{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T06:02:05Z","timestamp":1782280925560,"version":"3.54.5"},"posted":{"date-parts":[[2025,11,26]]},"group-title":"Neuroscience","reference-count":40,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,11,26]],"date-time":"2025-11-26T00:00:00Z","timestamp":1764115200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["AA029467"],"award-info":[{"award-number":["AA029467"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["GM130456"],"award-info":[{"award-number":["GM130456"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["DA035200"],"award-info":[{"award-number":["DA035200"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,11,26]]},"abstract":"<jats:label>1.<\/jats:label>\n                <jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Parabrachial nucleus (PBN) neurons expressing Calcitonin Gene-Related Peptide (CGRP) modulate fear\u2013 and anxiety-like behavior. It is unknown if PBN(CGRP) neurons play a role in anxiety during withdrawal from alcohol or after repeated stress. First, to investigate the anxiogenic role of activating PBN(CGRP) neurons in na\u00efve conditions,\n                  <jats:italic>Calca<\/jats:italic>\n                  <jats:sup>CRE<\/jats:sup>\n                  female and male mice expressing CRE-dependent hM3D(Gq) DREADDs in the PBN were tested on the elevated plus maze (EPM). PBN(CGRP) neurons drive phasic activity in the bed nucleus of the stria terminalis (BNST) that synchronizes to anxiety-like behavior. Therefore, a transsynaptic anterograde AAV-based strategy was used in C57BL\/6J female and male mice to activate BNST neurons innervated by the PBN projections (BNST\n                  <jats:sup>PBN<\/jats:sup>\n                  ) during EPM. Additionally, PBN(CGRP) neurons and CGRP-innervated BNST cells were measured in experimentally-na\u00efve CGRP-DTR\n                  <jats:sup>GFP<\/jats:sup>\n                  female and male mice, to investigate baseline sex-differences. Next, to investigate the impact of PBN(CGRP) inhibition on anxiety-like behavior following chronic intermittent ethanol vapor exposure (CIE),\n                  <jats:italic>Calca<\/jats:italic>\n                  <jats:sup>CRE<\/jats:sup>\n                  female and male mice expressing CRE-dependent hM4D(Gi) DREADDs in the PBN were tested in EPM during acute withdrawal. Additionally, mice were exposed to repeated forced swim stress (FSS) paired with PBN(CGRP) inhibition followed by testing in the novelty suppressed feeding task (NSFT), to investigate the role of PBN(CGRP) on anxiety-like behavior after stress in prolonged withdrawal. Activating PBN(CGRP) or BNST\n                  <jats:sup>PBN<\/jats:sup>\n                  neurons did not change behavior in EPM in either sex. Total PBN(CGRP) and CGRP-innervated BNST cells did not differ between females and males. In acute withdrawal, inhibiting PBN(CGRP) neurons decreased time in the open arm and increased time in the closed arm selectively in females. In prolonged withdrawal, inhibiting PBN(CGRP) neurons during FSS did not affect immobility in either sex, but did subsequently decrease approach frequency and average speed selectively in males. Altogether, the data report the anxiogenic effect of manipulating PBN(CGRP) neurons is stress and sex-specific during alcohol withdrawal.\n                <\/jats:p>","DOI":"10.1101\/2025.11.24.690162","type":"posted-content","created":{"date-parts":[[2025,11,26]],"date-time":"2025-11-26T20:15:16Z","timestamp":1764188116000},"source":"Crossref","is-referenced-by-count":1,"title":["Parabrachial Calcitonin Gene-Related Peptide Neurons Sex-Specifically Modulate Anxiety in Alcohol Withdrawal"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-6800-5162","authenticated-orcid":false,"given":"C.E.","family":"Van Doorn","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J.B.","family":"Tyree","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6267-4196","authenticated-orcid":false,"given":"A.A.","family":"Jaramillo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025112715550622000_2025.11.24.690162v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-12-405941-2.00019-5"},{"key":"2025112715550622000_2025.11.24.690162v1.2","doi-asserted-by":"publisher","DOI":"10.1124\/pharmrev.120.000083"},{"key":"2025112715550622000_2025.11.24.690162v1.3","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuropharm.2021.108757"},{"key":"2025112715550622000_2025.11.24.690162v1.4","doi-asserted-by":"publisher","DOI":"10.1016\/j.tins.2018.03.007"},{"key":"2025112715550622000_2025.11.24.690162v1.5","doi-asserted-by":"crossref","first-page":"649143","DOI":"10.3389\/fphar.2021.649143","article-title":"Efficacy