Naunyn-Schmiedeberg's Archives of Pharmacology

, Volume 368, Issue 5, pp 331–341 | Cite as

Different pattern of brain c-Fos expression following re-exposure to ethanol or sucrose self-administration environment

  • Krzysztof Wedzony
  • Eliza Koros
  • Anna Czyrak
  • Agnieszka Chocyk
  • Klaudia Czepiel
  • Katarzyna Fijal
  • Marzena Mackowiak
  • Artur Rogowski
  • Wojciech Kostowski
  • Przemyslaw BienkowskiEmail author
Original Article


Exposure of alcohol addicts to alcohol-related environmental cues may elicit alcohol-seeking behavior and lead to relapse to heavy drinking. The aim of the present study was to identify brain regions activated by alcohol (ethanol)-related stimuli in Wistar rats trained to lever press for 8% ethanol solution in operant self-administration cages. Ethanol self-administration was stabilized in a maintenance phase, which lasted for 30 days. c-Fos protein expression was used as a marker of neuronal activation.

Re-exposure to ethanol self-administration environment after 30-day but not after 24-h abstinence increased the number of Fos-positive nuclei in the thalamic paraventricular nucleus, granular insular cortex and medial prefrontal cortex. In general, no differences were found in c-Fos protein expression between the rats allowed to self-administer alcohol and the subjects exposed only to alcohol-related stimuli. In contrast, no increase in c-Fos immunoreactivity was observed in rats trained to lever press for sucrose solution and exposed to sucrose-related environmental stimuli after 30-day abstinence.

Taken together, these results suggest that at least some thalamo-cortical circuits become more responsive to ethanol-paired stimuli after prolonged abstinence and that ethanol- and sucrose-seeking behavior may be regulated by partially different neural mechanism(s).


Ethanol self-administration Extinction Reinstatement c-Fos Inducible transcription factors Brain mapping 



The study was supported by the State Committee for Scientific Research (grants no. 4P05A 08 317 and PBZ-KBN-033/P05/2002).


