• 한국가축번식학회지
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• 제22권3호
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• pp.277-286
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• 1998

The effect of MAP kinase activity on maturation of porcine oocytes was investigated. MAP kinase was detected by immunofluorescence staining in nucleus of oocytes just before entering GVBD (germinal vesicle breakdown)stage. In a Western blot with GV (germinal vesicle) from these oocytes (cultured for 25 hours), a shift of MAP kinase band a, pp.ared, suggesting an activated stage of the kinase. No activity was shown in the blot with GV isolated from, oocytes cultured for 0 hour. To confirm that activation of MAP kinase induce GVBD, we microinjected MAP kinase purified from matured oocytes starfish into the cytoplasm of oocytes in GV stage (cultured for 0 hour). The injected MAP kinase did not cause early a, pp.arance of GVBD. No oocytes showed GVBD state until 20 hours of culture. Activity of MAP kinase did not increase significantly after the injection. When the exogenous MAP kinase was injected into GV, GVBD was induced in about 20% of oocytes cultured for 5 to 10 hours. These results suggest that MAP kinase is traslocated to nucleus and function as a factor inducing GVBD.

• 한국발생생물학회지:발생과생식
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• 제23권1호
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• pp.1-9
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• 2019

The ability of oocytes to undergo normal fertilization and embryo development is acquired during oocyte maturation which is transition from the germinal vesicle stage (GV), germinal vesicle breakdown (GVBD) to metaphase of meiosis II (MII). Part of this process includes redistribution of inositol 1, 4, 5-triphosphate receptor (IP3R), a predominant $Ca^{2+}$ channel on the endoplasmic reticulum membrane. Type 1 IP3R (IP3R1) is expressed in mouse oocytes dominantly. At GV stage, IP3R1 are arranged as a network throughout the cytoplasm with minute accumulation around the nucleus. At MII stage, IP3R1 diffuses to the entire cytoplasm in a more reticular manner, and obvious clusters of IP3R1 are observed at the cortex of the egg. This structural reorganization provides acquisition of $[Ca^{2+}]_i$ oscillatory activity during fertilization. In this review, general properties of IP3R1 in somatic cells and mammalian oocyte are introduced.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2002년도 제37회 국제학술심포지움 및 추계학술대회
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• pp.67-68
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• 2002

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2002년도 제35회 학술심포지움 및 춘계학술대회
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• pp.78-78
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• 2002

• Asian-Australasian Journal of Animal Sciences
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• 제17권9호
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• pp.1227-1231
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• 2004

Pig oocytes contain high levels of lipids in the ooplasm, which reduces the visibility of the germinal vesicle (GV) under microscopic examination. Therefore, the purposes of this study were to investigate the effects of relative centrifugation force (RCF) on the visibility and maturation rates of the GV stage oocytes after centrifugation. In Experiment 1, cumulus-oocyte-complexes (COCs) were collected from slaughterhouse ovaries and randomly allocated to different RCFs (3,000 rpm: 970 g; 6,000 rpm: 3,900 g; or 10,000 rpm: 10,840 g) for 10 or 20 min. Percentages of visible GV were 76-79% in the oocytes centrifuged with 10,000 rpm, which were significantly higher (p<0.01) than those with 3,000 and 6,000 rpm. No significant differences in GV visibility were observed among oocytes with different lengths of centrifugation (p<0.05) regardless of the RCFs. In esperiment 2, the maturation rate of the oocyte was found significantly lower in the 20 min than in the 10 min group received 10,840 g of RCF (30 vs. 75%, p<0.05). In conclusion, the GV of porcine oocytes can be clearly visible by centrifugation at 10,840 g for 10 min without compromising their subsequent maturation rates and a longer centrifugation time (20 min) had no beneficial influence on the visibility of GV stage pig oocytes.

