Induction of maturation of Atlantic croaker oocytes by 17 alpha,20 beta,21-trihydroxy-4-pregnen-3-one in vitro: consideration of some biological and experimental variables.
We characterized the in vitro control of germinal vesicle breakdown (GVBD) by 17 alpha,20 beta,21-trihydroxy-4-pregnen-3-one (20 beta-S) in intact ovarian follicles of gonadotropin-primed Atlantic croaker. 20 beta-S-induced GVBD was determined in relation to ovarian (oocyte) morphology, duration of incubation, steroid metabolism, and interaction with other steroids. The rate of GVBD in vitro in the absence of exogenous steroid was positively correlated with initial stage of ovarian morphological development. Maximal responsiveness to 20 beta-S was seen in ovaries with oocytes showing the first signs of morphological maturation. Dose-response experiments with 20 beta-S and 17 alpha,20 beta-dihydroxy-4-pregnen-3-one (17 alpha,20 beta-P) over a range of incubation times yielded similar results for both steroids, suggesting that conversion of 17 alpha,20 beta-P to 20 beta-S is not required for 17 alpha,20 beta-P-induced GVBD. The ED50 of these steroids markedly decreased with increasing incubation times. Comparisons between patterns of follicular transformation of various radiolabelled steroids to 20 beta-S and their respective activities (using unlabelled steroids) in the GVBD bioassay suggested that, in addition to 17 alpha,20 beta-P, progesterone has some intrinsic maturational activity. However, the maturational effects of 11-deoxycortisol and pregnenolone may be explained by their conversion to 20 beta-S. For the first time in any vertebrate, we showed that the proposed maturation-inducing steroid (20 beta-S) is not significantly transformed to any extractable, potentially active metabolite by intact, maturing ovarian follicles. These findings strongly suggest that 20 beta-S is the terminal product of the MIS biosynthetic pathway in Atlantic croaker ovaries. Estradiol had no acute effects on 20 beta-S-induced GVBD. However, testosterone decreased and cortisol augmented the maturational activity of 20 beta-S. Excess progesterone reduced the activity of a maximally effective dose of 20 beta-S, but pregnenolone was without effect. The effects of these steroids on 20 beta-S-induced GVBD are discussed in relation to their possible interactions with 20 beta-S at the MIS receptor level.
Patiño
R
R
Marine Science Institute, University of Texas, Austin 78373-1267.
Thomas
P
P
D015065:17-Hydroxycorticosteroids; D000305:Adrenal Cortex Hormones / Q000378:metabolism; D000704:Analysis of Variance; D000818:Animals; D003350:Cortodoxone / Q000031:analogs & derivatives / Q000378:metabolism; D005260:Female; D005399:Fishes; D012739:Gonadal Steroid Hormones / Q000378:metabolism; D006908:Hydroxyprogesterones / Q000378:metabolism; D066298:In Vitro Techniques; D009866:Oogenesis / Q000502:physiology*; D006080:Ovarian Follicle / Q000378:metabolism; D013256:Steroids / Q000378:metabolism; D013997:Time Factors
D015065:17-Hydroxycorticosteroids; D000305:Adrenal Cortex Hormones; D012739:Gonadal Steroid Hormones; D006908:Hydroxyprogesterones; D013256:Steroids; C021012:17 alpha,20 beta-dihydroxypregn-4-en-3-one; C051954:17,20,21-trihydroxy-4-pregnen-3-one; D003350:Cortodoxone