{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,23]],"date-time":"2026-03-23T18:30:00Z","timestamp":1774290600760,"version":"3.50.1"},"reference-count":53,"publisher":"Public Library of Science (PLoS)","issue":"3","license":[{"start":{"date-parts":[[2026,3,23]],"date-time":"2026-03-23T00:00:00Z","timestamp":1774224000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:sec id=\"sec001\">\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      Excessive alcohol consumption is a global health issue and a leading cause of disease, disability, and mortality. This study aimed to determine the effects of a 24-hour ethanol exposure, post-exposure withdrawal (cessation of alcohol intake), and post-exposure withdrawal relief on the sensorimotor performance of the nematode\n                      <jats:italic>Caenorhabditis elegans<\/jats:italic>\n                      .\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec002\">\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>A modified kinetic chemotaxis assay (commonly referred as \u201cdiacetyl race\u201d) was conducted with worm populations subjected to three different doses of ethanol pre-exposure to assess the impact of ethanol on locomotion. Additionally, we employed lifespan, mobility, gene expression analysis and imaging assays to evaluate health status and molecular alterations occurring in the worms under different levels of ethanol exposure.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec003\">\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Wild-type, dopamine receptor mutant and serotonin biosynthesis null mutant worms presented different responses to ethanol in the kinetic chemotaxis assay. Furthermore, exposure to ethanol altered vesicle exocytosis in dopaminergic and serotonergic neurons and the expression of a panel of genes associated with stress responses. Additionally, 24-hour ethanol exposure differentially influenced the lifespan of wild-type and mutant worms.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec004\">\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>Different responses, which may be relevant to the pathogenesis of human alcohol use disorder, were observed in wild-type worms, a dopamine receptor mutant, and a serotonin biosynthesis null mutant in a variety of assays performed. Furthermore, we present a 3-step experimental model for drug tolerance, based on the well-established kinetic chemotaxis behavioral paradigm (\u201cdiacetyl race\u201d). This model provides new insights into the effects of alcohol in worms, particularly regarding the roles of dopamine and serotonin neurotransmission. Importantly, this model holds potential for investigating the effects of other addictive substances beyond alcohol.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1371\/journal.pone.0344966","type":"journal-article","created":{"date-parts":[[2026,3,23]],"date-time":"2026-03-23T17:42:38Z","timestamp":1774287758000},"page":"e0344966","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Specific genes of the dopaminergic (dop-3) and serotonergic (tph-1) pathways contribute to the effects of ethanol consumption in Caenorhabditis elegans"],"prefix":"10.1371","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5331-4244","authenticated-orcid":true,"given":"Teresa","family":"Rubio-Tom\u00e1s","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cynthia A.","family":"Hunn","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6936-0958","authenticated-orcid":true,"given":"G\u00e1bor","family":"Hajd\u00fa","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4057-7678","authenticated-orcid":true,"given":"Csaba","family":"S\u0151ti","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nektarios","family":"Tavernarakis","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Csaba","family":"Barta","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2026,3,23]]},"reference":[{"issue":"10347","key":"pone.0344966.ref001","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1016\/S0140-6736(22)00847-9","article-title":"Population-level