• Fisheries and Aquatic Sciences
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• v.4 no.2
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• pp.70-74
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• 2001

To examine the production of steroids with potential oocyte maturation-inducing activity in the spotted flounder, Verasper variegatus, we have incubated post-vitellogenic oocytes (0.82­0.95mm in diameters) with radiolabeled pregnenolone and $17\alpha-hydroxyprogesterone$. The resulting metabolites were analyzed by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). The two main metabolites (progestogens) found in both incubations co-migrated with $17\alpha-hydroxy$, $20\alpha-dihydroprogesterone$ $(17\alpha, 20\alpha OHP)$ and $17\alpha-hydroxy,\;$20\beta-dihydroprogesterone$ (17 a20{30HP). Additional chromatography by HPLC and TLC confirmed the presence of radioactive $17\alpha, 20\alpha OHP$ and a large amount of unknown metabolite. The present study did not reveal in vitro formation of $l7\alpha 20\beta OHP$. Although 1$l7\alpha 20\beta OHP$ was found in a small amount, the synthesis of this steroid suggests that it may play a role in regulating the oocyte maturation process in the spotted flounder.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2002.10a
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• pp.253-254
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• 2002

경골어류의 난소 성숙 과정은 여포층에서 생성ㆍ분비되는 성 스테로이드 호르몬에 의해 조절된다. 성 스테로이드 호르몬 중에서도 C2l-스테로이드가 난소 성숙 과정에 중요한 역할을 하는 것으로 알려져 있으며, 특히 20$\alpha$와 20$\beta$-hydroxy group을 가지고 있는 스테로이드, 17 $\alpha$-hyoxy, 20 $\alpha$-dihydroprogesterone (17 $\alpha$ 20 $\alpha$ OHP)과 17$\alpha$-hyoxy, 20$\beta$-dihydroprogesterone (17$\alpha$20$\beta$OHP)가 성숙유도에 가장 효과적인 것으로 보고하고 있다 (Canario and Scott, 1989; 1990). (중략)

• Korean Journal of Animal Reproduction
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• v.14 no.4
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• pp.273-280
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• 1990

This study was conducted to find out the changes of progesterone and 20$\alpha$-dihydroprogesterone(OHP) levels in the serum of female rats by progesterone-tube implantation during late pregnancy. One hundred and twenty rats, 10-13 weeks old, were offered for this experiment. Blood samples were taken from the rats on 18, 20, 21 and 22days of pregnancy. The rats were implanted with silicon tubes filled with pregesterone on day 15 of pregnancy. The progesterone and 20$\alpha$-OHP in serum were assayed by radioimmunoassay. The results obtained are summarized as follows : 1. The progesterone levels in rats of control group showed 126.36$\pm$20.19ng/ml on 18days and gradually decreased to 69.3$\pm$11.9, 29.2$\pm$1.8ng/ml 20 and 21 days. 2. The progesterone levels after progesterone-tube implantation showed 120.1$\pm$8.5, 59.01$\pm$3.1, 85.33$\pm$17.9 and 62.9$\pm$6.1ng/ml on 18, 20, 21 and 22days of pregnancy, respectively. 3. The 20$\alpha$-OHP levels in rats of control group showed 68.5$\pm$8.0ng/ml on 18 days and gradually increased to 139.9$\pm$3.7 and 141.4$\pm$6.7ng/ml on 21 and 22days. 4. The 20$\alpha$-OHP levles after progesterone-tube implantation showed 108.5$\pm$32.7 and 106.4$\pm$10.6ng/ml on 18 and 20days and increased to reach the peak level at 21days(225.01$\pm$9.9ng/ml) and rapidly decreased on 22days(85.9$\pm$10.7ng/ml).

• Journal of Aquaculture
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• v.11 no.1
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• pp.119-124
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• 1998

The relative effectiveness of C21-steroids and human chorionic gonadotropin(HCG) on maturation and ovulatin was investigated in vitro using the isolated oocytes or ovarian fragments from the sea bass, Lateolabrax japonicus. ${\alpha}$-hydroxy, 20${\beta}$-dihydroprogesterone(17${\alpha}$20${\beta}$OHP : 5, 50, 500, 1000ng/ml), 17${\alpha}$-hydroxy, 20${\alpha}$-dihydroprogesterone(17${\alpha}$20${\alpha}$OHP : 5, 50, 500, 1000ng/ml) and HCG (5, 50, 500IU/ml) were effective in inducing oocyte maturation, GVM (germinal vesicle migration) or GVBD(germinal vesicle breakdown), compared to control except 17${\alpha}$20${\beta}$OHP and 17${\alpha}$20${\alpha}$OHP at 5ng/ml. 17${\alpha}$20${\beta}$OHP showed the greatest effect on oocyte maturation at 50ng/ml. A combination of 17${\alpha}$20${\beta}$OHP(50ng/ml) and HCG(500IU/ml) led to a significant increase (p<0.05) in GVBD when compared with 17${\alpha}$20${\beta}$OHP or HCG alone. These findings suggest that the two in combination acts synergistically to induce GVBD. 17${\alpha}$20${\beta}$OHP (1~1000ng/ml) and HCG(1~500IU/ml) also induced ovulation in ovarian fragments at all concentrations used ; more effective at lower concentrations(1~50ng/ml or IU/ml). It was shown that HCG was more potent in inducting ovulatin than 17${\alpha}$20${\beta}$OHP.

