体细胞核替代生发泡重构卵母细胞减数分裂异常的机理研究

项目名称： 体细胞核替代生发泡重构卵母细胞减数分裂异常的机理研究

项目编号： No.31272444

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 谭景和

作者单位： 山东农业大学

项目金额： 88万元

中文摘要： 研究表明，卵母细胞减数分裂异常是导致不孕、自发流产和先天缺陷的主要原因之一。用体细胞核替代生发泡（GV）重构卵母细胞是研究减数分裂细胞周期调控和核质互作机理不可替代的模型，也是实现体细胞单倍体化，治疗由于卵母细胞或精子完全缺乏而导致不育的重要方法。然而，目前用体细胞核替代GV重构的卵母细胞还不能正常成熟和激活。因此，深入研究这种重构卵母细胞减数分裂异常的机理既有助于了解减数分裂，也有助于建立体细胞单倍体化技术。我们将在发现初级精母细胞核替代GV重构卵母细胞皮质不极化和MAPK在极体和卵之间分配不均的基础上，深入研究1）重构卵纺锤体迁移、皮质极化、染色体凝集及MAPK分配对大极体产生、极体退化和原核形成的影响；2）参与调控纺锤体迁移、染色体凝集/分离和分裂沟定位等相关分子在重构卵中的表达；3）通过RNAi和基因过表达研究相关分子在调控重构卵减数分裂中的作用，力争通过某些分子过表达使重构卵发育

中文关键词： 减数分裂；体细胞单倍体化；纺锤体迁移；染色体凝集；MAPK

英文摘要： Studies have demonstrated that abnormal oocyte meiosis is a major problem that causes apogeny, spontaneous abortion and birth defect. Nuclear transfer into prophase oocytes can be used as an approach not only to studying the cell cycle control and nuclear-cytoplasmic interactions during oocyte meiosis, but also to achieving somatic cell haploidization and treating sterility caused by complete absence of oocytes or spermatozoa. Up to date, however, normal maturation and activation have not been achieved in oocytes reconstructed by replacing germinal vesicles (GV) of oocytes with somatic cell nuclei. Therefore, mechanism studies on meiotic anomalies in oocytes manipulated to replace GV with somatic cell nuclei will contribute not only to our understanding of the mechanism of germ cell meiosis, but also to the establishment of techniques for somatic cell haploidization. Based on our findings that oocytes manipulated to replace GV with primary spermatocyte (PS) nuclei did not undergo cortical polarization and showed an unequal MAPK allocation with their polar bodies, this project will (i) study the effects of spindle migration, cortical polarization, chromosome condensation and MAPK allocation between oocyte and polar bodies on the generation of giant polar bodies, polar body degeneration and pronuclear formation af

英文关键词： Meiosis；Somatic cell haploidization；Spindle migration；Chromosome condensation；MAPK