• Clinical and Experimental Reproductive Medicine
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• v.37 no.4
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• pp.293-305
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• 2010

Ovarian hyperstimulation syndrome (OHSS) is a life-threatening iatrogenic complication of ovulation induction. Before ovarian stimulation, identification of patients vulnerable to developing OHSS is necessary. And ovarian stimulation should be started with low doses of gonadotropin or GnRH antagonist protocol. During monitoring of ovarian stimulation with risk of OHSS, coasting, low doses hCG and GnRH agonist for triggering ovulation are considered. If severe OHSS is predicted, cycle cancellation and cryopreservation of all embryos should be considered to reduce late-onset OHSS and morbidity. And metformin and dopamine agonist for reducing OHSS are being proposed as a prophylactic treatment for OHSS.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.59-59
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• 2003

ADAM은 metalloprotease/disintegrin domain을 가진 transmembrane glycoprotein으로서 지금까지 30종류 이상의 ADAM 및 10종류 이상의 ADAM-TS 단백질이 알려져 있다. 이들의 기능은 포유동물의 수정 시 sperm-egg binding과 fusion, myoblast fusion, integrin과의 결합 등에 직접 관여하거나, TNF-alpha 등의 생체신호전달물질이 세포로부터 분비될 때에 이들의 구조를 변화시켜 활성화시키는 효소로서의 작용, 그리고 dendritic cell differentiation 등에 관여하는 것으로 알려져 있다. 본 연구에서는 난소가 제거된 생쥐를 이용하여 자궁조직의 ADAM-8, -9, -10, -12, -15, -17 그리고 -TS1의 gene의 발현이 $17 \beta $-estradiol에 의하여 조절되는 지를 알아보았다. 생후 6 - 8주 된 암컷 생쥐의 난소를 제거하고, 2 주 후에 $17 \beta $-estradiol ($E_2$), progesterone ($P_4$) 혹은 이 둘 혼합액 ($E_2 + P_4$)을 sesame oil에 녹여 근육주사하였다. 2, 6, 12 시간 후 각각 자궁 조직을 얻고 유전자의 발현 양상을 알아보기 위하여 시료로부터 total RNA을 추출하여 역전사 중합효소반응 (RT-PCR)을 실시하였다. Densitometry를 이용, rpL7에 대한 ADAMS의 mRNA 발현 양을 상대적으로 분석하였다. 그 결과 ADAM-8과 -15는 6시간째에서, ADAM-10과 -TS1은 2시간째에서 sesame oil을 주사하거나 $P_4$만을 주사한 군보다 E$_2$를 주사한 군에서 mRNA의 양이 현저하게 증가하였고 ADAM-12는 2시간째에서 ADAM-17은 12시간째에서 sesame oil을 주사하거나 $P_$만을 주사한 군보다 E$_2$를 주사한 군에서 mRNA의 양이 현저하게 증가하였다. 이러한 결과로 미루어 ADAM-8, -10, -15 그리고 TS1은 progesterone에 의하여, ADAM-12와 17은 $17 \beta $-estradiol에 의하여 유전자의 발현이 upregulation 되는 것으로 생각되어진다.

• Development and Reproduction
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• v.4 no.2
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• pp.137-145
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• 2000

The regulatory mechanisms of the initiation and the formation of ovarian follicles during fetal stage of mammals are largely unknown. In addition to the gonadotropins secreted from pituitary, various growth factors, and steroid hormones are believed to be involved in the differentiation and initiation of growth of primordial follicles consisting of primordial germ cells migrated from yolk sac and streamed cells from mesonephric somatic cells. In human, primordial follicles that have already initiated differentiation at fetal stage undergo either folliculogenesis to ovulate or atresia after growth. Some of primordial follicles remain without growth for 50 years or longer. The objective of this paper is to review the mechanism of the formation, growth arrest, and initiation of primordial follicles in human fetal and neonatal ovaries.

