{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,6]],"date-time":"2026-05-06T18:06:46Z","timestamp":1778090806304,"version":"3.51.4"},"reference-count":40,"publisher":"Public Library of Science (PLoS)","issue":"3","license":[{"start":{"date-parts":[[2024,3,14]],"date-time":"2024-03-14T00:00:00Z","timestamp":1710374400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000065","name":"National Institute of Neurological Disorders and Stroke","doi-asserted-by":"publisher","award":["NS100469"],"award-info":[{"award-number":["NS100469"]}],"id":[{"id":"10.13039\/100000065","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Recent transcriptomic studies identified <jats:italic>Gucy2d<\/jats:italic> (encoding guanylate cyclase D) as a highly enriched gene within inhibitory dynorphin interneurons in the mouse spinal dorsal horn. To facilitate investigations into the role of the <jats:italic>Gucy2d+<\/jats:italic> population in somatosensation, <jats:italic>Gucy2d-cre<\/jats:italic> transgenic mice were created to permit chemogenetic or optogenetic manipulation of this subset of spinal neurons. <jats:italic>Gucy2d-cre<\/jats:italic> mice created via CRISPR\/Cas9 genomic knock-in were bred to mice expressing a cre-dependent reporter (either tdTomato or Sun1.GFP fusion protein), and the resulting offspring were characterized. Surprisingly, a much wider population of spinal neurons was labeled by cre-dependent reporter expression than previous mRNA-based studies would suggest. Although the cre-dependent reporter expression faithfully labeled ~75% of cells expressing <jats:italic>Gucy2d<\/jats:italic> mRNA in the adult dorsal horn, it also labeled a substantial number of additional inhibitory neurons in which no <jats:italic>Gucy2d<\/jats:italic> or <jats:italic>Pdyn<\/jats:italic> mRNA was detected. Moreover, cre-dependent reporter was also expressed in various regions of the brain, including the spinal trigeminal nucleus, cerebellum, thalamus, somatosensory cortex, and anterior cingulate cortex. Injection of AAV-CAG-FLEX-tdTomato viral vector into adult <jats:italic>Gucy2d-cre<\/jats:italic> mice produced a similar pattern of cre-dependent reporter expression in the spinal cord and brain, which excludes the possibility that the unexpected reporter-labeling of cells in the deep dorsal horn and brain was due to transient <jats:italic>Gucy2d<\/jats:italic> expression during early stages of development. Collectively, these results suggest that <jats:italic>Gucy2d<\/jats:italic> is expressed in a wider population of cells than previously thought, albeit at levels low enough to avoid detection with commonly used mRNA-based assays. Therefore, it is unlikely that these <jats:italic>Gucy2d-cre<\/jats:italic> mice will permit selective manipulation of inhibitory signaling mediated by spinal dynorphin interneurons, but this novel cre driver line may nevertheless be useful to target a broader population of inhibitory spinal dorsal horn neurons.<\/jats:p>","DOI":"10.1371\/journal.pone.0300282","type":"journal-article","created":{"date-parts":[[2024,3,14]],"date-time":"2024-03-14T17:25:41Z","timestamp":1710437141000},"page":"e0300282","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":2,"title":["Development and characterization of a Gucy2d-cre mouse to selectively manipulate a subset of inhibitory spinal dorsal horn interneurons"],"prefix":"10.1371","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7592-4617","authenticated-orcid":true,"given":"Elizabeth 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