Rab32在调控小鼠卵母细胞减数分裂成熟与受精中的作用及机制研究

Rab家族成员所介导的囊泡转运过程，通过其活性形式的动态转换，介导了胞质中营养物质和调控信号的传导，对于卵母细胞成熟、受精和早期胚胎发育具有重要意义。胞质成熟与卵母细胞质量密切相关，但胞质内组分复杂，调控过程和具体机制不详。我们前期研究发现，干扰Rab32的功能会影响细胞骨架装配及卵母细胞成熟过程，并导致异常受精。因此我们提出假说：Rab32 GTP酶可通过调控细胞骨架装配间接或直接影响细胞器分布与功能，并通过其转运特性影响特定蛋白的分布模式，最终影响卵母细胞成熟与受精过程。本课题将通过RNAi和点突变基因过表达等技术结合敲除鼠模型和挽救试验，重点探究：①Rab32对细胞骨架装配的动态调控机制；②Rab32对细胞器尤其是皮质颗粒的分布与功能的调控；③Rab32的效应因子与调节机制；④Rab32影响受精的分子机制。这将为深入揭示卵母细胞成熟与受精的动态调控过程奠定理论基础。

The vesicle transport process mediated by Rab family members mediates the transmission of nutrients and regulatory signals in the cytoplasm through the dynamic conversion of its active form, which is of great significance for oocyte maturation, fertilization and early embryo development. Cytoplasmic maturation is closely related to the quality of oocytes, but the cytoplasmic components are complex, and the regulation process and specific mechanisms are unknown. Our previous studies have found that Rab32 disruption will affect the cytoskeleton assembly and oocyte maturation process, and lead to abnormal fertilization. Therefore, we propose the hypothesis: Rab32 GTPase can indirectly or directly affect the distribution and function of organelles by regulating cytoskeleton assembly, and affect the distribution pattern of specific proteins through its transport characteristics, and ultimately affect the maturation and fertilization process of oocytes. This project will use RNAi and point mutation gene overexpression techniques combined with knockout mouse models and rescue experiments, focusing on these points: ①The dynamic regulation mechanism of Rab32 on cytoskeleton assembly; ②The regulation of Rab32 on the distribution and function of organelles, especially cortical granules; ③The effects of Rab32 factors and regulatory mechanism; ④The molecular mechanism of Rab32 affecting fertilization. This will lay a theoretical foundation for further revealing the dynamic regulation process of oocyte maturation and fertilization.