• Clinical and Experimental Reproductive Medicine
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• 제44권2호
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• pp.79-84
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• 2017

Objective: Optimizing in vitro maturation (IVM) media to achieve better outcomes has been a matter of interest in recent years. The aim of this prospective clinical trial was to investigate the effects of different media on the IVM outcomes of immature oocytes at the germinal vesicle (GV) stage. Methods: A total of 400 immature oocytes at the GV stage with normal morphology were retrieved from 320 infertile women aged $31{\pm}4.63years$ during stimulated intracytoplasmic sperm injection (ICSI) cycles. They were divided into groups of homemade IVM medium (I, n = 100), cleavage medium (II, n = 100), blastocyst medium (III, n = 100), and Sage IVM medium (IV, n = 100) and cultured for 24 to 48 hours at $37^{\circ}C$. ICSI was performed, and the rates of fertilization and embryo formation were compared across the four groups. Results: In the 400 retrieved GV oocytes, the total maturation rates showed significant differences in groups I to IV (55%, 53%, 78%, and 68%, respectively, p<0.001). However, there were no significant differences in the fertilization, embryo formation, or arrest rates of metaphase II oocytes across these groups. In all groups, GV maturation was mostly completed after 24 hours, with fewer oocytes requiring 48 hours to mature (p<0.01). Moreover, the rate of high-quality embryos was higher in group IV than in the other groups (p=0.01). Conclusion: The quality of the IVM medium was found to affect clinical IVM outcomes. Additionally, blastocyst medium may be a good choice in IVM/ICSI cycles as an alternative IVM medium.

• 한국발생생물학회지:발생과생식
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• 제15권3호
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• pp.223-230
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• 2011

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental contaminants derived from incomplete combustion of carbons and crude oil. In this study, we investigated the effects of benzo[a]pyrene (B[a]P), a representative PAHs on in vitro sex steroid hormone production and germinal vesicle breakdown (GVBD) using isolated oocytes of longchin goby (Chasmichthys dolichognathus) and chameleon goby (Tridentiger trigonocephalus). Oocytes in diameters of 0.8-0.9 (end vitellogenic stage) and 0.9-1.0 mm (germinal vesicle migratory stage) from longchin goby and 0.5 mm (fully vitellogenic stage) from chameleon goby were used. In GVBD assay, B[a]P at 10 nM stimulated GVBD in the oocytes of 0.8-0.9 mm from longchin goby. B[a]P at 1 nM stimulated GVBD in the oocytes with diameter 0.5 mm from chameleon goby. In steroid production from oocytes of longchin goby, B[a]P at 100 nM decreased testosterone (T) production, B[a]P at 1,000 nM increased estraiol-17 (J (E2) production and 10 and 100 nM increased $17,20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$) production in the oocytes with diameter 0.8-0.9 mm. B[a]P at 1,000 nM increased E2 production, 100 and 1,000 nM increased $17{\alpha}20{\beta}P$ production in the oocytes with diameter 0.9-1.0 mm. In steroid production of oocytes from chameleon goby, B[a]P at 1,000 nM increased $E_2$ production. B[a]P at 10 nM increased $17{\alpha}20{\beta}P$ production. In the ratio of $E_2$ to T ($E_2$/T), B[a]P at 100 and 1,000 nM increased $E_2$/T in the oocytes of longchin goby. B[a]P at 100 nM also increased $E_2$/T in the oocytes of chameleon goby. Taken together, these results suggest that B[a]P have not only weak estrogenic effects but progestogenic effects on oocyte maturation.

• 한국양식학회지
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• 제9권1호
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• pp.57-63
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• 1996

범가자미 난모세포의 최종성숙(GVBD=germinal vesicle breakdown) 유도를 위한 HCG (human chorionic gonadotropin)와 스테로이드 호르몬들의 효능 비교실험이 in vitro에서 이루어졌다. 실험에 사용한 스테로이드 호르몬, progestogens (progesterone, $17\alpha$-hydroxyprogesterone, $17\alpha$-hydroxy, $20\beta$-dihydroprogesterone)중에서는 $17\alpha$-hydroxy, $20\beta$-dihydroprogesterone ($17\alpha20{\beta}OHP$)이 매우 효과적으로 나타났으며, $17\alpha20{\beta}OHP$와 HCG와의 비교실험에서는 이들 모두 GVBD 유도에 효과적인 것으로 나타났다. 특히 $17\alpha20{\beta}OHP$는 저농도($10\~50$ ng/ml)에서도 효과적이며, HCG는 사용한 농도(50, 100, 500 IU/ml)사이에 유의한차를 보이지 않았다.

