Expression and distribution of cytoskeletal IFAP-300kD as an index of lens cell differentiation.

By their implication in the organization of the intermediate filament (IF) cytoskeleton, IF-associated proteins (IFAPs) can delineate subsets of the same IF type within a cell; moreover, they are proving useful as markers of the differentiation states of certain cells. For these reasons the expression of the vimentin-associated IFAP-300kD was investigated in the constantly differentiating cell lineage of the adult bovine lens. Immunofluorescence microscopy and immunoblot analysis were employed using a monoclonal anti-IFAP-300kD and a rabbit anti-lens vimentin. Cultures of adult lens epithelial cells were immunopositive for the IFAP. By double-label studies the IFAP-300kD pattern co-localized with that of the vimentin-type IF; moreover, the IFAP pattern co-distributed with that of both colchicine-sensitive and -insensitive IF systems. IFAP-300kD was also present in a co-distributing pattern with vimentin IF in fresh lens epithelial cells on whole mounts. There was a differential expression of the IFAP in the lens fiber cells in that those of the cortex exhibited the IFAP and vimentin IF, while both proteins were absent from the nuclear fiber cells. Furthermore, there was a differential distribution of the IFAP within the cortical fiber cells in that the IFAP localized only with a paramembranal subset of IF. Immunoblot analysis supported the presence of IFAP-300kD in the lens cytoskeletal fraction. IFAP-300kD thus identified a subset of vimentin IF whose location may have functional significance for the cortical fiber cell. The changes in the IFAP's expression and distribution pattern throughout lens cell differentiation in the adult organ suggest the usefulness of IFAP-300kD as a potential marker in studying lens cell differentiation in vitro.