, Volume 184, Issue 3–4, pp 328–338 | Cite as

β2-Subunit-containing nicotinic acetylcholine receptors are involved in nicotine-induced increases in conditioned reinforcement but not progressive ratio responding for food in C57BL/6 mice

  • Darlene H. Brunzell
  • Jessica R. Chang
  • Brandon Schneider
  • Peter Olausson
  • Jane R. Taylor
  • Marina R. Picciotto
Original Investigation



Nicotine administration potentiates conditioned reinforcement in rats, an effect that persists for weeks after chronic exposure. Little is known regarding the nicotinic receptor subtypes that may mediate this effect.


The purpose of this study was to determine whether β2-subunit-containing nicotinic acetylcholine receptors (β2*nAChRs) are necessary for lasting effects of nicotine on conditioned and primary reinforcement in mice.


β2 knockout (β2KO) and wild-type (WT) mice received 14 days of nicotine exposure (NIC, 200 μg/ml in 2% saccharin) or saccharin alone (SAC) in their drinking water. Five days later, mice received paired presentations of a conditioned stimulus (CS) with water unconditioned stimulus (US) or explicitly unpaired presentations of the CS and US during Pavlovian discriminative approach training. Training was followed by two conditioned reinforcement tests. Mice were subsequently tested for food-reinforced responding in the absence of explicit cues followed by a progressive ratio test.


During conditioned reinforcement testing, only mice in the paired condition showed increased responding in the CS-reinforced aperture over inactive apertures. WT-NIC mice showed enhanced conditioned reinforcement compared to WT-SAC animals. β2KO-SAC mice showed elevated conditioned reinforcement compared to WT-SAC subjects, but β2KO-NIC and β2KO-SAC mice did not differ in responding with conditioned reinforcement. Prior nicotine exposure did not alter food-reinforced responding but resulted in elevated break points for food in both genotypes.


These data show that nicotine exposure enhances conditioned reinforcement in mice and indicate that β2*nAChRs are necessary for nicotine-dependent enhancement of incentive aspects of motivation but not motivation for primary reinforcement measured by progressive ratio responding.


Learning Reward Motivation Mouse Associative learning Pavlovian Drug 



This work was supported by grants DA00436, DA14241, and AA15632 from the National Institutes of Health. We wish to thank Chris Kochevar and Elin Lof for experimental assistance and Natalie Tronson for help in modifying the behavioral software.


