摘要
目的探究沉默信息调节因子1(SIRT1)在小鼠生殖细胞发育中的功能。方法利用Cre-LoxP系统构建SIRT1雄性生殖细胞特异性敲除小鼠模型,所有小鼠均为C57BL/6背景。提取对照组(Con)小鼠和SIRT1雄性生殖细胞特异性敲除(cKO)小鼠睾丸总RNA,进行转录组测序分析。两组间比较采用t检验。结果 cKO小鼠睾丸组织SIRT1信使RNA水平(0.14±0.04)低于Con组(1.32±0.09),差异有统计学意义(t=26.144,P<0.01)。cKO小鼠睾丸组织SIRT1蛋白水平(0.28±0.05)低于Con组(1.01±0.06),差异有统计学意义(t=19.705,P<0.01)。转录组测序筛选出差异表达基因3 816个,基因本体(GO)功能富集分析显示,差异表达基因显著富集于转录调控、精子发生及精子活力等生物学功能;京都基因与基因组百科全书(KEGG)分析显示,差异表达基因主要涉及信号通路包括细胞因子-细胞因子受体相互作用、转化生长因子-β(TGF-β)信号通路、补体等。结论雄性生殖细胞特异性SIRT1敲除,导致小鼠睾丸中与精子发生、精子活力及转录调控相关关键基因表达发生变化,影响小鼠生育力。
Objective To explore the function of Silent information regulator 1(SIRT1)in the development of mouse germ cells.Methods Cre-LoxP system was used to construct a SIRT1 male germ cell-specific knockout mouse model,all mice were maintained on C57BL/6 J background.The total RNA from the testes of 2 control mice and 2 cKO mice was extracted,RNA-seq and analysis were performed.SPSS 22.0 software was used for analysis,and t-test was used for comparison between the two groups.Results SIRT1 mRNA level in cKOmice(0.14±0.04)was lower than that in Con group(1.32±0.09),and the difference was statistically significant(t=26.144,P<0.01).SIRT1 protein level in cKO mice(0.28±0.05)was lower than that in Con group(1.01±0.06),and the difference was statistically significant(t=19.705,P<0.01).A total of 3816 differentially expressed genes(DEGs)were screened.Gene Ontology(GO)enrichment analysis of DEGs found that DEGs were significantly enriched in protein binding,main sperm segments,spermatogenesis,and sperm motility,etc.Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis showed that DEGs mainly involved in cytokine-cytokine receptor interaction,transforming growth factor-β(TGF-β)signaling pathway,complement and coagulation cascade,etc.Conclusion Male germ cell-specific SIRT1 knockout leads to changes in the expression of key genes related to spermatogenesis,sperm motility and transcriptional regulation in the testis of mice,which affects the fertility of mice.
作者
梅宁华
杨菁
Mei Ninghua;Yang Jing(Reproductive Medical Center,Renmin Hospital,Wuhan University,Hubei Clinic Research Center for Assisted Reproductive Technology and Embryonic Development,Wuhan 430060,China)
出处
《中华实验外科杂志》
CAS
北大核心
2021年第11期2212-2215,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金(81571513、81771662、81771618)。
关键词
沉默信息调节因子1
转录组学
精子发生
睾丸发育
Silent information regulator 1
Transcriptomics
Spermatogenesis
Testicular development
