• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2004.10a
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• pp.68-68
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• 2004

• Development and Reproduction
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• v.23 no.2
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• pp.111-117
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• 2019

This study focused on the association of prostaglandins and a progestin, $17{\alpha}$, $20{\beta}P$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$) during the ovulation process in longchin goby, Chasmichthys dolichognathus. We performed several in vitro experiments using $850-920{\mu}m$ diameter oocytes which were at the migratory nucleus stage. With the $890-920{\mu}m$ diameter oocytes, no significant difference in ovulation was observed in any of the prostaglandins (PGE1, PGE2, and $PGF2{\alpha}$) treated groups although PGE2 and $PGF2{\alpha}$ at concentrations of 50 ng/mL increased ovulation slightly compared with controls; however, $17{\alpha}20{\beta}P$ production was stimulated with PGE1 alone at low concentrations (5 ng/mL). In $850{\mu}m$ diameter oocytes, $PGF2{\alpha}$ at concentrations of 50 and 500 ng/mL resulted in a significant increase in ovulation. $17{\alpha}20{\beta}P$ (50 ng/mL) alone had no observable effect on ovulation, but in the combined of $PGF2{\alpha}$ 50 or 500 ng/mL it caused the greatest effect on ovulation. The sensitivity of oocytes to the induction of ovulation varies between 850 and $890-920{\mu}m$, it appeared to vary depending on the migration status of nucleus. These results suggest that $PGF2{\alpha}$ (or combined of $17{\alpha}20{\beta}P$) was more potent in inducing ovulation of the longchin goby.

• Development and Reproduction
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• v.17 no.4
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• pp.421-426
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• 2013

In this study, oocyte steroidogenesis are investigated in relation to oocyte development in the starry flounder, Platichthys stellatus, a marine multiple spawner. Vitellogenic (0.52 and 0.55 mm oocyte diameter) and mature oocytes (0.63, 0.66 and 0.71 mm oocyte diameter) were incubated in vitro in the presence of $[^3H]17{\alpha}$-hydroxyprogesterone ($[^3H]17{\alpha}$-OHP) as a precursor. Steroid metabolites were extracted from the incubated media and oocytes, the extracts were separated and identified by thin-layer chromatography (TLC), high performance liquid chromatography (HPLC) and gas chromatographymass spectrometry (GC-MS). The major metabolites produced from $[^3H]17{\alpha}$-OHP were androgens [androstenedione ($A_4$) and testosterone (T)] and estrogens [$17{\beta}$-estradiol ($E_2$) and estrone ($E_1$)] and progestins [$17{\alpha},20{\alpha}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\alpha}P$) and $17{\alpha},20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$)] in vitellogenic and mature oocytes. The results from this study suggest the potential roles of $E_1$ in the oocytes with diameter 0.52-0.71 mm, $17{\alpha}20{\alpha}P$ and $17{\alpha}20{\beta}P$ at the oocytes of 0.63, 0.66 and 0.71 mm.

• Development and Reproduction
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• v.12 no.3
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• pp.275-281
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• 2008

Endocrine disrupting chemiclas (EDCs) such as bisphenol A (BPA) and nonylphenol (NP) have estrogenic activity and can alter reproduction in fish. In the present study, the effects of BPA and NP on in vitro steroid production from oocytes of maturation stage (oocyte diameter$\fallingdotseq$1.88 mm) from the greenling (Hexagrammos agrammus) were evaluated. Oocytes were incubated with different concentrations of BPA and NP (0.1, 1, 10, 100 and 1,000 ng/$m{\ell}$) in the presence or absence of 50 IU human chorionic gonadotropin (HCG) for 48 hours. After incubation, levels of $17{\alpha},\;20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}OHP$), estradiol-$17{\beta}(E_2)$ and testosterone (T) from incubated media were quantified by radioimmunoassay (RIA). In BPA treatment, 100 ng/$m{\ell}$ of BPA stimulated $E_2$ production regardless HCG supplement. Every concentration of BPA inhibited T production without HCG although 0.1 ng/$m{\ell}$ of BPA stimulated T production with HCG. In NP treatment, 10 ng/$m{\ell}$ of NP stimulated $17{\alpha}20{\beta}OHP$ and T production without HCG. 1 ng/$m{\ell}$ of NP inhibited $E_2$ production. Taken toghther, these results suggest that BPA might have weak estrogen-agonistic effect and NP has estrogenantagonistic effect at final oocyte maturation stage of H. agrammus.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.48 no.4
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• pp.483-488
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• 2015

