• Reproductive and Developmental Biology
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• v.31 no.2
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• pp.71-76
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• 2007

Previous studies have shown that BMI-1026 is a potent inhibitor of the cyclin-dependent kinases (cdk). In cell culture, the compound also arrests G2/M strongly and G1/S and S weakly. Two key kinases, cdk1 (p34cdc2 kinase) and mitogen-activated protein (MAP) kinase (erk1 and 2), perform crucial roles during oocyte maturation and, later, metaphase II (MII) arrest. In mammalian oocytes, both kinases are activated gradually around the time of germinal vesicle breakdown (GVBD) and maintain high activity in eggs arrested at metaphase II. In this study, we examined the effects of BMI-1026 on GVBD and MII arrest in mouse oocytes. BMI-1026 inhibited GVBD of immature oocytes and activated MII-arrested oocytes in a concentration-dependent manner, with more than 90% of oocytes exhibiting GVBD inhibition and MII activation at 100 nM This is approximately 500$\sim$1,000 times more potent than the activity reported for the cdk inhibitors roscovitine (${\sim}50{\mu}M$) and butyrolactone (${\sim}100{\mu}M$). Based on the results of previous in vitro kinase assays, we expected BMI-1026 to inhibit only cdk1 activation in oocytes and eggs, not MAP kinase. However, in our cell-based system, it inhibited the activity of both kinases. We also found that the effect of BMI-1026 is reversible. Our results suggest that BMI-1026 inhibits GVBD and activates MII-arrested oocytes efficiently and reversibly and that it also inhibits both cdk1/histone HI kinase and MAP kinase in mouse oocytes.

• Development and Reproduction
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• v.17 no.2
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• pp.141-147
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• 2013

Keeping the intact germinal vesicle (GV) is essential for maintaining the capacity of mammals including human. It is maintained by very complex procedures along with folliculogenesis and is a critical step for getting competent oocyte. So far, a few mechanisms involved in folliculogenesis are known but GV arrest mechanisms are largely unrevealed. Cyclic AMP, a adenosine derived substance, have been used as inhibitor of germinal vesicle breakdown as a putative oocyte maturation inhibitor. In this study, we examined the potency of adenosine as GV maintainer and a possible signaling mediator for that. A1, A2b, and A3 were detected in cumulus cells of cumulus enclosed-oocyte (CEO). Intact of germinal vesicle was not kept like in follicle but the spontaneous maturation was inhibited by exogenous adenosine. It is inhibited with concentration dependent manners. Intracellular calcium level of cumulus was extensively increased after adenosine treatment. Based on these results it is suggested that one of the pathway for GV arrest by adenosine and its receptors is calcium mediated signaling pathway in CEO.

• Korean Journal of Animal Reproduction
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• v.19 no.3
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• pp.171-179
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• 1995

This study was carried out to investigate the fertilizing ability of round spematids isolated from seminiferous tubules. A round spermatid was introduced into the perivitelline space of a mature oocyte using Leitz micromanipulators and then subjected to electrofusion. Electrofusion was induced by applying a single DC pulse of 90V with a duration of 60$\mu$sec using Model 611 Square Wave Stimulator(Phipps and Bird, U.S.A) in 0.3 M sucrose fusion medium containing 0.05mM CaCl2 and 0.1mM MgSO4, Oocyte pre-activation was conducted by exposure to a single DC(80V, 80$\mu$sec) pulse in electrofusion medium at 1 hour before electrofusion. The incidence of fusion with pre-activated oocytes(23.8%, 57/239) was higher than that with nonactivated oocytes(6.7%, 3/45). The most of electro-stimulated mouse oocytes cleaved regardless of the success or failure of fusion. Karyotyping of embryos that developed into blastocysts after exposure to the fusion pulse were performe. We found that blastocysts from the fused oocytes were diploid whereas blastocysts from the unfused oocytes were haploid. About 11.7 and 11.5% of fused and unfused oocytes were developmental potentials of fused and unfused oocytes. Therefore, these results suggest that the mouse mture oocyte can be fertilized by fusion with a round spermtid and subsequently developed normally.

