Porcine granulosa cell desensitization: prolonged FSH-responsive cAMP production in vitro

Granulosa cells from small (1-2 mm) follicles of porcine ovaries produced cAMP in response to follicle-stimulating hormone (FSH); they produced little cAMP in response to human chorionic gonadotropin (hCG) or to cholera toxin. Production, estimated by quantification of total cAMP (medium + cells) by radioimmunoassay, appeared to be episodic in the absence or presence of the phosphodiesterase inhibitor 1-methyl-3-isobutylxanthine (MIX) and continued for at least 20 h. Results of dual incubation experiments with thorough washing of cells between incubations indicated that cells remained responsive to FSH, to hCG, and to cholera toxin after prior exposure to homologous stimulator. Preincubation of cells for 3 h in the absence of stimulator enhanced FSH responsiveness. In experiments with heterologous stimulators in two incubations, first, preincubation with FSH enhanced subsequent hCG and cholera toxin responsiveness; and second, preincubation with hCG or cholera toxin did not affect FSH responsiveness. Desensitization of FSH-activated adenylate cyclase was never greater than 35%; the degree of attenuation was dependent on FSH concentration and duration of exposure to FSH. Cells perifused with medium containing MIX and a maximally effective concentration of FSH for 5 h released cAMP continuously. In summary, the results indicated that porcine granulosa cells do not become unresponsive to FSH after prolonged exposure to FSH in vitro.