽  
pp. 333-339 ◽  
Author(s):  
G Greenburg ◽  
E D Hay
Keyword(s):  
Epithelial Cells ◽  
Cell Polarity ◽  
Three Dimensional ◽  
Collagen Gel ◽  
Mesenchymal Cells ◽  
Apical Surface ◽  
Collagen Gels ◽  
Mesenchymal Transformation

This study of epithelial-mesenchymal transformation and epithelial cell polarity in vitro reveals that environmental conditions can have a profound effect on the epithelial phenotype, cell shape, and polarity as expressed by the presence of apical and basal surfaces. A number of different adult and embryonic epithelia were suspended within native collagen gels. Under these conditions, cells elongate, detach from the explants, and migrate as individual cells within the three-dimensional lattice, a previously unknown property of well-differentiated epithelia. Epithelial cells from adult and embryonic anterior lens were studied in detail. Elongated cells derived from the apical surface develop pseudopodia and filopodia characteristic of migratory cells and acquire a morphology and ultrastructure virtually indistinguishable from that of mesenchymal cells in vivo. It is concluded from these experiments that the three-dimensional collagen gel can promote dissociation, migration, and acquisition of secretory organelles by differentiated epithelial cells, and can abolish the apical-basal cell polarity characteristic of the original epithelium.

scholarly journals SOX7 suppresses endothelial-to-mesenchymal transitions by enhancing VE-cadherin expression during outflow tract development

2021 ◽  
Author(s):  
Xuechao Jiang ◽  
Tingting Li ◽  
Bojian Li ◽  
Wei Wei ◽  
Fen Li ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Migration ◽  
Molecular Mechanisms ◽  
Collagen Gel ◽  
Mesenchymal Cells ◽  
Outflow Tract ◽  
Negative Regulator ◽  
Mobility Shift ◽  
Mesenchymal Transition ◽  
Endocardial Cell

The endothelial-to-mesenchymal transition (EndMT) is a critical process that occurs during the development of the outflow tract (OFT). Malformations of the OFT can lead to the occurrence of conotruncal defects (CTD). SOX7 duplication has been reported in patients with congenital CTD, but its specific role in OFT development remains poorly understood. To decipher this, histological analysis showed that SOX7 was regionally expressed in the endocardial endothelial cells and in the mesenchymal cells of the OFT, where EndMT occurs. Experiments, using invitro collagen gel culture system, revealed that SOX7 was a negative regulator of EndMT that inhibited endocardial cell migration and resulted in decreased number of mesenchymal cells. Forced expression of SOX7 in endothelial cells blocked further migration and improved the expression of the adhesion protein vascular endothelial (VE)-cadherin. Moreover, a VE-cadherin knockdown could partly reverse the SOX7-mediated repression of cell migration. Luciferase and electrophoretic mobility shift assays demonstrated that SOX7 up-regulated VE-cadherin by directly binding to the gene’s promoter in endothelial cells. The coding exons and splicing regions of the SOX7 gene were also scanned in the 536 sporadic CTD patients and in 300 unaffected controls, which revealed four heterozygous SOX7 mutations. Luciferase assays revealed that two SOX7 variants weakened the transactivation of the VE-cadherin promoter. In conclusion, SOX7 inhibited EndMT during OFT development by directly upregulating the endothelial-specific adhesion molecule VE-cadherin. SOX7 mutations can lead to impaired EndMT by regulating VE-cadherin, which may give rise to the molecular mechanisms associated with SOX7 in CTD pathogenesis.

scholarly journals EFFECT OF COLLAGEN I GEL ON APOPTOSIS OF RAT HEPATIC STELLATE CELLS

2013 ◽  
Vol 56 (2) ◽  
pp. 73-79
Author(s):  
Lenka Bittnerová ◽  
Alena Jiroutová ◽  
Emil Rudolf ◽  
Martina Řezáčová ◽  
Jiří Kanta
Keyword(s):  
Hepatic Stellate Cells ◽  
Type I Collagen ◽  
Collagen Gel ◽  
Stellate Cells ◽  
Type I ◽  
Function Type ◽  
Fibrotic Liver ◽  
Normal Rat ◽  
And Function

Activated hepatic stellate cells (HSC) are a major source of fibrous proteins in cirrhotic liver. Inducing or accelerating their apoptosis is a potential way of liver fibrosis treatment. Extracellular matrix (ECM) surrounding cells in tissue affects their differentiation, migration, proliferation and function. Type I collagen is the main ECM component in fibrotic liver. We have examined how this protein modifies apoptosis of normal rat HSC induced by gliotoxin, cycloheximide and cytochalasin D in vitro and spontaneous apoptosis of HSC isolated from CCl4-damaged liver. We have found that type I collagen gel enhances HSC apoptosis regardless of the agent triggering this process.

