中文摘要：

[目的]本试验旨在研究猪卵母细胞成熟过程中细胞骨架的动态变化以及秋水仙碱对卵母细胞成熟及MⅠ期纺锤体的影响。[方法]采用间接免疫荧光结合激光共聚焦显微成像技术,检测微管蛋白（α-tubulin）等细胞骨架蛋白的表达与亚细胞定位,之后采用秋水仙碱处理猪卵母细胞,研究秋水仙碱对卵母细胞成熟及MⅠ期纺锤体结构的影响。[结果]猪卵母细胞α-tubulin在生发泡破裂（GVBD）之后开始组装,第一次减数分裂中期（MⅠ）形成典型的纺锤体结构,第一次减数分裂后期至末期（ATⅠ）则分布于两组染色体之间,第二次减数分裂中期（MⅡ）在染色体附近重新组装形成典型的纺锤体;微丝蛋白（F-actin）在GVBD之后富集在细胞外周皮质下区域,并分别于MⅠ期和MⅡ期在纺锤体紧邻的皮质区富集形成微丝帽。在此基础上,通过秋水仙碱处理MⅠ期卵母细胞后,第一极体（pbⅠ）排出率显著下降,且绝大多数卵母细胞纺锤体解聚,染色体排列紊乱;但秋水仙碱撤除后,微管蛋白可重新组成MⅠ期纺锤体结构,延长培养时间后,pbⅠ排出率恢复。[结论]猪卵母细胞成熟过程中细胞骨架的动态变化具有阶段性分布特点,秋水仙碱可以使猪卵母细胞MⅠ期纺缍体发生解聚,成熟受到抑制,但秋水仙碱对MⅠ期卵母细胞纺缍体结构及功能的影响在一定程度上具有可逆性。

英文摘要：

[Objectives]This experiment was conducted to study the dynamic changes of cytoskeleton during porcine oocyte meiotic maturation process,and the effect of colchicine on the MⅠ spindle structure and in vitro maturation of porcine oocytes. [Methods]Indirect immunofluorescence and colchicine treatment were used to examine the cytoskeleton dynamic changes and the effect of colchicine on the M Ⅰ spindle structure of porcine oocytes. [Results]The subcellular localization results showed that α-tubulin began to assemble near the chromosome after germinal vesicle breakdown（ GVBD）,and formed a typical spindle structure at M Ⅰstage,then microtubules were distributed at the spindle between the two group of chromosome at ATⅠperiod,and a typical spindle structure was formed at MⅡ stage. F-actin was distributed at the cell outer periphery cortex after GVBD,and enriched at cortical region adjacent the spindle and formed actin cap at MⅠ and MⅡ stage,respectively. When treated MⅠ stage oocytes was treated with colchicine,the meiotic spindle disappeared and the pbⅠ extrusion rate reduced significantly. However,the spindle assembly was restored,and the pbⅠ extrusion rate was also recovered after removal of colchicine. [Conclusions]The dynamic changes of the cytoskeleton were characterized by stage distribution during the maturation of porcine oocytes. Colchicine treatment resulted in the pig MⅠ stage oocytes spindle depolymerization,and in vitro maturation inhibition,however,the effect of colchicine on the structure and function of MⅠ oocyte spindle had a certain degree of reversibility.