Ubiquitously expressed genes have been implicated in a variety of specific behaviors, including responses to ethanol. However, the mechanisms that confer this behavioral specificity have remained elusive. Previously, we showed that the ubiquitously expressed small GTPase Arf6 is required for normal ethanol-induced sedation in adult Drosophila. Here, we show that this behavioral response also requires Efa6, one of (at least) three Drosophila Arf6 guanine exchange factors. Ethanol-naive Arf6 and Efa6 mutants were sensitive to ethanol-induced sedation and lacked rapid tolerance upon re-exposure to ethanol, when compared with wild-type flies. In contrast to wild-type flies, both Arf6 and Efa6 mutants preferred alcohol-containing food without prior ethanol experience. An analysis of the human ortholog of Arf6 and orthologs of Efa6 (PSD1-4) revealed that the minor G allele of single nucleotide polymorphism (SNP) rs13265422 in PSD3, as well as a haplotype containing rs13265422, was associated with an increased frequency of drinking and binge drinking episodes in adolescents. The same haplotype was also associated with increased alcohol dependence in an independent European cohort. Unlike the ubiquitously expressed human Arf6 GTPase, PSD3 localization is restricted to the brain, particularly the prefrontal cortex (PFC). Functional magnetic resonance imaging revealed that the same PSD3 haplotype was also associated with a differential functional magnetic resonance imaging signal in the PFC during a Go/No-Go task, which engages PFC-mediated executive control. Our translational analysis, therefore, suggests that PSD3 confers regional specificity to ubiquitous Arf6 in the PFC to modulate human alcohol-drinking behaviors.
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Edenberg HJ, Foroud T . Genetics and alcoholism. Nat Rev Gastroenterol Hepatol 2013; 10: 487–494.
Topper SM, Aguilar SC, Topper VY, Elbel E, Pierce-Shimomura JT . Alcohol disinhibition of behaviors in C. elegans. PLoS ONE 2014; 9: e92965.
Schuckit MA . Low level of response to alcohol as a predictor of future alcoholism. Am J Psychiatry 1994; 151: 184–189.
Newlin DB, Thomson JB . Alcohol challenge with sons of alcoholics: a critical review and analysis. Psychol Bull 1990; 108: 383.
King AC, de Wit H, McNamara PJ, Cao D . Rewarding, stimulant, and sedative alcohol responses and relationship to future binge drinking. Arch Gen Psychiatry 2011; 68: 389–399.
Dick DM, Jones K, Saccone N, Hinrichs A, Wang JC, Goate A et al. Endophenotypes successfully lead to gene identification: results from the collaborative study on the genetics of alcoholism. Behav Genet 2006; 36: 112–126.
Reich DE, Lander ES . On the allelic spectrum of human disease. Trends Genet 2001; 17: 502–510.
True WR, Xian H, Scherrer JF, Madden PA, Bucholz KK, Heath AC et al. Common genetic vulnerability for nicotine and alcohol dependence in men. Arch Gen Psychiatry 1999; 56: 655–661.
Heath AC, Bucholz KK, Madden PA, Dinwiddie SH, Slutske WS, Bierut L et al. Genetic and environmental contributions to alcohol dependence risk in a national twin sample: consistency of findings in women and men. Psychol Med 1997; 27: 1381–1396.
Prescott CA, Kendler KS . Genetic and environmental contributions to alcohol abuse and dependence in a population-based sample of male twins. Am J Psychiatry 1999; 156: 34–40.
Narayanan AS, Rothenfluh A . I believe I can fly!: Use of Drosophila as a model organism in neuropsychopharmacology research. Neuropsychopharmacology 2016; 41: 1439–1446.
Kaun KR, Devineni AV, Heberlein U . Drosophila melanogaster as a model to study drug addiction. Hum Genet 2012; 131: 959–975.
Lee H-G, Kim Y-C, Dunning JS, Han K-A . Recurring ethanol exposure induces disinhibited courtship in Drosophila. PLoS ONE 2008; 3: e1391.
Wolf FW, Rodan AR, Tsai LTY, Heberlein U . High-resolution analysis of ethanol-induced locomotor stimulation in Drosophila. J Neurosci 2002; 22: 11035–11044.
Scholz H, Ramond J, Singh CM, Heberlein U . Functional ethanol tolerance in Drosophila. Neuron 2000; 28: 261–271.
