Li Hongyi, Li Yuan, Wang Yuting, Liu Lijian, Cui Ji, Zhang Mao, Yan Yiming
College of Biology and Agriculture, Shaoguan University, Guangdong, China.
PLoS One. 2025 Jun 27;20(6):e0325260. doi: 10.1371/journal.pone.0325260. eCollection 2025.
Nanos2 plays a key role in self-renewing spermatogenic stem cells (SSCs) and maintains the stem cell state during spermatogenesis. Alleles of the Nanos2 gene knockout showed germline ablated but otherwise structurally normal. To identify the probable ceRNA regulator involved in the process of spermatogenesis by Nanos2, whole transcriptome sequencing was performed in the testes between Nanos2 knock out mice and wild type mice. Finally, a total of 8644 Differentially expressed (DE) mRNAs,180 DE miRNAs, 9538 DE lncRNA and 481 DE circRNAs were identified. Three of each RNAs were selected randomly and identified by real-time PCR to verify the accuracy of sequencing. GO and KEGG functional enrichment analyses revealed similar result of DE mRNAs and target of DE miRNAs/lncRNAs/ circRNAs, mainly involved in the generation, composition, and activity of sperm cells. Furthermore, the regulatory ceRNA network of miRNA(up)-circRNA-lncRNA-mRNA and miRNA(down)-circRNA-lncRNA-mRNA were constructed based on the common targeted miRNA.The results enable us to better understand the interaction of coding RNA and non coding RNA in regulating the generation of spermatogenic stem cells through Nanos2 pathway, and also provided novel insights into molecular mechanism of spermatogenesis.
Nanos2在自我更新的生精干细胞(SSCs)中起关键作用,并在精子发生过程中维持干细胞状态。Nanos2基因敲除的等位基因显示生殖系被消融,但结构上正常。为了鉴定Nanos2在精子发生过程中可能涉及的ceRNA调节因子,对Nanos2基因敲除小鼠和野生型小鼠的睾丸进行了全转录组测序。最终,共鉴定出8644个差异表达(DE)的mRNA、180个DE miRNA、9538个DE lncRNA和481个DE circRNA。随机选择每种RNA中的三个,并通过实时PCR进行鉴定,以验证测序的准确性。GO和KEGG功能富集分析揭示了DE mRNA以及DE miRNA/lncRNA/circRNA靶标的相似结果,主要涉及精子细胞的生成、组成和活性。此外,基于共同靶向的miRNA构建了miRNA(上调)-circRNA-lncRNA-mRNA和miRNA(下调)-circRNA-lncRNA-mRNA的调控ceRNA网络。这些结果使我们能够更好地理解编码RNA和非编码RNA在通过Nanos2途径调节生精干细胞生成中的相互作用,也为生精的分子机制提供了新的见解。