• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.5
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• pp.624-628
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• 1999

Annual plasma levels of vitellogenin and sex steroids were investigated in relation to the gonadal development for understanding the endocrine control of reproduction in spotted flounder, Verasper variegatus. The plasma vitellogenin level was highest, 6.36 mg/ml, in November when vitellogenesis was most active. The level, thereafter, decreased to 3.81 mg/ml in December with the initiation of spawning. On the other hand, estradiol-17 $\beta$ was highest, 2.7 ng/ml, in December, and rapidly decreased in January when spawning occurred. The decreased level of estradiol-17$\beta$, around 0.2 ng/ml, remained unchanged until May. The profiles of plasma testosterone were similar to those of estradiol-17$\beta$ in the fish, The plasma 17 $\alpha$-hydroxyprogesterone level was relatively low throughout the spawning period, but increased slightly with the initiation of ovarian development, In males, the plasma testosterone and 11-ketotestosterone were highest in December when spermiation actively proceeded, but rapidly decreased during the spawning period (January).

• Clinical and Experimental Reproductive Medicine
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• v.30 no.4
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• pp.309-316
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• 2003

연구목적: 스테로이드 계통의 에스트로겐 호르몬은 막 수용체와 결합하고 DNA에 부착되어, 자궁조직에서 발현되는 많은 유전자들의 발현 양상을 조절하는 것으로 알려져 있다. 본 연구에서는 난소를 제거한 생쥐 모델을 이용하여 에스트로겐에 의해 조절되는 전사 관련 유전자(transcription factor)들을 동정하고, early up-regulation gene으로 확인된 Fos related antigen (Fra1과 Fra2) 유전자의 발현 양상을 RT-PCR과 면역염색방법으로 살펴보았다. 연구재료 및 방법: 난소 절제술을 시행한 생쥐에 에스트로겐을 피하주사하고 2, 4, 6, 12시간 간격으로 자궁조직을 적출하였다. 대조군으로는 sesame oil만을 주사한 후 2시간째에 수획한 자궁조직을 사용하였으며, 시간대별로 채취한 자궁조직(n=4)에서 RT-PCR을 수행하였다. RT-PCR을 통해 early response gene으로 확인된 Fra1과 Fra2에 대한 에스트로겐의 영향을 살펴보기 위해 estrogen receptor antagonist인 ICI 182, 780을 주사하여 유전자 발현 양상의 변화를 살펴보았다. 또한, 자궁조직내에서의 단백질 발현 부위를 관찰하기 위해 면역조직화학염색을 실시하였다. 결 과: 생쥐 자궁조직에서 에스트로겐에 의해 발현 양상의 변화가 확인된 유전자는 early up-regulation genes (CREB2, Fra-1, 2, GATA5), late up-regulation gene (E2F1), no response genes (CREB1, ATF1, GLI3, E2F3), down-regulation genes (GLI2, E2F5, GATA-2, 3, 6) 등으로 구분할 수 있었다. 그 중 early up-regulation genes에 해당하는 Fra1과 Fra2 유전자는 ICI 182, 780에 의해 그 발현이 유의하게 감소되는 것을 확인하였다(p < 0.01). 이들 단백질은 생쥐 자궁조직의 상피세포층, 기질층, 근육층에서 고루 발현되었으며, 특히 근육층에서 강한 염색정도를 관찰할 수 있었다. 결 론: 이상의 결과를 통해 Fra1과 Fra2 유전자의 발현은 에스트로겐에 의해 조절됨을 알 수 있었으며, 이들의 강한 발현이 자궁조직의 근육층에서 관찰되어 이들의 기능에 대한 연구가 필요할 것으로 생각된다.

