• The Korean Journal of Zoology
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• v.35 no.2
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• pp.125-135
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• 1992

참개구리의 난자형성 단계에 따른 난황막의 구조적 변화와 막단백질의 변화에 대하여 연구함으로써 난자형성에 따른 난탈막의 기능적 분화의 가능성을 알아보고자 하T:다. 난황막의 구조적 변화는 난자형성에 따른 미세음모의 수와 모양의 변화로 확인되었다. 그 수가 초기에는 적으나 중기에 증가하고 알기에 다시 감소하며, 그 모양도 처음에는 킬고 가늘지만, 나중에는 짧고 굵어진다. 막단백질은 wheat germ agglutinin에 대해서만 특이하게 반응을 보이는 당단백질로서 작은 난모세포(직경 100-800 urn)와 큰 난모세포(직경 1500 urn)에서 다르게 나타난다. 분자량은 106 KD, 60 KD, 순 KD과 같이 어느 단계에서나 공통적으로 나타나는 단백질이 존재하는 반면에 특히 작은 난모세포의 난황막에서만 찾아 볼 수 있는 130 KD, 125 KD, 90 KD, 28 KD, 26KD과 같은 단겐 특이적으로 나타나는 막단백질도 있다. 이와 같은 결과로 미루어 보아 난황막 단백질은 난자형성단계에 따라 변화한다는 사실을 알 수 있다.

• Development and Reproduction
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• v.8 no.1
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• pp.35-42
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• 2004

Oocyte maturation refers to the process that prophase I arrested germinal vesicle(GV) drives the progression of meiosis to metaphase II(MII) to have the capacity for fertilization and embryo development. To better understand the molecular mechanism(s) involved in oocyte maturation, we identified differentially expressed genes(DEGs) between GV and MII mouse oocytes using a new innovative annealing control primer (ACP) technology. Using 20 ACPs, we successfully cloned 32 DEGs between GV and Mll oocytes, and 26 out of these 32 DEGs were functionally known genes. Four genes including Pscd2 were GV-specific, 10 genes including PKD2 and CSN3 were highly expressed in GV oocytes(GV-selective), and 12 genes including Diva were highly expressed in MII oocytes (MII-selective). Ail of the genes identified in this study were first reported in the oocyte expression using ACP system and especially, we could characterize the existence of PKD-CSW signaling pathwayin the mouse oocytes. Results of the present study would provide insight for studying molecular mechanisms regulating oocyte maturation.

• Clinical and Experimental Reproductive Medicine
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• v.33 no.3
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• pp.189-198
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• 2006

Objective: We previously described that Diva is highly expressed in matured metaphase II (MII) oocytes compared to immature germinal vesicle (GV) oocytes in mouse. We report here that the expression of Diva transcript as well as protein is oocyte-specific. To elucidate its physiological role in oocyte, the binding partner(s) of Diva has been identified by using immunoprecipitation (IP) followed by Mass Spectrometry. Methods: NIH/3T3 cells were transiently transfected for 24 h with either empty vector for control or FLAG-tagged mouse Diva construct, and IP was performed with anti-FLAG antibody. The immuno-isolated complexes were resolved by SDS-PAGE on a 12% gel followed by Coomassie Blue staining. For in-gel digestion, 15 bands of interest were excised manually and digested with trypsin. All mass spectra were acquired at a positive reflector mode by a 4700 Proteomics Analyzer (Applied Biosystems, Framingham, MA). Proteins were identified by searching the NCBI nonredundant database using MASCOT Peptide Mass Fingerprint software (Matrixscience, London). Results: Diva-associated complexes were formed in FLAG-tagged mouse Diva-overexpressed NIH/3T3 cells via IP using anti-FLAG-conjugated beads. Among the excised 15 bands, actin and actin-binding proteins such as tropomyosin, tropomodulin 3, and ${\alpha}$-actinin were identified. Binding between Diva and actin or tropomyosin was confirmed by IP followed by Western blot analysis. Both bindings were also detected endogenously in mouse ovaries, indicating that Diva works with actin and tropomyosin. Conclusions: This is the first report that immuno-isolated Diva-associated complexes are related to actin filament of the cytoskeletal system. When we consider the association of Diva with actin and tropomyosin, oocyte-specific Diva may play a role in modulating the cytoskeletal system during oocyte maturation.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.204-204
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• 2004

