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Reversal of follicular fluid inhibition of granulosa cell progesterone secretion by manipulation of intracellular cyclic AMP.

Fluid from small (1-2 mm) porcine follicles inhibits progesterone secretion, LH stimulation of cyclic AMP (cAMP) accumulation, adenylyl cycla.se activity and morphological luteinization of granulosa cells obtained from medium (3-5 mm) or large (6-12 mm) porcine follicles. The goal of the present experiments was to determine whether restoration of cAMP levels would reverse the inhibitory action of follicular fluid on progesterone secretion. Following 2-day incubations with either the inhibitory fluid from small follicles (SFFI) or noninhibitory fluid from large follicles (LFFI), the presence of phosphodiesterase inhibitors theophylline or 3-isobutylmethyl xanthine (MIX) restored LH stimulation of cAMP accumulation by SFFI-treated cells to 50% of that of the controls (i.e., LFFI-treated cells). LH and MIX together, but neither alone, fully restored progesterone secretion by SFFI-treated cells to amounts secreted by the controls. Analogs of cAMP, 8 bromo cyclic AMP (8 BrcAMP) and N2O6dibutyryl cyclic AMP (DBC), in combination with MIX, but not alone, also fully restored progesterone secretion by the inhibited cells. These data suggest that the follicular fluid does not alter the steroidogenic machinery of the granulosa cells directly, but influences it via inhibition of cAMP accumulation.