Abstract
Intact ovarian follicles, obtained from untreated and human chorionic gonadotropin (HCG) treated Japanese yellowtail Seriola quinqueradiata during different maturational stages, were incubated with radioactive [3H]pregnenolone, [ 3 H]17‐hydroxyprogesterone or [ 14 C] androstenedione and steroid metabolites identified by thin layer chromatography (TLC) followed by recrystallization to constant specific activity. In untreated late vitellogenic (0 h) follicles, androstenedione was the major product with smaller amounts of testosterone and oestradiol‐17α. In post‐vitellogenic (12 h post‐injection) intact follicles, androstenedione predominated, and although testosterone and oestradiol‐17α were not produced, there were small amounts of 17, 20β‐dihydroxy‐4‐pregnen‐3‐one (17,20β‐P) and 17,21‐dihydroxy‐4‐pregnene‐3, 20‐dione (11‐deoxycortisol). In HCG‐treated fish, a steroidogenic shift resulted in the disappearance of testosterone and oestradiol‐17 coinciding with the appearance of 17, 20β‐P. During early and late final oocyte maturation FOM (24 and 36 h post‐injection), there was a five‐ to seven‐fold increase in the production of 17, 20β‐P, whereas production of 11‐deoxycortisol remained almost the same. During FOM, in addition to 17,20β‐P, its 5β‐reduced metabolite, 17,20β‐dihydroxy‐5β‐pregnan‐3‐one (5β‐17,20β‐P) was synthesized, suggesting a decrease in maturation‐inducing 17,20β‐P activity. 17, 20β,21‐Trihydroxy‐4‐pregnen‐3‐one (20β‐S) was not synthesized by ovarian fragments in Japanese yellowtail at any maturational stage. The metabolites identified on TLC during FOM were tested to evaluate their maturation‐inducing activity in an in vitro bioassay. Of the steroids tested, 17,20β‐P was the most effective inducer of germinal vesicle breakdown (GVBD), followed by 5β‐17,20β‐P. Timely synthesis of 17,20β‐P immediately prior to and during FOM as well as its great potency in inducing GVBD in vitro supports the evidence for a physiological role of 17,20β‐P as a maturation‐inducing hormone in Japanese yellowtail.