조류에서는 포유류와 달리 암컷이 후대의 성을 결정하는 것으로 알려져 있다. 암컷은 일생동안 산란할 난자를 가지고 태어나며, 이들 난자가 배란되어 정자와 만나면서 수정됨과 동시에 닭의 성이 결정되어진다. 본 연구는 산란계가 어떤 특이적 경향으로 암 혹은 수(W, Z) 난자를 배란하는지의 여부를 인공수정 및 배양의 방법들을 이용하여 수행하였다. 본 연구는33개체의 37주령 Single Comb white Leghorn 암컷들을 사용하여 13주 (37∼50주령) 동안 연속적으로 수행하였다. 인공수정은 동종 동계의 수탉 정액을 이용하였으며, 발생이 완료된 후대의 성비 판별은 다음과 같은 방법으로 수행하였다. (중략)

• Clinical and Experimental Reproductive Medicine
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• v.34 no.2
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• pp.95-106
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• 2007

Objective: We previously identified differentially expressed genes (DEGs) between germinal vesicle (GV) and metaphase II (MII) mouse oocyte. The present study was accomplished as a preliminary study to elucidate the role of ribose 5-phosphate isomerase A (Rpia), the essential enzyme of the pentose phosphate pathway (PPP), in oocyte maturation. We observed expression of Rpia in the mouse and porcine oocytes. Methods: Expression pattern of the 11 MII-selective DEGs in various tissues was evaluated using RT-PCR and selected 4 genes highly expressed in the ovary. According to the oocyte-selective expression profile, we selected Rpia as a target for this study. We identified the porcine Rpia sequence using EST clustering technique, since it is not yet registered in public databases. Results: The extended porcine Rpia nucleotide sequence was submitted and registered to GenBank (accession number EF213106). We prepared primers for porcine Rpia according to this sequence. In contrast to the oocyte-specific expression in the mouse, Rpia was expressed in porcine cumulus and granulosa cells as well as in oocytes. Conclusion: This is the first report on the characterization of the Rpia gene in the mouse and porcine ovarian cells. Results of the present study suggest that the mouse and porcine COCs employ different mechanism of glucose metabolism. Therefore, the different metabolic pathways during in vitro oocyte maturation (IVM) in different species may lead different maturation rates. It is required to study further regarding the role of Rpia in glucose metabolism of oocytes and follicular cell fore exploring the regulatory mechanism of oocyte maturation as well as for finding the finest culture conditions for in vitro maturation.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.10 no.6
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• pp.1407-1413
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• 2009

The present study was conducted to gain insights into oocyte maturation and early embryo development, SELDI-TOF-MS was used to find the protein candidates that are specifically or prominently expressed in mouse oocytes at the in vitro matured metaphase II (MIl) and germinal vesicle (GV) stages. By using selected CM10 chip, found 16 candidates which were up-regulated in GV stage oocytes compared with in MIl stage oocytes, molecular weight are 8180 (2 candidates), 10226 (5 candidates), 15767 (5 candidates) and 16770 (4 candidates) Da respectively. And 29 candidates were higher in MIl than in GV stage oocytes, molecular weight are 10832 (3 candidates), 17744(8 candidates), 20122 (3 candidates), 22131 (3 candidates), 24857 (7 candidates) and 33507 (5 candidates) Da, respectively. All (45) candidate (0.2 and 1.0 % error tolerances) were performed real time RT-PCR analysis and further selected 13 more potential candidates.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2005.10a
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• pp.74-75
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• 2005

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2005.10a
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• pp.76-77
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• 2005

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2005.10a
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• pp.74-75
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• 2005

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2005.10a
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• pp.76-77
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• 2005