Aurora A对猪卵母细胞向胚胎转换过程中纺锤体结构的影响  

Effect of Aurora A on Spindle Structure During Porcine(Sus scrofa)Oocyte-to-embryo Transition

作　　者：崔盼盼[1] 杨笑柳 陈长超[1] 王令怡 孙嘉豪 靳莎莎[1] 芮荣[1] 剧世强[1] 

机构地区：[1]南京农业大学动物医学院,南京210095

出　　处：《农业生物技术学报》2016年第4期488-496,共9页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(No.31572589;No.31201967);江苏省自然科学基金(No.BK2012369);高等学校博士学科点专项基金(No.20130097110020);国家大学生创新性实验计划(No.201510307055)

摘　　要：极光激酶A(Aurora A)是一种参与体细胞中心体复制、纺锤体组装与胞质分裂等生物学事件的丝氨酸/苏氨酸蛋白激酶。为了探讨Aurora A在猪(Sus scrofa)卵母细胞向胚胎转换这一特殊时期的动态表达及其相关功能,本研究采用间接免疫荧光结合激光共聚焦显微成像技术检测Aurora A在转换过程中动态表达与亚细胞定位,并在此基础上通过特异性抑制实验分析Aurora A在该特殊转换时期的调节作用。结果表明,Aurora A在猪卵母细胞向胚胎转换的各个时期均有表达,并与α-微管蛋白(α-tubulin)具有相似的亚细胞定位;使用Aurora A特异性抑制剂MLN8054处理卵母细胞,卵母细胞的第一极体排出率呈浓度依赖性下降,且纺锤体异常率显著升高(P<0.05);将经MLN8054处理且排出第一极体的卵母细胞孤雌激活后,胚胎卵裂率与对照组相比无显著差异。结果表明,在猪卵母细胞向胚胎转换过程中,Aurora A的表达与α-tubulin分布密切相关,且呈现阶段性分布特点;Aurora A主要通过调节纺锤体组装及其稳定性参与猪卵母细胞成熟分裂的调控;MLN8054抑制Aurora A后,对所获得的第二次减数分裂中期(metaphaseⅡ,MⅡ)卵母细胞顺利向胚胎期转换并无显著影响。研究结果为进一步了解Aurora A在猪卵母细胞向早期胚胎转换这一特殊时期的调控作用提供了重要的实验依据。Aurora A is one of the serine/threonine protein kinase who participates in a series of biological events such as centrosome replication, spindle assembly and cytokinesis. The purpose of this study was to explore the dynamic expression and function of Aurora A during oocyte-to-embryo transition in porcine （Sus scrofa）. As Aurora A has an extremely close relationship with microtubules, our examination began with the expression and distribution of α-tubulin during the oocyte-to-embryo transition. Meanwhile, The dynamic expression and subcellular localization of Aurora A during oocyte-to-embryo transition were detected using indirect immunofluorescence method and laser confocal microscopy analysis. Then, the specific inhibition test was used to analyze the regulatory function of Aurora A in this particular transition stage. The results showed that Aurora A expressed in each period in the oocyte-to-embryo transition and had the similar subcellular localization with α-tubulin. At the metaphase I （M I ） and M II stage, α-tubulin assembled a typical spindle in the cortex of oocyte, and a similar distribution presented in the same place where Aurora A was. During the prophase of mitosis, α- tubulin gradually gathered and reticulated, and Aurora A also had a netlike distribution. Following, α-tubulin was translated from the symmetric and barrel shaped spindle in the central of embryo to migration towards poles of embryo, at the same time Aurora A had a similar trend from mitosis metaphase to anaphase. When oocytes were treated with a highly selective inhibitor （MLN8054） of Aurora A, the extrusion rate of first polar body was significantly reduced with the increasing concentrations of MLN8054 （P〈0.05）, and the percentage of abnormal spindle morphology, mainly including monopolar spindles and multipolar spindles, was significantly increased （P〈0.05）. What＇s more, the high frequencies of abnormal spindle and chromosome alignment varied with the different exposure times. When oocytes at M 11 sta

关 键 词：猪 极光激酶A 卵母细胞 胚胎 纺锤体 

分 类 号：S828.3[农业科学—畜牧学]