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Here, we demonstrate that LDT-NAc cholinergic projections undergo cocaine-induced adaptations and modulate cocaine-related behaviors. Using cell type-specific tracing, we show that LDT neurons preferentially innervate NAc medium spiny neurons and cholinergic interneurons. Large-scale\n                  <jats:italic>in vivo<\/jats:italic>\n                  recordings reveal that cocaine pre-exposure induces persistent alterations in both LDT and NAc neuronal dynamics and modifies responses to subsequent cocaine challenge. Remarkably, pre-exposure to cocaine triggers population-specific adaptations in the LDT, selectively enhancing excitability of LDT-NAc-projecting cholinergic neurons while reducing that of non-projecting cholinergic cells. Behaviorally, optogenetic activation of LDT-NAc cholinergic projections enhances cocaine conditioning, whereas their inhibition diminishes cocaine\u2019s reinforcing effects.\n                <\/jats:p>\n                <jats:p>Our findings identify LDT-NAc cholinergic inputs as key substrates of cocaine-induced plasticity, and critical mediators of cocaine\u2019s rewarding properties, introducing a novel component to addiction circuitry.<\/jats:p>","DOI":"10.1101\/2025.06.30.662309","type":"posted-content","created":{"date-parts":[[2025,7,4]],"date-time":"2025-07-04T18:00:16Z","timestamp":1751652016000},"source":"Crossref","is-referenced-by-count":1,"title":["Adaptive changes of cholinergic projections to the nucleus accumbens bidirectionally mediate cocaine reinforcing effects"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8195-4601","authenticated-orcid":false,"given":"LAA","family":"Aguiar","sequence":"first","affiliation":[{"name":"Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, Braga, Portugal"},{"name":"ICVS\/3B\u2019s\u2013PT Government Associate Laboratory, Braga\/Guimar\u00e3es, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5598-1948","authenticated-orcid":false,"given":"L","family":"Royon","sequence":"additional","affiliation":[{"name":"Institut de Pharmacologie Mol\u00e9culaire & Cellulaire, CNRS, UMR7275, Valbonne, France"},{"name":"Universit\u00e9 C\u00f4te d\u2019Azur, Nice, 06560, France"},{"name":"Inserm U1323"}]},{"given":"R","family":"Bastos-Gon\u00e7alves","sequence":"additional","affiliation":[{"name":"Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, Braga, Portugal"},{"name":"ICVS\/3B\u2019s\u2013PT Government Associate Laboratory, Braga\/Guimar\u00e3es, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9583-4830","authenticated-orcid":false,"given":"TTA","family":"Carvalho","sequence":"additional","affiliation":[{"name":"Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, Braga, Portugal"},{"name":"ICVS\/3B\u2019s\u2013PT Government Associate Laboratory, Braga\/Guimar\u00e3es, Portugal"}]},{"given":"E","family":"Teixeira","sequence":"additional","affiliation":[{"name":"Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, Braga, Portugal"},{"name":"ICVS\/3B\u2019s\u2013PT Government Associate Laboratory, Braga\/Guimar\u00e3es, Portugal"}]},{"given":"AC","family":"Faria","sequence":"additional","affiliation":[{"name":"Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, Braga, Portugal"},{"name":"ICVS\/3B\u2019s\u2013PT Government Associate Laboratory, Braga\/Guimar\u00e3es, 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