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However, the characteristics of pain-enhancing microglia, which are more prominent in males, remain poorly understood. Here, we reanalyzed a previously published single-cell RNA sequencing dataset and identified a microglial subpopulation that significantly increases in the spinal dorsal horn (SDH) of male mice following peripheral nerve injury. CC-chemokine ligand 4 (CCL4) was highly expressed in this subpopulation and its mRNA levels were increased in the SDH after partial sciatic nerve ligation (PSL) only in male mice. Notably, CCL4 expression was reduced in male mice following microglial depletion, indicating that microglia are the primary source of CCL4. Intrathecal administration of maraviroc, an inhibitor of the CCL4\u2013CC-chemokine receptor 5 (CCR5) signaling pathway, after PSL, significantly suppressed mechanical allodynia only in male mice. Furthermore, intrathecal administration of CCL4 induced mechanical allodynia in both sexes, accompanied by increased expression of c-fos, a neuronal excitation marker, in the SDH. These findings highlight a sex-biased difference in the gene expression profile of spinal microglia following peripheral nerve injury, with elevated CCL4 expression in male mice potentially contributing to pain exacerbation.<\/jats:p>","DOI":"10.3390\/cells14070484","type":"journal-article","created":{"date-parts":[[2025,3,24]],"date-time":"2025-03-24T13:48:20Z","timestamp":1742824100000},"page":"484","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Male-Dominant Spinal Microglia Contribute to Neuropathic Pain by Producing CC-Chemokine Ligand 4 Following Peripheral Nerve Injury"],"prefix":"10.3390","volume":"14","author":[{"given":"Fumihiro","family":"Saika","sequence":"first","affiliation":[{"name":"Department of Physiological Sciences, School of Pharmaceutical Sciences, Wakayama Medical University, Wakayama 640-8156, Japan"},{"name":"Faculty of Wakayama Health Care Sciences, Takarazuka University of Medical and Health Care, Wakayama 640-8392, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5518-7824","authenticated-orcid":false,"given":"Tetsuya","family":"Sato","sequence":"additional","affiliation":[{"name":"H.U. Group Research Institute G.K., Tokyo 197-0833, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takeru","family":"Nakabayashi","sequence":"additional","affiliation":[{"name":"H.U. Group Research Institute G.K., Tokyo 197-0833, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5747-5758","authenticated-orcid":false,"given":"Yohji","family":"Fukazawa","sequence":"additional","affiliation":[{"name":"Department of Anatomy, Kansai University of Health Sciences, Osaka 590-0482, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shinjiro","family":"Hino","sequence":"additional","affiliation":[{"name":"Department of Medical Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto 860-0811, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6872-5292","authenticated-orcid":false,"given":"Kentaro","family":"Suzuki","sequence":"additional","affiliation":[{"name":"Faculty of Life and Environmental Sciences, University of Yamanashi, Yamanashi 400-8510, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5401-8650","authenticated-orcid":false,"given":"Norikazu","family":"Kiguchi","sequence":"additional","affiliation":[{"name":"Department of Physiological Sciences, School of Pharmaceutical Sciences, Wakayama Medical University, Wakayama 640-8156, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,3,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1038\/35093019","article-title":"Molecular mechanisms of nociception","volume":"413","author":"Julius","year":"2001","journal-title":"Nature"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1016\/j.cell.2009.09.028","article-title":"Cellular and molecular mechanisms of pain","volume":"139","author":"Basbaum","year":"2009","journal-title":"Cell"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"2082","DOI":"10.1016\/S0140-6736(21)00393-7","article-title":"Chronic pain: An update on burden, best practices, and new advances","volume":"397","author":"Cohen","year":"2021","journal-title":"Lancet"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"17002","DOI":"10.1038\/nrdp.2017.2","article-title":"Neuropathic pain","volume":"3","author":"Colloca","year":"2017","journal-title":"Nat. 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