Identification and localization of talin in chick retinal pigment epithelial cells

Abstract

Retinal pigmented epithelial cells are adherent at their basal surface to Bruch's membrane and at their apical surface to the neural retina. We examined the expression and distribution of two proteins that are found in regions of cell-matrix interaction, talin and integrin. Talin is a 235-kDa cytoplasmic protein that has been localized to regions of cell-substrate adhesion. It binds to both integrin, a transmembrane glycoprotein complex, and to vinculin, a cytoskeletal protein. In the present study, we produced a polyclonal antibody to chicken gizzard talin. Using this antibody we showed by western blot analysis that talin is expressed by RPE cells and is found in the triton-soluble fraction. Talin was shown to co-localize with integrin and vinculin in the basal region of chick RPE cells isolated from 18-day-old chick embryos. Neither talin nor integrin was found in the apical processes or in the zonula adherens. Antibodies to vinculin showed staining both in the apical and basal regions of the RPE cells. The localization of integrin, talin and vinculin along the basal membrane suggests that this complex is important in the attachment of the RPE cells to the basement membrane. The distribution of integrin and talin was examined in primary cultures of RPE cells grown on permeable filters. In these cells, a polarized distribution of integrin and talin was not observed. This may suggest that the neural retina may be important for maintaining the differentiated state of the RPE cells.