• Reproductive and Developmental Biology
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• 제32권3호
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• pp.147-152
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• 2008

The aim of this study was to investigate what components of porcine epididymal fluid (pEF) influences the nuclear maturation of porcine germinal vesicle oocytes. Porcine cumulus-oocytes complexes from follicles were cultured in TCM 199 containing pEF. After 48 h cultures, oocytes were examined for evidence of GV breakdown, metaphase I, anaphase-telophase I, and metaphase II. Maturation rate of oocytes was significantly increased in media supplemented with 10% pEF during in vitro maturation (IVM) than in those without pEF. When lipid component of pEF was removed by treating n-heptane, no significant difference was observed in maturation of oocytes between n-heptane treatrment and intact pEF group. However, the proportion of oocytes reaching at metaphase II (M II) was significantly (p<0.05) decreased in the oocytes cultured in media containing trypsin-treated pEF compared to those in media with intact pEF. When porcine GV oocytes were matured in the medium supplemented with intact pEF or pEF heated at $56^{\circ}C$ and $97^{\circ}C$, rates of oocytes remained at GV stage were 11.7%, 29.4% and 42.0%, respectively. However, there were no difference in proportion of oocytes reaching at MII stage among intact pEF group and $56^{\circ}C$ group. Present study suggests that 1) pEF contains an enhancing component(s) for nuclear maturation in vitro of oocytes, 2) protein(s) of pEF may be capable to promote nuclear maturation in vitro, and 3) enhancing component for nuclear maturation may consist of two factors, which are responsible for germinal vesicle breakdown (GVBD) and promotion of MII stage.

• 한국양식학회지
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• 제11권1호
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• pp.119-124
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• 1998

농어의 난성숙과 배란유도를 위한 C하(21)-스테로이드와 HCG의 효능 비교실험이 in vitro에서 이루어졌다. 분리된 난모세포를 대상으로 17${\alpha}$-hydroxy, 20${\beta}$-dihydroprogesterone(17${\alpha}$20${\beta}$OHP), 17${\alpha}$-hydroxy, 20${\beta}$-dihydroprogesterone(17${\alpha}$20${\beta}$OHP : 5~100ng/ml)와 HCG(5~500IU/ml)를 사용하여 관찰한 난성숙 유도효과, 즉 GVM(germinal vesicle migration)과 GVBD(germinal vesicle breakdown)를 보면, 5ng/ml 농도의 17${\alpha}$20${\beta}$OHP와 17${\alpha}$20${\beta}$OHP를 제외한 모든 실험군에서의 반응은 대조군보다 효과가 있는 것으로 나타났으며, 특히 17${\alpha}$20${\beta}$OHP 50ng/ml 농도에서 가장 높은 성숙 유도효과를 보였다. GVBD 유도효과에 대한 17${\alpha}$20${\beta}$OHP(50ng/ml), HCG(50IU/ml) 그리고 이들 호르몬을 혼합처리한 17${\alpha}$20${\beta}$OHP+HCG에서는 혼합처리한 실험군이 가장 높은 반응을 보여 이들의 상호 보완적 작용을 관찰할 수 있었다. 난소조직을 대상으로 HCG와 17${\alpha}$20${\beta}$OHP를 사용하여 in vitro에서 농어의 배란유도효과를 관찰한 결과 사용한 모든 농도(17${\alpha}$20${\beta}$OHP : 1~1000ng/ml, HCG : 1~500IU/ml)에서 배란반응을 보였다. 특히 저농도 50ng/ml 또는 50IU/ml 이하로 처리하는 것이 더 효율적인 것으로 나타났다. 또한 HCG 처리가 17${\alpha}$20${\beta}$OHP보다 배란 반응에 더 민감하게 작용하는 것으로 생각된다.