To evaluate the effects of benzo[a]pyrene (B[a]P), one of polycyclic aromatic hydrocarbons (PAHs), on in vitro oocyte maturation (GVBD) and sex steroid hormone production, maturing oocytes (oocyte diameters=0.74, 0.88 and 0.93 mm) of the longchin goby, Chasmichthys dolichognathus were incubated with B[a]P (1, 10 and 100 ng/mL) for 24 hours. After incubation, the oocytes were fixed with clearing solution (ethanol:formalin:glacial acrtic acid=6:3:1). The location of the germinal vesicle was observed under low-power magnification using a dissecting microscope. Steroids in aliquots of the incubation media were extracted twice using five volumes of ethylacetate:cyclohexane (1:1). Then, the T, E2 and 17α20βP levels were measured by RIA. In oocytes 0.74 mm diameter (vitellogenic oocytes), B[a]P had no significant effect on GVBD at the concentrations tested. In oocytes 0.88 mm diameter (fully vitellogenic oocytes), B[a]P inhibited GVBD significantly at 1 and 100 ng/mL. T production was decreased and the ratio of E2/T was increased significantly at 1 and 10 ng/mL compared with control. In 0.93 mm diameter oocytes (germinal vesicle located near the center of oocytes), B[a]P induced GVBD significantly at 10 and 100 ng/mL and decreased the ratio of E2/T significantly at 1 and 10 ng/mL compared with control. These findings suggest that B[a]P has different sensitivity to the oocyte maturation according to the oocyte diameters.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental contaminants derived from incomplete combustion of carbons and crude oil. In this study, we investigated the effects of benzo[a]pyrene (B[a]P), a representative PAHs on in vitro sex steroid hormone production and germinal vesicle breakdown (GVBD) using isolated oocytes of longchin goby (Chasmichthys dolichognathus) and chameleon goby (Tridentiger trigonocephalus). Oocytes in diameters of 0.8-0.9 (end vitellogenic stage) and 0.9-1.0 mm (germinal vesicle migratory stage) from longchin goby and 0.5 mm (fully vitellogenic stage) from chameleon goby were used. In GVBD assay, B[a]P at 10 nM stimulated GVBD in the oocytes of 0.8-0.9 mm from longchin goby. B[a]P at 1 nM stimulated GVBD in the oocytes with diameter 0.5 mm from chameleon goby. In steroid production from oocytes of longchin goby, B[a]P at 100 nM decreased testosterone (T) production, B[a]P at 1,000 nM increased estraiol-17 (J (E2) production and 10 and 100 nM increased -dihydroxy-4-pregnen-3-one () production in the oocytes with diameter 0.8-0.9 mm. B[a]P at 1,000 nM increased E2 production, 100 and 1,000 nM increased production in the oocytes with diameter 0.9-1.0 mm. In steroid production of oocytes from chameleon goby, B[a]P at 1,000 nM increased production. B[a]P at 10 nM increased production. In the ratio of to T (/T), B[a]P at 100 and 1,000 nM increased /T in the oocytes of longchin goby. B[a]P at 100 nM also increased /T in the oocytes of chameleon goby. Taken together, these results suggest that B[a]P have not only weak estrogenic effects but progestogenic effects on oocyte maturation.

점망둑의 난모세포 성숙과정에서 생성되는 주요 성 스테로이드 호르몬, -스테로이드를 분석하고자 전구물질 ()를 성숙기 난모세포(난경 ) 배양초기에 첨가하여 24시간 배양하였다. 스테로이드 대사물질 분석과 동정은 thin layer chromatography와 gaschromatography-mass spectrometry로 이루어졌다. 로부터 생성된 주요 성 스테로이드 대사물질은 -hydroxy, -dihydroprogesterone ()와 -hydrox