The aim of this study was to investigate the effects of testosterone on basal and follicle-stimulating hormone (FSH)-induced progesterone production by cultured bovine granulosa cells. Granulosa cells were isolated from small (< 5 mm) and large (> 8 mm) follicles and cultured for 48 h in 1 ml of Medium-199 with different concentrations of FSH (0.1, 1, 10 and 35 mg/l). In addition, the combined effects of different amounts of testosterone (1 nmol-10 mumol) and 1 mg/l FSH for 48 h on progesterone production by granulosa cells of both groups of follicles were studied; progesterone production during the subsequent 24-h incubation period was evaluated in the absence of hormones. In a third experiment, granulosa cells were treated with 500 micrograms of dibutyryl-cAMP and 10 mumol of testosterone for 48 h. At the end of each incubation period, the progesterone content in the culture media was determined by a validated radioimmunoassay. Basal progesterone release during the 48-h incubation period was higher in granulosa cells from small as compared to cells from large follicles; in both groups of cells, progesterone production was stimulated maximally by 1 mg/l FSH. The treatment with 10 mumol of testosterone induced a decrease of progesterone production in both groups of cells, while lower amounts exerted an inhibitory effect only in cells from large follicles. Furthermore, 10 mumol of testosterone inhibited FSH-induced progesterone release, while lower dosages were ineffective. Dibutyryl-cAMP stimulated significantly the progesterone output by granulosa cells of both groups and testosterone was effective in suppressing this increase.(ABSTRACT TRUNCATED AT 250 WORDS)