• Korean Journal of Animal Reproduction
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• v.19 no.4
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• pp.323-329
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• 1996

In vitro cultrue systems for the growth of sma-II oocytes and for meiotic maturation are expected to provide a new source of a large population of oocytes as well as assistance in basic physiological studies of oogenesis. Mouse oocytes mid-growth phase can complete grovvth and acquire full developmental capacity in vitro. On the other hand, growing pig oocytes need some other factors. FSH at a low concentration maintains the viability of both oocytes and granulosa cells, and hypoxanthine promotes the meiotic competence of the oocytes during culture period. Considerable improvement in the culture systems for growth of pig oocytes, suggested from mouse studies, and for oocyte maturation could help to develop this technology in larger species.

• The Korean Journal of Zoology
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• v.38 no.4
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• pp.523-530
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• 1995

Specific affinity binding sites for atrial natriuretic peptide (ANP) were Investigated in the pig ovarian tissues by in vitro autoradiographic techniques. In the pig ovary, the highest binding sites for 12514abeiled rANP(l~28) were localized in the granulosa cell layer of the forncles. The binding sies on theca layer of the ovarian follicles were mainly localized in the external layer, but none was observed In the Internal layer. In the corpus luteum, the binding site was not observed. The specific bindings of 200 pM of l2Sl4abelled rANP(l~28) to granulosa and theca externa layers were reversed completely by excess concentration (1 ~4) of unlabelled rANP(l~28) but not by 10 ~ of unrelated peptides, human angiotensin II and arginine vasopressin. The binding was also displaced by 1 ~ of desiGIn18, Ser19, Gly2O, leu21, Gly22I ANP(4~2g) (C- ANF) as a spedfic ligand of the ANP clearance receptor. Therefore these results indicate ~hat the biological and the clearance ANP receptors exist in the theca externa and granulosa layer of the pig ovary, and suggest that the ANP receptors may be related with the regulatory lundion of the ovarian follicular development including oocyte maturation.

• Korean Journal of Animal Reproduction
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• v.16 no.4
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• pp.289-299
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• 1993

The polypeptide patterns of cellular and follicular components were analysed by SDS-PAGE and two dimensional(2-D)electrophoresis combined with isoelectric focusing (IEF) to establish protein profiles in each of the components in porcine follicles. Oocyte-cumulus complexes were cultured in M16+FCS+Gn at 39 in an atmosphere of 5% CO$_2$, in air for 35 h. At the end of the culture, the zona-free oocyte, ZP alone and cumulus cells were prepared and analysed either on 10% SDS-PAGE for the protein profile at the first dimensional gel or 2-D protein pattern. The amounts of each samples were determined for the visualization with Coomasie brilliant blue (CBB) or silver staining, thus giving useful information for the identification of specific proteins in the components or appropriate amount of samples for proper visualization. Oocyte showed 25 and 114 kd major protein band. Other minor components were additionally visualized with CBB on the same gel after silver staining procedure. Cumulus cells also showed specific proteins which is not present in the oocytes. The number of cumulus cell was proper to give major bands with CBB and additional minor bands with silver staining. To establish the degree of contamination from the remnant of the corona radiata to the ZP, zonae were differently prepared or analysed by SDS-PAGE.The preparation of the ZP in this study did not showed any contamination judged by the protein profile of the components. Also follicular fluid showed its specific protein profile without any significant differences among the different sizes of follicles. The established protein profile of each follicular component should be helpful for the identification and elimination of contaminated components, i. e., antigen preparation or immunological studies. The results also suggest that the preparation of each components in the study was appropriate and can be used for a further sensitive biochemical analysis in mammalian oocytes and early embryos.

• Journal of Animal Science and Technology
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• v.46 no.4
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• pp.571-584
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• 2004

It is known widely that granulosa cell apoptosis leads follicular atresia and macrophages exert their effects directly and/or indirectly from the initiation to the completion of follicular atresia by phagocytosis of apoptotic bodies and secretion of various cytokines. However, the site of initiation, propagation routes and the elimination methods of apoptotic bodies, and the time and methods of penetration of macrophages into the follicles are not known completely. Using pig(Yorkshire-breed) ovary, immunohistochemical studies with TUNEL for apoptotic bodies and pig macrophage monoclonal antibody 4E9 for macrophages, and light and transmission electron microscopic observations were performed. In the pig, follicular atresia began with the granulosa cell apoptosis, and the apoptosis of theca intema cells occured at the same time. The apoptosis of granulosa cells initiated randomly within the granulosa cell layer and propagated rapidly into the whole layer. Ultrastructura1ly, apoptotic granulosa cells showed characteristic pyknotic and deformed nucleus and intracytoplasmic vesicles. Apoptotic bodies were eliminated by intact granulosa cells and macrophages. Intact granulosa cells ingested apoptotic bodies transiently, soon after they fell into the apoptosis. Finally, apoptotic bodies and degenerated oocyte were phagocytosed by macrophages. Macrophages entered the ovarian follicle at the time of initiation of granulosa cell apoptosis, and migrated with the progression of apoptosis. By elimination of theca cells, macrophages contributed the completion of follicular atresia These results will provide valuable informations on the study of the interrelation between macrophage and ovarian follicular atresia.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 1999.10a
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• pp.41-41
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• 1999