• Korean Journal of Animal Reproduction
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• v.21 no.4
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• pp.363-371
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• 1997

The present study was carried out to investigate the effect of gonadotropin administration on blood ovarian steroid hormone in angora rabbit. Mature angora rabbits were primed for superovulation with PMSG 100IU. Eighty hours later, the rabbit were induced to ovulate with HCG 100IU. In exp 1, blood progesterone and estradiol of superovulated does were measured by radiommunoassay. Blood progesterone concentration at 93, 99, 102 and 114 hours after HCG injection were 12.9$\pm$0.5, 34.8$\pm$5.1, 12.2$\pm$2.7 and 43.4$\pm$5.8ng/ml, respectively. Mean progesterone concentration of blood collected at 99 and 114 hours after HCG injection(p<0.05). However, mean blood estradiol concentration was not changed. In exp 2, superovulated does were unilaterally ovariectomized at 96 hours after HCG injection. Blood progesterone concentration was tend to be decreased after ovariectomy. Nosignificant changes in blood estradiol concentration was observed after ovariectomy. In exp 3, superovulated does were bilaterally ovariectomized at 96 hours after HCG injection Ovariectomized does were treated with progesterone. Blood progesterone level in the rabbits treated, twice daily, with 5mg progesterone after ovariectomy was similiar to that in the superovulated intact rabbits. Blood estradiol concentration of the rabbits after bilateral ovariectomy was beyond detection range. Blood progesterone concentration was significantly decreased to 7.6$\pm$3.0ng/ml wi thin 3 hours after ovriectomy(p<0.05). However, that value was increased to 34.8$\pm$8.2ng/ml by 5 mg progesterone treatment and this elevated level was significatnly decreased to 7.3$\pm$2.4ng/ml at 12 hours after progesterone administration(p<0.05).

• Development and Reproduction
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• v.11 no.1
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• pp.21-29
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• 2007

Previously, we found progesterone receptor membrane component 2 (pgrmc2) was highly expressed in germinal vesicle (GV) stage oocytes. The present study was conducted to characterize the expression of pgrmc2, as well as pgrmc1, in the mouse gonads and embryos according to their developmental stages. We found that these membrane components were expressed in ovaries, testes, and embryos at various developmental stages in addition to oocytes. Progesterone-3-O-carboxymethyl oxime-BSA-fluorescein isothiocyanate (P4-BSA-FITC) was applied to visualize the presence of the progesterone receptor on mouse oocyte membrane, and we confirmed that immobilized progesterone is localized at surface of the oocyte. This is, at our knowledge, the first report regarding the expression of membrane component of progesterone receptor in the mouse oocytes, embryos, and gonads. The function and signal transduction pathway of progesterone receptor membrane components in oocytes requires further studies.

• Clinical and Experimental Reproductive Medicine
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• v.16 no.2
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• pp.119-130
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• 1989

본 연구는 생식주기중 폐쇄여포액내에서 생물학적， 면역학적 특성을 나타내는 GTH 의 변화를 조사하고 steroid hormone과의 상관관계를 조사하며 국부조절인자로서 의 GTH의 역 활을 조사하고자 하였다. 가임기간중 215개의 여포와 IVF과정에서 185개의 여포를 얻어 여포액내 GTH의 생물학적 또는 면역학적 활성을 측정하였다. Bioactive LH(bLH)는 생쥐의 Leydig cell-testosterone production assay, bFSH는 흰쥐의 Sertoli cell aromatase assay로 측정 하였 다. Immunological GTH(iLH , iFSH) 는 MaiaClone RIA , Delfia kits를 사용하였다. 여포액내 iLH, iFSH , ihCG 는 hyperstimulation에 의해 형성된 여포의 크기와는 무관하였다. 또 hMG, huFSH 의 처리와도 상관성이 없었다. T의 농도가 높은 여포액내의 iFSH는 현저히 낮았으며 E, P 가 고농도인 여포의 ihCG 양은 현저히 낮았다. 과배란이 유도된 난소의 여포액내 iLH는 LH specific RIA로 측정시 3mIU/ml 이하이었다. 생식주기중 여포액내 bLH, bFSH는 배란기에 현저히 증가 하였다. 혈청내 GTH B/I ratio는 엘정한 반면 여포액내 LH，FSH의 생물학적， 면역학적 활성은 미수정란을 가지거나 폐쇄된 여포내의 활성보다 현저하게 높았다. 위의 결과로 보아 여포액내 생식소자극호르온은 면역학적활성보다 높은 생물학적 활성을 가지며， 생리적 현상의 지표가 된다고 추론된다. 또한 steroid, bGTH는 여포의 선택， 폐쇄를 구분하는 지표로 사용가능하며， 여포가 폐쇄될때 여포액내 B/I ratio가 현저히 낮아지는 것으로 보아 GTH의 활성이 감소되는 것으로 판단된다.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.40 no.3
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• pp.153-159
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• 2007

