Nicotinic Acetylcholine Receptors Containing the α4 Subunit Modulate Alcohol Reward

Published:November 12, 2012DOI:


      Nicotine and alcohol are the two most co-abused drugs in the world, suggesting a common mechanism of action might underlie their rewarding properties. Although nicotine elicits reward by activating ventral tegmental area dopaminergic (DAergic) neurons via high-affinity neuronal nicotinic acetylcholine receptors (nAChRs), the mechanism by which alcohol activates these neurons is unclear.


      Because most high-affinity nAChRs expressed in ventral tegmental area DAergic neurons contain the α4 subunit, we measured ethanol-induced activation of DAergic neurons in midbrain slices from two complementary mouse models, an α4 knock-out (KO) mouse line and a knock-in line (Leu9′Ala) expressing α4 subunit-containing nAChRs hypersensitive to agonist compared with wild-type (WT). Activation of DAergic neurons by ethanol was analyzed with both biophysical and immunohistochemical approaches in midbrain slices. The ability of alcohol to condition a place preference in each mouse model was also measured.


      At intoxicating concentrations, ethanol activation of DAergic neurons was significantly reduced in α4 KO mice compared with WT. Conversely, in Leu9′Ala mice, DAergic neurons were activated by low ethanol concentrations that did not increase activity of WT neurons. In addition, alcohol potentiated the response to ACh in DAergic neurons, an effect reduced in α4 KO mice. Rewarding alcohol doses failed to condition a place preference in α4 KO mice, paralleling alcohol effects on DAergic neuron activity, whereas a sub-rewarding alcohol dose was sufficient to condition a place preference in Leu9′Ala mice.


      Together, these data indicate that nAChRs containing the α4 subunit modulate alcohol reward.

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        • Batel P.
        • Pessione F.
        • Maitre C.
        • Rueff B.
        Relationship between alcohol and tobacco dependencies among alcoholics who smoke.
        Addiction. 1995; 90: 977-980
        • Hurt R.D.
        • Offord K.P.
        • Croghan I.T.
        • Gomez-Dahl L.
        • Kottke T.E.
        • Morse R.M.
        • Melton 3rd, L.J.
        Mortality following inpatient addictions treatment. Role of tobacco use in a community-based cohort.
        JAMA. 1996; 275: 1097-1103
        • Rodd Z.A.
        • Melendez R.I.
        • Bell R.L.
        • Kuc K.A.
        • Zhang Y.
        • Murphy J.M.
        • McBride W.J.
        Intracranial self-administration of ethanol within the ventral tegmental area of male Wistar rats: Evidence for involvement of dopamine neurons.
        J Neurosci. 2004; 24: 1050-1057
        • Okamoto T.
        • Harnett M.T.
        • Morikawa H.
        Hyperpolarization-activated cation current (Ih) is an ethanol target in midbrain dopamine neurons of mice.
        J Neurophysiol. 2006; 95: 619-626
        • Tsai H.C.
        • Zhang F.
        • Adamantidis A.
        • Stuber G.D.
        • Bonci A.
        • de Lecea L.
        • Deisseroth K.
        Phasic firing in dopaminergic neurons is sufficient for behavioral conditioning.
        Science. 2009; 324: 1080-1084
        • Pidoplichko V.I.
        • DeBiasi M.
        • Williams J.T.
        • Dani J.A.
        Nicotine activates and desensitizes midbrain dopamine neurons.
        Nature. 1997; 390: 401-404
        • Maskos U.
        • Molles B.E.
        • Pons S.
        • Besson M.
        • Guiard B.P.
        • Guilloux J.P.
        • et al.
        Nicotine reinforcement and cognition restored by targeted expression of nicotinic receptors.
        Nature. 2005; 436: 103-107
        • McBride W.J.
        • Lovinger D.M.
        • Machu T.
        • Thielen R.J.
        • Rodd Z.A.
        • Murphy J.M.
        • et al.
