顺式高尔基体蛋白GM130影响小鼠卵母细胞减数不对称分裂的研究  

A cis-Golgi protein GM130 regulates asymmetric division in mouse oocyte

作　　者：张春晖[1] 钱卫平[1] 孙洪梅[1] 孟繁华[1] 

机构地区：[1]北京大学深圳医院生殖医学科,深圳518000

出　　处：《生殖医学杂志》2013年第8期596-601,共6页Journal of Reproductive Medicine

基　　金：国家自然科学基金(2012-81200438)

摘　　要：目的确定顺式高尔基体蛋白(GM130)对小鼠卵母细胞纺锤体迁移及减数不对称分裂的影响。方法免疫荧光法检测GM130在小鼠卵母细胞的定位,活细胞工作站延时拍摄系统捕捉纺锤体在降调GM130表达后的的动态变化。结果降调GM130的表达后,第一极体排出率下降,在排出的极体中,大极体的比率升高;但对微丝网络及微丝帽的影响不明显。向卵母细胞内注射β5-tubulin--GFP mRNA和GM130特定的寡聚核苷酸(morpholino,MO)的混合物后,用活细胞工作站延时拍摄系统证实了降调GM130会影响卵母细胞纺锤体的迁移,纺锤体单极拉长,并能与该侧的皮质接触,产生胞质分裂环,导致大极体排出。若拉长的纺锤体尚不能与皮质接触,则细胞阻滞在分裂中期I(MI期),没有极体排出。另外,降调GM130的表达也会影响磷酸化丝裂原活化蛋白激酶激酶1/2(p-MEK1/2)在纺锤体极的定位,用丝裂原活化蛋白激酶激酶(MEK)的抑制剂U0126处理MI期的卵母细胞则发现GM130不再在纺锤体两极聚集,而是散在分布于胞浆。结论 GM130可能参与丝裂原活化蛋白激酶(MAPK)信号通路在小鼠卵母细胞纺锤体迁移及不对称分裂中起重要作用。Objective： To identify the role of GM130 in regulating MI spindle migration and the meiotic asymmetric division. Methods： GM130 localizations were determined by immunofluorescence and confocal microscopy. Microtubule dynamics were filmed on Perkin Elmer precisely Ultra VIEW VOX confocal Imaging System. Results： A large proportion of oocytes extruded an abnormally large polar body （PB） in the GM130- MO injected group, and the enlarged PB contained a spindle with normal morphology in some cases. In many cases, the oocyes arrested at MI stage. It was observed by using live cell imaging that depletion of GM130 affected spindle migration and resulted in dongated spindle and large polar body extrusiom It was further found that depletion of GM130 blocked p-MEK1/2 accumulation at the spindle poles. And the results showed that GM130 detached from the spindle poles in oocytes treated with MEK specific inhibitor U0126. Conclusions： GM130 may cooperate with the MAPK pathway to play roles in spindle migration and asymmetric division during mouse oocyte maturation.

关 键 词：顺式高尔基体蛋白GM130 小鼠卵母细胞 减数不对称分裂 纺锤体 

分 类 号：R[医药卫生]