小分子化合物在水牛原始卵泡体外激活中的作用及其分子机制的研究

人造卵母细胞被《科学》评为2016年度十大科学突破之一，动物生殖生理研究也是生命科学研究中最具吸引力的领域之一。原始卵泡激活是动物卵泡发生的基础，mTOR通路与PI3K/Akt通路是原始卵泡激活的两条关键通路，具有平行和协同效应的特点，不存在上下游的级联关系，但是否存在主次之分仍不清楚。本研究拟在课题组从事水牛生殖生理研究近30 年的基础上，通过单独或联合添加mTOR和PI3K/Akt通路激活剂与抑制剂，比较不同添加浓度和配伍方式对水牛卵泡发育的影响，建立小分子化合物促进水牛原始卵泡体外激活的最佳方案；在此基础上，应用免疫组化、RNA-Seq、QRT-PCR和蛋白印迹技术，阐明小分子化合物调节水牛原始卵泡发育的分子机制。本研究将丰富水牛等单胎大家畜卵泡发育的分子机制理论，并为水牛优良种畜遗传资源的充分开发应用奠定理论基础。

As one of the top-ten scientific breakthroughs of 2016, artificial oocytes stand as one of the most attractive life-science research areas in the field of animal reproductive physiology. Activation of primordial follicles is the basis of follicular development where mTOR and PI3K/Akt pathways exist as two critical pathways for the activation of primordial follicles. These two pathways have parallel and synergistic characteristics,with no relationship between the upstream and downstream cascades. However, it remains unclear whether there is a primary and secondary point. This study follows nearly 30 years of buffalo reproductive physiology research as performed by our research group. The development of buffalo primordial follicles through mTOR and PI3K/Akt activation or inhibition were of interest in this study. This was accomplished through testing the concentration capability of agonists and antagonists of the aforementioned pathways.Various techniques including immunohistochemistry, RNA-Seq, QRT-PCR, and protein imprinting were employed to elucidate the molecular mechanisms of small molecular compounds regulating the development of buffalo primordial follicles. This study will enrich the molecular mechanism theory of follicular development of a single foetus of large livestock, such as buffalo, and establish a theoretical foundation for the full development and application of excellence in breeding livestock genetic resources of buffalo.