Rho is present and phosphorylated in avian corneal epithelia cultured in the presence of ECM molecules.
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In Avian corneal epithelia, previous studies have shown that extracellular matrix (ECM) molecules, fibronectin (FN) and collagen (COL), reorganize the actin cortical mat (ACM). This reorganization is accompanied by changes in tyrosine phosphorylation of the actin associated proteins, focal adhesion kinase (FAK) and paxillin (PAX). Phosphorylation inhibitors, herbimycin A and genistein, partially blocked FAK and PAX phosphorylation. In cell culture systems, Rho, a member of the Ras superfamily, is involved in regulating cell adhesion and actin cytoskeletal reorganization. The purpose of this study was to analyze Rho expression and phosphorylation in intact Avian corneal epithelia upon stimulation with FN or COL. 8 day chick corneal epithelia were isolated without basal lamina (-BL) and cultured for 0,5,10, 15, 30, 60 and 120 min. in control media, 50 g/ml FN or 100 g/ml COL. Cell extracts were prepared for western blotting. Preliminary studies show that Rho protein is present in epithelia stimulated with either FN or COL. Phosphorylation studies show that there is a high molecular weight shift in Rho which increases over the time sequence examined. This increase in phosphorylated Rho is more prominent in epithelia stimulated by FN. This suggests that there is modulation of Rho expression and phosphorylation in our system. We conclude that Rho phosphorylation may be one of the downstream signal transduction pathways induced in the Avian corneal epithelia stimulated with ECM molecules.
