小鼠体外发育与体内发育成熟卵子胞浆成熟度和细胞骨架比较  

Ooplasm Maturation and Cytoskeleton Are Less Optimum in Oocytes from In Vitro Growth Follicles

作　　者：彭宇洪[1] 庄广伦[1] 陈文芳[2] 李涛[1] 周灿权[1] 

机构地区：[1]中山大学附属第一医院生殖中心,广东广州510080 [2]中山大学附属第一医院肾内科,广东广州510080

出　　处：《中山大学学报（医学科学版）》2005年第3期304-307,共4页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广东省科技攻关基金资助项目(2003A3020305)

摘　　要：【目的】体外培养小鼠窦前卵泡得到成熟卵子,比较其与体内发育成熟卵子直径、谷胱甘肽含量、皮质颗粒分布以及纺锤体结构的差异,评估胞浆成熟度对卵子发育潜能的影响。【方法】选择出生10d小鼠,分离窦前卵泡,培养后得到326个MII期卵子作为体外发育组。8～10周雌鼠经孕马血清和绒毛膜促性腺激素注射得到244个MII卵子作为体内发育组。酶法检测卵子谷胱甘肽含量,免疫荧光染色,共聚焦显微镜成像显示纺锤体形态和皮质颗粒分布。【结果】体外发育成熟卵子直径(69.6±5.7)μm,明显小于体内发育成熟卵子直径(84.2±3.0)μm;体外发育成熟卵子谷胱甘肽含量(4.3±0.7)pmol明显低于体内发育成熟卵子(6.1±1.0)pmol;50个体外发育成熟卵子中18个(36%)皮质颗粒周边分布,40个体内发育成熟卵子中36个(90%)皮质颗粒周边分布(P<0.05);14个(28%)体外发育卵子出现无皮质颗粒区,34个(85%)体内发育成熟卵子出现无皮质颗粒区(P<0.05);120个体外发育成熟卵子中46个(38.3%)显示正常桶状纺锤体,84个体内发育成熟卵子中80个(95.2%)显示正常桶状纺锤体(P<0.05)。【结论】体外发育成熟卵子胞浆成熟度没有达到体内发育成熟卵子水平。Objective]To acquire mature oocytes from in vitro preantrum follicles and compare the oocyte diameter, glutathione content, cortical granulae distribution and spindle appearance between in vitro growth oocytes and in vivo growth oocytes so as to evaluate ooplasm maturation's effect on oocyte competence. [Methods]10-day-old mouse pre-antrum follicles were cultured in medium drops; To staine the acquired MII oocytes with fluorescence conjugated α-tubulin antibody and lectin.To image the spindles and cortical granulae with confocal microscope; To detecte gultathione content by enzyme method. [Results] A total of 326 oocytes was acquired after in vitro culture; The diameter of in vitro growth oocytes was 69.6±5.7 μm, which was significantly smaller than that of in vivo growth oocytes 84.2±3.0 μm; The glutathione content of in vitro growth oocytes and in vivo growth oocytes was 4.3±0.7pmol and 6.1±1.0 pmol respectively(the difference was significant); 18/50 of in vitro growth oocytes and 36/40 of in vivo growth oocytes formed cortical granulae monolayer ring, and 14/50 of in vitro growth oocytes and 34/40 of in vivo growth oocytes formed cortical granule-free domain, difference was significant; 46/120 of in vitro growth oocytes and 80/84 of in vivo growth oocytes formed barrel-shaped spindle(the difference was significant).[Conclusion]MII oocytes achieved from in vitro culture of mouse pre-antrum follicles bears less optimum ooplasm maturation. To a certain extent, the oocyte cytoskeleton is abnormal.

关 键 词：卵泡 组织培养 微管蛋白 皮质颗粒 谷胱甘肽 

分 类 号：Q954.43[生物学—动物学]