• The Korean Journal of Zoology
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• v.10 no.1
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• pp.1-9
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• 1967

본 실험은 4 일간의 성주기가 뚜렷한 성체인 golden hamster로부터 미성숙인 난자를 적출하여 화학적배양액내에서 그의 성숙을 유도시키고 성주기에 따른 미성숙 난자의 성렬율을 관찰하는 것을 주요 목적으로 하여 행하여졌다. 한 개의 난소로부터 5-6개의 미성숙 난자를 적출하여 5% bovine serum albumin이 섞인 TC Medium 199에 넣어서$CO_2$-incubator를 이용하여 6 시간내지 24시간 동안 37$^{\circ}C$를 유지해가며, 배양액이 직접 공기와접촉하는 것을 막기 위해 유동성인 paraffin oil로 배양액을 덮고서 배양을 완수했다. 실험의 결과를 다음과 같이 완수했다. 1. Hamster의 난자의 성숙분렬을 유발시키는데 가장 적합한 배양액은 BMOC, Eagle's Medium , Waymouth's Medium 그리고 TC Medium 199의 네가지 가운데 TC Medium199이었다. 즉 이 배양액을 이용했을 때 난자가 높은 성렬도를 보여주었다. 2. 5% bovine serum alumn을 TC Medium 199에 섞어주었을 때 미성숙 난자가 성순분렬을 유기하는 율이 가장 높았다. 3. 발정기에 있는 난소로부터 얻은 난자가 가장 현저하게 높은 율로 성숙분렬을 보여주었다. 반대로 발정후기의 난자는 배양 시작 후 단시간 내에 대부분이 퇴화하였으며 발정기에 가까워 갈수록 퇴화율이 줄어들었다. 이것은 노포가 성숙분렬을 억제하는 물질을 생성하리라고 여겨지고 있긴 \ulcorner만, 동시에 이 노포는 또 한편 난자의 배란뒤에까지도 생명력을 유지할수 있는 능력을 발정기에 이르기까지의 기간동안 난자에게 부여하며, 난자는 이 능력을 배란전까지 축적하게 되며 이 때문에 노포로부터 유리되어 나온 난자가 장기간 그 생명력을 유지해나가는 것이라고 추정된다. 4. Paraffin oil 로 공기를 차단하여서 배양한 난자나 혹은 watch glass를 이용하여 공기와 접촉시킨 난자에서나 모든 비슷한 율로 성숙분렬을 보여주었다. 이로 보아 조작이 까다로운 paraffin oil을 이용하는 방법보다는 손쉽게 배양할 수 있는 watch glass 의 방법이 오히려 유용하다고 할 수 있다.

• Journal of Embryo Transfer
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• v.16 no.2
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• pp.99-106
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• 2001

This study was undertaken to compare the efficiency of recovery rate and development rate of follicular oocytes collected either by aspiration or by slicing method. The follicular oocytes collected by the two methods matured in TCM199 supplemented with 10% steer serum at 39$^{\circ}C$ in a humidified atmosphere of 5% $CO_2$in air. After 22 h of culture, the oocytes were inseminated with frozen-thawed semen (2$\times$10$^{6}$ sperm/ml of final concentration) prepared with Percoll-density gradient in IVF-TALP medium for 16 h. Later, sets of 15 presumptive zygotes were transferred into 50 $\mu$L, droplets of CR1aa medium. On day 4 of the culture, embryos were transferred to TCM199 until day 9. The percentages of nuclear maturation to pre-metaphase II in the oocytes collected by aspiration are significantly (P<0.05) higher than that by slicing (83% vs. 62%, respectively). The mean number of oocytes recovered by slicing per ovary is significantly (P<0.05) higher than that by aspiration (15.1 vs. 6.7, respectively). Although the rates of cleavage and development to blastocyst of oocytes collected b)\\\\`aspiration are significantly (P<0.05) higher than that by slicing, the number of transferable embryos obtained by slicing method is significantly (P<0.05) higher than that by aspiration. From the results. we may conclude that slicing method is better than aspiration method for obtaining large number of transferable embryos per ovary.

