Institute of Reproductive Medicine, Medical School, Nantong University, Nantong 226019, China.
Experimental Animal Center, Nantong University, Nantong 226001, China.
Int J Mol Sci. 2022 Aug 3;23(15):8627. doi: 10.3390/ijms23158627.
Many testis-specific lncRNAs are highly expressed in late spermatogenesis, especially in spermiogenesis. However, their functions and the underlying mechanisms in male fertility are largely unknown. Here, we screened two highly expressed lncRNAs, 1700101O22Rik (O22Rik) and NONMMUG030480.1 (NM480) in testes, to investigate the roles in spermatogenesis using lncRNA knockout (KO) mouse generated by CRISPER/Cas9 technology. Both testis-specific lncRNAs were mainly expressed from secondary spermatocytes to round spermatids, suggesting that they might be involved in spermiogenesis. Phenotypic analysis showed that the deletion of O22Rik or NM480 did not affect the development of testis and epididymis or spermatogenesis. These results were confirmed in both young and middle-aged male mice. In addition, there was no significant difference in sperm morphology and other parameters including concentration and motility between wild type (WT) and KO mice. Fertility tests showed that litter size was significantly lower in O22Rik KO mice compared with WT controls. Although O22Rik did not exert dramatic roles in spermatogenesis, on molecular levels, its surrounding gene expression was disturbed significantly. Gm32773 was decreased; however, Gm32828 was increased in KO mice. In conclusion, lncRNA O22Rik and NM480 are not individually essential for spermatogenesis in mice.
许多睾丸特异性 lncRNA 在晚期精子发生中高度表达,尤其是在精子发生中。然而,它们在男性生育力中的功能和潜在机制在很大程度上尚不清楚。在这里,我们筛选了两种在睾丸中高度表达的 lncRNA,1700101O22Rik(O22Rik)和 NONMMUG030480.1(NM480),使用 CRISPER/Cas9 技术生成的 lncRNA 敲除(KO)小鼠来研究它们在精子发生中的作用。这两种睾丸特异性 lncRNA 主要从次级精母细胞表达到圆形精子细胞,表明它们可能参与精子发生。表型分析表明,O22Rik 或 NM480 的缺失不影响睾丸和附睾的发育或精子发生。这些结果在年轻和中年雄性小鼠中均得到证实。此外,野生型(WT)和 KO 小鼠之间的精子形态和其他参数(包括浓度和活力)没有显著差异。生育力测试表明,O22Rik KO 小鼠的产仔数明显低于 WT 对照组。尽管 O22Rik 在精子发生中没有发挥显著作用,但在分子水平上,其周围基因的表达受到了显著干扰。Gm32773 减少;然而,KO 小鼠中的 Gm32828 增加。总之,lncRNA O22Rik 和 NM480 对于小鼠的精子发生并不是必不可少的。