Role of vitronectin in embryonic rat endocardial cell migration in vitro

1992 ◽  
Vol 268 (1) ◽  
pp. 41-49 ◽  
Author(s):  
Hiroshi Sumida ◽  
Harukazu Nakamura ◽  
Mineo Yasuda
Keyword(s):  
Cell Migration ◽  
Endocardial Cell

scholarly journals Virus-Induced Cell Motility

1998 ◽  
Vol 72 (2) ◽  
pp. 1235-1243 ◽  
Author(s):  
Christopher M. Sanderson ◽  
Michael Way ◽  
Geoffrey L. Smith
Keyword(s):  
Cell Migration ◽  
Cell Motility ◽  
Uv Light ◽  
Cell Metabolism ◽  
Cell Movement ◽  
Time Lapse ◽  
Late Gene Expression ◽  
Infected Cells ◽  
And Function

ABSTRACT Many viruses induce profound changes in cell metabolism and function. Here we show that vaccinia virus induces two distinct forms of cell movement. Virus-induced cell migration was demonstrated by an in vitro wound healing assay in which infected cells migrated independently into the wound area while uninfected cells remained relatively static. Time-lapse microscopy showed that the maximal rate of migration occurred between 9 and 12 h postinfection. Virus-induced cell migration was inhibited by preinactivation of viral particles with trioxsalen and UV light or by the addition of cycloheximide but not by addition of cytosine arabinoside or rifampin. The expression of early viral genes is therefore necessary and sufficient to induce cell migration. Following migration, infected cells developed projections up to 160 μm in length which had growth-cone-like structures and were frequently branched. Time-lapse video microscopy showed that these projections were formed by extension and condensation of lamellipodia from the cell body. Formation of extensions was dependent on late gene expression but not the production of intracellular enveloped (IEV) particles. The requirements for virus-induced cell migration and for the formation of extensions therefore differ from each other and are distinct from the polymerization of actin tails on IEV particles. These data show that poxviruses encode genes which control different aspects of cell motility and thus represent a useful model system to study and dissect cell movement.

Integrin expression and differentiation in transformed human epidermal cells is regulated by fibroblasts

1992 ◽  
Vol 103 (3) ◽  
pp. 755-763 ◽  
Author(s):  
P. Kaur ◽  
W.G. Carter
Keyword(s):  
Human Fibroblasts ◽  
Collagen Gel ◽  
Integrin Expression ◽  
Human Papillomavirus Type 16 ◽  
3T3 Fibroblasts ◽  
Extracellular Matrix Components ◽  
Alternate Mechanism ◽  
Vitro Model ◽  
And Function

Normal human foreskin keratinocytes (HFKs) and transformed HFKs (FEPE1L-8 cells) generated by the introduction of cloned human papillomavirus type 16 sequences were compared for the expression and function of a family of adhesion receptors termed integrins. Initially, cells were examined in conventional monolayer cultures. FEPE1L-8s expressed integrins alpha 1 beta 1, alpha 2 beta 1, alpha 3 beta 1, alpha 5 beta 1, alpha 6 beta 4 and beta 1 at comparable levels to HFKs. Further, these receptors were fully functional in mediating specific interactions with exogenously supplied ligands. However, FEPE1L-8s exhibited decreased synthesis of a number of extracellular matrix components, including laminin, fibronectin and epiligrin, compared to normal HFKs, which may be an alternate mechanism for regulating adhesion. Subsequently, organotypic cultures (OCs), which provide a suitable in vitro model system for the ordered stratification and differentiation of keratinocytes, were used to study the regulation of integrins and various epidermal markers in normal and transformed cells. OCs consisted of keratinocytes plated on a collagen gel containing primary human fibroblasts, grown at an air-medium interface. Unlike normal HFKs, the transformed FEPE1L-8 cells exhibited (a) disorganized stratification and limited differentiation capacity, (b) invasion into the collagen gel, and (c) unregulated expression of alpha 3 beta 1 and alpha 2 beta 1, and under-expression of alpha 6 beta 4 integrins. Ordered stratification and spatial regulation of integrin expression could be induced in the FEPE1L-8s by substituting Swiss 3T3 fibroblasts in the collagen gel. Further data indicate that the human fibroblasts induce the transformed HFKs to invade into the collagen gel.(ABSTRACT TRUNCATED AT 250 WORDS)

Abstract 315: Surgical Pen Dye Inhibits Cell Migration From Saphenous Vein Explants

2014 ◽  
Vol 34 (suppl_1) ◽  
Author(s):  
Shinsuke Kikuchi ◽  
Richard D Kenagy ◽  
Alexander W Clowes
Keyword(s):  
Cell Migration ◽  
Crystal Violet ◽  
Primary Response ◽  
Alternative Methods ◽  
Vein Grafts ◽  
Significant Difference ◽  
Vein Bypass ◽  
Fetal Bovine ◽  
And Function

Markers containing dyes such as crystal violet (CAS 548-62-9) are routinely used on the adventitia of vein bypass grafts to avoid twisting during placement. Since little is known about how these dyes affect vein graft healing and function, we determined the effect of crystal violet on cell migration, a primary response to injury after grafting. Human saphenous veins were split into adventitia and media after removal of endothelium. Cell migration from 2.5 square mm explants in DMEM with 20% fetal bovine serum was measured daily for 8 days as either 1) % migration positive explants (≥ 1 cell/explant), which only measures migration, or 2) the number of cells/explant, which measures a combination of migration and proliferation. Dye was extracted from explants with ethanol and quantified by spectrophotometry. Cell migration by both methods was significantly less from blue, compared to non-blue, adventitial explants (P<.05, mean±SEM, N=11 veins with 15-30 explants/condition; Figures A and B). No medial explants were visibly blue, and there was no significant difference in migration of cells from medial explants of blue and non-blue sections of vein (data not shown). Blue adventitial explants had 65.9±8.0 ng dye/explant compared to 2.1±1.3 for non-blue explants (mean±SEM, N=7-11 veins). Dye applied in vitro to either adventitial or medial explants dose-dependently inhibited migration (IC50=21.5 ng/explant). Conclusion: Crystal violet is toxic and inhibits venous adventitial cell migration; alternative methods should be considered for marking vein grafts.

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