• Development and Reproduction
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• v.12 no.1
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• pp.107-112
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• 2008

To investigate the $C_{21}$-steroids produced from maturating oocytes in the longchin goby, Chasmichthys dolichognathus, the oocytes ($0.74{\sim}0.97\;mm$) were incubated with radiolabeled $17{\alpha}$-hydroxyprogesterone ($^3H-17{\alpha}OHP$) for 24 hours. The resulting metabolites were analyzed by thin layer chromatography and identified by gas chromatography-mass spectrometry. Two $C_{21}$-steroids, $17{\alpha}$-hydroxy, $20{\alpha}$-dihydroprogesterone ($17{\alpha}20{\alpha}P$) and $17{\alpha}$-hydroxy, $20{\beta}$-dihydroprogesterone ($17{\alpha}20{\beta}P$), were converted from $^3H-17{\alpha}OHP$ in the maturing oocytes. These two main metabolites were detected at 0.80 mm diameter oocytes or greater. In addition, the effects of these metabolites on in vitro germinal vesicle breakdown (GVBD) were tested. The sensitivity of oocytes to the induction of GVBD was greater at $17{\alpha}20{\beta}P$ than $17{\alpha}20{\alpha}P$. This result showed that $17{\alpha}20{\beta}P$ is a major maturation inducing steroid (MIS) in longchin goby, suggesting $17{\alpha}20{\alpha}P$ may play a role in regulating the oocyte maturation process.

• Journal of Aquaculture
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• v.9 no.1
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• pp.57-63
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• 1996

The relative effectiveness of steroids and human chorionic gonadotropin (HCG) on germinal vesicle breakdown (GVBD) was investigated in vitro using the isolated oocytes (folliculated oocytes) from the spotted halibut, Verasper variegatus. Among the steroids tested, $17\alpha-hydroxy,\;20\beta-dihydroprogesterone\;(17\alpha20{\beta}OHP)$ was more effective than progesterone (P4) and $17\alpha-hydroxyprogesterone\;(17{\alpha}OHP$). In comparing $17\alpha,\;20{\beta}OHP$ with HCG, both $17\alpha20{\beta}OHP$ and HCG were effective in inducing GVBD at all concentrations : $17\alpha20{\beta}OHP$ was effective even at lower concentrations ($10\~50$ ng/ml). HCG at three concentrations used (50, 100, 500 IU/ml) showed no significant differences in inducing GVBD.

• Korean Journal of Animal Reproduction
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• v.16 no.2
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• pp.133-139
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• 1992

The study was conducted to find out the changes of progesterone and 20$\alpha$-dihydroprogesterone (20$\alpha$-OHP) levels during the oestrous cycle in Korean native goats. 4 cyclic goats were offered for this experiment. Blood samples were taken from jugular vein on day 0, 1, 3, 5, 7, 9, 11, 13, 15, 17 and 19 during the oestrous cycle, then the next oestrous day. The serum levels of progesterone and 20$\alpha$-OHP were measured by radioimmunoassay. The progesterone concentration in seurm of the cyclic goats was 0.29$\pm$0.06ng/ml on the first day of oestrous (day 0), increased to 5.29$\pm$0.73ng/ml on day 9, reached to a peak level of 5.73$\pm$0.61ng/ml on day 13, and thereafter decreased to 0.35$\pm$0.30ng/ml on day 1. The serum level of 20$\alpha$-OHP during the oestrous cycle was 0.42$\pm$0.33ng/ml on day 0 and then decreased to 0.28$\pm$0.01ng/ml on day 5. This basal level was maintained until day 13, increased gradually, and reached a peak level of 0.62$\pm$0.05ng/ml on day 19. From the above results, it was suggested that the enzyme 20$\alpha$-hydroxysteroid dehydrogenase(20$\alpha$-HSD) catalyzing the conversion of progesterone to a biologically inactive steroid, 20$\alpha$-OHP, should be active in the luteal cells during the oestrous cycle in Korean native goats.