        Serotonin-3 receptors in the actions of alcohol, alcohol reinforcement, and alcoholism.
        Alcohol Clin Exp Res. 2004; 28: 257-267
        • Dopico A.M.
        • Lovinger D.M.
        Acute alcohol action and desensitization of ligand-gated ion channels.
        Pharmacol Rev. 2009; 61: 98-114
        • Albuquerque E.X.
        • Pereira E.F.
        • Alkondon M.
        • Rogers S.W.
        Mammalian nicotinic acetylcholine receptors: From structure to function.
        Physiol Rev. 2009; 89: 73-120
        • Tapper A.R.
        • Nashmi R.
        • Lester H.A.
        Neuronal nicotinic acetylcholine receptors and nicotine dependence.
        in: Madras B.K. Colvis C.M. Pollock J.D. Rutter J.L. Shurtleff D. Zastrow M.V. Cell Biology of Addiction. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY2006: 179-191
        • Laviolette S.R.
        • van der Kooy D.
        The neurobiology of nicotine addiction: Bridging the gap from molecules to behaviour.
        Nat Rev Neurosci. 2004; 5: 55-65
        • Larsson A.
        • Edstrom L.
        • Svensson L.
        • Soderpalm B.
        • Engel J.A.
        Voluntary ethanol intake increases extracellular acetylcholine levels in the ventral tegmental area in the rat.
        Alcohol Alcohol. 2005; 40: 349-358
        • Cardoso R.A.
        • Brozowski S.J.
        • Chavez-Noriega L.E.
        • Harpold M.
        • Valenzuela C.F.
        • Harris R.A.
        Effects of ethanol on recombinant human neuronal nicotinic acetylcholine receptors expressed in Xenopus oocytes.
        J Pharmacol Exp Ther. 1999; 289: 774-780
        • Zuo Y.
        • Kuryatov A.
        • Lindstrom J.M.
        • Yeh J.Z.
        • Narahashi T.
        Alcohol modulation of neuronal nicotinic acetylcholine receptors is alpha subunit dependent.
        Alcohol Clin Exp Res. 2002; 26: 779-784
        • Blomqvist O.
        • Engel J.A.
        • Nissbrandt H.
        • Soderpalm B.
        The mesolimbic dopamine-activating properties of ethanol are antagonized by mecamylamine.
        Eur J Pharmacol. 1993; 249: 207-213
        • Blomqvist O.
        • Ericson M.
        • Engel J.A.
        • Soderpalm B.
        Accumbal dopamine overflow after ethanol: Localization of the antagonizing effect of mecamylamine.
        Eur J Pharmacol. 1997; 334: 149-156
        • Ericson M.
        • Blomqvist O.
        • Engel J.A.
        • Soderpalm B.
        Voluntary ethanol intake in the rat and the associated accumbal dopamine overflow are blocked by ventral tegmental mecamylamine.
        Eur J Pharmacol. 1998; 358: 189-196
        • Hendrickson L.M.
        • Zhao-Shea R.
        • Tapper A.R.
        Modulation of ethanol drinking-in-the-dark by mecamylamine and nicotinic acetylcholine receptor agonists in C57BL/6J mice.
        Psychopharmacology (Berl). 2009; 204: 563-572
        • Chi H.
        • de Wit H.
        Mecamylamine attenuates the subjective stimulant-like effects of alcohol in social drinkers.
        Alcohol Clin Exp Res. 2003; 27: 780-786
        • Steensland P.
        • Simms J.A.
        • Holgate J.
        • Richards J.K.
        • Bartlett S.E.
        Varenicline, an alpha4beta2 nicotinic acetylcholine receptor partial agonist, selectively decreases ethanol consumption and seeking.
        Proc Natl Acad Sci U S A. 2007; 104: 12518-12523
        • McKee S.A.
        • Harrison E.L.
        • O’Malley S.S.
        • Krishnan-Sarin S.
        • Shi J.
        • Tetrault J.M.
        • et al.
        Varenicline reduces alcohol self-administration in heavy-drinking smokers.