Devineni AV, Heberlein U . Preferential ethanol consumption in Drosophila models features of addiction. Curr Biol 2009; 19: 2126–2132.
Peru y Colón de Portugal RL, Ojelade SA, Penninti PS, Dove RJ, Nye MJ, Acevedo SF et al. Long-lasting, experience-dependent alcohol preference in Drosophila. Addic Biol 2014; 19: 392–401.
Grotewiel M, Bettinger JC . Drosophila and Caenorhabditis elegans as discovery platforms for genes involved in human alcohol use disorder. Alcoholism Clin Exp Res 2015; 39: 1292–1311.
Ojelade SA, Jia T, Rodan AR, Chenyang T, Kadrmas JL, Cattrell A et al. Rsu1 regulates ethanol consumption in Drosophila and humans. Proc Natl Acad Sci USA 2015; 112: E4085–E4093.
Rothenfluh A, Cowan CW . Emerging roles of actin cytoskeleton regulating enzymes in drug addiction: actin or reactin’? Curr Opin Neurobiol 2013; 23: 507–512.
Peru y Colón de Portugal RL, Acevedo SF, Rodan AR, Chang LY, Eaton BA, Rothenfluh A . Adult neuronal Arf6 controls ethanol-induced behavior with Arfaptin downstream of Rac1 and RhoGAP18B. J Neurosci 2012; 32: 17706–17713.
Song J, Khachikian Z, Radhakrishna H, Donaldson JG . Localization of endogenous Arf6 to sites of cortical actin rearrangement and involvement of Arf6 in cell spreading. J Cell Sci 1998; 111: 2257–2267.
Lebeda RA, Johnson SK, Stewart MI, Haun RS . Sequence, genomic organization, and expression of the human ADP-Ribosylation Factor 6 (ARF6) gene: a class III ARF. DNA Cell Biol 2004; 22: 737–741.
D'Souza-Schorey C, Chavrier P . ARF proteins: roles in membrane traffic and beyond. Nature Rev Mol Cell Biol 2006; 7: 347–358.
Acevedo SF, Peru y Colón de Portugal RL, Gonzalez DA Rodan AR, Rothenfluh A . S6 Kinase reflects and regulates ethanol-induced sedation. J Neurosci 2015; 35: 15396–15402.
Neasta J, Ben Hamida S, Yowell Q, Carnicella S, Ron D . Role for mammalian target of rapamycin complex 1 signaling in neuroadaptations underlying alcohol-related disorders. Proc Natl Acad Sci USA 2010; 107: 20093–20098.
Barak S, Liu F, Ben Hamida S, Yowell QV, Neasta J, Kharazia V et al. Disruption of alcohol-related memories by mTORC1 inhibition prevents relapse. Nat Neurosci 2013; 16: 1111–1117.
Santos, dos G, Schroeder AJ, Goodman JL, Strelets VB, Crosby MA, Thurmond J et al. FlyBase: introduction of the Drosophila melanogaster release 6 reference genome assembly and large-scale migration of genome annotations. Nucleic Acids Res 2015; 43: D690–D697.
Wu C, Jin X, Tsueng G, Afrasiabi C, Su AI . BioGPS: building your own mash-up of gene annotations and expression profiles. Nucleic Acids Res 2016; 44: D313–D316.
Huang J, Zhou W, Dong W, Watson AM, Hong Y . Directed, efficient, and versatile modifications of the Drosophila genome by genomic engineering. Proc Natl Acad Sci USA 2009; 106: 8284–8289.
Rothenfluh A, Threlkeld RJ, Bainton RJ, Tsai LTY, Lasek AW, Heberlein U . Distinct behavioral responses to ethanol are regulated by alternate RhoGAP18B isoforms. Cell 2006; 127: 199–211.
Morgan M, Hibell B, Andersson B, Bjarnason T, Kokkevi A, Narusk A . The ESPAD study: implications for prevention. Drugs 2009; 6: 243–256.
Viner RM, Taylor B . Adult outcomes of binge drinking in adolescence: findings from a UK national birth cohort. J Epidemiol Commun Health 2007; 61: 902–907.
Schumann G, Loth E, Banaschewski T, Barbot A, Barker G, Büchel C et al. The IMAGEN study: reinforcement-related behaviour in normal brain function and psychopathology. Mol Psychiatry 2010; 15: 1128–1139.
Bierut LJ, Agrawal A, Bucholz KK, Doheny KF, Laurie C, Pugh E et al. A genome-wide association study of alcohol dependence. Proc Natl Acad Sci USA 2010; 107: 5082–5087.
