{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T14:05:54Z","timestamp":1785420354419,"version":"3.56.0"},"posted":{"date-parts":[[2025,4,20]]},"group-title":"Neuroscience","reference-count":65,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,4,20]],"date-time":"2025-04-20T00:00:00Z","timestamp":1745107200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/publicdomain\/zero\/1.0\/"}],"funder":[{"award":["ZIA-MH002386"],"award-info":[{"award-number":["ZIA-MH002386"]}],"id":[{"id":"https:\/\/ror.org\/04xeg9z08","id-type":"ROR","asserted-by":"publisher"}]},{"award":["ZIA-MH002497-35"],"award-info":[{"award-number":["ZIA-MH002497-35"]}],"id":[{"id":"https:\/\/ror.org\/04xeg9z08","id-type":"ROR","asserted-by":"publisher"}]},{"award":["ZIA-DA000633"],"award-info":[{"award-number":["ZIA-DA000633"]}],"id":[{"id":"https:\/\/ror.org\/00fq5cm18","id-type":"ROR","asserted-by":"publisher"}]},{"award":["ZIA-DK075168-03"],"award-info":[{"award-number":["ZIA-DK075168-03"]}],"id":[{"id":"https:\/\/ror.org\/00adh9b73","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,4,20]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  Immediate-early gene (IEG) induction after administration of amphetamine or cocaine has been used to trace the signaling pathways that mediate neuronal plasticity required for the short- and long-term behavioral effects of these psychostimulants. We recently reported that a novel cyclic AMP (cAMP)-dependent Rap guanine nucleotide exchange factor-2 (RapGEF2)-ERK signaling pathway is required for Egr-1 induction in D1 medium-spiny neurons (MSNs) of the nucleus accumbens (NAc) after cocaine treatment, and that its deletion from the NAc neurons attenuates cocaine-induced locomotor sensitization and conditioned place preference (CPP). However, the cell type-specific neuronal mechanisms underlying this effect remain unclear. In this study, we used Cre-LoxP technology and a novel Cre-amplifier transgene to generate conditional RapGEF2 knockout mice targeting D1-MSNs and investigated the functional role of RapGEF2 in cocaine reward. Deletion of RapGEF2 in D1-MSNs blocked cocaine-induced ERK phosphorylation (p-ERK) and Egr-1 induction. D1-MSN-specific RapGEF2 deletion did not affect intravenous cocaine self-administration, nor did it affect Fos induction by cocaine, prompting us to examine more closely the role of metabotropic (cAMP-dependent) signaling to IEGs after cocaine administration. We used a battery of D1-MSN-specific genetic interventions targeting cAMP signaling, including Drd1-Cre::Rap1A\/B\n                  <jats:sup>fl\/fl<\/jats:sup>\n                  mice, and AAV injection of a Cre-dependent catalytically active phosphodiesterase (PDE4D3-cat) or a Cre-dependent protein kinase A-inhibitor (PKI) in the NAc of Drd1-Cre mice, to explore further the underlying cAMP dependence of IEG induction by acute and chronic cocaine administration, and the cAMP sensors required. Rap1 is reported as a necessary component for both RapGEF2- and PKA-dependent ERK activation, but a requirement for Rap in Fos induction by cocaine has not been examined. Deletion of Rap1A\/B in D1-MSNs blocked cocaine-induced p-ERK and Egr-1 expression, but not c-Fos, supporting the idea that the RapGEF2-Rap1-ERK pathway specifically regulates Egr-1, not c-Fos, expression. D1-specific expression of the PDE4D3-cat ablated up-regulation of both Egr-1 and Fos in NAc after cocaine administration, demonstrating that induction of both IEGs requires cAMP elevation in D1-MSNs. Notably, inhibition of PKA activity via AAV-mediated expression of PKI-alpha in D1-MSNs blocked both c-Fos and Egr-1 induction. Thus, acute or chronic cocaine administration activates at least two cAMP-dependent signaling pathways in D1-MSNs: a PKA-Fos pathway and a RapGEF2-ERK-Egr-1 pathway. The finding that PKA also activates the ERK-Egr-1 signaling pathway by convergence on Rap1, and concomitantly activates c-Fos independently of Rap1, may underlie selective effects of metabotropic activation of RapGEF2 and PKA activation by cAMP on cocaine-dependent behaviors in mice.\n                <\/jats:p>","DOI":"10.1101\/2025.04.15.648980","type":"posted-content","created":{"date-parts":[[2025,4,23]],"date-time":"2025-04-23T10:55:46Z","timestamp":1745405746000},"source":"Crossref","is-referenced-by-count":0,"title":["Cocaine-induced immediate-early gene expression in the nucleus accumbens: roles of separate cAMP sensors"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0593-5940","authenticated-orcid":false,"given":"Hai-Ying","family":"Zhang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tabinda","family":"Salman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guo-Hua","family":"Bi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0706-1581","authenticated-orcid":false,"given":"Sunny Z.","family":"Jiang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charles R.","family":"Gerfen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6991-2898","authenticated-orcid":false,"given":"Andrew","family":"Lutas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenqin","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6482-8104","authenticated-orcid":false,"given":"Zheng-Xiong","family":"Xi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7524-944X","authenticated-orcid":false,"given":"Lee E.","family":"Eiden","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025052504044499000_2025.04.15.648980v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/j.nlm.2016.07.010"},{"key":"2025052504044499000_2025.04.15.648980v1.2","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.1039-08.2008"},{"key":"2025052504044499000_2025.04.15.648980v1.3","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.2284-07.2007"},{"key":"2025052504044499000_2025.04.15.648980v1.4","doi-asserted-by":"publisher","DOI":"10.3389\/fnbeh.2017.00112"},{"key":"2025052504044499000_2025.04.15.648980v1.5","doi-asserted-by":"publisher","DOI":"10.3389\/fncel.2022.979078"},{"key":"2025052504044499000_2025.04.15.648980v1.6","first-page":"56","article-title":"A PKA activity sensor for quantitative analysis of endogenous GPCR signaling via 2-photon FRET-FLIM imaging","volume":"5","year":"2014","journal-title":"Frontiers in pharmacology"},{"key":"2025052504044499000_2025.04.15.648980v1.7","doi-asserted-by":"publisher","DOI":"10.1016\/0306-4522(96)00145-5"},{"key":"2025052504044499000_2025.04.15.648980v1.8","doi-asserted-by":"publisher","DOI":"10.1038\/nn1579"},{"key":"2025052504044499000_2025.04.15.648980v1.9","doi-asserted-by":"publisher","DOI":"10.1016\/j.biopsych.2009.03.014"},{"key":"2025052504044499000_2025.04.15.648980v1.10","doi-asserted-by":"publisher","DOI":"10.1038\/sj.npp.1301014"},{"key":"2025052504044499000_2025.04.15.648980v1.11","unstructured":"Franklin, K. 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