• Journal of Embryo Transfer
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• v.26 no.4
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• pp.251-256
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• 2011

Presently, the effect of 0.5 mM dibutyryl cAMP (dbcAMP)-supplemented maturation medium during different incubation time on meiotic arrest (germinal vesicle) and resumption (metaphase II) of porcine oocytes and embryonic development of porcine oocytes following in vitro fertilization (IVF) or parthenogenetic activation (PA) was determined. Porcine cumulus oocyte complexes (COCs) were cultured in 0.5 mM dbcAMP for 17, 22, 27, or 42 h, and an additional 22 h without 0.5 mM dbcAMP. The nuclear status was examined at each time point. Oocytes cultured from 39~49 h displayed more than 80% meiotic resumption. More than 85 % of meiotic arrest was presented at 17~22 h. Oocytes were cultured for 22 h with 0.5 mM dbcAMP and additional 22 h without dbcAMP to assess developmental potential following IVF or PA. There were no significant differences in blastocyst rates among the dbcAMPIVF, IVF, dbcAMP-PA, and PA groups, although cleavage rate of IVF group was significantly higher than those of dbcAMP-PA, and PA groups. In conclusion, 0.5 mM dbcAMP influenced meiotic maturation of porcine oocytes depending on incubation time of oocyte, although embryonic development was not improved in both IVF and PA.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.11
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• pp.1477-1486
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• 2009

Female germ cell meiotic divisions are typically asymmetric, giving rise to two daughter cells with different sizes. Spindle movements including spindle migration from the oocyte center to the cortex and spindle rotation from parallel to perpendicular (typically in the mouse) at the cortex are crucial for these asymmetric divisions and therefore are crucial for gamete production. Different regulatory mechanisms for spindle movements have been determined in different species and a wide variety of different molecular components and processes that are involved in spindle movements have also been identified in different species. Here, we review the current state of knowledge as well as our understanding of mechanisms for spindle movements in different systems with focus on three main aspects: microtubules (MT), microfilaments (MF) and molecules associated with cytoskeletal organization as well as molecules that are not directly related to the cytoskeleton. How they might interact or function independently during female meiotic divisions in different species is discussed in detail.

• Reproductive and Developmental Biology
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• v.39 no.2
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• pp.29-36
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• 2015

Pigs are considered an ideal source of human disease model due to their physiological similarities to humans. However, the low efficiency of in vitro embryo production (IVP) is still a major barrier in the production of pig offspring with gene manipulation. Despite ongoing advances in the associated technologies, the developmental capacity of IVP pig embryos is still lower than that of their in vivo counterparts, as well as IVP embryos of other species (e.g., cattle and mice). The efficiency of IVP can be influenced by many factors that affect various critical steps in the process. The previous relevant reviews have focused on the in vitro maturation system, in vitro culture conditions, in vitro fertilization medium, issues with polyspermy, the utilized technologies, etc. In this review, we concentrate on factors that have not been fully detailed in prior reviews, such as the oocyte morphology, oocyte recovery methods, denuding procedures, first polar body morphology and embryo quality.

• Reproductive and Developmental Biology
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• v.38 no.2
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• pp.63-70
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• 2014

Plasminogen activators (PAs) are serine proteases that convert plasminogen to plasmin. Two type of PAs are urokinase-type PA (uPA) and tissue-type PA (tPA). Plasminogen is present in most extracellular fluids. PAs play in various reproductive processes including implantation, ovulation and fertilization. In the spermatozoa, PAs and PAIs play a role in sperm motility and fertilization. PAs in the sertoli cell are stimulated spermatozoa maturation and sperm activation through the phospholipase A2. The oocyte maturation is the process for fertilization and implantation. PAs in cumulus-oocyte complexes (COCs) are related to oocyte maturation by protein kinase A and C. In the ovulatory process, PAs activity are changed and it are related to reducing the tensile strength of ovarian follicle wall. The uterine environment is important for reproduction and the uterus undergo tissue remodeling. In the uterus and oviduct of mammals, expression and activity of PAs are changed during estrous cycle. Thus, expression and activity of PAs are concerned to many reproductive functions. Therefore, PAs seem to important factor of regulator in reproductive events.

