• Korean Journal of Fisheries and Aquatic Sciences
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• v.39 no.5
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• pp.391-397
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• 2006

This study correlated changes in estradiol-l7$\beta$ ($E_2$), testosterone (T), 17$\alpha$,20-dihydroxy-4-pregnen-3-one (DHP), and vitellogenin (VTG) levels with changes in the gonadosomatic index (GSI) and ovarian histology during the annual reproductive cycle of the wild marbled sole, Limanda yokohamae. Synchronous oocyte development occurs in this fish. Ovary maturity was classified into four periods, based on histological observations: the spawning (December to February), post-spawning (February to April), recovery (May to August), and vitellogenic (September to November) periods. Seasonal changes in the GSI were inversely correlated with water temperatures and reflected the degree of ovarian maturity. Plasma VTG levels were correlated with changes in the GSI, which increased from September to a peak in January, and levels remained comparatively high until February. Estradiol-17$\beta$ was at baseline levels (<0.11 ng/mL) during the spring and summer, and peaked rapidly (1.55$\pm$0.445 ng/mL) from October to January. Plasma T and DHP levels had a similar profile; they rose markedly during the spawning period and remained low (or were not detectable) from spring through autumn. These data indicate that changes in plsama steroid hormones and VTG levels are correlated with the annual ovarian activity of the marbled sole. Based on these results and published reports, it appears that in this species DHP is the most important maturation-inducing steroid and that T is also related to final maturation.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.31 no.4
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• pp.599-607
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• 1998

To clarify annual reproductive cycle of Korean dark sleeper, Odontobutis platycephala (Iwata et Jeon), we examined the seasonal changes of gonadosomatic index (GSI), the proportional frequency of oocyte development stages in the ovary and the changes of sex steroid hormone levels in blood from December 1995 to November 1997. In July and August, GSI was 0.35 to 0.72 and most oocytes in the ovary were chromatin-nucleolus stage and perinucleolar stage (proportional frequency: $87\%\~96\%$). In September, GSI was 1.20 $\pm$ 0.12, some oocytes in the ovary were yolk vesifle stage (proportional frequency: $22.8\%$) and vitellogenic stage which appeared very rarely(proportional frequency: $2.2\%$). GSI increased gradually from October and reached 4.59± 0.61 to December. During this period, oocytes of vitellogenic stage increased slightly (proportional frequency in December: $22.1\%$). In January, GSI was 4.32 $\pm$ 0.72 but the proportional frequency of oocytes in vitellogenic stage increased (proportional frequency: $51.2\%$). from February, GSI was increased sharply and reached to 10.51 $\pm$ 1.04 in March, the highest value throughout the year and the proportional frequency of oocytes in vitellogenic stage also reached the highest levels (proportional frequency: $60\%$). From April, GSI was gradually decreased and fell down to 1.11 $\pm$ 0.35 in June. During this period, the proportional frequency of mature oocytes was the highest in April (proportional frequency of mature oocyte stage: $40\%$ in April, $12\%$ May, $5\%$ June) throughout the year, and atretic ovarian follicles were appeared. The blood level of estradiol-17$\beta$ ($E_2$), which stimulates the hepatic synthesis and secretion of vitellogenin, was $0.84{\pm}0.20\;ng/m{\ell}$ in August, and thereafter was not changed until December. from January, it increased sharply and reached the highest level of $ 2.85{\pm}0.35\;ng/m{\ell}$ in March throughout the year, but fell to $0.14{\pm}0.02\;ng/m{\ell}$ in July(P<0.05), 17$\alpha$-hydroxprogesterone(17$\alpha$-OHP) was the peak $13.37{\pm}0.52ng/m{\ell}$ in March, but no significant changes in other period(below $3ng/m{\ell}$, P<0.05). 17$\alpha$, 20$\beta$-dihydroxy-4-pregnen-3-one(17$\alpha$, 20$\beta$-P), which was known as the final maturation inducing hormone in teleost, was $0.74{\pm}0.09ng/m{\ell}$ in April and $0.54{\pm}0.07ng/m{\ell}$ in May, but no significant changes in other period (below $0.26\;ng/m{\ell}$, p<0.05). Taken together these results, the annual reproductive cycle of O. platycephala divided into 4 periods as follows: 1) ripe and spawning period from April to June, main spawning period was from April to May, 2) Resting period from July to August, 3) Growing period from September to December, 4) Maturing period from January to March. Moreover, It was showed that the changes of sex steroid hormone in blood played a important roles in the annual reproductive cycle of O. platycephala.