• Journal of Radiation Protection and Research
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• v.23 no.2
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• pp.89-96
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• 1998

To assess the radioprotective effects of follicle stimulating hormone (FSH) on ovarian follicles, 3 week-old female mice were irradiated with 8.33 Gy of ${\gamma}$-ray (group R) and followed by 5 IU ip-injection of FSH (group RF). For control groups, 5 IU of saline (group C) or 5 IU of FSH (group F) was ip-injected. Ovaries were collected 0h, 6h, 12h, 14, 2d, 4d, and 8d after irradiation or saline/FSH injection, and followed by fixation in neutral buffered formalin for routine histochemistry. Immunohistochemistry was used to assess the status of follicles and DNA fragmentation was analyzed by agarose gel electrophoresis for total DNA. Staining specific for apoptotic follicles showed high intensity at 6h and 12h in group R and RF On the other hand, staining specific for proliferating follicles showed noticeably high intensity at 8d in group R and Rf. DNA fragmentation of 185bp increased with time in all experimental groups. Especially 370bp appeared at 6h in group R, then disappeared after 1d. In case of group RF, it appeared at 12h and disappeared after 1d. From the above results, the irradiated antral follicles become completely disappeared from 4d to 8d, and then new follicles started to grow again at 8d. FSH had delaying or suppressing effects on follicular atresia after irradiation. In addition, it became clear that radiation-induced follicular atresia was mediated by granulosa cell apoptosis.

• Development and Reproduction
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• v.7 no.2
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• pp.119-126
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• 2003

In the previous study, we obtained list of differentially expressed genes between postnatal day 1 and day 5 mouse ovaries using suppression subtractive hybridization(SSH) and found that MT Oansposon-like element, clone MTi7(MTi7) was one of the highly expressed genes in the day 5 mouse ovary(Park et at., 2002). Results of in situ hybridization and RNA interference revealed that the expression of MTi7 is oocyte-specific in the ovary and may be involved in the regulation of oocyte maturation(Park et at., 2003). At present, MTi7 sequence has been known only in the mouse. Therefore, the present study was accomplished 1) to identify MTi7 sequence in the other mammalian species, such as bovine, porcine, rat, and human, and 2) to evaluate the flanking sequence of the mouse MTi7 since it has transposon characteristics. Using ovarian cDNAs derived from low different species, we cloned and identified new MTi7 sequence showing a high degree of sequence homology with the mouse MTi7(87∼98%). By using inverse PCR, we found that the mouse MTi7 may intercalated the beta-carotene 15, 15'-monooxygenase(Bcdo) gene and/or serine protease inhibitor, Kunitz Dpe I(Spint 1) gene. By finding the MTi7 sequences in the other mammalian species and the flanking gene of the MTi7 in mouse, it is expected to reveal the role(s) of MTi7 in the oogenesis as well as folliculogenesis in the near future.

• The Korean Journal of Zoology
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• v.39 no.3
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• pp.266-272
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• 1996

Experiments were carried out to ascertain whether gonadotropin or a phorbol ester (12-O-tetradecanoyl phorbol-13-acetate, TPA) induces oocyte ovulation and stimulates prostaglandin synthesis by Rana ovarian follicles in vitro. Rana nigromaculata collected from underground in spring were utilized for the present experiment. Treatment of frog pituitary homogenate (FPH) or TPA to ovarian fragments in culture induced oocyte ovulation in a dose dependent manner and stimulated prostaglandin F2a (PGF$_2$$\alpha$ synthesis. Both treatruents were more effective in inducing the ovulation and PGF$_2$$\alpha$ secretion by the follicles obtained in May than those in April. A Protein kinase C inactivator, 1-(5-isoquinolinyl-sulfonyl)-2-methyl-piperazine (H-7), or cyclooxygenase inhibitor, indomethacin (IM) suppressed the FPH- or TPA-induced PGF$_2$$\alpha$ production, but IM failed to suppress the FPH- or TPA-induced ovulation. Time course of oocyte ovulation and PGF$_2$$\alpha$ secretion by FPH and TPA treatments were very similar to each other. FPH stimulated progesterone secretion by the follicle but TPA failed to do so. Taken together, the data presented here suggest that protein kinase C (PKC) in follicle play a role in the ovulation process of Rana nigromaculata, probably via prostaglandin synthesis.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.2
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• pp.151-163
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• 2003