  1. Arthur D, Levin ED (2002) Chronic inhibition of alpha4beta2 nicotinic receptors in the ventral hippocampus of rats: impacts on memory and nicotine response. Psychopharmacology (Berl) 160:140–145CrossRefGoogle Scholar
  2. Bancroft A, Levin ED (2000) Ventral hippocampal alpha4beta2 nicotinic receptors and chronic nicotine effects on memory. Neuropharmacology 39:2770–2778CrossRefPubMedGoogle Scholar
  3. Benwell ME, Balfour DJ (1992) The effects of acute and repeated nicotine treatment on nucleus accumbens dopamine and locomotor activity. Br J Pharmacol 105:849–856PubMedGoogle Scholar
  4. Benwell ME, Balfour DJ, Anderson JM (1988) Evidence that tobacco smoking increases the density of (−)-[3H]nicotine binding sites in human brain. J Neurochem 50:1243–1247PubMedCrossRefGoogle Scholar
  5. Benwell ME, Balfour DJ, Khadra LF (1994) Studies on the influence of nicotine infusions on mesolimbic dopamine and locomotor responses to nicotine. Clin Investig 72:233–239CrossRefPubMedGoogle Scholar
  6. Brody AL, Mandelkern MA, London ED, Childress AR, Lee GS, Bota RG, Ho ML, Saxena S, Baxter LR Jr, Madsen D, Jarvik ME (2002) Brain metabolic changes during cigarette craving. Arch Gen Psychiatry 59:1162–1172CrossRefPubMedGoogle Scholar
  7. Brunzell DH, Russell DS, Picciotto MR (2003) In vivo nicotine treatment regulates mesocorticolimbic CREB and ERK signaling in C57Bl/6J mice. J Neurochem 84:1431–1441CrossRefPubMedGoogle Scholar
  8. Buisson B, Bertrand D (2001) Chronic exposure to nicotine upregulates the human (alpha)4((beta)2 nicotinic acetylcholine receptor function. J Neurosci 21:1819–1829PubMedGoogle Scholar
  9. Burns LH, Robbins TW, Everitt BJ (1993) Differential effects of excitotoxic lesions of the basolateral amygdala, ventral subiculum and medial prefrontal cortex on responding with conditioned reinforcement and locomotor activity potentiated by intra-accumbens infusions of d-amphetamine. Behav Brain Res 55:167–183CrossRefPubMedGoogle Scholar
  10. Cadoni C, Di Chiara G (2000) Differential changes in accumbens shell and core dopamine in behavioral sensitization to nicotine. Eur J Pharmacol 387:R23–R25CrossRefPubMedGoogle Scholar
  11. Cador M, Taylor JR, Robbins TW (1991) Potentiation of the effects of reward-related stimuli by dopaminergic-dependent mechanisms in the nucleus accumbens. Psychopharmacology (Berl) 104:377–385CrossRefGoogle Scholar
  12. Caggiula AR, Donny EC, White AR, Chaudhri N, Booth S, Gharib MA, Hoffman A, Perkins KA, Sved AF (2001) Cue dependency of nicotine self-administration and smoking. Pharmacol Biochem Behav 70:515–530CrossRefPubMedGoogle Scholar
  13. Caggiula AR, Donny EC, Chaudhri N, Perkins KA, Evans-Martin FF, Sved AF (2002a) Importance of nonpharmacological factors in nicotine self-administration. Physiol Behav 77:683–687CrossRefPubMedGoogle Scholar
  14. Caggiula AR, Donny EC, White AR, Chaudhri N, Booth S, Gharib MA, Hoffman A, Perkins KA, Sved AF (2002b) Environmental stimuli promote the acquisition of nicotine self-administration in rats. Psychopharmacology (Berl) 163:230–237CrossRefGoogle Scholar
  15. Chiamulera C (2005) Cue reactivity in nicotine and tobacco dependence: a “multiple-action” model of nicotine as a primary reinforcement and as an enhancer of the effects of smoking-associated stimuli. Brain Res Brain Res Rev 48:74–97CrossRefPubMedGoogle Scholar
  16. Cohen C, Perrault G, Griebel G, Soubrie P (2005) Nicotine-associated cues maintain nicotine-seeking behavior in rats several weeks after nicotine withdrawal: reversal by the cannabinoid (CB1) receptor antagonist, rimonabant (SR141716). Neuropsychopharmacology 30:145–155CrossRefPubMedGoogle Scholar
