细胞松弛素B浓度对小鼠卵母细胞去核效果的影响

Effect of Cytochalasin B on Mouse Oocyte Enucleating

目的 探讨细胞松弛素 B浓度对小鼠卵母细胞去核效果的影响。方法 常规超排 KM小鼠获卵母细胞 ,将有明显第一极体的卵母细胞放入含有 5× 10 - 3、1× 10 - 2 、2× 10 - 2 g/ L CB的成熟培养液中孵育 15 min,采用盲吸法去核 ,去核后的卵母细胞放入含 5× 10 - 3g/ L Hoechst33342的成熟培养液中染色 15 min,在荧光显微镜下检查去核的效率。结果 CB组去核率可达 88.2 % ,而对照组去核成功率仅达 2 9.7% ,两者的去核效果统计学上有显著差异 ;同时 CB的浓度过高会影响去核的效果 ,使卵母细胞的破溃率升高。结论 CB有利于去核 ,同时卵胞膜可以借助磷脂双层的游动性而很快得到修复 ;高浓度的 CB对卵胞膜作用过于强烈 ,细胞骨架严重破坏 ,磷脂双层结构的弹性和粘性减弱 ,修复能力降低 ,影响胚胎的构建。

AIM To study the effect of cytochalasin B on the mouse oocyte enucleating.METHODS The oocytes were gotten from superovulated KM mice.The oocytes which have first polar body were cultured in M16 medium supplemented with 5×10^(-3),1×10^(-2),2×10^(-2) g/L CB respectively for 15 min.Then these oocytes were enucleated and cultured in M16 medium supplemented with 5×10^(-3) g/L Hoechst33342 for 15 min.At last we checked these oocytes with fluorescence microscope and got enucleation rate.RESULTS 1×10^(-2) g/L CB enucleation rate was 88.2% and control was 29.7%.High concentration CB(2×10^(-2) g/L) can reduce enucleation rate.CONCLUSION Appropriate concentration of CB is important in oocyte enucleating because it can raise cell flexibility with relaxing cytoskeleton and increase the fluidity of cytomembrane phospholipid bilayer.