• The Korea Swine Journal
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• v.16 no.3 s.175
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• pp.120-124
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• 1994

• Journal of Embryo Transfer
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• v.18 no.1
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• pp.1-14
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• 2003

This study was conducted to investigate that the immature and mature oocytes of porcine can be cryopreserved by vitrification. Oocytes were centrifuged to polarize the cytoplasmic lipid droplets. The lipids were removed from cytoplasm by micromanipulation. Delipated oocytes were centrifuged after being preincubated with cytochalasin B(CB) fer 10 min, and lipid droplets were removed. Centrifuged oocytes were treated with CB and centrifuged to polorize lipid droplets but not delipated and control oocytes is not-treatment. Oocytes of three types were vitrified in electron microscope(EM) grids. The results of survival, maturation and cleavage rates were as follows. 1 The survival rates of immature oocytes were 15.1%, 0% and 0% in the Delipated, Centrifuged and Control after vitrification, respectively, and its rate of Delipated wassignificantly higher than Centrifuged and Control(P<.01). 2. The survival rates of mature oocytes were 12.21%, 0% and 0% in the Delipated, Centrifuged and Control after vitrification, respectively, and its rate of Delipated was significantly higher than Centrifuged and Control(P<.01). 3 The maturation rates of immature oocytes were 37.5% and 68.9% for metaphase II in the Delipated after vitrification and Non-vitrification, respectively, and its rate of Non-vitrification was significantly higher than Delipated after vitrification(P<.01). 4. The cleavage rates of immature oocytes were 12.5%, 0%, 0% and 56.1% in the Delipated, Centrifuged, Control after vitrification and Non-vitrification, respectively. It's rate of Delipated was higher than Centrifuged and Control, but there were no significant difference, and its rate of Non-vitrification was significantly higher than Delipated, Centrifuged and Control(P<.05). 5 The cleavage rates of mature oocytes were 25.0%, 0%, 0% and 67.9% in the Delipated, Centrifuged, Control after vitrification and Non-vitrification, respectively. It's rate of Delipated was higher than Centrifuged and Control, but there were no significant difference, and its rate of Non-vitrification was significantly higher than Delipated, Centrifuged and Control(P<.05).

• Journal of Embryo Transfer
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• v.20 no.3
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• pp.207-215
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• 2005

본 연구는 돼지 난포란의 동결보존 후 생존성과 난자의 활성화 처리에 따른 체외발생율과 이를 이용한 핵 이식배의 체외발생율을 조사하였다. 활성화 처리된 배는 $5\%$ FBS가 첨가된 NCSU 23 배양액으로 $38.5^{\circ}C$, $5\%\;CO_2$와 $95\%$ air의 조건으로 배양하였다. 1. 난포란을 EDS와 $5\%$ PVP로 동결 후 $10\%$ FBS가 첨가된 NCSU 23 배양액으로 $0{\~}10$시간 배양했을 때 체외발생율은 $36.0\%$로서 대조군인 비동결 난포란의 체외발생율 $46.0\%$에 비해 낮았다. 2. Ethanol과 cyclojexamide로 처리 후 42 및 46시간 배양한 배의 분할율은 각각 $33.3\%$, $36.0\%$ 및 $27.1\%$, $30.0\%$로서 대조군의 $8.8\%$, $11.4\%$에 비해 높게 나타났다. 3. 동결 및 비동결 난포란을 이용한 핵이식 배의 융합율과 발생율 간에는 유의한 차이가 없었다. 4. Ethanol과 cyclojexamide로 활성화 처리한 난자를 이용하여 재구축한 핵 이식배의 발생율은 $2.8\%$, $5.3\%$ 및 $1.5\%$, $2.9\%$로서 대조군의 $0.0\%$, $0.0\%$에 비해 높은 발생율을 나타냈다.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.224-224
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• 2004

Plasminogen activators (PAs)는 자궁분비액, 난포액, 정장물질 등을 포함한 여러 가지 세포외 분비액(extracellular fluids) 및 plasma에 풍부하게 존재하는 세포외 전구효소인 plasminogen을 plasmin으로 전환시키는 단백질분해효소이다. PAs는 섬유소용해, 배란, 착상 및 수정을 포함한 다양한 생리적인 과정에서 중요한 역할을 수행하는 것으로 알려져 있다. 따라서, 본 연구는 돼지의 체외수정 과정시 정자와 난자에서의 plasminogen activators activity의 변화를 SDS-PAGE와 zymography를 이용하여 검토하였다. (중략)

• Proceedings of the KSAR Conference
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• 2002.06a
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• pp.44-44
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• 2002

본 연구는 돼지의 미성숙 난포란을 체외에서 성숙, 수정시킨 후, 생산된 체외 수정란을 NCSU 23 체외배양액에 항산화제(NAC, ebselen 및 glutathione) 와 성장 인자 (EGF, PDGF)의 첨가가 돼지 체외 수정란의 체외 발육에 미치는 영향을 검토하였다. NAC을 0, 0.5, 1.0 및 2.0 mL 첨가하여 체외배양한 결과, 상실배기 이상 발육된 체외발육성적이 1.0mL과 2.0 mM 첨가구가 여타구보다 유의하게 높은 체외발육성적을 나타냈다.(p〈0.05). (중략)