  17. Collins AC, Romm E, Wehner JM (1990) Dissociation of the apparent relationship between nicotine tolerance and up-regulation of nicotinic receptors. Brain Res Bull 25:373–379CrossRefPubMedGoogle Scholar
  18. Cunningham ST, Kelley AE (1992) Opiate infusion into nucleus accumbens: contrasting effects on motor activity and responding for conditioned reward. Brain Res 588:104–114CrossRefPubMedGoogle Scholar
  19. Di Ciano P, Everitt BJ (2004) Contribution of the ventral tegmental area to cocaine-seeking maintained by a drug-paired conditioned stimulus in rats. Eur J Neurosci 19:1661–1667CrossRefPubMedGoogle Scholar
  20. Donny EC, Chaudhri N, Caggiula AR, Evans-Martin FF, Booth S, Gharib MA, Clements LA, Sved AF (2003) Operant responding for a visual reinforcer in rats is enhanced by noncontingent nicotine: implications for nicotine self-administration and reinforcement. Psychopharmacology (Berl) 169:68–76CrossRefGoogle Scholar
  21. Due DL, Huettel SA, Hall WG, Rubin DC (2002) Activation in mesolimbic and visuospatial neural circuits elicited by smoking cues: evidence from functional magnetic resonance imaging. Am J Psychiatry 159:954–960CrossRefPubMedGoogle Scholar
  22. Epping-Jordan MP, Watkins SS, Koob GF, Markou A (1998) Dramatic decreases in brain reward function during nicotine withdrawal. Nature 393:76–79CrossRefPubMedGoogle Scholar
  23. Epping-Jordan MP, Picciotto MR, Changeux JP, Pich EM (1999) Assessment of nicotinic acetylcholine receptor subunit contributions to nicotine self-administration in mutant mice. Psychopharmacology (Berl) 147:25–26CrossRefGoogle Scholar
  24. Fuchs RA, Evans KA, Parker MC, See RE (2004) Differential involvement of the core and shell subregions of the nucleus accumbens in conditioned cue-induced reinstatement of cocaine seeking in rats. Psychopharmacology (Berl) 176:459–465CrossRefGoogle Scholar
  25. Grabus SD, Martin BR, Batman AM, Tyndale RF, Sellers E, Damaj MI (2005) Nicotine physical dependence and tolerance in the mouse following chronic oral administration. Psychopharmacology (Berl) 178:183–192CrossRefGoogle Scholar
  26. Grady SR, Meinerz NM, Cao J, Reynolds AM, Picciotto MR, Changeux JP, McIntosh JM, Marks MJ, Collins AC (2001) Nicotinic agonists stimulate acetylcholine release from mouse interpeduncular nucleus: a function mediated by a different nAChR than dopamine release from striatum. J Neurochem 76:258–268CrossRefPubMedGoogle Scholar
  27. Harmer CJ, Phillips GD (1998) Enhanced appetitive conditioning following repeated pretreatment with d-amphetamine. Behav Pharmacol 9:299–308PubMedCrossRefGoogle Scholar
  28. Hatfield T, Han JS, Conley M, Gallagher M, Holland P (1996) Neurotoxic lesions of basolateral, but not central, amygdala interfere with Pavlovian second-order conditioning and reinforcer devaluation effects. J Neurosci 16:5256–5265PubMedGoogle Scholar
  29. Hitchcott PK, Harmer CJ, Phillips GD (1997) Enhanced acquisition of discriminative approach following intra-amygdala d-amphetamine. Psychopharmacology (Berl) 132:237–246CrossRefGoogle Scholar
  30. Ito R, Robbins TW, Everitt BJ (2004) Differential control over cocaine-seeking behavior by nucleus accumbens core and shell. Nat Neurosci 7:389–397CrossRefPubMedGoogle Scholar
  31. Jentsch JD, Taylor JR (1999) Impulsivity resulting from frontostriatal dysfunction in drug abuse: implications for the control of behavior by reward-related stimuli. Psychopharmacology (Berl) 146:373–390CrossRefGoogle Scholar
  32. King SL, Marks MJ, Grady SR, Caldarone BJ, Koren AO, Mukhin AG, Collins AC, Picciotto MR (2003) Conditional expression in corticothalamic efferents reveals a developmental role for nicotinic acetylcholine receptors in modulation of passive avoidance behavior. J Neurosci 23:3837–3843PubMedGoogle Scholar
