细胞松弛素B对猪卵母细胞去核的影响  

Effect of Cytochalasin B on Porcine Oocyte Enucleate

作　　者：朱秀萍[1] 朱淑文[1] 张菁[1] 华修国[1] 唐峰[1] 

机构地区：[1]上海交通大学农业与生物学院,上海201101

出　　处：《上海交通大学学报（农业科学版）》2005年第3期253-255,共3页Journal of Shanghai Jiaotong University(Agricultural Science)

基　　金：上海市科委国际合作交流项目(015407005)

摘　　要：适宜的细胞松弛素B(CB)浓度可提高卵母细胞的成功去核率。将有明显第一极体排出的卵母细胞放入含有3.5×10-3、5×10-3、6.5×10-3、7.5×10-3mg.ml-1CB的NCSU37成熟培养液中孵育30min,采用盲吸法去核,将去核后的卵母细胞置入含5×10-3g.ml-1的Hoechst33342培养液中染色15min,用荧光显微镜检验去核率。结果表明:CB处理组去核率显著高于对照组(P<0.05)。不同浓度梯度处理组中,CB浓度过低(3.5×10-3mg.ml-1)或过高(7.5×10-3mg.ml-1)都使去核成功率(54.44%,51.46%)下降,溃破率(28.21%,34.11%)增高,与5×10-3、6.5×10-3mg.ml-1组的去核成功率(75.83%,73.83%)、溃破率(9.86%,11.62%)差异显著(P<0.05),且5×10-3mg.ml-1组的去核成功率高于6.5×10-3mg.ml-1组,差异不显著(P>0.05)。证明5×10-3mg.ml-1浓度的CB有利于第一极体和核质的吸出,同时可降低细胞膜撕裂,胞质破溃流出,获得较高的去核效率。The proper concentration of cytochalasin B improves the enucleation rate of oocytes successfully. The porcine oocytes with first polar were cultured in NCSU - 37 supplemented with 3.5 ×10^-3、5 ×10^-3,6. 5 ×10^-3,7. 5 ×10^-3 mg· ml^-1 CB respectively for 30min. Then these oocytes were enucleated and exposure to medium supplemented with 5 ×10^-3g· ml^-1 Hoechst33342 for 15min. The treated oocytes were checked with fluorescence microscope and evaluated the enucleation rate. The results show that the enucleation rate of CB treating group is significantly higher than those in control group（ P 〈0.05） . Among the CB treating groups, the low concentration CB （3.5 ×10^-3 mg· ml^-1） and the high （3.5 ×10^-3 mg· ml^-1） can reduce enucleation rate（54.44%, 51.46% ） and enhance the debacle rate （9.86%, 11.62% ）, which are significantly different from the enocleation rate （75.83%, 73.83% ） and debacle rate（9. 86%, 11.62% ） with 5 ×10^-3, 6. 5 ×10^-3mg· ml^-1 CB treatments（ P 〈0. 05） . The treatment with the concentration of 5 ×10^-3 mg· ml^-1 can receive the higher enucleation rate than that of 6.5 ×10^-3mg· ml^-1 CB, which were insignificant differences（ P〉 0.05） . The conclusion can be got that the 5 ×10 -3 mg· ml^-1 CB are contribute to the extraction of first polar as well as karyoplasm and can avoid the avulsion of membrane, prevent the cytoplasmic from effusing. The efficiency of porcine oocytes enucleating can be improved.

关 键 词：细胞松弛素B 猪卵母细胞 去核 

分 类 号：Q954.4[生物学—动物学]