{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T15:41:24Z","timestamp":1785426084643,"version":"3.56.0"},"reference-count":45,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2026,5,26]],"date-time":"2026-05-26T00:00:00Z","timestamp":1779753600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Northwest A&F University"},{"id":[{"id":"https:\/\/ror.org\/0051rme32","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Bingtuan Science and Technology Program","award":["2026YD015-01"],"award-info":[{"award-number":["2026YD015-01"]}]},{"name":"Populus euphratica Talents Research Startup Fund for Introduced Talents","award":["TDZKBS202682"],"award-info":[{"award-number":["TDZKBS202682"]}]},{"name":"Key Research and Development Program of Shaanxi Province","award":["2025NC-YBXM-110"],"award-info":[{"award-number":["2025NC-YBXM-110"]}]},{"name":"Key Research and Development Program of Shaanxi Province","award":["2025NC-YBXM-113"],"award-info":[{"award-number":["2025NC-YBXM-113"]}]},{"name":"Dong\u2019e black donkey industry integration and development of key technology innovation and demonstration","award":["2021TZXD012"],"award-info":[{"award-number":["2021TZXD012"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Testicular development and spermatogenesis are critical for male reproduction, but their molecular mechanisms in Dezhou donkeys remain understudied. This study used single-cell RNA sequencing (scRNA-seq) to analyze testicular tissues from Dezhou donkeys at juvenile (2 months), pre-pubertal (12 months), and mature (24 months) stages. A total of 24,606 high-quality cells were profiled, constructing a comprehensive single-cell transcriptional atlas. Unsupervised clustering identified nine major cell types: three germ cell subtypes (spermatogonia, spermatocytes, spermatids) and six somatic cell subtypes (Leydig cells, Sertoli cells, peritubular muscle cells, macrophages, endothelial cells, T cells). Key marker genes (AMH, TNP1, UTF1, ZMYND10) were validated by immunofluorescence. Pseudotemporal trajectory analysis revealed sequential germ cell differentiation (spermatogonia \u2192 spermatocytes \u2192 spermatids) and Sertoli cell maturation (immature \u2192 mature), while Leydig cells and peritubular muscle cells shared common progenitors. CellChat analysis identified critical ligand\u2013receptor pairs in BMP, IGF, WNT, and FSH pathways, which regulate testicular development. This study provides the comprehensive single-cell transcriptional map of Dezhou donkey testicular development, elucidating key molecular mechanisms of germ and somatic cell maturation. The findings offer valuable insights into donkey reproductive biology, supporting breeding improvement and male infertility research.<\/jats:p>","DOI":"10.3390\/ani16111621","type":"journal-article","created":{"date-parts":[[2026,5,26]],"date-time":"2026-05-26T16:17:58Z","timestamp":1779812278000},"page":"1621","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Single-Cell RNA Sequencing Reveals Dynamic Transcriptional Landscape of Testicular Maturation in Dezhou Donkeys"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0235-7030","authenticated-orcid":false,"given":"Zhaofei","family":"Wang","sequence":"first","affiliation":[{"name":"Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, Xianyang 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Yu","sequence":"additional","affiliation":[{"name":"National Engineering Research Center for Gelatin-Based Traditional Chinese Medicine, Dong-E-E-Jiao Co., Ltd., No. 78, E-Jiao Street, Dong\u2019e Country, Liaocheng 252201, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6230-9885","authenticated-orcid":false,"given":"Guiqin","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Agriculture and Biology, Liaocheng University, 1 Hunan Road, Liaocheng 252000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Halima","family":"Jafari","sequence":"additional","affiliation":[{"name":"Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, Xianyang 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cong","family":"Li","sequence":"additional","affiliation":[{"name":"Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, Xianyang 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ge","family":"Yang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, Xianyang 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chuzhao","family":"Lei","sequence":"additional","affiliation":[{"name":"Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, Xianyang 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruihua","family":"Dang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, Xianyang 712100, China"},{"name":"College of Animal Science and Technology, Tarim University, Alar 843300, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1097\/MED.0000000000000339","article-title":"Testicular growth and development in puberty","volume":"24","author":"Koskenniemi","year":"2017","journal-title":"Curr. 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