Homocysteine and Cardiac Neural Crest Cell Cytoskeletal Proteins in the Chick Embryo

Abstract

Elevated serum homocysteine (Hcys) is correlated with cardiovascular disease and with embryonic malformations related to neural crest cells (NCCs). We predicted Hcys may alter the balance of actin networks, stress fibers and focal adhesions, altering migration. We cultured neural tube explants in control and Hcys-treated medium and visualized actin, α-actinin, vinculin, filamin, and LIM3 protein in NCCs migrating outward. In Hcys, phalloidin-stained actin in stress fibers was brighter, and vinculin was more abundant in focal adhesions and lamellipodia. α-actinin and LIM3 were also enhanced around nuclei and in focal adhesions, and α-actinin also in filopodia. Filamin was unchanged. Hcys caused more spreading and migration of NCCs, but not more cell-cell adhesions. The findings support our hypothesis that Hcys adjusts NCCs for greater adhesion and migration. Its effect on LIM3 suggests it may modulate signaling that adjusts the cytoskeleton for enhanced migration, leading to mistimed and defective development of target tissues.

Effect of tricalcium silicate cements in gene expression of COL1A1, MAPK’s, and NF-kB, and cell adhesion in primary teeth’ pulp fibroblasts

“…Cell adhesion was determined through focal adhesions (FA), which are dynamically assembled and disassembled by the cells 27 . Vinculin is one of the essential and best characterized FA proteins; it is a membrane-cytoskeleton protein located on FA, involved in the anchoring of integrin molecules to the actin cytoskeleton 27,49 .Vinculin is a major regulator of cell adhesion 48 ; it contributes to the stability of focal adhesion under high forces by regulating the contractile stress generation. Intregin β1 constitutes a group of transmembrane proteins belonging to the family of adhesion molecules 50 .…”

Anastrozole Promotes Implantation by Altering the Expression of Paxillin and FAK in Rat Luminal Uterine Epithelium

“…Table II. A description of the scoring system for the focal adhesion protein expression and localization in the confocal images among treatment groups adopted from previous studies (Englund et al, 2001;Mwakikunga et al, 2011). Table III.…”

“…Other studies have provided evidence to suggest that the structural links between actin filaments and integrins are regulated in at least some cell types, and it would not be unreasonable to hypothesize that agents known to alter cell behavior may do so by affecting the expression or function of actin binding proteins such as paxillin, and signaling molecules such as FAK which are recruited to focal adhesions (Kaneko et al, 2009;Mwakikunga et al, 2011). FAs are dynamically assembled and disassembled by cells including uterine luminal epithelial cells (Hosie et al, 2008;Kaneko et al, 2008Kaneko et al, , 2011 and these mechanisms are likely governed by the modulation of cross-talk between their constituent proteins, which influence their gene expression and morphology.…”

Focal Adhesion Proteins, Vinculin and Integrin β5, During Early Pregnancy in Rat Uterine Epithelial Cells: Anastrozole Favors their Normal Distribution

“…Using standard procedures, samples for immunofluorescence and confocal microscopy were fixed in 10 % buffered formalin for 48 hours (Karaer et al), then placed in an automatic tissue processor (Shandon Citadel 1000, Labotek, South Africa) in which samples were incubated in a series of 70 %, 95 %, 95 %, 95 %, 100 %, 100 %, 100 % ethanol, then in chloroform and finally in paraffin Table II. Description of the scoring system for the focal adhesion protein expression and localization in the confocal images among treatment groups adopted from previous studies (Englund et al, 2001;Mwakikunga et al, 2011).…”

“…Moreover, the implantation of blastocysts into the endometrial stroma cells is regulated by the Rho GTPases RhoA, Rac 1 and Cdc42 (Grewal et al, 2010). the structural links between actin filaments and integrins are regulated in at least some cell types, and it would not be unreasonable to hypothesize that agents known to alter cell behavior may do so by affecting the expression or function of actin binding proteins such as vinculin, talin, paxillin and integrins that are recruited to focal adhesions (Mwakikunga et al, 2011). Focal adhesions are dynamically assembled and disassembled by cells, including uterine luminal epithelial cells (Hosie et al, 2008;Kaneko et al, 2008Kaneko et al, , 2011, and these mechanisms are likely governed by the modulation of cross-talk between their constituent proteins, which influences their gene expression and morphology.…”