Steroidogenesis in isolated rat granulosa cells--changes during follicular maturation

Steroid release was investigated in granulosa cells isolated from rat ovarian follicles at different maturation stages. Immature rats were treated with 10 IU of pregnant mare's serum gonadotrophin (PMSG) to induce follicular growth and maturation. Granulosa cells were isolated and subsequently incubated for 4 h in the absence or presence of follicle stimulating hormone (FSH), luteinizing hormone (LH) or forskolin. The concentrations of progesterone (P), 20 alpha-dihydroprogesterone (20 alpha DHP), testosterone (T) and oestradiol (E2) in the media were determined by radioimmunoassay. During maturation, basal release of T increased markedly at late dioestrus. At pro-oestrus, concomitant with a sharp rise in E2 and P release, T release returned to previously low levels. Addition of FSH or LH stimulated P release at late dioestrus and pro-oestrus but had no effect on E2 or T levels. Basal 20 alpha DHP levels remained low until a sharp rise at mid pro-oestrus. A stimulatory effect of gonadotrophins on 20 alpha DHP was first seen at this stage. The adenylate cyclase stimulator forskolin resembled FSH in all its effects. In summary, the present study demonstrates that maturing, but not preovulatory, granulosa cells have an increased capacity to release T during a relatively short period. It is suggested that this T peak could be of great importance for the further progress of follicular maturation.