不同因素对小鼠MⅡ期卵母细胞纺锤体的影响

Effects of Different Factors on Spindle of Mouse MⅡ Oocytes

[目的]研究在不同条件下小鼠MⅡ期卵母细胞纺锤体的变化，为卵母细胞纺锤体的研究提供参考依据。[方法]在不同条件下处理小鼠MⅡ期卵母细胞，通过免疫细胞化学方法和激光共聚焦扫描显微镜观察其纺锤体变化。[结果]卵母细胞在25℃、20min时纺锤体异常，25℃、30min或4℃、10min时纺锤体缺失；5、10、20和40μg/ml细胞松弛素B（CB）作用2h未对纺锤体产生影响；卵母细胞在20μmol／L秋水仙素中作用1h纺锤体异常，40μmol／L时作用1h纺锤体缺失；在4％多聚甲醛中4℃条件下，卵母细胞纺锤体结构以固定时间为1～12h效果较好。[结论]小鼠MⅡ期卵母细胞纺锤体对外界环境变化敏感，体外操作时控制外部环境是保持卵母细胞纺锤体正常生理功能的关键。

[ Objective ] The research aimed to study the effects of different factors on spindle of mouse MⅡ ooeytes, and provide references for the oocyte spindle. [Method] The mouse MⅡ oocytes were treated with different conditions,and their spindles were observed by immunoeytochemical method and laser seaming eonfoeal microscopy. [Result] The spindles of oocytes were abmnnal at 25℃ for 20 rain,and disappeared at 25 ℃ for 30 rain or at 4℃for 10 rain. 5,10,20 and 40μg/ml Cytoehalasin B （CB） for 2 h had no effects on the spindles of the ooeytes. The spindles of oecytes were abnormal in 20 μmol/L eolehieines for 1 h,and disappeared in 40μmol/L eolchieines for 1 h.The structure of spindles were perfect for 1 - 12 h when the oocytes were fixed in 4% parafonnaldehyde. [ Conclusion] The spindle of the mouse MII ooeytes is sensitive to environmental ehanges, and the control of envirolunental factors is crucial for nonmal funetion of the spindle in vitro manipulation.