We studied oocyte steroidogenesis in relation to oocyte development in the greenling, Hexagrammos otakii, a marine multiple spawner. Vitellogenic and mature oocytes were incubated in vitro in the presence or absence of $[^3H]-17\;{\alpha}-hydroxyprogesterone$ as a precursor. The major metabolites were androgens [androstenedione $(A)_4)$ and testosterone (T)] and estrogens [$17\;{\beta}-estradiol\;(E_2)$ and estrone ($E_1$)] in vitellogenic oocytes. The metabolic rate of T was lower in 1.08 to 12-mm oocytes, while that of $E_2$ increased with oocyte size. The endogenous productions of T, $E_2$ and 17 ${\alpha}-hydroxy$, 20 ${\beta}-dihydroprogesterone\;(17{\alpha}20{\beta}OHP)$ were quantified using a radioimmunoassay in the non-precursor group. The endogenous levels of T and $E_2$ were highest in 1.08 to 12-mm oocytes and $17{\alpha}20{\beta}OHP$ was produced only in 1.90 to 95-mm oocytes. The relationship between oocyte size and steroidogenesis showed that 1.08 to 12-mm oocytes are full vitellogenic following induction of the maturation process. Moreover, $17{\alpha}20{\beta}OHP$ acts as a maturation inducing hormone in H. otakii.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.37 no.3
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• pp.192-196
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• 2004

The in vitro effect of bisphenol A (BPA) on ovarian steroidogenesis of the longchin goby (Chasmichthys dolichognathus) was investigated. Oocytes taken during the maturing phase (vitellogenic, fully vitellogenic or germinal vesicle breakdown stage) were incubated with BPA (100 ng/mL) in the presence of exogenous precursor $^{3}H-17\alpha\;hydroxyprogesterone\;(^{3}H-17\alphaOHP).$ Steroids were extracted from the media and the isolated oocytes, and the extracts were separated and identified by thin layer chromatography and gas chromatography-mass spectrometry. The identities of the major metabolites were progestogens $[17{\alpha}-hydroxy,20{\alpha}-dihydroprogesterone\;(17{\alpha}20{\alpha}OHP)\;and\;17{\alpha}-hydrxy,20{\beta}-dihydroprogesterone\;(17{\alpha}20{\beta}OHP),$ androgens [androstenedione (A4) and testosterone (T)] and estrogens [estrone $(E_1)\;and\;estradiol-17{\beta}(E_2)].$ BPA treatment inhibited production of estrogens in all the maturing phases and progestogens in the germinal vesicle migrating stage. Percentage yield of estrogens was decreased with increased yield of androgens. In conclusion, BPA had an inhibitory effect on the conversion of $^3H-17\alphaOHP$ to estrogens and progestogens. These results demonstrate that BPA can act either estrogenic or anti-estrogenic effects.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.1
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• pp.15-22
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• 2000

Objective: To elucidate the location of leptin and receptors of ovary specimens obtained from patients undergoing hysterectomy by immunohistochemical staining and to determine the effect of leptin on the steroidogenesis of cultured granulosa cells. Method: In the culturing process of the granulosa cells, FSH (1 IU/ml)and leptin (50 ng/ml), IGF-I (50 ng/ml) was administered to each study group (Group I: FSH; Group II: FSH, leptin; Group III: FSH, IGF-I; Group IV: FSH, IGF-I, leptin), and the levels of estradiol, progesterone, androstenedione in the culture media was measured by radioimmunoassay. Statistical analysis was conducted by one-way ANOVA with Scheffe test. Results: The results showed that leptin and leptin receptors were both found to be strongly stained in granulosa and theca cells, and also in some interstitial cells. Leptin receptors were also observed in cultured granulosa cells. While there was no statistically significant difference in the androstnedione concentrations between the groups, estradiol concentrations was significantly decreased in Group IV ($2202.0{\pm}151.14$ pg/ml) compared to Group III ($2859.0{\pm}122.6$ pg/ml), and progesterone concentrations were also significantly decreased in Group II($4696.3{\pm}190.6$ ng/ml) and Group IV ($4517{\pm}206.78$ ng/ml) compared to Group III($5546.0{\pm}179.5$ ng/ml). Conclustion: The study result of this study suggest that leptin is directly involved in the regulation of ovarian functions, in particular steroidogenesis.