• Korean Journal of Animal Reproduction
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• v.16 no.2
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• pp.141-147
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• 1992

The present study was conducted to find out the changes of progesterone and 20$\alpha$-dihydroprogesterone(20$\alpha$-OHP) levels during the gestation period in Korean native goats. 4 pregnant goats were offered for this experiment. Blood samples were taken from jugular vein on Day 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140 and 145 of the pregnant goats. The serum levels of progesterone and 20$\alpha$-OHP were measured by radioimmunoassay. The progesterone level in goat serum during the gestaton was low as 2.94$\pm$0.21 at 5 days, and then increased to 4.51$\pm$0.25ng/ml at 10 days of gestation and increased greatly from Days 60 and reached a peak level of 7.21$\pm$0.58ng/ml at 100 days of gestation, and thereafter decreased to 6.01$\pm$0.57, 5.26$\pm$0.64ng/ml on Days 130, 140 of gestation, and decreased to 4.05$\pm$0.52ng/ml on Days 145 of pregnancy. The serum level of 20$\alpha$-OHP during the gestation was low as 0.3~0.4ng/ml in the early stage of pregnancy, and increased gradually and increased to 0.85$\pm$0.06ng/ml, 0.97$\pm$0.08ng/ml on Days 90 or 100, and then increased to 1.18$\pm$0.18, 1.25$\pm$0.21ng/ml on Days 140 or 145 of gestation. While the serum levels of progesterone during the luteal regression decreased, the 20$\alpha$-OHP increased continuously. From the above results, it was concluded that the enzyme 20$\alpha$-hydroxysteroid dehydrogenase (20$\alpha$-HSD) catalyzing theconversion of progesterone to a biologically inactive steroid, 20$\alpha$-OHP was active properly in the luteal cells what the levels of progesterone decreased and the levels of 20$\alpha$-OHP increased during the late pregnancy in Korean native goats.

• Korean Journal of Animal Reproduction
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• v.16 no.2
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• pp.149-155
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• 1992

The present study was conducted to find out the changes of progesterone and 20$\alpha$-dihydroprogesterone(20$\alpha$-OHP) levels before and after parturition, 4 pluriparous goats were offered for this experiment. Blood samples were taken from jugular vein on Days, 5, 3, 2 and 1 before parturition, the day of parturition, 1, 3, 5, 7 and 9 after parturition. The blood samples were centraifuged and stored at -2$0^{\circ}C$ until hormone assay. The serum levels of progesterone and 20$\alpha$-OHP were measrued by radioimmunoassay. The changes of serum progesterone level during peripartum period were characterized as a remarkable decrease. The progesterone level was 4.05$\pm$0.52ng/ml on 56 days before parturition and decreased to 2.24$\pm$0.38ng/ml on 1 day before parturition and 0.79$\pm$0.09ng/ml on the day of parturition and the basal level was maintained through 9 days of postpartum period. The serum level of 20$\alpha$-OHP during the peripartum period was 1.25$\pm$0.21ng/ml on 5 days before paturition and increased to 1.32$\pm$0.25 on 3 days and 1.59$\pm$0.24ng/ml on 1 day before parturition, and reached a peak level of 1.78$\pm$0.25ng/ml just prior to parturition and then decreased greatly to 0.31$\pm$0.03ng/ml on 1 day postpartum and the basal level was remained until 9 days postpartum. The high serum level of 20$\alpha$-OHP, which was peak just prior to parturition, was maintained for 2 days following the onset of remarkable decrease in the serum level of progesterone. From the above results, it was concluded that the enzyme 20$\alpha$-hydroxysteroid dehydrogenase (20$\alpha$-HSD) catalyzing the conversion of progesterone to a biologically inactive steroid, 20$\alpha$-OHP was active properly in the luteal cells in Korean native goats.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.3
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• pp.342-347
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• 2003

The enzyme $20{\alpha}$-hydroxysteroid dehydrogenase ($20{\alpha}$-HSD) catabolizes progesterone to $20{\alpha}$-dihydroprogesterone ($20{\alpha}$-OHP), and is appeared in rat corpora luteal and placenta. A polled samples of 10-15 placental and ovarian tissues collected from each individual rat were subjected to measurement of $20{\alpha}$-HSD activity. A $20{\alpha}$-HSD activity in the cytosol fraction was based on the generation of NADPH. In this study, it is designed to study cytosolic $20{\alpha}$-HSD activity in rat ovarian and placenta during pregnancy, and its relationship to embryonic mortality. It was found that from days 5 to 18 of pregnancy the $20{\alpha}$-HSD activities steady by decreased but at parturition time rapidly increased in ovary. On the other hand, placental cytosolic $20{\alpha}$-HSD activities were high detected from days 8 to 10 of pregnancy, not detectable from days 11 to 20 of pregnancy, but again very high at the time of parturition. Analysis of DEAE column chromatography revealed that two different types of $20{\alpha}$-HSD (HSD-1 and HSD-2) were found with similar activity in the placental cytosol on day 10 of pregnancy. The number of fetuses on day 10 of pregnancy was 15.4 and decreased significantly to 12.9 on day 12. The results suggested that expression of $20{\alpha}$-HSD in the placental tissues seems to be related the number of fetal survived in the specific time (days 11 and 12) which spontaneous fetal loss occurs.