颗粒细胞自噬通过调控糖酵解影响卵母细胞减数分裂成熟及早期胚胎发育的机制研究

The regulation of oocyte maturation is a hot spot in reproductive biology. Recent studies indicated that granulosa cell could provide energy substracts to meet the energy needs of oocyte maturation through activating cell glycolysis, which suggests glycolysis plays an important role in regulating oocyte quality. Our previously results demonstrated that the induction of granlusa cell autophagy was closely related with glycolysis level in mouse antral follicles, however, it is still unkown that whether granulosa cell autophagy could affect oocyte quality through glycolysis pathway. In this project, we investigate the role of granulosa cell autophagy in regulating oocyte meiotic maturation and the development of early embryo by using conditional knockout model and oocyte-granulosa cell coculture system. We detect the molecular phenotype of glycolysis and investigate the interaction role between autophagy and glycolysis by using glycolysis inhibitor, mass spectrometry and RNA interference, which helps to understand the molecular mechanism of autophagy in regulating oocyte meiotic maturation and the development of early embryo through glycolysis pathway. Overall, this project provides new insights into oocyte maturation, which could further help to improve the development of target drug for female ovulatory disorders.

卵母细胞成熟质量的调控是当前生殖生物领域的研究热点。颗粒细胞糖酵解被证实可以为卵母细胞的成熟提供能量代谢底物，是影响卵母细胞成熟质量的重要因素。我们前期发现，小鼠有腔卵泡中颗粒细胞自噬的发生与其糖酵解水平的稳定密切相关，然而颗粒细胞自噬能否通过糖酵解影响卵母细胞成熟质量，目前尚不清楚。本项目拟以颗粒细胞条件性敲除自噬小鼠和卵母细胞-颗粒细胞共培养为模型，分别观察卵母细胞成熟以及早期胚胎发育指标变化情况以评价颗粒细胞自噬对卵母细胞成熟质量的影响；检测颗粒细胞自噬激活糖酵解的分子表型并通过抑制剂、蛋白质谱及RNA干涉等手段探究颗粒细胞自噬与糖酵解的互作机制，明确颗粒细胞自噬通过调控糖酵解影响卵母细胞减数分裂成熟及早期胚胎发育的分子机制。本项目不仅为卵母细胞成熟的机制研究提供新思路，也为女性排卵障碍相关疾病靶向药物的研发提供新参考。

卵母细胞成熟质量调控是生殖生物学、胚胎工程领域中的关键环节。本项目依托前期工作基础，围绕卵泡发育过程中颗粒细胞与卵母细胞对话机制，重点探究了颗粒细胞中的自噬过程对于卵母细胞的生长发育的重要意义。利用体外颗粒细胞-卵母细胞共培养模型，我们研究颗粒细胞自噬对于卵母细胞成熟及后续胚胎发育的具体影响，并发现自噬有助于改善卵母细胞的核成熟以及受精能力。进一步，基于颗粒细胞对于卵母细胞支撑作用的本质，我们运用转录组学、代谢组学探究并发现了颗粒细胞自噬能够显著影响代谢相关基因通路。再次，我们运用分子生物学、细胞生物学等手段，具体的研究了自噬调控颗粒细胞能量代谢的分子机制，筛选到1个重要的生物分子标记。在人类临床卵泡液样本及猪的卵泡中，该分子标志物与卵母细胞成熟质量具有高相关性。总体来说，项目的研究结果系统阐明了卵泡发育过程中，颗粒细胞自噬的具体作用，揭示了自噬通过调控颗粒细胞能量代谢从而影响卵母细胞成熟质量的分子机制，为临床上治疗女性排卵障碍等疾病提供了一定的干预策略。