• Clinical and Experimental Reproductive Medicine
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• v.22 no.2
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• pp.155-161
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• 1995

The present investigation has been undertaken to elucidate the differentiation mechanism the uterus which is the environment of the embryo development, by demonstrating the role of ovarian steroids hormone in the decidualization of the pseudopregnant rat uterus. To determine the effect of ovarine steroids and artificial stimulation (trauma) on the differenciation of the uterine endometrium and decidualization for implantation, attempt was made to measure concentrations of serum estradiol($E_2$), progesterone($P_4$) and nuclear $P_4$ receptor in the traumatized and non-traumatized uterine tissue of the pseudopregnant rat. The results obtained are as followings : The concentration of serum $E_2$ on day 9(implantation stage) was similar in both of intact pseudopregnant rat(47.63pg/ml) and normal pregnant rat(40.71pg/ml). And among the treated groups, $E_2$ concentration was highest in the $E_2$ treated group in comparision with intact control group(relative value; 73.27%). The concentration of serum $P_4$ was also highest in the $P_4$ treated group(23.12pg/ml). Relative value of $P_4$ treated group in comparision with intact group(24.88pg/ml) was 92.93%. The nuclear $P_4$ receptor levels in the artificial traumatized groups were higher compared with the non-traumatized control groups. This study, therefore, clearly demonstrates that the methods for inducing pseudopregnant (vagina tapping;120/min) and inducing decidualization(oil injection; 0.1ml/uterine horn) appear to be effective, $P_4$ appears to be effective in the differenciation of the uterine endometrial tissue for the implantation process. Concentration of serum $P_4$ seems to be well correlated with the level of the nuclear $P_4$ receptor during the early embryo development. These results seem to be well correlated with ALPase activities in the normal and pseudopregnant rat uterus shown in the previous study.

• Development and Reproduction
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• v.13 no.4
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• pp.329-339
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• 2009

Cortisol is present in high concentration in the ovary and its receptor is expressed in the ovarian cells. Moreover, cortisol is known to have a role in steroid synthesis and cell metabolism in human granulosa and lutein cells. However, little is known of the role of cortisol presenting in high concentration in the follicles after LH surge on the granulosa-lutein cells. Therefore, the this study we evaluated the apoptosis and the production of progesterone $(P_4)$ and estradiol $(E_2)$ in the granulosa-lutein cells that are obtained during oocyte-retrieval after treatment with 5, 50, and $500{\mu}g/m\ell$ cortisol and 1 IU/$m\ell$ FSH. Results of DNA fragment analysis and TUNEL assay demonstrated that DNA fragmentation and the rate of apoptotic cells were increased in a dose-dependent manner showing a significant increase in 50 and $500{\mu}g/m\ell$ cortisol treated cells. We found, however, that FSH did not suppress the apoptosis of the cells induced by cortisol. In the results of chemiluminescence assay for $P_4$ and $E_2$, $P_4$ production was decreased by cortisol treatment, whereas $E_2$ was not changed. We also demonstrated that FSH did not inhibit the suppressive effect of GnRH on $P_4$ production as the result of apoptosis. The present study suggests that cortisol of high concentration could cause the apoptosis of human granulosa-lutein cells by suppressing the production of $P_4$. However, we need more studies to elucidate the mechanism by which cortisol induces apoptosis in human granulosa-lutein cells in view of the fact that our results are inconsistent with previous reported data.