and Safety of Monoclonal Antibody Against Calcitonin Gene-Related Peptide or Its Receptor for Migraine: A Systematic Review and Network Meta-analysis","volume":"12","year":"2021","journal-title":"Front Pharmacol"},{"issue":"5","key":"2025112715550622000_2025.11.24.690162v1.6","doi-asserted-by":"crossref","first-page":"669","DOI":"10.1097\/ACO.0000000000001553","article-title":"The potential role of calcitonin gene-related peptide antagonists for the management of hangover headaches","volume":"38","year":"2025","journal-title":"Curr Opin Anaesthesiol"},{"key":"2025112715550622000_2025.11.24.690162v1.7","doi-asserted-by":"crossref","unstructured":"Narouze S. CGRP Antagonists as a Preventive Treatment for Hangover Headaches. Annals Of Headache Medicine Journal. 2025.","DOI":"10.30756\/ahmj.2025.14.01"},{"key":"2025112715550622000_2025.11.24.690162v1.8","doi-asserted-by":"crossref","first-page":"12559","DOI":"10.1038\/srep12559","article-title":"Calcitonin gene-related peptide pre-administration acts as a novel antidepressant in stressed mice","volume":"5","year":"2015","journal-title":"Sci Rep"},{"issue":"1","key":"2025112715550622000_2025.11.24.690162v1.9","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1038\/s42003-024-05937-9","article-title":"CGRP causes anxiety via HP1gamma-KLF11-MAOB pathway and dopamine in the dorsal hippocampus","volume":"7","year":"2024","journal-title":"Commun Biol"},{"key":"2025112715550622000_2025.11.24.690162v1.10","doi-asserted-by":"publisher","DOI":"10.1007\/s12031-009-9201-z"},{"key":"2025112715550622000_2025.11.24.690162v1.11","doi-asserted-by":"crossref","first-page":"911105","DOI":"10.3389\/fphar.2022.911105","article-title":"Unpredictable Sound Stress Model Causes Migraine-Like Behaviors in Mice With Sexual Dimorphism","volume":"13","year":"2022","journal-title":"Front Pharmacol"},{"key":"2025112715550622000_2025.11.24.690162v1.12","doi-asserted-by":"publisher","DOI":"10.1038\/nn.4101"},{"key":"2025112715550622000_2025.11.24.690162v1.13","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.5274-10.2011"},{"key":"2025112715550622000_2025.11.24.690162v1.14","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbr.2013.01.024"},{"key":"2025112715550622000_2025.11.24.690162v1.15","doi-asserted-by":"publisher","DOI":"10.1101\/lm.032482.113"},{"key":"2025112715550622000_2025.11.24.690162v1.16","doi-asserted-by":"publisher","DOI":"10.1007\/s00429-018-01825-1"},{"key":"2025112715550622000_2025.11.24.690162v1.17","doi-asserted-by":"publisher","DOI":"10.1002\/cne.20697"},{"key":"2025112715550622000_2025.11.24.690162v1.18","doi-asserted-by":"publisher","DOI":"10.1016\/S0149-7634(96)00023-1"},{"key":"2025112715550622000_2025.11.24.690162v1.19","doi-asserted-by":"crossref","unstructured":"Bowen AJ , Chen JY , Huang YW , Baertsch NA , Park S , Palmiter RD . Dissociable control of unconditioned responses and associative fear learning by parabrachial CGRP neurons. Elife. 2020;9.","DOI":"10.7554\/eLife.59799"},{"key":"2025112715550622000_2025.11.24.690162v1.20","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2015.05.057"},{"key":"2025112715550622000_2025.11.24.690162v1.21","doi-asserted-by":"crossref","first-page":"100247","DOI":"10.1016\/j.ynstr.2020.100247","article-title":"BNST transient activity associates with approach behavior in a stressful environment and is modulated by the parabrachial nucleus","volume":"13","year":"2020","journal-title":"Neurobiol Stress"},{"key":"2025112715550622000_2025.11.24.690162v1.22","doi-asserted-by":"crossref","unstructured":"Pyeon GH , Cho H , Chung BM , Choi JS , Jo YS . Parabrachial CGRP neurons modulate active defensive behavior under a naturalistic threat. Elife. 