School of Life Science, Bengbu Medical College, Bengbu, P. R. China.
NHC Key Laboratory of Male Reproduction and Genetics, Guangdong Provincial Reproductive Science Institute (Guangdong Provincial Fertility Hospital), Guangzhou, P. R. China.
Poult Sci. 2022 Jul;101(7):101930. doi: 10.1016/j.psj.2022.101930. Epub 2022 Apr 27.
Long noncoding RNAs (lncRNAs) have been demonstrated to regulate reproduction in mammals. Our previous study revealed that the expression level of lncRNA-Gm2044 was obviously elevated in nonobstructive azoospermia with spermatogonial arrest. Here, a transgenic mouse model of lncRNA-Gm2044 in spermatogonia using the Stra8 promoter was constructed to explore the roles of upregulated lncRNA-Gm2044 in male fertility. Testicular morphology and fertility weren't affected in transgenic mice expressing lncRNA-Gm2044. However, overexpression of lncRNA-Gm2044 in spermatogonia partially impaired spermatogenesis in the transgenic mice. Then, transcriptome sequencing was executed to find the potential signaling pathway repressing spermatogenesis in germ cells of lncRNA-Gm2044 transgenic mice. Through quantitative analysis of differentially expressed genes, 442 upregulated mRNAs and 147 downregulated mRNAs were displayed in male germ cells of Gm2044-transgenic mice (Gm2044-Tg) compared with non-transgenic mice (Non-Tg). Using gene ontology (GO) analysis, differentially expressed genes were shown to play vital roles in RNA_metabolic_process, Central_element, Enzyme_binding, and Intracellular_bridge. Using Kyoto encyclopedia of genes and genomes (KEGG) analysis, differentially expressed genes were shown to participate in RNA_transport, Cell_cycle, Renin-angiotensin_system, and Chemokine_signaling_pathway. Gene Set Enrichment Analysis (GSEA) revealed that Acrosome_assembly and Sperm_plasma_membrane were involved in the overexpression of lncRNA-Gm2044 blocking spermatogenesis. Furthermore, some of the most differentially expressed mRNAs were verified by RT-qPCR. In addition, we determined that the lncRNA-Gm2044 has no ability to translate into peptides by the bioinformatics method and molecular experiment. Thus, lncRNA-Gm2044 is a novel molecular target for the diagnosis and treatment of male infertility.
长链非编码 RNA(lncRNA)已被证明在哺乳动物的生殖中起调节作用。我们之前的研究表明,lncRNA-Gm2044 在生精细胞阻滞的非梗阻性无精子症中的表达水平明显升高。在这里,我们构建了使用 Stra8 启动子在生精细胞中转录 lncRNA-Gm2044 的转基因小鼠模型,以探讨上调的 lncRNA-Gm2044 在男性生育力中的作用。lncRNA-Gm2044 转基因小鼠的睾丸形态和生育力没有受到影响。然而,lncRNA-Gm2044 在生精细胞中的过表达部分损害了转基因小鼠的精子发生。然后,进行转录组测序以找到抑制 lncRNA-Gm2044 转基因小鼠精母细胞中精子发生的潜在信号通路。通过差异表达基因的定量分析,与非转基因小鼠(Non-Tg)相比,Gm2044 转基因小鼠(Gm2044-Tg)的雄性生殖细胞中显示出 442 个上调的 mRNAs 和 147 个下调的 mRNAs。通过基因本体论(GO)分析,差异表达基因在 RNA 代谢过程、中央元件、酶结合和细胞内桥中发挥重要作用。通过京都基因与基因组百科全书(KEGG)分析,差异表达基因参与 RNA 转运、细胞周期、肾素-血管紧张素系统和趋化因子信号通路。基因集富集分析(GSEA)表明,顶体组装和精子质膜参与了 lncRNA-Gm2044 过表达阻断精子发生。此外,一些差异表达最明显的 mRNAs 通过 RT-qPCR 进行了验证。此外,我们通过生物信息学方法和分子实验确定 lncRNA-Gm2044 没有翻译为肽的能力。因此,lncRNA-Gm2044 是男性不育症诊断和治疗的新分子靶点。