risks of alcohol consumption by amount, geography, age, sex, and year: a systematic analysis for the Global Burden of Disease Study 2020","volume":"400","author":"GBD 2020 Alcohol Collaborators","year":"2022","journal-title":"Lancet"},{"issue":"6","key":"pone.0344966.ref002","doi-asserted-by":"crossref","first-page":"1423","DOI":"10.1111\/j.1530-0277.1995.tb01002.x","article-title":"Mutations affecting sensitivity to ethanol in the nematode, Caenorhabditis elegans","volume":"19","author":"PG Morgan","year":"1995","journal-title":"Alcohol Clin Exp Res"},{"issue":"6","key":"pone.0344966.ref003","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1038\/sj.tpj.6500440","article-title":"The concentration-dependent effects of ethanol on Caenorhabditis elegans behaviour","volume":"7","author":"PH Mitchell","year":"2007","journal-title":"Pharmacogenomics J"},{"issue":"5","key":"pone.0344966.ref004","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0010422","article-title":"A differential role for neuropeptides in acute and chronic adaptive responses to alcohol: behavioural and genetic analysis in Caenorhabditis elegans","volume":"5","author":"P Mitchell","year":"2010","journal-title":"PLoS One"},{"issue":"3","key":"pone.0344966.ref005","doi-asserted-by":"crossref","first-page":"1445","DOI":"10.1534\/genetics.116.193102","article-title":"Behavioral deficits following withdrawal from chronic ethanol are influenced by SLO channel function in Caenorhabditis elegans","volume":"206","author":"LL Scott","year":"2017","journal-title":"Genetics"},{"key":"pone.0344966.ref006","first-page":"1","article-title":"A Transparent window into biology: A primer on Caenorhabditis elegans","volume-title":"WormBook: the online review of C. elegans biology","author":"AK Corsi","year":"2015"},{"issue":"5396","key":"pone.0344966.ref007","doi-asserted-by":"crossref","first-page":"2012","DOI":"10.1126\/science.282.5396.2012","article-title":"Genome sequence of the nematode C. elegans: a platform for investigating biology","volume":"282","author":"C. elegans Sequencing Consortium","year":"1998","journal-title":"Science"},{"key":"pone.0344966.ref008","author":"O Hobert","year":"2013"},{"issue":"6","key":"pone.0344966.ref009","doi-asserted-by":"crossref","first-page":"655","DOI":"10.1016\/S0092-8674(03)00979-6","article-title":"A central role of the BK potassium channel in behavioral responses to ethanol in C. elegans","volume":"115","author":"AG Davies","year":"2003","journal-title":"Cell"},{"key":"pone.0344966.ref010","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1895\/wormbook.1.40.1","article-title":"Ethanol","author":"SL McIntire","year":"2010","journal-title":"WormBook"},{"issue":"5","key":"pone.0344966.ref011","doi-asserted-by":"crossref","first-page":"853","DOI":"10.1111\/j.1530-0277.2008.00639.x","article-title":"Effects of developmental exposure to ethanol on Caenorhabditis elegans","volume":"32","author":"JR Davis","year":"2008","journal-title":"Alcohol Clin Exp Res"},{"issue":"1","key":"pone.0344966.ref012","doi-asserted-by":"crossref","first-page":"29984","DOI":"10.1371\/journal.pone.0029984","article-title":"Caenorhabditis elegans battling starvation stress: low levels of ethanol prolong lifespan in L1 larvae","volume":"7","author":"PV Castro","year":"2012","journal-title":"PLoS One"},{"key":"pone.0344966.ref013","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1016\/j.copbio.2019.12.011","article-title":"Caenorhabditis elegans as a model system for human diseases","volume":"63","author":"M Markaki","year":"2020","journal-title":"Curr Opin Biotechnol"},{"issue":"5","key":"pone.0344966.ref014","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1101\/gr.10.5.703","article-title":"Identification of novel human genes evolutionarily conserved in Caenorhabditis elegans by comparative proteomics","volume":"10","author":"CH Lai","year":"2000","journal-title":"Genome