Gap junctional intercellular communication (GJIC) and the expression of gap junction proteins (connexins) may be involved in growth regulation and neoplastic transformation. The mechanisms of connexin gene regulation in normal and neoplastic tissues are poorly understood. In this study, the glucocorticoids, dexamethasone and hydrocortisone, enhanced fluorescent dye-coupling in primary cultured rat hepatocytes and MH1C1 rat hepatoma cells. Other types of steroids (beta-estradiol, testosterone, aldosterone and progesterone) had no effect. Northern blot, Western blot, nuclear run-on and immunohistochemical analyses showed that glucocorticoids enhanced the expression of connexin32 in these cells in a dose- and time-dependent fashion. Connexin26 expression was also enhanced slightly by dexamethasone in hepatocytes, but not MH1C1 cells. Connexin43 expression in these cells was not affected by steroids. In WB-F344 rat liver epithelial cells, which were highly coupled and expressed high levels of connexin43 and no detectable connexin32 or connexin26, dexamethasone had no effect on coupling or connexin expression. These results indicate that dye-coupling and the expression of connexin32 and connexin26, but not connexin43, were upregulated by glucocorticoids in a cell-specific manner. These effects on GJIC and connexin expression may be involved in the induction of hepatic differentiation and inhibition of growth.