高寒环境相关调节因子对牦牛卵母细胞成熟及附植前不同来源胚胎发育调控的作用机制

高寒物种可调节相关因子使自身生理环境平衡，这些因子可与哺乳动物卵母细胞成熟和胚胎发育分泌的多种细胞因子结合，调控自身的发育。本项目拟以高原牦牛为研究对象，通过基因芯片和实时荧光定量PCR检测牦牛卵母细胞、附植前体内胚胎、体外受精胚胎、核移植胚胎发育各阶段HIF-1α、HSP70及Ref-1的表达差异，筛选相关调控因子；添加相关因子或应用RNAi 技术分别诱导或抑制 HIF-1α、HSP70或Ref-1的表达，分析与发育相关促生长因子、凋亡蛋白的调控作用，评估卵母细胞成熟及胚胎的发育质量；构建卵母细胞成熟和胚胎生长的相似高寒环境，分析高寒调节因子的表达差异及与胚胎发育之间的关系，阐明其对高寒环境适应性及三种因子之间的作用关系。本项目有助于通过调控高寒因子信号通路克服高寒环境的影响，改善牦牛辅助繁殖技术，提高牦牛繁殖效率和经济利益。

Many factors relate with high altitude were regulated by animals living in high altitude for maintain balance of physiological environment, which combined with the factors secreted during in vitro maturation of oocytes and embryonic development, and the development of themselves was regulated. The yak will be as object of study in this project, and the expression of HIF-1α, HSP70, Ref-1 and some factors relate with development were detected with methods of cDNA microarrays and real time PCR during in vitro maturation of oocytes and development of preimplantation embryo from in vivo, in vitro fertilization and nuclear transfer. The regulatory factors was screened; The expression of HIF-1α, HSP70 and Ref-1were inhibited or induced through add some ralation factors or using the method of RNAi, the role of cytokines and apoptin was analysed during in vitro maturation of oocytes and embryonic development, the quality of oocytes and embryo was detected using methods of immunofluorescence and electron microscope; The environment which similar to plateau will be establishment through changed temperature and concentration of oxygen, the expression of HIF-1α, HSP70 and Ref-1 was analysed, and the quality of oocytes and embryo was evaluated，the role of them to oocytes and embryo in plateau environment will be clarified. This project will be help to enhance the assisted reproductive technology, and improve economic benefits and the efficiency of reproduction of yak. which achieved through regulate and control the expression of HIF-1α, HSP70 and Ref-1.efficiency of reproduction of yak,