        Biol Psychiatry. 2009; 66: 185-190
        • Kamens H.M.
        • Andersen J.
        • Picciotto M.R.
        Modulation of ethanol consumption by genetic and pharmacological manipulation of nicotinic acetylcholine receptors in mice.
        Psychopharmacology (Berl). 2010; 208: 613-626
        • Hendrickson L.M.
        • Zhao-Shea R.
        • Pang X.
        • Gardner P.D.
        • Tapper A.R.
        Activation of alpha4low asterisk nAChRs is necessary and sufficient for varenicline-induced reduction of alcohol consumption.
        J Neurosci. 2010; 30: 10169-10176
        • Hendrickson L.M.
        • Gardner P.
        • Tapper A.R.
        Nicotinic acetylcholine receptors containing the alpha4 subunit are critical for the nicotine-induced reduction of acute voluntary ethanol consumption.
        Channels (Austin). 2011; 5: 124-127
        • National Research Council
        Guide for the Care and Use of Laboratory Animals. National Academy Press, Washington, DC1996
        • Liu L.
        • Zhao-Shea R.
        • McIntosh J.M.
        • Gardner P.D.
        • Tapper A.R.
        Nicotine persistently activates ventral tegmental area dopaminergic neurons via nicotinic acetylcholine receptors containing alpha4 and alpha6 subunits.
        Mol Pharmacol. 2012; 81: 541-548
        • Gibb S.L.
        • Jeanblanc J.
        • Barak S.
        • Yowell Q.V.
        • Yaka R.
        • Ron D.
        Lyn kinase regulates mesolimbic dopamine release: implication for alcohol reward.
        J Neurosci. 2011; 31: 2180-2187
        • Zhao-Shea R.
        • Liu L.
        • Soll L.G.
        • Improgo M.R.
        • Meyers E.E.
        • McIntosh J.M.
        • et al.
        Nicotine-mediated activation of dopaminergic neurons in distinct regions of the ventral tegmental area.
        Neuropsychopharmacology. 2011; 36: 1021-1032
        • Tapper A.R.
        • McKinney S.L.
        • Nashmi R.
        • Schwarz J.
        • Deshpande P.
        • Labarca C.
        • et al.
        Nicotine activation of alpha4low asterisk receptors: sufficient for reward, tolerance, and sensitization.
        Science. 2004; 306: 1029-1032
        • Pons S.
        • Fattore L.
        • Cossu G.
        • Tolu S.
        • Porcu E.
        • McIntosh J.M.
        • et al.
        Crucial role of alpha4 and alpha6 nicotinic acetylcholine receptor subunits from ventral tegmental area in systemic nicotine self-administration.
        J Neurosci. 2008; 28: 12318-12327
        • Ross S.A.
        • Wong J.Y.
        • Clifford J.J.
        • Kinsella A.
        • Massalas J.S.
        • Horne M.K.
        • et al.
        Phenotypic characterization of an alpha 4 neuronal nicotinic acetylcholine receptor subunit knock-out mouse.
        J Neurosci. 2000; 20: 6431-6441
        • Fonck C.
        • Cohen B.N.
        • Nashmi R.
        • Whiteaker P.
        • Wagenaar D.A.
        • Rodrigues-Pinguet N.
        • et al.
        Novel seizure phenotype and sleep disruptions in knock-in mice with hypersensitive alpha 4low asterisk nicotinic receptors.
        J Neurosci. 2005; 25: 11396-11411
        • Rodd Z.A.
        • Bell R.L.
        • Melendez R.I.
        • Kuc K.A.
        • Lumeng L.
        • Li T.K.
        • et al.
        Comparison of intracranial self-administration of ethanol within the posterior ventral tegmental area between alcohol-preferring and Wistar rats.
        Alcohol Clin Exp Res. 2004; 28: 1212-1219
        • Cunningham C.L.
        • Gremel C.M.
        • Groblewski P.A.
        Drug-induced conditioned place preference and aversion in mice.