• Journal of Aquaculture
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• v.18 no.3
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• pp.167-172
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• 2005

Sex reversal to a functional male of sevenband grouper $(41.0{\pm}1.3cm\;TL,\;1.4{\pm}0.1kg\;BW)$ was induced by $17{\alpha}-methyltestosterone\;(MT,\;0.5\sim2.0mg/kg\;BW)$ implantation from March 17 to May 12,2002. Gonad of control group was composed of genial cells and peri-nucleolus oocyte during the experimental period. Gonad of fish treated with 0.5 mg MT/kg BW had peri-nucleolus oocytes, spermatogonia, spermatids and spermatozoa at the late stages of spermatogenesis, while the fish group treated with 1.0 and 2.0 mg MT/kg BW contained spermatoza in the efferent duct. Sperm were obtained from the experimental groups treated with a dose of $1.0{\sim}2.0mg$ MT/kg BW. In the MT treated groups, testosterone and 11-ketotestosterone levels were higher than those in the control group during the $2{\sim}6$ weeks of the experimental period (P<0.05). $Estradiol-17{\beta}$ was detected from fish in the experimental fish.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.34-34
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• 2001

본 연구의 목적은 한우 난포발달에 있어서 난포액 및 anti-inhibin serum의 생리적 역할을 검토하기 위해 수행하였다. Anti-inhibin serum(AI)은 항원으로서 porcine inhibin-$\alpha$-subunit 19~32의 peptide를 사용하여 adjuvant 용액을 1:3의 비율로 혼합하여 앙고라종 토끼 5두(체중 2.5kg)에게 주 2회 간격으로 접종 후 얻어진 항혈청을 사용하였다. 난포액(bFF; bovine follicular fluid)은 도축장에서 도축되는 한우 난소로부터 직경 1.0cm 이하의 난포로부터 회수하여 스테로이드를 제거하기 위해 10% chacoal so lution(50 mg/$m\ell$, Norrit-A, Fisher Sci., USA)을 처리하여 45분간 배양후 원심분리후 상층액을 회수하여 실험에 공시하였다. 공시동물은 1산후 정상적으로 발정주기가 반복되는 한우암소 9두를 난소감정후 황체를 확인하여 PGF2$\alpha$제제(lutylase. USA)를 주사하여 발정을 유기한 다음, 난소의 first wave가 시작되는 시기인 배란직후 12시간째부터 4일간 일일 2회 5 $m\ell$씩 총 8회 40 $m\ell$의 AI와 bFF를 각각 경정맥으로 주사하였으며 대조구로서 생리식염수를 주사하였으며 채혈 및 정맥주사를 용이하게 하기 위해 경정맥에 카테타를 설치하여 6시간간격으로 총 200시간까지 채혈하였으며 초음파진단기를 이용하여 난포의 발달을 검토하였다. 채혈후 혈중Inhibin, progesterone 및 Estrad iol-17$\beta$농도의 분석은 RIA 및 ELISA법으로 분석하여 얻어진 결과는 다음과 같다. 혈중 Progesterone농도는 대조구와 AI처리구에서는 배란후 68시간째부터 유의적으로 증가하기 시작하였으나, bFF처리구에서는 배란후 68시간부터 170시간까지 대조구에 비해 유의적으로 낮은 농도를 나타내었다. 이에 반해 혈중 Estradiol-17$\beta$농도는 대조구의 경우 bFF처리구와 비슷한 수준으로 배란후부터 낮은 농도를 유지하였으나, AI 처리구는 배란후 36시간이후부터 108시간까지 유의적으로 높은 수준을 유지하였다가 그 이후 감소되었다. 한편, 혈중 Inhibin농도는 전 구간에서 배란후 84시간까지 불규칙한 농도를 보이다가 bFF처리구에서는 배란후 84시간부터 유의적으로 증가하였다. 배란후 72시간째에 초음파진단기를 이용하여 난소의 난포발달을 조사한 결과 , 대조구와 bFF처리구에 비해 AI처리구에서 발달난포가 유의적으로 많은 것을 확인하였다. 이상과 같은 결과로, Anti-inhibin serum은 한우 자체에서 분비하는 Inhibin을 특이하게 억제하여 Inhibin에 의해 억제되는 FSH분비가 촉진됨으로써 난포발달과 estrogen의 농도가 촉진되는 것으로 사료되어 anti-inhibin serum이 한우의 과배란유기 효과가 있는 것으로 사료된다.