Res"},{"key":"pone.0344966.ref015","doi-asserted-by":"crossref","first-page":"285268","DOI":"10.3389\/fnhum.2017.00484","article-title":"An update on the role of serotonin and its interplay with dopamine for reward","volume":"11","author":"AG Fischer","year":"2017","journal-title":"Front Hum Neurosci"},{"issue":"1","key":"pone.0344966.ref016","doi-asserted-by":"crossref","first-page":"114","DOI":"10.1038\/npp.2010.165","article-title":"The roles of dopamine and serotonin in decision making: evidence from pharmacological experiments in humans","volume":"36","author":"RD Rogers","year":"2011","journal-title":"Neuropsychopharmacology"},{"key":"pone.0344966.ref017","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1371\/journal.pgen.1009346","article-title":"Increased dopaminergic neurotransmission results in ethanol dependent sedative behaviors in Caenorhabditis elegans","volume":"17","author":"P Pandey","year":"2021","journal-title":"PLoS Genetics"},{"key":"pone.0344966.ref018","article-title":"Role of dopamine in responsiveness to acute ethanol exposure in Caenorhabditis elegans","volume":"2025","author":"D Gayadin","year":"2025","journal-title":"MicroPubl Biol"},{"issue":"3","key":"pone.0344966.ref019","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0092965","article-title":"Alcohol disinhibition of behaviors in C. elegans","volume":"9","author":"SM Topper","year":"2014","journal-title":"PLoS One"},{"issue":"6","key":"pone.0344966.ref020","doi-asserted-by":"crossref","first-page":"578","DOI":"10.1111\/j.1601-183X.2009.00513.x","article-title":"Ethanol preference in C. elegans Jungsoo","volume":"8","author":"J Lee","year":"2009","journal-title":"Genes Brain Behav"},{"issue":"5","key":"pone.0344966.ref021","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1038\/nrd2031","article-title":"Finding function in novel targets: C. elegans as a model organism","volume":"5","author":"T Kaletta","year":"2006","journal-title":"Nat Rev Drug Discov"},{"key":"pone.0344966.ref022","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1251\/bpo79","article-title":"Using C. elegans to screen for targets of ethanol and behavior-altering drugs","volume":"6","author":"AG Davies","year":"2004","journal-title":"Biol Proced Online"},{"issue":"5","key":"pone.0344966.ref023","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1159\/000514858","article-title":"Caenorhabditis elegans as an in vivo model to assess amphetamine tolerance","volume":"95","author":"D Torres Valladares","year":"2020","journal-title":"Brain Behav Evol"},{"issue":"9","key":"pone.0344966.ref024","doi-asserted-by":"crossref","first-page":"2941","DOI":"10.1534\/g3.118.200374","article-title":"Acute effects of drugs on caenorhabditis elegans movement reveal complex responses and plasticity","volume":"8","author":"M Spensley","year":"2018","journal-title":"G3 Genes|Genomes|Genetics"},{"issue":"3","key":"pone.0344966.ref025","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1016\/j.vascn.2004.04.002","article-title":"Caenorhabditis elegans as model system for rapid toxicity assessment of pharmaceutical compounds","volume":"50","author":"M Dengg","year":"2004","journal-title":"J Pharmacol Toxicol Methods"},{"issue":"1","key":"pone.0344966.ref026","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1093\/genetics\/77.1.71","article-title":"The genetics of Caenorhabditis elegans","volume":"77","author":"S Brenner","year":"1974","journal-title":"Genetics"},{"issue":"1","key":"pone.0344966.ref027","doi-asserted-by":"crossref","first-page":"1412","DOI":"10.1038\/s41467-017-01752-5","article-title":"Semisynthetic fluorescent pH sensors for imaging exocytosis and endocytosis","volume":"8","author":"M Martineau","year":"2017","journal-title":"Nat Commun"},{"issue":"3","key":"pone.0344966.ref028","doi-asserted-by":"crossref","first-page":"1013","DOI":"10.1534\/genetics.115.185025","article-title":"Small heat shock proteins are novel common determinants of alcohol and nicotine sensitivity in Caenorhabditis elegans","volume":"202","author":"JR