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However, the extent of interneuron heterogeneity and how this diversity relates to functional aspects of movement remain unclear. Here, through a focus on mouse spinal V1 interneurons, we show that loss of the V1 transcription factor En1 selectively disrupts the frequency of rhythmic locomotor output but does not disrupt flexion\/extension limb movement, thereby decoupling two key functional roles ascribed to this neuronal population. To investigate the cellular basis of these deficits, we generated a single-nucleus transcriptomic atlas of V1 interneurons across postnatal development. Our analysis reveals age-dependent transcriptional changes while also demonstrating that their core molecular taxonomy perdures into adulthood. Notably, En1 deficiency selectively perturbed a single subset of V1\n                    <jats:sup>Pou6f2<\/jats:sup>\n                    interneurons, thereby identifying a possible cellular substrate for influencing locomotor speed. Beyond serving as a molecular resource, our study highlights how deep neuronal profiling provides an entry point for understanding the multifunctional nature of heterogeneous interneuron populations.\n                  <\/jats:p>","DOI":"10.1038\/s41467-026-76522-3","type":"journal-article","created":{"date-parts":[[2026,8,10]],"date-time":"2026-08-10T15:24:46Z","timestamp":1786375486000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Transcriptomic analysis of spinal V1 interneurons informs their multifunctional role in motor output"],"prefix":"10.1038","volume":"17","author":[{"given":"Alexandra J.","family":"Trevisan","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katie","family":"Han","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Phillip D.","family":"Chapman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anand S.","family":"Kulkarni","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer M.","family":"Hinton","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ines","family":"Klein","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5598-0876","authenticated-orcid":false,"given":"Cody","family":"Ramirez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alfonso","family":"Lavado","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4244-8925","authenticated-orcid":false,"given":"Graziana","family":"Gatto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mariano I.","family":"Gabitto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4096-8601","authenticated-orcid":false,"given":"Vilas","family":"Menon","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-4780-5348","authenticated-orcid":false,"given":"Jay B.","family":"Bikoff","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,8,10]]},"reference":[{"key":"76522_CR1","doi-asserted-by":"publisher","first-page":"507","DOI":"10.1038\/nrn2608","volume":"10","author":"M Goulding","year":"2009","unstructured":"Goulding, M. 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