Androgenic stimulation of progesterone production by granulosa cells from preantral ovarian follicles: further in vitro studies using replicate cell cultures.
The influence of testosterone (T) on progesterone (P) production by isolated rat ovarian granulosa cells was studied in vitro using a new replicate culture technique. Preantral granulosa cells from ovaries of estrogen-primed hypophysectomized immature female rats were cultured in the presence of graded concentrations of T, diethylstilbestrol (DES), cyproterone acetate (CPA), flutamide and the hydroxylated derivative of flutamide, Sch 16423. The accumulation of P in medium collected from granulosa cell cultures was measured by specific radioimmunoassay. Maximal P production by cultured granulosa cells was attained during the second day of culture and declined markedly thereafter. The presence of 10(-9), 10(-8) or 10(-7)M T elicited increases in P production 2.4, 8 and 11 times that of controls, respectively, during the initial 48 h of culture. Each concentration of T elicited enhanced P production within the first 24 h of culture. Granulosa cells cultured in control medium for 2 days did not respond to 10(-7)M T during the subsequent 3 days. DES at a high concentration in the medium (10(-5)M) markedly suppressed the response to 10(-9) and 10(-8)M T. At a lower concentration (10(-9)M) DES significantly enhanced the stimulatory effect of 10(-9)M T but did not alter the response to higher concentrations of T. Neither high nor low concentrations of DES influenced P production in response to 10(-7) M T. The stimulatory effects of T on P production were suppressed in the presence of a 100-fold molar excess of the anti-androgens, CPA or Sch 16423. The present data indicate that androgenic stimulation of P production by preantral granulosa cells is a specific receptor mediated event which is modulated by the presence of estrogen in vitro. It is suggested that androgen-responsive P production is a functional capacity which granulosa cells acquire at a very early stage of hormonal differentiation and may be of physiological consequence in the intraovarian control of follicular maturation in vivo.
D000818:Animals; D002478:Cells, Cultured; D003534:Cyproterone / Q000494:pharmacology; D004054:Diethylstilbestrol / Q000494:pharmacology; D005260:Female; D005485:Flutamide / Q000031:analogs & derivatives / Q000494:pharmacology; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism*; D006080:Ovarian Follicle / Q000378:metabolism*; D010053:Ovary / Q000187:drug effects / Q000378:metabolism; D011374:Progesterone / Q000096:biosynthesis*; D051381:Rats; D013739:Testosterone / Q000494:pharmacology*
D013739:Testosterone; D011374:Progesterone; D004054:Diethylstilbestrol; D005485:Flutamide; D003534:Cyproterone