Concurrent LH and forskolin action on adenylate cyclase activation and progesterone synthesis in corpora lutea from pregnant ewes.

The present communication documents LH- and forskolin-induced activation of adenylate cyclase (AC) system and progesterone synthesis in corpora lutea from pregnant ewes. The activation of AC in plasma membranes by LH or forskolin was amplified by Gpp(NH)p. These results suggest that regulatory nucleotide component (Ns) of the AC complex is required for forskolin. Simultaneous addition of maximal concentrations of forskolin (10(-4) M), Gpp(NH)p (10(-4) M) and LH (10(-7) M) led to greater than additive (i.e. synergistic) responses: the experimental value was 4.71 +/- 0.19 nmoles cAMP/mg of membrane protein, whereas the theoretical additive effect was 3.17 +/- 0.10 nmoles/mg of membrane protein (p less than 0.001). These data reveal that more Ns or C component is being activated in these cells when combined treatments with these agents are applied. In intact cells maximum stimulatory concentrations of forskolin or LH caused similar increase in progesterone production with similar time courses. In striking contrast, the exposure of the luteal cells to LH and forskolin simultaneously led to a decrease in progesterone synthesis as early as 1h30 (40%, p less than 0.001). Thus, the synergism observed between LH and forskolin on the stimulation of plasma membranes AC activity did not occur in steroidogenesis. The AC responses in crude plasma membranes form these cells to different stimulants were enhanced (i.e. 15%, p less than 0.2 for Gpp(NH)p, 33%, p less than 0.01 for LH plus Gpp(NH)p and 52%, p less than 0.01 for forskolin). These findings suggest that an early desensitization of the AC system cannot explain the impaired steroidogenic response observed.