We studied oocyte steroidogenesis in the blackfin flounder Glyptocephalus stelleri as a region-specific species, in the East Sea of Korea during the spawning season. Maturing oocytes (0.76, 0.82, 0.88, and 0.91 mm in oocyte diameter) were incubated in vitro in the presence of [$^3H$] $17{\alpha}$-hydroxyprogesterone ($[^3H]17{\alpha}$-OHP) as a precursor. Steroid metabolites were extracted from the incubated medium and oocytes, and the extracts were separated and identified by thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC) and gas chromatographymass spectrometry (GC/MS). The major metabolites produced from $[^3H]17{\alpha}$-OHP were androgens [androstenedione (A4) and testosterone (T)] and estrogens [$17{\beta}$-estradiol (E2) and estrone (E1)] and progestins [$17{\alpha},20{\alpha}$-dihydroxy-4-pregen-3-one ($17{\alpha}20{\alpha}P$) and $17{\alpha}20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$)] in maturing oocytes. The metabolic rate of $17{\alpha}20{\beta}$ was elevated (29.04%) in oocytes measuring 0.88 mm (nucleus migration stage following the induction of germinal vesicle breakdown), but was very low in oocytes measuring 0.76, 0.82, and 0.91 mm (0.42, 0.67, and 2.62%, respectively). From these results, we suggest that $17{\alpha}20{\beta}P$ acts as a maturation-inducing steroid in the blackfin flounder.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.5
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• pp.431-438
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• 2000

To clarify the annual reproductive cycle in a rockfish, Sebastes schlegeli, monthly changes in gonadosomatic index (GSI), hepatosomatic index (HSI) and histological feature of gonads and plasma levels of sex steroid hormones ($estradiol-l7{\beta},\;17{\alpha},\;20{\beta}-dihydroxy-4-pregnen-3-one,\;testosterone\;and\;11-ketotestosterone$) were investigated. The annual reproductive cycle in females could be divided into 5 periods as follows: 1) recovery period (June to September): serum level of $estradiol-l7{\beta}$ increased gradually; 2) vitellogenesis period (Septemer to february) : vitellogenic oocytes were obsewed, GSI sustained high value, and serum level of $estradiol-l7{\beta}$ increased; 3) gestation period (February-April): developing larva showed in the ovary, and serum levels of $17{\alpha},\;20{\beta}-dihydroxy-4-pregnen-3-one$ and testosterone increased; 4) partrition period (April to May) : larva were delivered, and value of GSI and serum levels of hormones decreased rapidly; 5) resting period (May to June) : value of GSI and serum levels of $estradiol-l7{\beta}$ and testosterone remained low. The annual reproductive cycle in males could be divided into 6 periods; 1) early maturation period (April to June): value of GSI and serum levels of hormones incresed gradually, cyst of spermatogonia incresed in number, and a small number of cyst of spermatocyte was observed; 2) mid-maturation perid (June to September); value of GSI and serum levels of hormones increased, and germ cells in many cysts were undergoing active sperrnatogenesis; 3) late maturation period (September to November) : value of GSI and serum levels of hormones remained high and spermatozoa were released into the lumina of the seminal lobules; 3) spermatozoa dischaging period (Nobember to December) : the lumina of the seminal lobules were enlarged and filled with mature spermatozoa; 4) degeneration period (December to Februauy)i value of GSI decresed and cyst of spermatocyte were decresed in number; 5) resting period (December to April) : no histological changes of testes were observed, and value of GSI and serum levels of hormones remained low. In November, the lumina of the seminal lobules were filled with mature spermatozoa and sperm masses were present in the ovarian cavity. Thus, copulation in this species occurred in November and December.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.2
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• pp.203-210
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• 1997

The aim of this study was to determine the effect of steroids and human chorionic gonadotropin (HCG) on in vitro maturation and ovulation of oocyte in Pseudobagrus fulvidraco. Oocytes were incubated in the media Leibovitz L15 supplemented with the various concentration of $17\alpha,\;20\beta-dihydroxy-4-pregnen-3-one(17\alpha20{\beta}OHP),\;17\alpha-hydroxyprogesterone(17{\alpha}OHP),\;progesterone(P_4),\;estradiol-17\beta(E_2)and\;HCG$. After 60 hours incubation, the maturation ability of oocyte was assessed by the appearance of germinal vesicle breakdown (GVBD). GVBD was significantly enhanced by the addition of $17\alpha20{\beta}OHP,\;17{\alpha}OHP,\;P_4\;and\;HCD(P<0.05)$. The highest CVBD was observed when $17\alpha20{\beta}OHP$ and HCG were supplemented to media. When oocytes were cultured for 16 hours in media containing $10\~1,000\;ng/ml\;17\alpha20{\beta}OHP,\;17{\alpha}OHP\;and\;P_4$, the rate of GVBD in oocytes cultured in the medium supplemented with 100 ng/ml $17\alpha20{\beta}OHP(65\%)$ was significantly higher than that with $17{\alpha}OHP\;(40\%)\;and\;P_4(35\%)$. The efforts of $17\alpha20{\beta}OHP$ and HCG on GVBD were assessed by various concentration of these hormones. When oocytes were cultured for 60 hours in various media containing $1\~1,000\;ng/ml\;17{\alpha}20{\beta}OHP\;or\;5\~1,000\;IU/ml$ HCG, the GVBD of oocytes was significantly increased in the medium with $10\~100\;ng/ml\;17\alpha20{\beta}OHP$ and 500 IU/ml HCT. When oocytes were cultured in the various media supplemented with $1\~1,000\;ng/ml\;17\alpha20{\beta}OHP\;or\;5\~1,000\;IU/ml$ HCG for 60 hours, the media with $1\~100\;ng/ml\;17\alpha20{\beta}OHP\;or\;50\~1,000IU/ml$ HCG significantly increased in the rate of ovulation. However supplementation with $1,000\;ng/ml\;17\alpha20{\beta}OHP$or 5 IU/ml HCG did not improve the rate of ovulation compared to controls. This results indicate that supplementation of steroid and HCG except $E_2$ can improve the in vitro maturation and ovulation of oocyte in P. fulvidrac; HCG and $17\alpha20{\beta}OHP$ may be more effective than other steroids on oocyte maturation and ovulation in P. fulvidraco.