Gabriel SB, Schaffner SF, Nguyen H, Moore JM, Roy J, Blumenstiel B et al. The structure of haplotype blocks in the human genome. Science 2002; 296: 2225–2229.
Reich DE, Schaffner SF, Daly MJ, McVean G, Mullikin JC, Higgins JM et al. Human genome sequence variation and the influence of gene history, mutation and recombination. Nat Genet 2002; 32: 135–142.
Barrett JC, Fry B, Maller J, Daly MJ . Haploview: analysis and visualization of LD and haplotype maps. Bioinformatics 2005; 21: 263–265.
Purcell S, Neale B, Todd-Brown K, Thomas L, Ferreira MAR, Bender D et al. PLINK: a tool set for whole-genome association and population-based linkage analyses. Am J Hum Genet 2007; 81: 559–575.
Ojelade SA, Acevedo SF, Kalahasti G, Rodan AR, Rothenfluh A . RhoGAP18B isoforms act on distinct Rho-Family GTPases and regulate behavioral responses to alcohol via cofilin. PLoS ONE 2015; 10: e0137465.
Naassila M, Ledent C, Daoust M . Low ethanol sensitivity and increased ethanol consumption in mice lacking adenosine A2A receptors. J Neurosci 2002; 22: 10487–10493.
Moore MS, DeZazzo J, Luk AY, Tully T, Singh CM, Heberlein U . Ethanol intoxication in Drosophila: genetic and pharmacological evidence for regulation by the cAMP signaling pathway. Cell 1998; 93: 997–1007.
Devineni AV, McClure K, Guarnieri D, Corl A, Wolf F . The genetic relationships between ethanol preference, acute ethanol sensitivity, and ethanol tolerance in Drosophila melanogaster. Fly 2011; 5: 191–199.
Pohl JB, Baldwin BA, Dinh BL, Rahman P, Smerek D, Prado FJ III et al. Ethanol preference in Drosophila melanogaster is driven by its caloric value. Alcoholism Clin Exp Res 2012; 36: 1903–1912.
Xu S, Chan T, Shah V, Zhang S, Pletcher SD, Roman G . The propensity for consuming ethanol in Drosophila requires rutabaga adenylyl cyclase expression within mushroom body neurons. Genes Brain Behav 2012; 11: 727–739.
Skeeles LE, Fleming JL, Mahler KL, Toland AE . The impact of 3′UTR variants on differential expression of candidate cancer susceptibility genes. PLoS ONE 2013; 8: e58609.
Brown JB, Boley N, Eisman R, May GE, Stoiber MH, Duff MO et al. Diversity and dynamics of the Drosophila transcriptome. Nature 2014; 512: 393–399.
Donaldson JG, Jackson CL . ARF family G proteins and their regulators: roles in membrane transport, development and disease. Nature Rev Mol Cell Biol 2011; 12: 362–375.
Su AI, Wiltshire T, Batalov S, Lapp H, Ching KA, Block D et al. A gene atlas of the mouse and human protein-encoding transcriptomes. Proc Natl Acad Sci USA 2004; 101: 6062–6067.
Robbins TW, Arnsten A . The neuropsychopharmacology of fronto-executive function: monoaminergic modulation. Annu Rev Neurosci 2009; 32: 267–287.
Lu YL, Richardson HN . Alcohol, stress hormones, and the prefrontal cortex: a proposed pathway to the dark side of addiction. Neuroscience 2014; 277: 139–151.
Jasinska AJ, Chen BT, Bonci A, Stein EA . Dorsal medial prefrontal cortex (MPFC) circuitry in rodent models of cocaine use: implications for drug addiction therapies. Addic Biol 2015; 20: 215–226.
Volkow ND, Koob GF, McLellan AT . Neurobiologic advances from the brain disease model of addiction. N Engl J Med 2016; 374: 363–371.
Sanda M, Kamata A, Katsumata O, Fukunaga K, Watanabe M, Kondo H et al. The postsynaptic density protein, IQ-ArfGEF/BRAG1, can interact with IRSp53 through its proline-rich sequence. Brain Res 2009; 1251: 7–15.
Hernández-Deviez DJ, Roth MG, Casanova JE, Wilson JM . ARNO and ARF6 regulate axonal elongation and branching through downstream activation of phosphatidylinositol 4-phosphate 5-kinase alpha. Mol Biol Cell 2004; 15: 111–120.
