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대한생식의학회지 제29권 제4호 2010년
생쥐 난자의 초자화동결 시 Cytochalasin B가 Cytoskeleton에 미치는 영향
경상대학교 의과대학 산부인과학교실, 경상대학교병원 불임 유전 연구실
백원영, 최원준,김세나,이종학
The Effect of Cytochalasin B on Cytoskeletal Stability of Mouse Oocyte Frozen by Vitrification
Paik WY
Obstetrics and Gynecology, Gyeongsang National Univ, Lab of Infertility and Genetics, Gyeongsang
Objective: The purpose of this study was to evaluate the effect of Cytochalasin B (CCB) on the cytoskeletal stability of mouse oocyte frozen by vitrification. Methods: Mouse oocytes retrieved from cycle stimulated by PMSG and hCG were treated by CCB and then vitrified in EFS-30. These oocytes were placed onto an EM grid and submerged immediately in liquid nitrogen. Thawing of the oocytes was carried out at room temperature for 5 seconds, then the EM grid was placed into 0.75 M, 0.5 M and 0.25 M sucrose at 37℃ for 3 minutes, each. These oocytes were fixed in 4% formaldehyde for an hour and then washed in PPB for 15 minutes 3 times, then incubated in PPB containing anti-tubulin monoclonal antibody at 4℃ overnight. And then, the oocytes were incubated with FITC-conjugated anti-mouse IgG and propidium iodide (PI) for 45 minutes. Pattern of microtubules and microfilaments of oocytes were evaluated with a confocal microscope. Results: The rate of oocytes containing normal microtubules and microfilaments was significantly decreased after vitrification. The rate of oocyte containing normal microtubules in CCB treated group was higher than those in non-treated group (53.7% vs. 48.9%), but the difference was not significant. The rate of oocyte containing normal microfilaments in CCB treated group was significantly higher than those in non-treated group (64.5% vs. 38.3%, p<0.05). Conclusion: Microfilaments stability could be improved by CCB treatment prior to vitrification. It is suggested that CCB treatment prior to vitrification improve stability of cytoskeleton and then increase success rate in IVF-ET program using vitrification and thawing oocyte.
키워드 : Cytochalasin B, Vitrification, Cytoskeleton, Microtubule, Microfilament
교신저자 : wypaik@gshp.gsnu.ac.kr
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