• Development and Reproduction
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• v.16 no.1
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• pp.39-45
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• 2012

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 mm in average diameter) in the presence of exogenous steroid precursor ($[^3H]17{\alpha}$-hydroxyprogesterone). Steroids were extracted, separated and identified by thin layer chromatography. The major metabolites produced were androstenedione, estradiol-$17{\beta}$, estrone and progestogens. Progestogen metabolites in the oocytes of 0.50 mm were more abundant than those of 0.41 mm. Also, we investigated the $in$ $vitro$ effects of human chorionic gonadotropin (HCG; 5, 50 and 500 $IU/m{\ell}$), $17{\alpha},20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$) and $17{\alpha},20{\beta}$-trihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}21P$; 5, 50 and 500 $ng/m{\ell}$, respectively) on oocyte maturation. In the oocytes of 0.41 mm, treatment with 50 IU HCG stimulated GVBD ($55.30{\pm}1.20%$) compared with controls ($32.41{\pm}3.13%$, $p$<0.05). In the oocytes of 0.50 mm, treatment of $17{\alpha}20{\beta}P$ (50 and 500 $ng/m{\ell}$) stimulated GVBD ($50.13{\pm}2.52$ and $51.77{\pm}5.91%$, respectively) compared with controls ($36.81{\pm}2.89%$, $p$<0.05). Treatment with 500 IU HCG also stimulated GVBD ($49.59{\pm}5.15%$) compared with controls ($p$<0.05). Taken together, these results suggested that both HCG and $17{\alpha}20{\beta}P$ were effective on in vitro oocyte maturation and $17{\alpha}20{\beta}P$ may act as a maturation inducing hormone in blacktip grouper.

• Development and Reproduction
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• v.16 no.4
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• pp.279-287
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• 2012

We investigated the changes in plasma sex steroid hormones, testosterone (T), estradiol-$17{\beta}$ ($E_2$), 17,$20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$), 11-ketotestosterone (11KT) and cortisol levels from ribbed gunnel, Dictyosoma burgeri in associated with annual reproductive cycle. The gonadosomatic index (GSI) of females increased from November, peaked in February and decreased rapidly from March. The GSI of males also increased from November, peaked in January and then decreased gradually. In females, $E_2$ levels increased and remained high from December to February. The levels of T showed a similar tendency and correlated ($r_s$=0.898, p<0.01) with $E_2$ levels. The levels of $17{\alpha}20{\beta}P$ increased rapidly in February ($4.78{\pm}1.01ng/ml$) and peaked in July ($5.08{\pm}0.65ng/ml$). Cortisol level was peaked in March and correlated with $17{\alpha}20{\beta}P$ levels ($r_s$=0.696, p<0.01). In males, the levels of T was peaked in January and then decreased rapidly. The levels of 11KT were remained high from October to January. On the other hand, the levels of $17{\alpha}20{\beta}P$ fluctuated during reproductive cycle. These results suggest that plasma sex steroids in ribbed gunnels have annual periodicity, and that cortisol may involve in maturation of females.

• Development and Reproduction
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• v.15 no.3
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• pp.223-230
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• 2011

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 $17,20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$) 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 $17{\alpha}20{\beta}P$ 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 $E_2$ production. B[a]P at 10 nM increased $17{\alpha}20{\beta}P$ production. In the ratio of $E_2$ to T ($E_2$/T), B[a]P at 100 and 1,000 nM increased $E_2$/T in the oocytes of longchin goby. B[a]P at 100 nM also increased $E_2$/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.