• Archives of Pharmacal Research
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• v.20 no.6
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• pp.525-528
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• 1997

Steroid $9{\alpha}$.-hydroxylase is a key enzyme system in steroid nucleus degradation in company with ${\Delta}$-dehydrogenase. To examine $9{\alpha}$-hydroxylase activity during microbial transformation of steroids, 9(11)-dehydro-$17{\alpha}$-methyl-testosterone was adopted as a stable substrate for preventing the rupture of steroid nucleus. Using Nocardia restrictus ATCC 14887 capable of introducing a $9{\alpha}$-hydroxyl group into steroids, $9{\alpha}$,$11{\alpha}$-oxido-$17{\beta}$-hydroxy-$17{\alpha}$-methyl-4-androstene-3-one and $9{\alpha}$-hydroxyl group into steroids,$9{\alpha}$,$11{\alpha}$-oxido-$17{\beta}$-hydroxy-$17{\alpha}$-methyl-1,4-androstadiene-3- one were obtained. These microbiologically transformed products could be used as reference compounds in the enzyme assay.

• 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.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.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.

• 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.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.6
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• pp.638-642
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• 2001

To investigate the production of $C_{21}$-steroids during the spawning period of spotted sea bass, Lateolabrax maculatus, we have incubated maturing and ovulating follicles with radiolabeled pregnenolone and $17\alpha$-hydroxyprogesterone for 24 hours. The resulting metabolites were analyzed by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC), When maturing follicles ($700\sim800{\mu}m$ in diameters) were incubated with radiolabeled precursors, $C_{21}$-metabolites were corticosteroids and $17\alpha$-hydroxy, $20\beta$-dihydroprogesterone ($17\alpha20\beta OHP$). When ovulation follicles ($1,000\sim1,150{\mu}m$ in diameters) were incubated with radiolabeled precursors, the major $C_{21}$-metabolites were $17\alpha20\beta OHP$, $17\alpha$,$20\beta$, 21-trihydroxy-4-pregnen-3-one ($17\alpha20\beta21P$), and corticosterone. Additional chromatography by TLC and HPLC confirmed the presence of radioactive $17\alpha20\beta OHP$ in the maturing follicles, and $17\alpha20\beta OHP$,$17\alpha20\beta21P$ and corticosterone in ovulating follicles. Although $17\alpha20\beta OHP$ was found in a small peak, the synthesis of this steroid suggests that it may play a role in regulating the oocyte maturation process. Whereas ovulation is regulated by both $17\alpha20\beta OHP$ and $17\alpha20\beta21P$ in the spotted sea bass. In addition, an unusual finding was the biosynthesis of corticosterone. Whether this production is responsible for the ovulation, and is an area requiring continued research.

• 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.

• Journal of the Korean Chemical Society
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• v.19 no.3
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• pp.174-178
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• 1975

The intermediate, 17${\alpha}$-methyl-4-aza-5${\alpha}$-androstan-17${\beta}$-ol(Ⅸ) required for the synthesis of 4-(${\beta}$-guanidinoethyl)-17${\alpha}$-methyl-4-aza-5${\alpha}$-androstan-17${\beta}$-ol(V) was obtained through a reaction of 17${\alpha}$-methyl-3,5-seco-4-norandrostan-17${\beta}$-ol-5-on-3-oic acid(VI) with ammonium hydroxide followed by two reductions(platinum dioxide with hydrogen and lithium aluminium hydride). Condensation of Ⅸ with chloroacetonitrile under anhydrous condition, followed by reduction of the nitrile with lithium aluminium hydride gave 4-(${\beta}$-aminoethyl)-17${\alpha}$-methyl-4-aza-5${\alpha}$-androstan-17${\beta}$-ol(XI). The reaction of XI with 2-methyl-2-thiopseudourea or 3,5-dimethylpyrazole-1-carboxamidine, or cyanamide provided the title compound, V. Relaxation of the nictitating membrane, in the absence of mydriasis, is considered to be evidence of adrenergic neurone blockade. Thus the test compound(V) resembles that of the classical adrenergic neurone blocking agents.

• 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.

• 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)