Steroidogenic shift by cultured ovarian follicles of Rana dybowskii at breeding season

The steroid secretory activity of cultured ovarian follicles of Rana dybowskii and the activities of relevant steroidogenic enzymes were examined during the natural hibernation period (October-February). Enzyme activities were measured indirectly by monitoring the conversion of exogenous substrates to products by isolated follicles. Follicles were incubated for 6 h in amphibian Ringer in the presence or absence of frog pituitary homogenate (FPH, 0.1 pituitary/2 ml) and/or various steroid precursors. Progesterone (P4), 17 alpha-hydroxyprogesterone (17 alpha-OHP) or testosterone (T) secreted by the follicles into the medium were measured by RIA. In the presence of FPH, high levels of P4 were produced by follicles at the early and mid-hibernation period (695 and 898 pg/follicle, respectively) whereas markedly elevated levels of P4 were produced during late hibernation (1,393 pg/follicle) (just prior to or breeding season, February). In contrast, high levels of T were produced by the follicles early in hibernation (1,206 pg/follicle) while negligible levels were produced in late hibernation (69 pg/follicle). Higher levels of 17 alpha-OHP were produced by follicles at early and mid hibernation (594 and 705 pg/follicle, respectively) than in later hibernation (221 pg/follicle). Addition of exogenous pregnenolone markedly increased P4 levels in a dose-dependent manner when added to follicles at early and late stages of hibernation: FPH addition further enhanced conversion of pregnenolone. Similarly, addition of androstenedione (AD) increased T levels in a dose-dependent manner by these follicles. However, following addition of exogenous P4, less 17 alpha-OHP was produced by follicles collected in late hibernation as compared to those collected earlier.(ABSTRACT TRUNCATED AT 250 WORDS)