  33. King SL, Caldarone BJ, Picciotto MR (2004) Beta2-subunit-containing nicotinic acetylcholine receptors are critical for dopamine-dependent locomotor activation following repeated nicotine administration. Neuropharmacology 47(Suppl 1):132–139CrossRefPubMedGoogle Scholar
  34. Lesage MG, Burroughs D, Dufek M, Keyler DE, Pentel PR (2004) Reinstatement of nicotine self-administration in rats by presentation of nicotine-paired stimuli, but not nicotine priming. Pharmacol Biochem Behav 79:507–513CrossRefPubMedGoogle Scholar
  35. Lindgren JL, Gallagher M, Holland PC (2003) Lesions of basolateral amygdala impair extinction of CS motivational value, but not of explicit conditioned responses, in Pavlovian appetitive second-order conditioning. Eur J Neurosci 17:160–166CrossRefPubMedGoogle Scholar
  36. Mackintosh N (1974) The psychology of animal learning. Academic, New YorkGoogle Scholar
  37. Marubio LM, Gardier AM, Durier S, David D, Klink R, Arroyo-Jimenez MM, McIntosh JM, Rossi F, Champtiaux N, Zoli M, Changeux JP (2003) Effects of nicotine in the dopaminergic system of mice lacking the alpha4 subunit of neuronal nicotinic acetylcholine receptors. Eur J Neurosci 17:1329–1337CrossRefPubMedGoogle Scholar
  38. Niaura R, Abrams D, Demuth B, Pinto R, Monti P (1989) Responses to smoking-related stimuli and early relapse to smoking. Addict Behav 14:419–428CrossRefPubMedGoogle Scholar
  39. Nooney JM, Peters JA, Lambert JJ (1992) A patch clamp study of the nicotinic acetylcholine receptor of bovine adrenomedullary chromaffin cells in culture. J Physiol 455:503–527PubMedGoogle Scholar
  40. Olausson P, Jentsch JD, Taylor JR (2003) Repeated nicotine exposure enhances reward-related learning in the rat. Neuropsychopharmacology 28:1264–1271CrossRefPubMedGoogle Scholar
  41. Olausson P, Jentsch JD, Taylor JR (2004a) Nicotine enhances responding with conditioned reinforcement. Psychopharmacology (Berl) 171:173–178CrossRefGoogle Scholar
  42. Olausson P, Jentsch JD, Taylor JR (2004b) Repeated nicotine exposure enhances responding with conditioned reinforcement. Psychopharmacology (Berl) 173:98–104CrossRefGoogle Scholar
  43. Parkinson JA, Olmstead MC, Burns LH, Robbins TW, Everitt BJ (1999) Dissociation in effects of lesions of the nucleus accumbens core and shell on appetitive pavlovian approach behavior and the potentiation of conditioned reinforcement and locomotor activity by d-amphetamine. J Neurosci 19:2401–2411PubMedGoogle Scholar
  44. Perkins KA, Gerlach D, Vender J, Grobe J, Meeker J, Hutchison S (2001) Sex differences in the subjective and reinforcing effects of visual and olfactory cigarette smoke stimuli. Nicotine Tob Res 3:141–150CrossRefPubMedGoogle Scholar
  45. Picciotto MR, Zoli M, Lena C, Bessis A, Lallemand Y, Le Novere N, Vincent P, Pich EM, Brulet P, Changeux JP (1995) Abnormal avoidance learning in mice lacking functional high-affinity nicotine receptor in the brain. Nature 374:65–67CrossRefPubMedGoogle Scholar
  46. Picciotto MR, Zoli M, Rimondini R, Lena C, Marubio LM, Pich EM, Fuxe K, Changeux JP (1998) Acetylcholine receptors containing the beta2 subunit are involved in the reinforcing properties of nicotine. Nature 391:173–177CrossRefPubMedGoogle Scholar
  47. Pratt WE, Kelley AE (2004) Nucleus accumbens acetylcholine regulates appetitive learning and motivation for food via activation of muscarinic receptors. Behav Neurosci 118:730–739PubMedCrossRefGoogle Scholar
  48. Rice ME, Cragg SJ (2004) Nicotine amplifies reward-related dopamine signals in striatum. Nat Neurosci 7:583–584CrossRefPubMedGoogle Scholar
