Cell-to-cell communication in a differentiating ovine lens culture system

Abstract

Purpose: This study was performed to determine whether the junctions between both the epithelial and the differentiating fiber-like cells of ovine lens cultures, like gap junctions in other tissues, exhibit cell-to-cell communication that is inhibited by n-octanol, and to determine whether lens connexins and the fiber cell membrane proteins MP20 and MP26 are expressed by these ovine lens cell cultures.

Methods: Cells were injected with Lucifer yellow CH to measure cell-to-cell communication. Antibodies to connexin-related lens membrane protein MP70, connexin 43 (Cx43), and connexin 46 (Cx46) and to membrane proteins MP20 and MP26 were used to immunofluorescently label lens cultures and probe Western blots of membranes isolated from lens cultures.

Results: Both epithelial cells and differentiating clear cells exhibited cell-to-cell transfer of Lucifer yellow that was inhibited by n-octanol. Although a Cx43 antibody immunofluorescently labeled small plaques between the epithelial cells, an MP70 antibody labeled large plaques as well as small punctate areas of the differentiating fiber-like cells. It is interesting that Cx43 and MP70 were frequently present in the same plaques at cell interfaces between epithelial cells as well as some of the larger plaques on the differentiating fiber-like cells. Cx46 and MP70 antibodies labeled the same plaques in membranes of differentiating fiber-like cells and late-stage epithelial cells. The electrophoretic mobility of all three connexin proteins was modified after treatment with alkaline phosphatase. Immunohistochemical staining of these differentiating regions and Western immunoblotting of purified membranes derived from differentiated cultures also showed the presence of MP20 and MP26.

Conclusions: The different cell types in the ovine lens culture exhibit gap junction-mediated cell-to-cell communication that is likely effected by one or more of the connexin proteins.