• Clinical and Experimental Reproductive Medicine
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• v.16 no.1
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• pp.1-7
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• 1989

배양중인 생쥐 난자의 성숙에 미치는 배양액 내 calcim의 영향을 알아보기 위하여 1.71mM $Ca^{2+}$을 처리한 배양액과 $Ca^{2+}$이 존재하지 않는 배양액에서 난자를 배양하여 불꽃 원자 흡수 분광 광도계를 이용하여 배양된 난자의 $Ca^{2+}$농도를 측정하였다. 1) $Ca^{2+}$처리한 배양액에서 배양된 cumulus-cell이 제거된 (denuded oocyte)난자들은 시간이 지남에 따라 $Ca^{2+}$농도가 높게 나타났고, 2) $Ca^{2+}$처리하지 않은 배양액에서는 denuded난자는 3시간째 배양될때 부터 $Ca^{2+}$양이 줄어 들었다. Cumulus-enclosed난자는 $Ca^{2+}$존재하에서는 GVBD가 일어났다고 생각되는 4시간까지 계속 증가를 보인 반면 $Ca^{2+}-free$에서는 배양되지 않은 난자와 거의 차이가 없게 나타났다. 3) 핵막붕괴가 일어난 후부터 15시간까지 배양시컸을 때에는 CEO와 denuded oocyte에서 공히 $Ca^{2+}$의 농도는 다시 증가된 상태로 계속 되었다. 이런 결과로 미루어 보아 난자 성숙시 부터 성숙과정이 끝날때가지 exteral $Ca^{2+}$이 요구되고 있음을 증명해 주고있다. 그러나 이러한 세포질내의 $Ca^{2+}$및 bound calcium이 난자 성숙시부터 어떤 역활을 하고 있는 기작에 대해서는 좀 더 연구가 있어야겠다.

• Journal of the Korean Magnetic Resonance Society
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• v.9 no.1
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• pp.29-37
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• 2005

Intra-cellular drug uptake in Xenopus laevis oocyte has been elucidated using localized MR spectroscopy (MRS) and PFG NMR techniques at a 600 $MH_z$(Bruker, 14.1 T) NMR spectrometer. The localized MRS has been done with a homemade probe, and shows the intra-cellular uptake of nicotinamide. The self-diffusion of the molecule in Xenopus oocyte was obtained by PFG NMR technique. The measured data are well fitted with a linear combination of two exponential functions, which shows that there are two types of drug molecules, intra-and extra-cellular molecules. Diffusion coefficients of intra- and extra-cellular drug molecules are 3.7 $\times$ $10^{-11}$ $\m^{2}/s$and 6.4 $\times$ $10^{-10}$ $\m^{2}/s$, respectively. In the weighting factors there is shown that about 5% of drug molecule is inside the cells. These techniques can be used for drug screening in molecule-, cell-, and tissue-based preclinical test.

• Korean Journal of Ichthyology
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• v.22 no.1
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• pp.56-60
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• 2010

The Cobitis hankugensis-Iksookimia longicorpa complex from Korea has been presumed to be a unisexual lineage, originating from C. hankugensis and I. longicorpa and having almost all females. Recently, it was confirmed that the complex consisted of one diploid and two triploid complexes. From observation of their oocyte envelopes, three forms could be classified: a villous projection in I. longicorpa, a granular one in C. hankugensis and two triploid complexes, and a granule with villous one in the diploid complex. Even within the same granular projection, they showed specific features in length and density number from each other. These architectures are first observed in cobitid complexes and may playa role in identification of diploids and triploids.