Chen Zhen, Liu Cong, Qu Wei, Han Yan, Zhu Xiaoyu, Li Zejia, Ma Dupeng, Huang Mengya, Gong Weihao, Sun Qi, Lei Junhao, Guo Rui, Luo Mengcheng
Provincial Key Laboratory of Developmentally Originated Disease, TaiKang Center for Life and Medical Sciences, School of Basic Medical Sciences, Wuhan University, Wuhan, China.
Frontier Science Center for Immunology and Metabolism, Hubei Key Laboratory of Cell Homeostasis, College of Life Sciences, Taikang Center for Life and Medical Sciences, Wuhan University, Wuhan, China.
Andrology. 2025 Jul;13(5):1236-1250. doi: 10.1111/andr.13774. Epub 2024 Oct 3.
The PWWP domain-containing proteins are involved in chromatin-associated biological processes, including transcriptional regulation and DNA repair, and most of them are significant for gametogenesis and early embryonic development in mammals. PWWP3A, one of the PWWP domain proteins, is a reader of H3K36me2/H3K36me3 and a response factor to DNA damage. However, the physiological role of PWWP3A in spermatogenesis and fertility remains unclear.
The goal of this study was to explore the function and mechanism of PWWP3A in the process of spermatogenesis.
We generated V5-Pwwp3a KI mice and PWWP3A polyclonal antibody to observe the localization of PWWP3A in vivo. Meanwhile, Pwwp3a KO mice was used to explore the function in spermatogenesis.
We reported that PWWP3A is a predominant expression in the testis of mice. During spermatogenesis, PWWP3A exhibits the temporal expression from early-pachytene to the round spermatids. The results of spermatocyte spreading and immunostaining showed that PWWP3A aggregated on the XY body, which then diffused as the XY chromosome separated at late-diplotene. Although the depletion of PWWP3A had no obvious reproductive defects in young male mice, there were observed morphological abnormalities in sperm heads. Immunoprecipitation demonstrated the interaction of PWWP3A with DNA repair proteins SMC5/6; however, PWWP3A deficiency did not result in any meiotic defects. Notably, the testes of aged male Pwwp3a KO mice displayed pronounced degeneration, and were characterized by the presence of vacuolated seminiferous tubules. Furthermore, RNA-seq analysis revealed an upregulation in the expression of genes which may be involving in immunoregulatory and inflammatory response pathways in aged Pwwp3a KO mice with testicular degeneration.
Our study showed that PWWP3A was highly enriched in the mouse testis, and the Pwwp3a KO mice were fertile. However, the aged Pwwp3a KO male mice displayed testicular atrophy that may be due to changes in the immune micro-environment or abnormal repair of DNA damage.
含PWWP结构域的蛋白质参与染色质相关的生物学过程，包括转录调控和DNA修复，其中大多数对哺乳动物的配子发生和早期胚胎发育具有重要意义。PWWP3A是PWWP结构域蛋白之一，是H3K36me2/H3K36me3的识别蛋白和DNA损伤的反应因子。然而，PWWP3A在精子发生和生育中的生理作用仍不清楚。
本研究旨在探讨PWWP3A在精子发生过程中的功能及机制。
我们构建了V5-Pwwp3a基因敲入小鼠和PWWP3A多克隆抗体，以观察PWWP3A在体内的定位。同时，利用Pwwp3a基因敲除小鼠探讨其在精子发生中的功能。
我们报道PWWP3A在小鼠睾丸中呈优势表达。在精子发生过程中，PWWP3A表现出从粗线期早期到圆形精子细胞的时序表达。精母细胞铺展和免疫染色结果显示，PWWP3A聚集在XY体上，随后随着XY染色体在双线期后期分离而扩散。虽然PWWP3A的缺失在年轻雄性小鼠中没有明显的生殖缺陷，但观察到精子头部存在形态异常。免疫沉淀证明PWWP3A与DNA修复蛋白SMC5/6相互作用；然而，PWWP3A缺陷并未导致任何减数分裂缺陷。值得注意的是，老年雄性Pwwp3a基因敲除小鼠的睾丸出现明显退化，其特征是生精小管出现空泡化。此外，RNA测序分析显示，在睾丸退化的老年Pwwp3a基因敲除小鼠中，可能参与免疫调节和炎症反应途径的基因表达上调。
我们的研究表明，PWWP3A在小鼠睾丸中高度富集，Pwwp3a基因敲除小鼠具有生育能力。然而，老年Pwwp3a基因敲除雄性小鼠出现睾丸萎缩，这可能是由于免疫微环境的改变或DNA损伤修复异常所致。
