小鼠极体重组卵母细胞的体外受精与发育  

Fertilization In vitro and Development of the Mouse Oocytes Recombined by First Polar Bodies

作　　者：王公金[1] 刘文华[2] 谭晓东[2] 徐小波[1] 范必勤[1] 赵伟[1] 

机构地区：[1]江苏省农业科学院畜牧研究所,南京210014 [2]南京师范大学生命科学学院,南京210097

出　　处：《农业生物技术学报》2008年第1期77-81,共5页Journal of Agricultural Biotechnology

基　　金：江苏省自然科学基金(No.BK2006165)资助

摘　　要：分别以昆明系、ICR系、C57BL/6-Tg(CAG-EGFP)C14-Y01-FM131Osb系(即转绿色荧光蛋白基因小鼠,简称荧光标记小鼠)和无标记C57BL/6J系小鼠(Mus musculus)为试验动物,探索其卵母细胞第一极体核重组卵母细胞的生殖功能。超数排卵后采集小鼠卵丘卵母细胞复合体(COCs),利用链霉蛋白酶酶解法分离出第一极体(PbⅠ);以形态学和台盼蓝染色相结合的方法对不同温度保存条件下COCs中PbI进行活力鉴定和分级;采用显微操作术将PbⅠ核供体注射到去核卵母细胞中,进一步观察重组卵母细胞体外受精和体外培养发育能力。结果表明,注射hCG后12～14h回收的PbⅠ活力最佳,4℃条件下保存48h后PbⅠ仍保持活性;117枚昆明系和ICR系小鼠重组卵母细胞经体外受精获得13枚2-细胞胚胎,38枚荧光标记小鼠的卵母细胞中有3枚卵裂成2-细胞胚胎。初步证明小鼠第一极体重组卵母细胞可体外受精和体外早期发育,为发掘和利用家畜母本基因资源提供了参考依据。The reproductive function of three strains of mice （Mus musculus） （Kunming, ICR, C57BL/6-Tg （CAG-EGFP） C 14-Y01-FM 131Osb and C57BL/6J lines） oocytes recombined by the nuclei of first polar body （Pb Ⅰ） was explored, respectively. Cumulus oocyte complexes （COCs） from the mouses were collected after superovulation, then PbIs were obtained from the COCs by 2% pronase treatment. The survival PbIs under different temperature were identified and stapled by the methods of morphology and trypanblau staining, later the contents including nuclei and a little cytoplasm ofPb I were injected into the oocyte whose nuclei was enucleated by micromanipulation, and the recombined oocytes were fertilized and cultured in vitro in order to observe its ability of further development. The results showed that vigor of Pb Ⅰ was in the best status within 12 to 14 h after superovulation, and maintained in 48 h under 4℃. Thirteen out of 117 recombined oocytes from Kunming and ICR mice developed into 2-cell embryos, and 3 out of 38 recombined oocytes from C57BL/6-Tg （CAG-EGFP）C 14-Y01-FM 131Osb mice were cleavage as 2-cell embryos. The experiments confirmed that the mouse oocytes recombined by the nuclei ofPb Ⅰ could be normally maintained in vitro fertilization and in vitro development, this has offered valuable reference for further utilization of gene resources for other farm animals.

关 键 词：小鼠 第一极体 重组卵母细胞 体外发育 

分 类 号：S185[农业科学—农业基础科学]