  49. Robbins TW, Everitt BJ (2002) Limbic-striatal memory systems and drug addiction. Neurobiol Learn Mem 78:625–636CrossRefPubMedGoogle Scholar
  50. Robbins TW, Watson BA, Gaskin M, Ennis C (1983) Contrasting interactions of pipradrol, d-amphetamine, cocaine, cocaine analogues, apomorphine and other drugs with conditioned reinforcement. Psychopharmacology (Berl) 80:113–119CrossRefGoogle Scholar
  51. Robinson TE, Berridge KC (2003) Addiction. Annu Rev Psychol 54:25–53CrossRefPubMedGoogle Scholar
  52. Rose JE, Tashkin DP, Ertle A, Zinser MC, Lafer R (1985) Sensory blockade of smoking satisfaction. Pharmacol Biochem Behav 23:289–293CrossRefPubMedGoogle Scholar
  53. Schultz W (2002) Getting formal with dopamine and reward. Neuron 36:241–263CrossRefPubMedGoogle Scholar
  54. Sparks JA, Pauly JR (1999) Effects of continuous oral nicotine administration on brain nicotinic receptors and responsiveness to nicotine in C57Bl/6 mice. Psychopharmacology (Berl) 141:145–153CrossRefGoogle Scholar
  55. Stein EA, Pankiewicz J, Harsch HH, Cho JK, Fuller SA, Hoffmann RG, Hawkins M, Rao SM, Bandettini PA, Bloom AS (1998) Nicotine-induced limbic cortical activation in the human brain: a functional MRI study. Am J Psychiatry 155:1009–1015PubMedGoogle Scholar
  56. Tapper AR, McKinney SL, Nashmi R, Schwarz J, Deshpande P, Labarca C, Whiteaker P, Marks MJ, Collins AC, Lester HA (2004) Nicotine activation of alpha4* receptors: sufficient for reward, tolerance, and sensitization. Science 306:1029–1032CrossRefPubMedGoogle Scholar
  57. Taylor JR, Horger BA (1999) Enhanced responding for conditioned reward produced by intra-accumbens amphetamine is potentiated after cocaine sensitization. Psychopharmacology (Berl) 142:31–40CrossRefGoogle Scholar
  58. Taylor JR, Jentsch JD (2001) Repeated intermittent administration of psychomotor stimulant drugs alters the acquisition of Pavlovian approach behavior in rats: differential effects of cocaine, d-amphetamine and 3,4-methylenedioxymethamphetamine (“Ecstasy”). Biol Psychiatry 50:137–143CrossRefPubMedGoogle Scholar
  59. Taylor JR, Robbins TW (1984) Enhanced behavioural control by conditioned reinforcers following microinjections of d-amphetamine into the nucleus accumbens. Psychopharmacology (Berl) 84:405–412CrossRefGoogle Scholar
  60. Tiffany ST, Carter BL (1998) Is craving the source of compulsive drug use? J Psychopharmacol 12:23–30PubMedCrossRefGoogle Scholar
  61. Waters AJ, Shiffman S, Bradley BP, Mogg K (2003) Attentional shifts to smoking cues in smokers. Addiction 98:1409–1417CrossRefPubMedGoogle Scholar
  62. Waters AJ, Shiffman S, Sayette MA, Paty JA, Gwaltney CJ, Balabanis MH (2004) Cue-provoked craving and nicotine replacement therapy in smoking cessation. J Consult Clin Psychol 72:1136–1143CrossRefPubMedGoogle Scholar
  63. Wustenberg DG, Grunewald B (2004) Pharmacology of the neuronal nicotinic acetylcholine receptor of cultured Kenyon cells of the honeybee, Apis mellifera. J Comp Physiol A Neuroethol Sens Neural Behav Physiol 190:807–821CrossRefPubMedGoogle Scholar
  64. Yin X, Cui W, Hu G, Wang H (2004) Desensitization of alpha7 nicotinic receptors potentiated the inhibitory effect on M-current induced by stimulation of muscarinic receptors in rat superior cervical ganglion neurons. J Neural Transm (in press). DOI 10.1007/s0070200402606Google Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Darlene H. Brunzell
    • 1
  • Jessica R. Chang
    • 1
  • Brandon Schneider
    • 1
  • Peter Olausson
    • 1
  • Jane R. Taylor
    • 1
  • Marina R. Picciotto
    • 1
  1. 1.Department of Psychiatry, Division of Molecular PsychiatryYale University School of MedicineNew HavenUSA

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