        Nat Protoc. 2006; 1: 1662-1670
        • Tzschentke T.M.
        Measuring reward with the conditioned place preference (CPP) paradigm: Update of the last decade.
        Addict Biol. 2007; 12: 227-462
        • Chatterjee S.
        • Steensland P.
        • Simms J.A.
        • Holgate J.
        • Coe J.W.
        • Hurst R.S.
        • et al.
        Partial agonists of the α3β4low asterisk neuronal nicotinic acetylcholine receptor reduce ethanol consumption and seeking in rats.
        Neuropsychopharmacology. 2011; 36: 603-615
        • Lester H.A.
        • Fonck C.
        • Tapper A.R.
        • McKinney S.
        • Damaj M.I.
        • Balogh S.
        • et al.
        Hypersensitive knockin mouse strains identify receptors and pathways for nicotine action.
        Curr Opin Drug Discov Devel. 2003; 6: 633-639
        • Yang K.
        • Hu J.
        • Lucero L.
        • Liu Q.
        • Zheng C.
        • Zhen X.
        • et al.
        Distinctive nicotinic acetylcholine receptor functional phenotypes of rat ventral tegmental area dopaminergic neurons.
        J Physiol. 2009; 587: 345-361
        • Bechtholt A.J.
        • Cunningham C.L.
        Ethanol-induced conditioned place preference is expressed through a ventral tegmental area dependent mechanism.
        Behav Neurosci. 2005; 119: 213-223
        • Rezvani A.H.
        • Slade S.
        • Wells C.
        • Petro A.
        • Lumeng L.
        • Li T.K.
        • et al.
        Effects of sazetidine-A, a selective alpha4beta2 nicotinic acetylcholine receptor desensitizing agent on alcohol and nicotine self-administration in selectively bred alcohol-preferring (P) rats.
        Psychopharmacology (Berl). 2010; 211: 161-174
        • Larsson A.
        • Jerlhag E.
        • Svensson L.
        • Soderpalm B.
        • Engel J.A.
        Is an alpha-conotoxin MII-sensitive mechanism involved in the neurochemical, stimulatory, and rewarding effects of ethanol?.
        Alcohol. 2004; 34: 239-250
        • Larsson A.
        • Svensson L.
        • Soderpalm B.
        • Engel J.A.
        Role of different nicotinic acetylcholine receptors in mediating behavioral and neurochemical effects of ethanol in mice.
        Alcohol. 2002; 28: 157-167
        • Ericson M.
        • Molander A.
        • Lof E.
        • Engel J.A.
        • Soderpalm B.
        Ethanol elevates accumbal dopamine levels via indirect activation of ventral tegmental nicotinic acetylcholine receptors.
        Eur J Pharmacol. 2003; 467: 85-93
        • Kuzmin A.
        • Jerlhag E.
        • Liljequist S.
        • Engel J.
        Effects of subunit selective nACh receptors on operant ethanol self-administration and relapse-like ethanol-drinking behavior.
        Psychopharmacology (Berl). 2009; 203: 99-108
        • Moroni M.
        • Zwart R.
        • Sher E.
        • Cassels B.K.
        • Bermudez I.
        alpha4beta2 nicotinic receptors with high and low acetylcholine sensitivity: Pharmacology, stoichiometry, and sensitivity to long-term exposure to nicotine.
        Mol Pharmacol. 2006; 70: 755-768
        • Grady S.R.
        • Salminen O.
        • Laverty D.C.
        • Whiteaker P.
        • McIntosh J.M.
        • Collins A.C.
        • Marks M.J.
        The subtypes of nicotinic acetylcholine receptors on dopaminergic terminals of mouse striatum.
        Biochem Pharmacol. 2007; 74: 1235-1246
        • Kamens H.M.
        • Hoft N.R.
        • Cox R.J.
        • Miyamoto J.H.
        • Ehringer M.A.
        The alpha6 nicotinic acetylcholine receptor subunit influences ethanol-induced sedation.
        Alcohol. 2012; 46: 463-471