mTOR通路对蒙古马血睾屏障建立的调控作用及其机制

血睾屏障的建立是哺乳动物性成熟过程中较早事件之一，作为机体内重要免疫屏障，是精子正常生成必需的。目前血睾屏障调控研究集中在啮齿模式动物，且主要关注成体睾丸血睾屏障动态开合机理，对初情期阶段血睾屏障初建机制研究不多。前期我们RNAseq技术测序对比初情期前后蒙古马睾丸转录组表达发现曾报道的成体鼠血睾屏障动态变化相关mTOR通路多个因子在初情期后有显著增高。由此我们认为mTOR通路不仅如其他实验证明对成体血睾屏障动态变化重要，可能对马性成熟阶段血睾屏障初建也有重要作用。所以本课题将延续前期工作通过免疫组化/荧光法细致鉴定蒙古马睾丸内mTOR通路和血睾屏障初建相关分子之间时空表达关系；通过组织细胞实验解析mTOR对屏障初建的调控作用；体外支持细胞实验向下确定mTOR介导屏障初建主要作用点和向上探寻睾酮等潜在mTOR上游因子，结合体内外数据以期阐明较完整的mTOR通路介导蒙古马血睾屏障初建机制。

The establishment of the blood-testis barrier is one of the earlier events in sexual maturation in mammals. As an important immune barrier in the body, it is necessary for the normal spermtogenesis. At present, studies on the regulation of blood-testis barrier mainly focus on rodent animal models, and largely on the dynamic opening-closing mechanism of the blood testis barrier in adult testes. Our previous RNAseq for the comparison of testes transcriptomes between prepubertal and postpubertal Mongolian stallions showed that between the two groups, there were significant expressional differences in the mTOR pathway-related factors, which are reported as main regulators in the dynamic changes of the blood-testis barrier of adult mice, and these changes well corresponded to the testis state of before and after barrier formation. We speculate that the mTOR pathway play an important role not only in the dynamics of adult blood-testis barrier, as evidenced by other experiments, but also in barrier initial formation. Therefore, this project will further identify the spatiotemporal relationship between the mTOR pathway factors and the blood-testis barrier establishment-related molecules with the method of immuno-histochemistry/fluorescence, and then prove the regulatory function of mTOR on the barrier establishment via histochemistry and cell biology experiments, which followed by the experiments to identify the downtream factors, on which mTOR mainly exerts effects to regulate barrier establishment as well as the mTOR upstream factors such as testosterone that control barrier establishment through mTOR, so that finally figure out a more complete and detailed molecular process of the mTOR signaling pathway mediated blood-testis barrier establishment in Mongolian horses.