Effects of Steroid Hormones on In Vitro GVBD and Oocyte Steroidogenesis in Blacktip Grouper, Epinephelus fasciatus
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To verify the sex steroids which are involved in oocyte maturation of the blacktip grouper, Epinephelus fasciatus, we incubated vitellogenic oocytes (0.41 and 0.50 ㎜ in average diameter) in the presence of exogenous steroid precursor ([3H]17α-hydroxyprogesterone). Steroids were extracted, separated and identified by thin layer chromatography. The major metabolites produced were androstenedione, estradiol-17β, estrone and progestogens. Progestogen metabolites in the oocytes of 0.50 ㎜ were more abundant than those of 0.41 ㎜. Also, we investigated the in vitro effects of human chorionic gonadotropin (HCG; 5, 50 and 500 IU/㎖), 17α,20β-dihydroxy-4-pregnen-3-one (17α20βP) and 17α,20β-trihydroxy-4-pregnen-3-one (17α20β21P; 5, 50 and 500 ng/㎖, respectively) on oocyte maturation. In the oocytes of 0.41 ㎜, treatment with 50 IU HCG stimulated GVBD (55.30±1.20%) compared with controls (32.41±3.13%, p<0.05). In the oocytes of 0.50 ㎜, treatment of 17α20βP (50 and 500 ng/㎖) stimulated GVBD (50.13±2.52 and 51.77±5.91%, respectively) compared with controls (36.81±2.89%, p<0.05). Treatment with 500 IU HCG also stimulated GVBD (49.59±5.15%) compared with controls (p<0.05). Taken together, these results suggested that both HCG and 17α20βP were effective on in vitro oocyte maturation and 17α20βP may act as a maturation inducing hormone in blacktip grouper.
Abstract
To verify the sex steroids which are involved in oocyte maturation of the blacktip grouper, Epinephelus fasciatus, we incubated vitellogenic oocytes (0.41 and 0.50 ㎜ in average diameter) in the presence of exogenous steroid precursor ([3H]17α-hydroxyprogesterone). Steroids were extracted, separated and identified by thin layer chromatography. The major metabolites produced were androstenedione, estradiol-17β, estrone and progestogens. Progestogen metabolites in the oocytes of 0.50 ㎜ were more abundant than those of 0.41 ㎜. Also, we investigated the in vitro effects of human chorionic gonadotropin (HCG; 5, 50 and 500 IU/㎖), 17α,20β-dihydroxy-4-pregnen-3-one (17α20βP) and 17α,20β-trihydroxy-4-pregnen-3-one (17α20β21P; 5, 50 and 500 ng/㎖, respectively) on oocyte maturation. In the oocytes of 0.41 ㎜, treatment with 50 IU HCG stimulated GVBD (55.30±1.20%) compared with controls (32.41±3.13%, p<0.05). In the oocytes of 0.50 ㎜, treatment of 17α20βP (50 and 500 ng/㎖) stimulated GVBD (50.13±2.52 and 51.77±5.91%, respectively) compared with controls (36.81±2.89%, p<0.05). Treatment with 500 IU HCG also stimulated GVBD (49.59±5.15%) compared with controls (p<0.05). Taken together, these results suggested that both HCG and 17α20βP were effective on in vitro oocyte maturation and 17α20βP may act as a maturation inducing hormone in blacktip grouper.
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