Luteal cell suspensions obtained by enzymatic digestion of pregnant cow corpus luteum were found to be heterogenous and mainly made up of two types of cells of different sizes. The large cells (37 micrometers, average diameter) could be separated from the small ones (18 micrometers, average diameter) by sedimentation at unit gravity in a gradient of Ficoll-bovine serum albumin. A comparative in-vitro study of the synthesis of progesterone by the two types of cells indicated striking differences between them. The average content and the synthesis of progesterone in the absence and presence of a saturating dose of bovine LH after incubation for 2 h were 0.07, 0.12 and 6.9 pg/cell for the small cells and 0.65, 2 and 10 pg/cell for the large ones. Moreover, the sensitivity to low concentrations of LH was 100 to 1000 times higher for the small cells than for the large ones. Oestradiol-17 beta at concentrations ranging from 5 X 10(-10) to 5 X 10(-4) mol/l exerted a dose-dependent inhibition on the stimulation of LH in both cell types. These results suggest a possible involvement of both cell types in the synthesis of progesterone in vivo with a greater contribution by the small cells to stimulation induced by LH. Moreover, it appears that small cell suspensions could be a useful model system for in-vitro studies of the control of the synthesis of progesterone in cow corpus luteum.
A highly sensitive and specific radioimmunoassay was developed to measure 19-nortestosterone (NT), which allowed us to demonstrate this steroid in the plasma of pregnant women. Plasma NT was detectable throughout gestation, reaching values of 12 to 60 pg/ml in the 3rd trimester. These results were validated by gas chromatography-mass spectrometry. No NT was detectable in the plasma of 12 normal men and 12 nonpregnant women in the mid follicular phase of their cycle (detection limit 4 pg/ml). Our results are compatible with current concepts concerning the possible involvement of 19-norsteroids in an accessory biosynthetic pathway for estrogen in the placenta.
Accumulating evidence has shown the ovary of mammals to contain an intrinsic renin-angiotensin system that has been ascribed an autocrine-paracrine role. The present study in the female rabbit ovary investigated the putative in vitro action of angiotensin II (A II) on basal and gonadotropin-induced steroidogenesis. Ovarian follicles from immature female rabbits treated with pregnant mare's serum gonadotropin (PMSG) were dissected out and a complete separation of the theca interna from the granulosa layer was performed, to demonstrate that A II affects separately the two individual cellular components of the follicular wall. We could show that theca is a source of estradiol whose production under human chorionic gonadotropin (hCG) stimulation was reduced by A II. At the same time, A II increased the in vitro hCG-stimulated secretion of testosterone by theca. In granulosa, A II decreased hCG-stimulated aromatization of androstenedione to estradiol but did not alter the release of hCG-stimulated progesterone production. These results suggest that A II could induce locally an increase in follicular fluid androgen/estrogen ratio and possibly participate in causing atresia.
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