State Key Laboratory of Reproductive Medicine and Offspring Health, Center for Reproductive Medicine, Institute of Women, Children and Reproductive Health, Shandong University, Jinan, Shandong, 250012, China.
National Research Center for Assisted Reproductive Technology and Reproductive Genetics, Shandong University, Jinan, Shandong, 250012, China.
Adv Sci (Weinh). 2024 Aug;11(31):e2403866. doi: 10.1002/advs.202403866. Epub 2024 Jun 18.
The progression of spermatogenesis along specific developmental trajectories depends on the coordinated regulation of pre-mRNA alternative splicing (AS) at the post-transcriptional level. However, the fundamental mechanism of AS in spermatogenesis remains to be investigated. Here, it is demonstrated that CWF19L2 plays a pivotal role in spermatogenesis and male fertility. In germline conditional Cwf19l2 knockout mice exhibiting male sterility, impaired spermatogenesis characterized by increased apoptosis and decreased differentiated spermatogonia and spermatocytes is observed. That CWF19L2 interacted with several spliceosome proteins to participate in the proper assembly and stability of the spliceosome is discovered. By integrating RNA-seq and LACE-seq data, it is further confirmed CWF19L2 directly bound and regulated the splicing of genes related to spermatogenesis (Znhit1, Btrc, and Fbxw7) and RNA splicing (Rbfox1, Celf1, and Rbm10). Additionally, CWF19L2 can indirectly amplify its effect on splicing regulation through modulating RBFOX1. Collectively, this research establishes that CWF19L2 orchestrates a splicing factor network to ensure accurate pre-mRNA splicing during the early steps of spermatogenesis.
精子发生沿着特定的发育轨迹进行，这取决于转录后水平上的前体 mRNA 可变剪接 (AS) 的协调调节。然而，精子发生中 AS 的基本机制仍有待研究。本文证明 CWF19L2 在精子发生和雄性生育力中起着关键作用。在生殖系条件性 Cwf19l2 敲除小鼠中观察到雄性不育，精子发生受损，特征为凋亡增加，分化的精原细胞和精母细胞减少。发现 CWF19L2 与几种剪接体蛋白相互作用，参与剪接体的正确组装和稳定性。通过整合 RNA-seq 和 LACE-seq 数据，进一步证实 CWF19L2 直接结合并调节与精子发生（Znhit1、Btrc 和 Fbxw7）和 RNA 剪接（Rbfox1、Celf1 和 Rbm10）相关基因的剪接。此外，CWF19L2 可以通过调节 RBFOX1 间接放大其对剪接调控的影响。总之，这项研究确立了 CWF19L2 协调一个剪接因子网络，以确保在精子发生的早期步骤中准确的前体 mRNA 剪接。
