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In this study, we performed single-cell RNA and spatial transcriptome sequencing to investigate the cellular composition of testicular and epididymal tissues and examined their gene expression characteristics. In the epididymis, we found that epididymal epithelial cells display a region specificity of gene expression in different epididymal segments, including the beta-defensin family genes. In particular,\n                    <jats:italic>Defb15<\/jats:italic>\n                    ,\n                    <jats:italic>Defb18<\/jats:italic>\n                    ,\n                    <jats:italic>Defb20<\/jats:italic>\n                    ,\n                    <jats:italic>Defb25<\/jats:italic>\n                    and\n                    <jats:italic>Defb48<\/jats:italic>\n                    are specific to the caput;\n                    <jats:italic>Defb22<\/jats:italic>\n                    ,\n                    <jats:italic>Defb23<\/jats:italic>\n                    and\n                    <jats:italic>Defb26<\/jats:italic>\n                    to the corpus;\n                    <jats:italic>Defb2<\/jats:italic>\n                    and\n                    <jats:italic>Defb9<\/jats:italic>\n                    to the cauda of the epididymis. To confirm this, we performed mRNA fluorescence in situ hybridisation (FISH) targeting certain exon region of beta-defensin genes, and found some of their expression matched the sequencing results and displayed a close connection with epididimosome marker gene\n                    <jats:italic>Cd63<\/jats:italic>\n                    . In addition, we paid attention to the Sertoli cells and Leydig cells in the testis, along with fibroblasts and smooth muscle cells in the epididymis, by demonstrating their gene expression profile and spatial information. Our study provides a single-cell and spatial landscape for analysing the gene expression characteristics of testicular and epididymal environments and has important implications for the study of spermatogenesis and sperm maturation.\n                  <\/jats:p>","DOI":"10.1186\/s12964-024-01637-3","type":"journal-article","created":{"date-parts":[[2024,5,14]],"date-time":"2024-05-14T04:02:19Z","timestamp":1715659339000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Single-cell and spatial transcriptomic investigation reveals the spatiotemporal specificity of the beta-defensin gene family during mouse sperm maturation"],"prefix":"10.1186","volume":"22","author":[{"given":"Guoliang","family":"Zhang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuanchao","family":"Sun","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Minkai","family":"Guan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mengmeng","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shiduo","family":"Sun","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,5,14]]},"reference":[{"key":"1637_CR1","doi-asserted-by":"publisher","first-page":"69","DOI":"10.1111\/j.1439-0272.2005.00656.x","volume":"37","author":"S Grunewald","year":"2005","unstructured":"Grunewald S, Paasch U, Glander HJ, Anderegg U. 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