Johnson","year":"2016","journal-title":"Genetics"},{"issue":"2","key":"pone.0344966.ref029","doi-asserted-by":"crossref","first-page":"102241","DOI":"10.1016\/j.xpro.2023.102241","article-title":"Protocols for treating C. elegans with pharmacological agents, osmoles, and salts for imaging and behavioral assays","volume":"4","author":"L Wang","year":"2023","journal-title":"STAR Protoc"},{"key":"pone.0344966.ref030","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/j.bbrc.2021.06.006","article-title":"Guanylate cyclases link serotoninergic signaling to modulate ethanol-induced food intake in C. elegans","volume":"567","author":"X Wang","year":"2021","journal-title":"Biochem Biophys Res Commun"},{"issue":"4","key":"pone.0344966.ref031","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method","volume":"25","author":"KJ Livak","year":"2001","journal-title":"Methods"},{"issue":"11","key":"pone.0344966.ref032","doi-asserted-by":"crossref","first-page":"1840","DOI":"10.1111\/j.1530-0277.2012.01799.x","article-title":"Ethanol metabolism and osmolarity modify behavioral responses to ethanol in C. elegans","volume":"36","author":"JT Alaimo","year":"2012","journal-title":"Alcohol Clin Exp Res"},{"issue":"42","key":"pone.0344966.ref033","doi-asserted-by":"crossref","first-page":"17504","DOI":"10.1073\/pnas.1108673108","article-title":"Caenorhabditis elegans selects distinct crawling and swimming gaits via dopamine and serotonin","volume":"108","author":"A Vidal-Gadea","year":"2011","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"4","key":"pone.0344966.ref034","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1016\/j.chom.2009.09.001","article-title":"Tissue-specific activities of an immune signaling module regulate physiological responses to pathogenic and nutritional bacteria in C. elegans","volume":"6","author":"RP Shivers","year":"2009","journal-title":"Cell Host Microbe"},{"key":"pone.0344966.ref035","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1016\/B978-0-12-800511-8.00007-1","article-title":"Visualizing Neural Function","volume-title":"Guid to Res Tech Neurosci","author":"M Carter","year":"2015"},{"issue":"7","key":"pone.0344966.ref036","doi-asserted-by":"crossref","first-page":"107117","DOI":"10.1016\/j.isci.2023.107117","article-title":"A proton-inhibited DEG\/ENaC ion channel maintains neuronal ionstasis and promotes neuronal survival under stress","volume":"26","author":"D Petratou","year":"2023","journal-title":"iScience"},{"issue":"1561","key":"pone.0344966.ref037","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1098\/rstb.2010.0276","article-title":"The first long-lived mutants: discovery of the insulin\/IGF-1 pathway for ageing","volume":"366","author":"C Kenyon","year":"2011","journal-title":"Philos Trans R Soc B Biol Sci"},{"issue":"6","key":"pone.0344966.ref038","doi-asserted-by":"crossref","first-page":"1036","DOI":"10.1016\/j.cub.2023.01.059","article-title":"Increased alcohol dehydrogenase 1 activity promotes longevity","volume":"33","author":"A Ghaddar","year":"2023","journal-title":"Curr Biol"},{"issue":"10","key":"pone.0344966.ref039","doi-asserted-by":"crossref","DOI":"10.1111\/acel.13214","article-title":"Exogenous ethanol induces a metabolic switch that prolongs the survival of Caenorhabditis elegans dauer larva and enhances its resistance to desiccation","volume":"19","author":"D Kaptan","year":"2020","journal-title":"Aging Cell"},{"key":"pone.0344966.ref040","doi-asserted-by":"crossref","first-page":"1031161","DOI":"10.3389\/fragi.2023.1031161","article-title":"Periodic ethanol supply as a path toward unlimited lifespan of Caenorhabditis elegans dauer larvae","volume":"4","author":"X Zhang","year":"2023","journal-title":"Front Aging"},{"key":"pone.0344966.ref041","author":"M Bilodeau","year":"2021"},{"issue":"4","key":"pone.0344966.ref042","doi-asserted-by":"crossref","first-page":"600","DOI":"10.1016\/j.ygeno.2003.10.008","article-title":"Ethanol-response genes and their regulation analyzed by a microarray and comparative