Concomitant alterations of desmosomes, adhesiveness, and diffusion through gap junction channels in a rat ovarian transformation model system

Abstract

Gap junctional intercellular communication (GJIC), desmosomes, and cell movement were evaluated in a rat ovarian epithelial cell model system which consisted of an immortalized clonal cell line (SIGC), a pSV3neotransfected clonal derivative (SV-SIGC), and a nude mouse SV-SIGC-tumor-derived cell line (T-SV-SIGC). Complementary ultrastructural, indirect immunofluorescence, and Western blot data identified a relatively small loss of desmosomes and associated cytokeratins in SV-SIGC compared to SIGC but a near total loss in T-SV-SIGC. SIGC and SV-SIGC migrated outward from monolayer-coated microcarrier beads as epithelial sheets, whereas in T-SV-SIGC there was dissociation and migration of individual fibroblastoid cells. GJIC was assessed by fluorescence recovery after photobleaching (gap FRAP) and equations based on Fick's first law of diffusion were derived to quantitatively compare GJIC of systems with different recovery equilibria after photobleaching. Taken together the data suggested that GJIC in SIGC was quantitatively reduced to similar levels by various conditions associated with reduced cell-cell adhesiveness including transformation to T-SV-SIGC, mitosis, and culture in low calcium medium. These results supported linkage between changes in desmosomal adhesiveness, cell movement, and GJIC.