RNA结合蛋白介导的转录后调控对精原干细胞维持至关重要，RNA结合蛋白也易与RNA在细胞质聚集形成P granule和Stress granule, 聚集可调控RNA代谢和信号转导等，参与细胞稳态维持和应激。哺乳动物精子发生对热激尤其敏感，RNA结合蛋白聚集参与该过程的研究还十分有限；Dnd1是生殖细胞特异RNA结合蛋白，该蛋白缺失导致原始生殖细胞凋亡，引发不育。Dnd1对胚胎期生殖细胞内聚集形成P granule是必须的，预实验结果显示，Dnd1在出生后小鼠睾丸中主要表达在精原干细胞。本研究以探讨Dnd1介导的蛋白聚集对精原干细胞维持和应激的作用和分子机理为出发点，离体条件下，通过体外培养的精原干细胞模型，在体条件下，用条件性基因敲除模型，结合免疫沉淀结合蛋白质组学、蛋白结构分析和RNAi等方法研究Dnd1介导的蛋白聚集对精子发生的影响，为精子发生的机理提供重要的理论支撑。

Post-transcriptional regulation mediated by RNA binding proteins plays crucial roles in spermatogonial stem cell (SSC) maintenance. RNA binding proteins interact with RNAs and prone to form aggregations named P granule or Stress granule, these non-membrane units in cytoplasm are implicated in mRNA metabolic pathways or signal transduction and are involved in cell homeostasis and stress responses. Heat stress has detrimental effect on mammalian spermatogenesis, however, whether and how protein aggregation pathway involved in this process is largely unknown; Dnd1 is a germ cell specific protein, loss of Dnd1 leads to primordial germ cell lost and infertility. Our previous study also showed that Dnd1 is required for P granule formation in embryonic germ cells. Preliminary data found Dnd1 mainly expressed in spermatogonial stem cell population in postnatal testis. In this study, we are planning to analyze functions and underlying mechanisms in SSC homeostasis and stress response, which mediated by Dnd1 protein aggregation. We will use 1) in vitro cultured SSC and 2) conditional knockout mouse as research model, and also use multiple methods like immuno-precipitation, proteomics, RNAi, protein structure analysis and testis phenotype analysis to uncover the molecular mechanisms in spermatogenesis.