genomic approach in the nematode Caenorhabditis elegans","volume":"83","author":"JY Kwon","year":"2004","journal-title":"Genomics"},{"issue":"4","key":"pone.0344966.ref043","doi-asserted-by":"crossref","first-page":"757","DOI":"10.1016\/j.bbrc.2011.08.053","article-title":"Beneficial and harmful effects of alcohol exposure on Caenorhabditis elegans worms","volume":"412","author":"X Yu","year":"2011","journal-title":"Biochem Biophys Res Commun"},{"issue":"4","key":"pone.0344966.ref044","doi-asserted-by":"crossref","first-page":"483","DOI":"10.1111\/j.1474-9726.2007.00303.x","article-title":"Serotonin receptors antagonistically modulate Caenorhabditis elegans longevity","volume":"6","author":"H Murakami","year":"2007","journal-title":"Aging Cell"},{"issue":"6769","key":"pone.0344966.ref045","doi-asserted-by":"crossref","first-page":"560","DOI":"10.1038\/35000609","article-title":"Food and metabolic signalling defects in a Caenorhabditis elegans serotonin-synthesis mutant","volume":"403","author":"JY Sze","year":"2000","journal-title":"Nature"},{"key":"pone.0344966.ref046","doi-asserted-by":"crossref","first-page":"8828","DOI":"10.1038\/ncomms9828","article-title":"Metabotropic GABA signalling modulates longevity in C. elegans","volume":"6","author":"L Chun","year":"2015","journal-title":"Nat Commun"},{"key":"pone.0344966.ref047","doi-asserted-by":"crossref","DOI":"10.7554\/eLife.06259","article-title":"A gene-expression-based neural code for food abundance that modulates lifespan","volume":"4","author":"EV Entchev","year":"2015","journal-title":"Elife"},{"issue":"1","key":"pone.0344966.ref048","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1186\/s12915-022-01272-9","article-title":"Dopamine D1- and D2-like receptors oppositely regulate lifespan via a dietary restriction mechanism in Caenorhabditis elegans","volume":"20","author":"Y Jiang","year":"2022","journal-title":"BMC Biol"},{"issue":"4","key":"pone.0344966.ref049","doi-asserted-by":"crossref","first-page":"689","DOI":"10.1007\/s12038-016-9652-7","article-title":"Reserpine requires the D2-type receptor, dop-3, and the exoribonuclease, eri-1, to extend the lifespan in C. elegans","volume":"41","author":"K Saharia","year":"2016","journal-title":"J Biosci"},{"issue":"7","key":"pone.0344966.ref050","doi-asserted-by":"crossref","first-page":"1827","DOI":"10.3390\/molecules26071827","article-title":"Increased stress resistance and lifespan in Chaenorhabditis elegans wildtype and knockout mutants-implications for depression treatment by medicinal herbs","volume":"26","author":"J Na\u00df","year":"2021","journal-title":"Molecules"},{"issue":"8","key":"pone.0344966.ref051","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pgen.1001084","article-title":"Extracellular dopamine potentiates mn-induced oxidative stress, lifespan reduction, and dopaminergic neurodegeneration in a BLI-3-dependent manner in Caenorhabditis elegans","volume":"6","author":"A Benedetto","year":"2010","journal-title":"PLoS Genet"},{"key":"pone.0344966.ref052","first-page":"1","article-title":"C. elegans chemotaxis assay","volume":"74","author":"O Margie","year":"2013","journal-title":"J Vis Exp"},{"issue":"2","key":"pone.0344966.ref053","doi-asserted-by":"crossref","DOI":"10.1002\/cpz1.70106","article-title":"Enhancing reproducibility in chemotaxis assays for Caenorhabditis elegans","volume":"5","author":"L Cesar","year":"2025","journal-title":"Curr Protoc"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0344966","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,23]],"date-time":"2026-03-23T17:42:51Z","timestamp":1774287771000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0344966"}},"subtitle":[],"editor":[{"given":"Efthimios M. C.","family":"Skoulakis","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2026,3,23]]},"references-count":53,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2026,3,23]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0344966","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,23]]}}}