{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T06:04:31Z","timestamp":1782281071022,"version":"3.54.5"},"posted":{"date-parts":[[2025,10,23]]},"group-title":"Neuroscience","reference-count":68,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,10,23]],"date-time":"2025-10-23T00:00:00Z","timestamp":1761177600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000038","name":"Natural Sciences and Engineering Research Council of Canada","doi-asserted-by":"crossref","award":["Michael E Hildebrand 315915"],"award-info":[{"award-number":["Michael E Hildebrand 315915"]}],"id":[{"id":"10.13039\/501100000038","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000065","name":"National Institute of Neurological Disorders and Stroke","doi-asserted-by":"crossref","award":["Ariel Levine 1ZIA NS003153"],"award-info":[{"award-number":["Ariel Levine 1ZIA NS003153"]}],"id":[{"id":"10.13039\/100000065","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,5,20]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  Despite being essential mediators of pain processing, the molecular identity of N-methyl-D-aspartate receptor (NMDAR) subtypes in nociceptive dorsal horn circuits is poorly understood, especially between sexes and in humans. Given the importance of GluN2 subunits in shaping NMDAR function and plasticity, we investigated the expression and localization of specific GluN2 NMDAR variants in the dorsal horn of viable spinal cord tissue from male and female rodents and human organ donors. Analysis of single-cell\/nuclei sequencing datasets and quantitative reverse transcriptase polymerase chain reactions (qRT-PCR) revealed that the GluN2A (\n                  <jats:italic>GRIN2A<\/jats:italic>\n                  ) and GluN2B (\n                  <jats:italic>GRIN2B<\/jats:italic>\n                  ) subunits are robustly expressed in dorsal horn neurons of mice, rats and humans, with moderate expression of GluN2D (\n                  <jats:italic>GRIN2D<\/jats:italic>\n                  ). Immunohistochemistry (IHC) with antigen retrieval demonstrated that GluN2A, GluN2B, and GluN2D proteins are all preferentially localized to the superficial dorsal horn of both adult rats and humans, which is conserved between males and females. Surprisingly, we found that these GluN2 NMDAR subunits are enriched in the lateral superficial dorsal horn in rats but not in humans, while presynaptic and neuronal markers are symmetrically distributed across the rat mediolateral axis. A dramatic shift in localization of GluN2A to the lateral superficial dorsal horn was observed across later postnatal development (PD21-PD90) in both male and female rats, with a corresponding change in synaptic NMDAR currents. This discovery of changes in NMDAR subunit distribution during maturation and between species will shed light on the physiological roles of NMDARs and their potential as therapeutic targets for pain.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>SIGNIFICANCE STATEMENT<\/jats:title>\n                  <jats:p>We used complementary single-cell\/nuclei analysis, immunostaining, quantitative reverse transcriptase polymerase chain reactions, RNAscope in situ hybridization, and electrophysiological approaches to compare the relative expression of N-methyl-D-aspartate receptor (NMDAR) GluN2 subunits in dorsal horn spinal cord pain circuits of mouse, rat, and human spinal cord tissue. Through these comparisons, we find that the transcripts and proteins of the GluN2A, GluN2B, and GluN2D NMDAR subunits are robustly expressed in superficial dorsal horn neurons, with conserved expression across sex but important differences in expression and localization patterns across late development and between species. These discoveries shed light on the physiological roles of NMDARs and their utility as potential therapeutic targets for pain.<\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2025.10.22.683915","type":"posted-content","created":{"date-parts":[[2025,10,23]],"date-time":"2025-10-23T09:15:43Z","timestamp":1761210943000},"source":"Crossref","is-referenced-by-count":1,"title":["Expression and localization of NMDA receptor GluN2 subunits in dorsal horn pain circuits across sex, species, and late postnatal development"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-9608-3895","authenticated-orcid":false,"given":"Katherine","family":"Griffiths","sequence":"first","affiliation":[{"name":"Department of Neuroscience, Carleton University, Ontario, Canada"},{"name":"Neuroscience Program, Ottawa Hospital Research Institute, Ontario, 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S.","family":"David","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Carleton University, Ontario, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Santina","family":"Temi","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Carleton University, Ontario, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica","family":"Parnell","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Carleton University, Ontario, Canada"},{"name":"Neuroscience Program, Ottawa Hospital Research Institute, Ontario, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christopher","family":"Rudyk","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Carleton University, Ontario, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julia","family":"Bursey","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Carleton University, Ontario, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeffrey L.","family":"Krajewski","sequence":"additional","affiliation":[{"name":"Lilly Research Laboratories, Indianapolis, Indiana, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeff S.","family":"McDermott","sequence":"additional","affiliation":[{"name":"Lilly Research Laboratories, Indianapolis, Indiana, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7215-4928","authenticated-orcid":false,"given":"Annemarie","family":"Dedek","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Carleton University, Ontario, Canada"},{"name":"Neuroscience Program, Ottawa Hospital Research Institute, Ontario, Canada"},{"name":"School of Pharmacy, University of Waterloo, Ontario, 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