• Toxicological Research
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• v.20 no.3
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• pp.205-211
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• 2004

Concern that some chemicals in our environment may affect human health by disrupt-ing normal endocrine function has prompted a research on interactions of environmental contaminants with steroid hormone receptor. Toxaphene and chlordane are among the 12 persistent organic pollutants identified by the United Nations Environment Programme as requiring urgent attention. We compared the estrogenic activity of two organochlorine pesticides, toxaphene and chlordane, at estrogen receptor a (ER$\alpha$) and estrogen receptor $\beta$ (ER$\beta$). Human hepatoma cells (HepG2) were transiently transfected with rat ER$\alpha$ or ER$\beta$ plus an estrogen-responsive complement C3-luciferase (C3-Luc) reporter gene. After transfection, cells were treated with various concentrations of toxaphene and chlordane to investigate agonism or antagonism of these chemicals. Both toxaphene and chlordane were potent agonists in HepG2 cells for ER$\alpha$. In contrast, these chemicals had a minimal agonist activity with ER$\beta$ and almost abolished 17$\beta$-estradiol-induced ER$\beta$-mediated activity. Therefore, toxaphene and chlordane behaved as an ER$\alpha$ agonist and an ER$\beta$ antagonist with estrogen-responsive reporter plasmid C3-Luc, and exposure to these organochlorine pesticides could have a crictical effect on normal endocrine function.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.2
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• pp.282-290
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• 1997

Hepatocytes of Anguilla japonica have been prepared using a collagenase perfusion technique. The isolated cells attached efficiently to fibronectin-coated culture dishes and subsequently formed monolayers in serum-free medium. These cultures maintained in appropriate medium at least for 10 days with minimal cell loss. The effects of estradiol and pituitary hormones on vitellogenin (Vg) synthesis were examined in primary hepatocyte culture of the immature eels. In fish, as in other oviparous vertebrates, estrogen is a major inducer of Vg synthesis. However, $estradiol-17\beta(E_2)$ alone was insufficient to induce Vg synthesis in cultures of eel hepatocytes. Combination of $E_2$ with growth hormone (GH) and/or prolactin (PRL) markedly stimulated Vg synthesis. Even in cultures exposed to $E_2$ or precultured without hormones for 8 days, $E_2$ alone could not fully induce Vg synthesis. The synthesis of Vg was dramatically increased when hepatocytes were cultured in medium supplemented with $E_{2}+GH+PRL$ for 6 days. At this point, even though GH and/or PRL were eliminated from the medium, Vg synthesis was not influenced by these factors during culture of further 3 days. These results indicate that pituitary hormones, in particular GH and PRL, play important roles in the regulation of Vg synthesis in primary cultures of eel hepatocytes.

