Activation of soluble guanylyl cyclase inhibits estradiol production and cyclic AMP accumulation from cultured rat granulosa cells.

OBJECTIVE: To demonstrate the expression of soluble guanylyl cyclase (sGC) alpha and beta subunits in rat granulosa cells and determine the effects sGC activation on levels of cyclic GMP (cGMP), E2, and cAMP. DESIGN: Basic research study. SETTING: University research laboratory. ANIMAL(S): Estrogen-treated immature Sprague-Dawley female rats from which primary cell culture of granulosa cells was obtained. INTERVENTION(S): Functionally immature rat granulosa cells were incubated for 48 hours with media alone, FSH, or FSH plus YC-1, a specific activator of sGC. MAIN OUTCOME MEASURE(S): Expression of sGC alpha and beta subunits was determined by immunoblot analysis. Media concentrations of E2, cAMP, and cGMP were measured by radioimmunoassays. RESULT(S): Immunoblot analysis of granulosa cells revealed the expression of sGC alpha and beta subunits. While cGMP accumulation was low in cells incubated with media alone or with FSH, cotreatment with FSH plus YC-1 increased cGMP levels approximately five-fold. Incubation of cells with FSH stimulated E2 production in a dose-dependent manner. However, cotreatment of cells with FSH plus YC-1 significantly decreased E2 concentrations at all doses of FSH tested. Similarly, while FSH increased cAMP accumulation from granulosa cells, cotreatment with YC-1 markedly inhibited FSH-stimulated cAMP levels. CONCLUSION(S): These findings demonstrate the expression of sGC subunits in rat granulosa cells and indicate that activation of sGC increases cGMP levels, which are associated with inhibition of FSH-stimulated E2 production and cAMP accumulation.

Roles of cyclic GMP in modulating ovarian functions.

The production of a viable oocyte is dependent upon the critical influences of gonadotrophins on follicular development, granulosa cell maturation, ovulation, and luteinization. While the effects of LH and FSH are due in large part to cyclic AMP-dependent signalling mechanisms, it is clear that a number of other factors modulate the actions of gonadotrophins on the ovary via activation of alternative signalling pathways. In this regard, recent studies indicate that the second messenger guanosine 3',5'-cyclic monophosphate (cGMP) mediates a wide range of influences on the ovary. Nitric oxide (NO) is a major regulator of cGMP production via its action on soluble guanylyl cyclase, while natriuretic peptides activate receptors with intrinsic guanylyl cyclase activities. In addition, other factors known to influence ovarian functions are now recognized to act via NO/cGMP pathways. This report will review these previous findings and present new data demonstrating the inhibitory influence of cGMP on cAMP-stimulated LH receptor expression in cultured granulosa cells.