2025;14.","DOI":"10.7554\/eLife.101523"},{"issue":"9","key":"2025112715550622000_2025.11.24.690162v1.23","doi-asserted-by":"crossref","first-page":"e70268","DOI":"10.1111\/ejn.70268","article-title":"Ex Vivo Modulation of the BNST by Parabrachial CGRP Projections Is Decreased After a History of Stress-Induced Anxiety","volume":"62","year":"2025","journal-title":"Eur J Neurosci"},{"issue":"2","key":"2025112715550622000_2025.11.24.690162v1.24","first-page":"88","article-title":"The role of CGRP and CALCA T-692C single-nucleotide polymorphism in psoriasis vulgaris","volume":"70","year":"2015","journal-title":"Pharmazie"},{"issue":"2","key":"2025112715550622000_2025.11.24.690162v1.25","first-page":"249","article-title":"Calcitonin gene-related peptide (CGRP) content and CGRP receptor binding sites in discrete forebrain regions of alcohol-preferring vs","volume":"690","year":"1995","journal-title":"nonpreferring rats, and high alcohol-drinking vs. low alcohol-drinking rats. Brain Res"},{"key":"2025112715550622000_2025.11.24.690162v1.26","doi-asserted-by":"crossref","first-page":"158","DOI":"10.3389\/fncel.2019.00158","article-title":"Predisposition to Alcohol Drinking and Alcohol Consumption Alter Expression of Calcitonin Gene-Related Peptide, Neuropeptide Y, and Microglia in Bed Nucleus of Stria Terminalis in a Subnucleus-Specific Manner","volume":"13","year":"2019","journal-title":"Front Cell Neurosci"},{"key":"2025112715550622000_2025.11.24.690162v1.27","doi-asserted-by":"publisher","DOI":"10.1017\/S1461145799001558"},{"issue":"2","key":"2025112715550622000_2025.11.24.690162v1.28","doi-asserted-by":"crossref","first-page":"e13366","DOI":"10.1111\/adb.13366","article-title":"Sex-specific negative affect-like behaviour and parabrachial nucleus activation induced by BNST stimulation in adult mice with adolescent alcohol history","volume":"29","year":"2024","journal-title":"Addict Biol"},{"key":"2025112715550622000_2025.11.24.690162v1.29","doi-asserted-by":"publisher","DOI":"10.1038\/nature12596"},{"key":"2025112715550622000_2025.11.24.690162v1.30","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuron.2013.01.030"},{"key":"2025112715550622000_2025.11.24.690162v1.31","doi-asserted-by":"publisher","DOI":"10.1111\/j.1530-0277.1993.tb00731.x"},{"key":"2025112715550622000_2025.11.24.690162v1.32","doi-asserted-by":"crossref","unstructured":"Samuels BA , Hen R . Novelty-Suppressed Feeding in the Mouse. Mood and Anxiety Related Phenotypes in Mice. Neuromethods 2011. p. 107\u201321.","DOI":"10.1007\/978-1-61779-313-4_7"},{"key":"2025112715550622000_2025.11.24.690162v1.33","doi-asserted-by":"publisher","DOI":"10.1016\/J.NEURON.2016.11.045"},{"key":"2025112715550622000_2025.11.24.690162v1.34","doi-asserted-by":"publisher","DOI":"10.1038\/s41386-018-0257-8"},{"key":"2025112715550622000_2025.11.24.690162v1.35","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.2158-19.2020"},{"key":"2025112715550622000_2025.11.24.690162v1.36","doi-asserted-by":"crossref","unstructured":"Zhu YB , Wang Y , Hua XX , Xu L , Liu MZ , Zhang R , et al. PBN-PVT projections modulate negative affective states in mice. Elife. 2022;11.","DOI":"10.7554\/eLife.68372"},{"key":"2025112715550622000_2025.11.24.690162v1.37","doi-asserted-by":"crossref","unstructured":"Claes M , De Groef L , Moons L . The DREADDful Hurdles and Opportunities of the Chronic Chemogenetic Toolbox. Cells. 2022;11(7).","DOI":"10.3390\/cells11071110"},{"key":"2025112715550622000_2025.11.24.690162v1.38","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuron.2016.01.040"},{"issue":"6","key":"2025112715550622000_2025.11.24.690162v1.39","first-page":"e15483","article-title":"Differential Synovial CGRP\/RAMP1 Expression in Men and Women With Knee Osteoarthritis","volume":"13","year":"2021","journal-title":"Cureus"},{"key":"2025112715550622000_2025.11.24.690162v1.40","doi-asserted-by":"publisher","DOI":"10.3389\/fnmol.2022.928587"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2025.11.24.690162","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T05:09:33Z","timestamp":1782277773000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2025.11.24.690162"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,26]]},"references-count":40,"URL":"https:\/\/doi.org\/10.1101\/2025.11.24.690162","relation":{"is-preprint-of":[{"id-type":"doi","id":"10.3389\/fphar.2025.1750956","asserted-by":"subject"}]},"subject":[],"published":{"date-parts":[[2025,11,26]]},"subtype":"preprint"}}