THE INFLUENCE OF CALCIUM IONOPHORE AND ACTIVATORS OF PROTEIN KINASE-C ON STEROID-PRODUCTION BY PREOVULATORY OVARIAN FOLLICLES OF THE GOLDFISH

Steroidogenesis in teleost fish, as in other vertebrate groups, is mediated by the activation of adenylate cyclase. For the present studies, calcium ionophore A23187 and either phorbol 12-myristate 13-acetate (PMA) or 1-oleoyl-2-acetylglycerol (OAG) were used to investigate the possible roles that changes in intracellular calcium content and protein kinase C activation play in steroid production by goldfish preovulatory ovarian follicles incubated in vitro. While ineffective alone, PMA (1.6 - 400 nM) and OAG (25 - 100 ♂g/ml) exhibited classical synergism with A23187 (1.0 - 10 μM), leading to increased testosterone production. The magnitude of these responses was at least ten fold lower than that obtained with human chorionic gonadotropin (hCG), forskolin, or dibutryl cyclic adenosine 3', 5'-monophosphate. Testosterone production stimulated by hCG and forskolin was blocked by addition of PMA but not OAG. Unlike PMA, the inactive phorbol ester 4α-phorbol 12, 13-dideconate did not influence basal or stimulated testosterone production. A23187 had a biphasic effect on stimulated testosterone production: a dosage of 0.25 or 1.0 μM potentiated the action of submaximally effective dosages of hCG or forskolin on testosterone production; a higher dosage of 4 μM inhibited stimulated testosterone production by up to 50%. In conclusion, these studies suggest that, in addition to the adenylate cyclase second messenger system, changes in intracellular calcium and activation of protein kinase C may modulate steroidogenesis in goldfish ovarian follicles.