HDAC8在小鼠卵母细胞减数分裂中的作用及其机制的研究
毕业论文关键词：HDAC8；纺锤体组装；染色体排列；整倍体；γ-tubulin
Study of the role and mechanism of HDAC8 during mouse oocyte meiosis
Abstract: HDAC8 is a classⅠhistone deacetylase that has been found to widely distribute in cytoplasm and involved in numerous biological processes via its non-histone substrates. However, the role of this enzyme in oocyte meiosis remains unclear. Here, in this study we have found, by immunofluorescent analysis, that HDAC8 localizes at spindle poles and dynamically participates in the regulation of microtubule organization during mouse oocyte meiosis. Depletion of HDAC8 by RNAi results in severe malformated spindle microtubules and impaired kinetochore-microtubule attachments in MⅠeggs, consequently leading to the misalignment of chromosomes and generation of aneuploidy. Analogously, these abnormities of spindle/chromosome are also observed in oocytes treated with HDAC8-selective inhibitor PCI-34051. What's more, we have also found that HDAC8 is required for the right localization of γ-tubulin. Taken together, these results indicate that HDAC8 positively functions in spindle assembley and chromosome assortment, thus ensuring the generation of normal euploidy in mouse oocytes.
Key words: HDAC8；spindle assembly；chromosome alignment；euploidy；γ-tubulin
目 录
摘要1
关键词1
Abstract1
Key words 1
引言2
1材料与方法2
1.1实验动物2
1.2实验抗体2
1.3 卵母细胞采集与培养3
1.4 HDAC8的抑制3
1.5 siRNA显微注射3
1.6 免疫荧光与激光共聚焦显微镜检测3
1.7 Western Blotting4
1.8 染色体铺片5
1.9 数据统计与分析5
2 结果与分析5
2.1 HDAC8定位于小鼠卵母细胞纺锤体两极 5
2.2 HDAC8的敲低导致卵母细胞纺锤体形态及染色体排列的异常 6
2.3 HDAC8的敲低导致卵母细胞动粒-微管结构的破坏与非整倍体的产生 8
2.4 HDAC8活性的抑制引起卵母细胞纺锤体形态及染色体排列的异常 9
2.5 HDAC8的敲低引起小鼠卵母细胞γ-tubulin定位的紊乱 10
3 讨论 12
致谢 13
参考文献 13
附录A 英文缩略词15
附录B 相关学术论文的发表16
HDAC8在小鼠卵母细胞减数分裂中的作用及其机制的研究
引言
在哺乳动物中，卵母细胞经过两次减数分裂使染色体数目减半。同源染色体的分离发生于第一次减数分裂，而姐妹染色单体则在第二次减数分裂分离至两个子细胞中[1]。卵母细胞的两次减数分裂过程受到精细的调控，以保证成熟分裂的正常进行和遗传物质的分配；减数分裂的异常，如纺锤体形态的畸变，很可能导致非整倍体的产生[2,3]。而且，两次减数分裂的异常还存在着一定的相关性和促发性[4]。子代染色体数量的异常（增多或减少）往往和遗传疾病以及癌症的发生息息相关[5,6]。因此，减数分裂的调控对于正常后代的产生显得尤为重要。