연구목적: Estrogen은 포유류의 생리주기와 착상과정에서 중요한 조절인자로 작용한다. 본 연구에서는 난소 절제된 생쥐의 자궁에서 estrogen에 의해 직접 또는 간접적으로 조절되어 발현하는 유전자를 분석하고자 하였다. 연구재료 및 방법: 생후 8주된 생쥐의 양쪽 난소를 절제하고 14일 동안 회복기간이 지난 후, estrogen (300 ng/mouse)을 피하로 주사하였다. Estrogen 주사 후 6, 12시간째 자궁을 적출하여 cDNA microarray와 laser capture microdissection (LCM) 기술을 이용하여 estrogen에 의해 조절되는 유전자의 시공간적인 발현 양상을 조사하였다. 결 과: Estrogen 주사 후 6시간째에는 조사된 전체 유전자 가운데 0.9% (증가 22, 감소 49), 12시간째에는 8.4% (증가 351, 감소 287)에 해당되는 유전자가 두 배 이상 증가 혹은 감소하는 결과를 보였다. 또한 일부 증감된 유전자를 선택한 후 LCM 기술을 이용하여 시공간적인 발현양상을 확인한 결과 자궁내막상피세포에서만 estrogen에 의해 유전자의 발현이 증가되는 일부 유전자를 선별하였다. 결 론: 이상의 결과들을 종합해보면 1) estrogen에 의해 조절되는 유전자의 수나 증감의 정도는 12시간 이후에 더 많고, 크게 조절되며, 2) 유전자의 조절부위가 자궁의 특이적인 세포층에서 시공간적으로 조절됨을 의미한다. 이러한 유전자의 정보는 생리주기 또는 착상과정의 분자생물학적 기작을 이해하는 데 도움이 될 것이다.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.12
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• pp.1930-1939
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• 2013

It has been reported that Chungkukjang, one of Korean traditional fermented soybean products, may improve hypertension, diabetes, and hyperlipidemia. In this study, we sought to investigate the immunoenhancing effects of Chungkukjang in ovariectomized mice. For the first period, female SLC ddy mice were either sham-operated (Sham; n=27) or ovariectomized (OVX; n=27). As a basal diet, ovariectomized mice were fed low-calcium diet for faster induction of osteoporosis for six weeks, and those in the Sham group were fed AIN-76 diet. For the second period, half of the OVX group (n=9) and the Sham group (n=9) were fed a Chungkukjang-based diet (CKJ); whereas the other half (OVX; n=9/ Sham; n=9) were fed a casein-based diet (CSI) for 8 weeks. After a second period, we collected the blood via heart puncture and measured the splenocytes proliferation, T lymphocyte subsets by flowcytometry, and levels of serum cytokines (IL-2, IL-4, IL-6, IL-10, IFN-${\gamma}$ and TNF-${\alpha}$) by ELISA assay. The OVX+CKJ group showed higher splenocytes proliferation, higher ratio of CD4/CD8, and lower levels of IL-6 and TNF-${\alpha}$ cytokines compared to the OVX+CSI group. The Sham+CKJ group showed cytokine productions, such as higher levels of IL-10 and IFN-${\gamma}$, and lower levels of IL-6 and TNF-${\alpha}$ compared to the Sham+CSI group. The result of this study suggests that Chungkukjang may lower the proinflammatory cytokine levels in both the OVX and Sham groups. In addition, Chungkukjang could make a balance of T cell subset proliferations and enhance the splenocyte proliferations in the OVX group.