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While GlyT2 inhibitors hold clinical promise, the specialization of spatially distributed GlyT2 neuronal populations across the brain remains incomplete. Here, we crossed GlyT2\u2010iCre with Ai9 to generate GlyT2\u2010iCre\u2010tdTomato mice, enabling anatomical mapping and functional interrogation of GlyT2 circuits. Here, we generated GlyT2\u2010iCre\u2010tdTomato mice through crossbreeding GlyT2\u2010iCre and Ai9 lines, enabling anatomical mapping and functional interrogation of GlyT2 circuits. Systematic quantification revealed sex\u2010 and region\u2010specific neuronal distributions: non\u2010estrus females exhibited significantly higher GlyT2\u2010tdTomato neuron densities than males in 12 brain regions spanning thalamus (anterodorsal, anteroventral dorsomedial thalamic nucleus), midbrain (ventral periaqueductal gray, precommissural nucleus of the lateral lemniscus, superior and lateral parabrachial nucleus), and hindbrain (pontine nucleus, reticular oralis, reticulotegmental nucleus of the pons, rostromedial tegmental nucleus, solitary nucleus, interpolar part of spinal trigeminal nucleus), whereas males surpassed females in ventral posteromedial nuclei and lateral cerebellar regions. No sex differences were detected in 17 regions, including thalamic nuclei (anteromedial and ventral medial nucleus), posterior hypothalamus, substantia nigra, ventral nucleus of the lateral lemniscus, the nucleus of the central acoustic tract, ventral periolivary nucleus, paralemniscal nucleus, cerebellum, lateral periaqueductal gray, superior colliculus, inferior colliculus, medial parabrachial nucleus, and isthmic reticular formation. Spatial alignment with\n                    <jats:italic>GlyT2mRNA<\/jats:italic>\n                    data from the GlyT2\u2010ISH atlas validated model specificity, establishing this resource as a foundation for investigating circuit\u2010specific GlyT2 functions. These findings not only delineate organizational principles of glycinergic networks but also highlight sex as a critical variable in GlyT2 population architecture\u2014a consideration essential for developing precision therapeutics targeting glycinergic dysfunction.\n                    <jats:boxed-text content-type=\"graphic\" position=\"anchor\">\n                      <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" mimetype=\"image\/png\" position=\"anchor\" specific-use=\"enlarged-web-image\" xlink:href=\"graphic\/jnc70135-gra-0001-m.png\">\n                        <jats:alt-text>image<\/jats:alt-text>\n                      <\/jats:graphic>\n                    <\/jats:boxed-text>\n                  <\/jats:p>","DOI":"10.1111\/jnc.70135","type":"journal-article","created":{"date-parts":[[2025,7,22]],"date-time":"2025-07-22T04:15:04Z","timestamp":1753157704000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Sex\u2010Specific Distribution of Glycine Transporter 2 in the Central Nervous System of Adult GlyT2\u2010iCre\u2010tdTomato Mouse Brain"],"prefix":"10.1111","volume":"169","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2085-4540","authenticated-orcid":false,"given":"Zhenhua","family":"Jiang","sequence":"first","affiliation":[{"name":"Department of Clinical Immunology Xijing Hospital, Fourth Military Medical University  Xi'an China"},{"name":"Department of Anesthesiology and Perioperative Medicine Xijing Hospital, Fourth Military Medical University  Xi'an China"},{"name":"Department of Pain Medicine Xijing Hospital, Fourth Military Medical University  Xi'an China"},{"name":"Key Laboratory of 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