Hernández-Deviez DJ, Casanova JE, Wilson JM . Regulation of dendritic development by the ARF exchange factor ARNO. Nat Neurosci 2002; 5: 623–624.
Graveley BR, Brooks AN, Carlson JW, Duff MO, Landolin JM, Yang L et al. The developmental transcriptome of Drosophila melanogaster. Nature 2011; 471: 473–479.
The modENCODE Consortium, Roy S, Ernst J, Kharchenko PV, Kheradpour P, Negre N et al. Identification of functional elements and regulatory circuits by Drosophila modENCODE. Science 2010; 330: 1787–1797.
Whelan R, Conrod PJ, Poline J-B, Lourdusamy A, Banaschewski T, Barker GJ et al. Adolescent impulsivity phenotypes characterized by distinct brain networks. Nat Neurosci 2012; 15: 920–925.
Dietz DM, Sun H, Lobo MK, Cahill ME, Chadwick B, Gao V et al. Rac1 is essential in cocaine-induced structural plasticity of nucleus accumbens neurons. Nat Neurosci 2012; 15: 891–896.
Toda S, Shen H-W, Peters J, Cagle S, Kalivas PW . Cocaine increases actin cycling: effects in the reinstatement model of drug seeking. J Neurosci 2006; 26: 1579–1587.
We thank the Bloomington stock center, and Yang Hong (University of Pittsburgh) for fly strains, and Michael Buszczak, Helmut Krämer and Rothenfluh lab members for helpful discussions. This work was supported by the NIH (T32 fellowships DA007290 to DAG and JHP, F32 AA021340 to SAO, K08 DK091316 to ARR, R01 AA019526 and R21 AA022404 to AR), the European Union-funded FP6 Integrated Project IMAGEN (Reinforcement-related behavior in normal brain function and psychopathology; LSHM-CT-2007–037286), the FP7 projects IMAGEMEND (602450) and MATRICS (603016), the Innovative Medicine Initiative Project EU-AIMS (115300-2), the European Research Council Award ‘STRATIFY’ as well as the Medical Research Council Programme Grant ‘Developmental pathways into adolescent substance abuse’ (93558). Further support was provided by the Swedish Funding Agency FORMAS, the MRC-ICMR Newton project ‘Consortium on Vulnerability to Externalizing Disorders and Addictions’ (c-VEDA) (MR/N000390/1), the National Institute for Health Research (NIHR) Biomedical Research Centre at South London and Maudsley NHS Foundation Trust and King’s College London, the German Bundesministerium für Bildung und Forschung (BMBF grants 01GS08152; 01EV0711; eMED SysAlc 01ZX1311A; Forschungsnetz AERIAL), the NIH (R01 MH085772-01A1), as well as the NIH-BD2K (Big Data to Knowledge) grant U54 EB020403–ENIGMA Center for Worldwide Medicine, Imaging and Genomics. AR was also supported by an Effie Marie Cain Scholarship in Biomedical Research from UT Southwestern Medical Center Dallas.
DAG, JHP, SAO, SFA and AR conceived, performed and analyzed the Drosophila experiments. SD, TB, CB, ALWB, PJC, HF, BI, ML, JM, TP, MS, GS and the IMAGEN consortium acquired the human data. TJ, BX and GS analyzed the human data. DAG, TJ, JHP, JLH, ARR, GS and AR wrote the paper.
TB served in an advisory or consultancy role for Hexal Pharma, Lilly, Medice, Novartis, Otsuka, Oxford Outcomes, PCM Scientific, Shire and Viforpharma. TB received conference attendance support and conference support or speaker’s fees from Lilly, Medice, Novartis and Shire. TB is/has been involved in clinical trials conducted by Shire and Viforpharma. JG has received research funding from AstraZeneca, Eli Lilly & Co., Janssen-Cilag and Bristol-Myers Squibb, and speaker’s fees from AstraZeneca, Janssen-Cilag and Bristol-Myers Squibb. The remaining authors declare no conflict of interest.
Supplementary Information accompanies the paper on the Molecular Psychiatry website
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Gonzalez, D., Jia, T., Pinzón, J. et al. The Arf6 activator Efa6/PSD3 confers regional specificity and modulates ethanol consumption in Drosophila and humans. Mol Psychiatry 23, 621–628 (2018). https://doi.org/10.1038/mp.2017.112
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