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In the testis of <jats:italic>Drosophila<\/jats:italic>, the terminal epithelium region is involved in spermatid release and maturation, while its functional diversity and regulatory mechanism remain poorly understood. In this study, we performed single\u2010cell RNA\u2010sequencing analysis (scRNA\u2010seq) to characterize the transcriptomes of terminal epithelium in <jats:italic>Drosophila<\/jats:italic> testes at 2\u2010, 10 and 40\u2010Days. Terminal epithelium populations were defined with <jats:italic>Metallothionein A<\/jats:italic> (<jats:italic>MtnA<\/jats:italic>) and subdivided into six novel sub\u2010cell clusters (EP0\u2013EP5), and a series of marker genes were identified based on their expressions. The data revealed the functional characteristics of terminal epithelium populations, such as tight junction, focal adhesion, bacterial invasion, oxidative stress, mitochondrial function, proteasome, apoptosis and metabolism. Interestingly, we also found that disrupting genes for several relevant pathways in terminal epithelium led to male fertility disorders. Moreover, we also discovered a series of age\u2010biased genes and pseudotime trajectory mediated state\u2010biased genes during terminal epithelium aging. Differentially expressed genes during terminal epithelium aging were mainly participated in the regulation of several common signatures, e.g. mitochondria\u2010related events, protein synthesis and degradation, and metabolic processes. We further explored the <jats:italic>Drosophila<\/jats:italic> divergence and selection in the functional constraints of age\u2010biased genes during aging, revealing that age\u2010biased genes in epithelial cells of 2\u2009Days group evolved rapidly and were endowed with greater evolutionary advantages. scRNA\u2010seq analysis revealed the diversity of testicular terminal epithelium populations, providing a gene target resource for further systematic research of their functions during aging.<\/jats:p>","DOI":"10.1111\/acel.14057","type":"journal-article","created":{"date-parts":[[2023,12,4]],"date-time":"2023-12-04T05:51:15Z","timestamp":1701669075000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Single\u2010cell transcriptome characteristics of testicular terminal epithelium lineages during aging in the <i>Drosophila<\/i>"],"prefix":"10.1111","volume":"23","author":[{"given":"Xia","family":"Chen","sequence":"first","affiliation":[{"name":"Department of Obstetrics and Gynecology, Nantong First People's Hospital Affiliated Hospital 2 of Nantong University; Medical School of Nantong University, Nantong University  Nantong Jiangsu China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yujuan","family":"Qi","sequence":"additional","affiliation":[{"name":"Clinical Center of Reproductive Medicine, Xuzhou Central Hospital Xuzhou Clinical School of Xuzhou Medical University  Xuzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiuru","family":"Huang","sequence":"additional","affiliation":[{"name":"Institute of Reproductive Medicine Medical School of Nantong University, Nantong University  Nantong China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chi","family":"Sun","sequence":"additional","affiliation":[{"name":"Department of Geriatrics Affiliated Hospital of Nantong University, Nantong University  Nantong China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanli","family":"Zheng","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Nantong First People's Hospital Affiliated Hospital 2 of Nantong University; Medical School of Nantong University, Nantong University  Nantong Jiangsu China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li","family":"Ji","sequence":"additional","affiliation":[{"name":"Institute of Reproductive Medicine Medical School of Nantong University, Nantong University  Nantong China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yi","family":"Shi","sequence":"additional","affiliation":[{"name":"Institute of Reproductive Medicine Medical School of Nantong University, Nantong University  Nantong 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Clinical School of Xuzhou Medical University  Xuzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhifeng","family":"Gu","sequence":"additional","affiliation":[{"name":"Department of Rheumatology Affiliated Hospital of Nantong University, Nantong University  Nantong China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3109-4102","authenticated-orcid":false,"given":"Jun","family":"Yu","sequence":"additional","affiliation":[{"name":"Institute of Reproductive Medicine Medical School of Nantong University, Nantong University  Nantong 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