• Journal of Embryo Transfer
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• v.1 no.1
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• pp.59-68
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• 1986

산양의 발정주기의 생리와 조절에 관한 최근 연구 보고들과 한국 재래산양의 대한 이들 연구 결과를 모아 고찰하였다. 온대지역에서 산양의 번식계절은 분명하나 한국재래산양의 경우는 비번식계절이 비교적 짧거나 계절적 무발정율이 낮은 편이며, 비번식계절에는 기능적 황체가 유지되지 않는다. 한국재래산양의 발정주기와 발정지속기간은 타품종과 비슷하며, 발정지속기간의 반복력은 매우 낮으며, 발정주기중의 progesterone과 estradiol-17$\beta$의 소장을 규명하였다. 한국재래산양의 경우 황체기에 PGF2 1-3mg을 투여하면 항체퇴행을 유기할 수 있으나 주기중의 투여시기에 따라 투여후 발정개시 시간에 차이가 있었다. 그리고 pro-gesterone priming후 PGF$\alpha$를 투여함으로써 효과적인 발정동기화를 이룰 수 있었다. 분만후 1개월경과 유상후 12일경에 발정이 재귀되었으며, 첫 발정 후 80%의 산양에서 5-7일의 단발정주기를 나타냈으며, 대부분의 과배란유기 산양에서도 단발정주기를 나타냈는데 이는 황체의 조기퇴행 때문이었다. 초발정후 3-5일에 GnRH를 연속 주사하면 단발정주기의 발생율이 유의적으로 감소되었다.

• Journal of Korean Society on Water Environment
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• v.26 no.6
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• pp.948-953
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• 2010

In this study, degradation of estrone (E1) and $17{\alpha}$-ethynylestradiol (EE2) by gamma-irradiation and subsequent reduction of estrogenic activity as a function of absorbed dose were conducted using the yeast two-hybrid assay. Relative potency of E1 and EE2 compared to estrogenic activity of $17{\beta}$-estradiol (E2) was found to be 0.0144 and 0.1605, respectively. More than 90% of E1 and EE2 (both $5.0{\times}10^{-6}M$) was removed at an absorbed dose of 5 kGy, but more than 40% of estrogenic activity still remained. The addition of $TiO_2$ catalyst appeared to improve the removal efficiency of E1 and decrease estrogenic activity while there was no significant effect for EE2. Additionally, the calculated estrogenic activity of E1 and EE2 based on a regression model was well correlated with the observed activity.

• Journal of Aquaculture
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• v.9 no.1
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• pp.19-23
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• 1996

Hormonal induction of sex reversal was examined by using sex steroid hormones in serranid fish, Epinephelus septemfasciatus. Young fish were collected from the coastal area of Cheju Island, and reared for 2 years before fish were used for the experiments. Without any hormonal treatment, gonads of fish ($1,000\~2,800$ g in body weight) were occupied by oocytes of the perinucleolus stage and bundles of protogonial cells in the area of germinal epithelium. When the induction of sex reversal was attempted by daily oral administration of $17\alpha$-methyltestosterone (0.5 mg/kg fish) for 90 days, active spermatogenesis was induced, and spermatogonia and spermatocytes and spermatids were appeared in all gonads we examined. However, after daily, oral treatment of $17\beta$-estradiol (0.5 mg/kg fish) to. 50 days with the following injection of human chorionic gonadotrophin ($1,000\~1,500$ IU/kg fish) mature oocytes were not induced in fish gonad.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.47 no.6
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• pp.882-887
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• 2014

We studied oocyte steroidogenesis in blacktip grouper Epinephelus fasciatus ovarian follicles during vitellogenesis. Vitellogenic oocytes with average diameters of 0.45, 0.48 and 0.50 mm were incubated in vitro in the presence of $[^3H]17{\alpha}$-hydroxyprogesterone as a precursor. The steroid metabolites were analyzed using thin layer chromatography (TLC), high performance liquid chromatography (HPLC), and gas chromatography-mass spectrometry (GC/MS). The major metabolites in the vitellogenic oocytes were androstenedione ($A_4$), testosterone (T), estradiol-$17{\beta}$ ($E_2$), and estrone ($E_1$). The metabolites of androgen ($A_4$ and T) were higher in the 0.50-mm oocytes than in the 0.45- and 0.48-mm oocytes, while the estrogen metabolites (E2 and E1) were lower in the 0.50-mm oocytes. These results suggest that 0.50-mm oocytes are fully vitellogenic following initiation of the maturation process.