二酰甘油激酶DGKη在雄性减数分裂过程中的功能及机制研究

减数分裂异常会导致非梗阻性无精子症(NOA)。深入探讨减数分裂异常的机制对改善男性生殖健康具有重大的意义。前期对临床NOA患者测序发现了DGKH的移码突变，而DGKH编码的二酰甘油激酶η（DGKη）在睾丸中优势表达，又因DGK是催化二酰甘油（DAG）产生磷脂酸（PA）的关键酶，负责细胞内信号传导，故推测DGKη在精子发生过程中可能发挥着重要功能，然而目前却并未有相关报道。因此，我们构建了Dgkh敲除小鼠来探究DGKη在精子发生过程中的功能与机制，前期研究结果发现DGKη缺失导致雄鼠不育，精母细胞分裂期阻滞，脂质组检测发现DGKη缺失雄鼠睾丸中DAG蓄积，PA减低，提示DGKη通过调节精母细胞局部DAG浓度或PA浓度，参与减数分裂进程。本项目拟通过深入分析Dgkh敲除小鼠表型，结合体外细胞系模型，寻找相互作用分子，全面阐明DGKη参与精母细胞核内微环境调节减数分裂进程的调控机制与调控网络。

Abnormal meiosis can lead to non-obstructive azoospermia (NOA). In-depth exploration of the mechanism of meiosis errors is important for improving male reproductive health and fertility. In the early stage，sequencing of clinical NOA patients found a frameshift mutation of DGKH. The diacylglycerol kinase η (DGKη) encoded by DGKH is extremely abundant in the testis, and as DGK plays an important role in intracellular signal transduction which catalyzes the phosphorylation of diacylglycerol (DAG) to produce phosphatidic acid (PA). It is speculated that DGKη may play an important role in the process of spermatogenesis, however, there is no relevant report yet. Therefore, we constructed Dgkh knockout mice to explore the function and mechanism of DGKη in the process of spermatogenesis. The previous study found that the lack of DGKη led to male infertility and the arrest of spermatocyte division. Lipidomic revealed that DAG accumulated and PA decreased in the testes of male mice lacking DGKη，which suggested that DGKη participates in the process of meiosis by regulating the local concentration of DAG or PA in spermatocytes. This project intends to comprehensively clarify the regulatory mechanism and regulatory network of DGKη involved in the microenvironment of spermatocyte nucleus which to regulate the process of meiosis by thoroughly analyze the phenotype of Dgkh knockout mice, combine with in vitro cell line models, and search for interacting molecules.