小鼠睾丸支持细胞参与精子发生障碍的单细胞转录组研究  

Single-cell transcriptome study of testicular Sertoli cells involved in defective spermatogenesis in mice

作　　者：凌晓辉[1] 刘利敏[1] 陈佳鸿[2] 陈志云[1] Ling Xiaohui;Liu Limin;Chen Jiahong;Chen Zhiyun(Reproductive Medicine Centre,Huizhou Municipal Central Hospital,Huizhou 516001,Guangdong,China;Department of Urology,Huizhou Municipal Central Hospital,Huizhou 516001,Guangdong,China)

机构地区：[1]惠州市中心人民医院生殖中心,广东惠州516001 [2]惠州市中心人民医院泌尿外科,广东惠州516001

出　　处：《中国男科学杂志》2024年第4期38-47,共10页Chinese Journal of Andrology

基　　金：广东省医学科学技术研究基金(B2021317);惠州市科技研发计划重点项目(2022CZ010415)。

摘　　要：目的精子发生过程复杂,需要睾丸内细胞之间复杂的相互作用,但是支持细胞参与精子发生的机制仍有待探讨。方法从高通量基因表达数据库(GEO:GSE113293),收集了正常精子发生的野生型C57BL/6J雄性小鼠和4种不同突变小鼠(敲除Mlh3、Hormad1、Cul4a;Cnp转基因)的单细胞RNA测序数据,进而对支持细胞差异基因进行基因功能富集分析(GO)、京都基因和基因组百科全书(KEGG)富集通路分析、蛋白-蛋白相互作用(PPI)网络分析、转录因子调控分析(SCENIC)及基因集变异分析(GSVA)。结果本研究鉴定出9种生精细胞亚群(未分化精原细胞、分化精原细胞、细线期/偶线期、粗线期、双线期、圆形精子细胞、长形精子细胞及精子)、2种体细胞群(间质细胞和支持细胞)和1种未知类型的细胞群。与正常精子发生模型相比,小鼠精子发生缺陷模型的支持细胞有276个基因表达上调,234个基因表达下调。GO功能富集分析显示,支持细胞的差异基因在精子发生及能量代谢等过程中显著富集。KEGG富集分析显示,支持细胞在核糖体、糖酵解、氧化磷酸化等途径显著富集。PPI网络分析鉴定出20个hub基因,均属于核糖体基因。GSVA分析显示支持细胞中上调的基因与Notch信号通路密切相关。结论支持细胞相关的的hub基因及Notch信号通路参与精子发生的过程,本研究为精子发生障碍导致不育症提供新的视角。Objective The process of spermatogenesis is complex involving multiple regulatory mechanisms and requiring complex interactions between germ cells and somatic cells.The mechanisms of Sertoli cells involving in spermatogenesis remain to be explored.Methods We collected single-cell RNA sequencing data from a high-throughput gene expression database(GEO:GSE113293)from wild-type C57BL/6J male mice with normal spermatogenesis and four different mutant mouse strains(knockout Mlh3,Hormad1,and Cul4a;Cnp transgene).Then bioinformatics analyses such as gene function enrichment analysis(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG)enriched pathway analysis,protein-protein interaction(PPI)network analysis,transcription factor regulation analysis(SCENIC),and gene set variation analysis(GSVA)on the Sertoli cells were further indentified.Results We identified nine subtypes of spermatogenic cell clusters(undifferentiated spermatogonia,differentiating spermatogonia,leptotene/zygotene,pachytene,diplotene,round spermatids,elongating spermatids,and spermatozoa),and two types of somatic cell clusters(Leydig cells,and Sertoli cells),and one unknown type of cell cluster.Compared to the normal spermatogenesis model,276 genes were upregulated and 234 genes were downregulated in the Sertoli cells of the mouse spermatogenesis defect model.GO analysis showed that differential genes in Sertoli cells were significantly enriched in spermatogenesis and energy metabolism.KEGG enrichment analyses showed that the homogeneous ribosomal pathway,glycolytic pathway,and oxidative phosphorylation pathway were significantly enriched in Sertoli cells.Protein-protein interaction(PPI)network analysis of differentially expressed genes identified 20 hub genes,all of which belonged to ribosomal genes,and GSVA analysis revealed that the upregulated genes in Sertoli cells were closely related to the Notch signaling pathway.Conclusion The hub genes and the Notch signaling pathway in Sertoli cells are involved in the process of spermatogenesis.This study provi

关 键 词：睾丸支持细胞 精子发生 单细胞分析 序列分析 RNA 小鼠 

分 类 号：R321.1[医药卫生—人体解剖和组织胚胎学] R318.04[医药卫生—基础医学]