  1. Abrams DB, Monti PM, Carey KB, Pinto RP, Jacobus SI (1988) Reactivity to smoking cues and relapse: two studies of discriminant validity. Behav Res Ther 26:225–233PubMedGoogle Scholar
  2. Ahmed SH, Koob G (1997) Cocaine- but not food-seeking behavior is reinstated by stress after extinction. Psychopharmacology 132:289–295PubMedGoogle Scholar
  3. Augustine JR (1996) Circuitry and functional aspects of the insular lobe in primates including humans. Behav Brain Res 22:229–244CrossRefGoogle Scholar
  4. Bachtell RK, Wang Y-M, Freeman P, Risinger F, Ryabinin AE (1999) Alcohol drinking produces brain region-selective changes in expression of inducible transcription factors. Brain Res 847:157–165CrossRefPubMedGoogle Scholar
  5. Bienkowski P, Kostowski W, Koros E (1999a) The role of drug-paired stimuli in extinction and reinstatement of ethanol-seeking behavior in the rat. Eur J Pharmacol 374:315–319CrossRefPubMedGoogle Scholar
  6. Bienkowski P, Kostowski W, Koros E (1999b) Ethanol-reinforced behavior in the rat: effects of naltrexone. Eur J Pharmacol 374:321–327PubMedGoogle Scholar
  7. Bienkowski P, Koros E, Kostowski W, Bogucka-Bonikowska A (2000) Reinstatement of ethanol seeking in rats: behavioral analysis. Pharmacol Biochem Behav 66:123–128CrossRefPubMedGoogle Scholar
  8. Brown EE, Robertson GS, Fibiger HC (1992) Evidence for conditional neuronal activation following exposure to a cocaine-paired environment: role of forebrain limbic structures. J Neurosci 12:4112–4121PubMedGoogle Scholar
  9. Buczek Y, Le AD, Wang A, Stewart J, Shaham Y (1999) Stress reinstates nicotine seeking but not sucrose solution seeking in rats. Psychopharmacology 144:183–188PubMedGoogle Scholar
  10. Chang SL, Patel NA, Romero AA (1995) Activation and desensitization of Fos immunoreactivity in the rat brain following ethanol administration. Brain Res 679:89–98CrossRefPubMedGoogle Scholar
  11. Chaudhuri A (1997) Neural activity mapping with inducible transcription factors. Neuroreport 8:iii–viiGoogle Scholar
  12. Childress AR, Mozley PD, McElgin W, Fitzgerald J, Reivich M, O’Brien C (1999) Limbic activation during cue-induced cocaine-craving. Am J Psychiatry 156:11–18PubMedGoogle Scholar
  13. Ciccocioppo R, Angeletti S, Weiss F (2001a) Long-lasting resistance to extinction of response reinstatement induced by ethanol-related stimuli: role of genetic ethanol preference. Alcohol Clin Exp Res 25:1414–1419Google Scholar
  14. Ciccocioppo R, Sanna PP, Weiss F (2001b) Cocaine-predictive stimulus induces drug-seeking behavior and neural activation in limbic brain regions after multiple months of abstinence: reversal by D1 antagonists. Proc Nat Acad Sci USA 98:1976–1981CrossRefPubMedGoogle Scholar
  15. Di Chiara G (1999) Drug addiction as dopamine-dependent associative learning disorder. Eur J Pharmacol 375:13–30PubMedGoogle Scholar
  16. Divac I, Mogensen J, Petrovic-Minic B, Zilles K, Regidor J (1993) Cortical projections of the thalamic mediodorsal nucleus in the rat. Definition of the prefrontal cortex. Acta Neurobiol Exp 53:425–429Google Scholar
  17. Drummond DC (2000) What does cue-reactivity have to offer clinical research? Addiction 95:S129–S144PubMedGoogle Scholar
  18. Drummond DC, Glautier S (1994) A controlled trial of cue exposure treatment in alcohol dependence. J Consult Clin Psychol 62:809–817CrossRefPubMedGoogle Scholar
  19. Drummond DC, Cooper T, Glautier S (1990) Conditioned learning in alcohol dependence: implications for cue exposure treatment. Br J Addict 85:725–743PubMedGoogle Scholar
  20. Erdtmann-Vourliotis M, Mayer P, Riechert U, Hollt V (1999) Acute injection of drugs with low addictive potential (delta(9)-tetrahydrocannabinol, 3,4-methylenedioxymethamphetamine, lysergic acid diamide) causes a much higher c-fos expression in limbic brain areas than highly addicting drugs (cocaine and morphine). Mol Brain Res 71:313–324CrossRefPubMedGoogle Scholar
  21. Garavan H, Pankiewicz J, Bloom A, Cho J-K, Sperry L, Ross TJ, Salmeron BJ, Risinger R, Kelley D, Stein EA (2000) Cue-induced cocaine craving: neuroanatomical specificity for drug users and drug stimuli. Am J Psychiatry 157:1789–1798PubMedGoogle Scholar
  22. Gawin FH, Kleber HD (1986) Abstinence symptomatology and psychiatric diagnosis in cocaine abusers. Clinical observations. Arch Gen Psychiatry 43:103–113Google Scholar
  23. George MS, Anton RF, Bloomer C, Teneback C, Drobes DJ, Lorberbaum JP, Nahas Z, Vincent DJ (2001) Activation of prefrontal cortex and anterior thalamus in alcoholic subjects on exposure to alcohol-specific cues. Arch Gen Psychiatry 58:345–352PubMedGoogle Scholar
  24. Grant S, London E, Newlin DB, Villemagne DL, Liu X, Contoreggi C, Phillips RL, Kimes AL, Margolin A (1996) Activation of memory circuits during cue-elicited cocaine craving. Proc Natl Acad Sci USA 93:12040–12045PubMedGoogle Scholar
  25. Grimm JW, Shaham Y, Hope BT (2002) Effect of cocaine and sucrose withdrawal period on extinction behavior, cue-induced reinstatement, and protein levels of the dopamine transporter and tyrosine hydroxylase in limbic and cortical areas in rats. Behav Pharmacol 13:379–388PubMedGoogle Scholar
  26. Hitzemann B, Hitzemann R (1997) Genetics ethanol and the Fos response: a comparison of the C57BL/6 J and DBA/2 J inbred mouse strains. Alcohol Clin Exp Res 21:1497–1507Google Scholar
  27. Hotsenpiller G, Horak BT, Wolf ME (2002) Dissociation of conditioned locomotion and Fos induction in response to stimuli formerly paired with cocaine. Behav Neurosci 116:634–645CrossRefPubMedGoogle Scholar
  28. Kaczmarek L (2000) Gene expression in learning processes. Acta Neurobiol Exp 60:419–424Google Scholar
  29. Kanaka TS, Balasubramaniam V (1978) Stereotactic cingulumotomy for drug addiction. Appl Neurophysiol 41:86–92PubMedGoogle Scholar
  30. Katner SN, Magalong JG, Weiss F (1999) Reinstatement of alcohol-seeking behavior by drug-associated discriminative stimuli after prolonged extinction in the rat. Neuropsychopharmacology 20:471–479Google Scholar
  31. Kilts CD, Schweitzer JB, Quinn CK, Gross RE, Faber TL, Muhammad F, Ely TD, Hoffman JM, Drexler KP (2001) Neural activity related to drug craving in cocaine addiction. Arch Gen Psychiatry 58:334–341PubMedGoogle Scholar
  32. Liste L, Guerra MJ, Caruncho H, Labandeira-Garcia JL (1997) Treadmill running induces striatal Fos expression via NMDA glutamate and dopamine receptors. Exp Brain Res 115:458–468PubMedGoogle Scholar
  33. Maas LC, Lukas SE, Kaufman MJ, Weiss RD, Daniels SL, Rogers VW, Kukes TJ, Renshaw PF (1998) Functional magnetic resonance imaging of human brain activation during cue-induced cocaine craving. Am J Psychiatry 155:124–126PubMedGoogle Scholar
  34. Mackowiak M, Chocyk A, Fijal K, Czyrak A, Wedzony K (1999) c-Fos proteins, induced by the serotonin receptor agonist DOI, are not expressed in 5-HT2A positive cortical neurons. Mol Brain Res 71:358–363CrossRefPubMedGoogle Scholar
  35. McGregor IS, Saharov T, Hunt GE, Topple AN (1999) Beer consumption in rats: the influence of ethanol content, food deprivation, and cocaine. Alcohol 17:47–56CrossRefPubMedGoogle Scholar
  36. Monti PM, Rohsenow DJ, Hutchison KE, Swift RM, Mueller TI, Colby SM, Brown RA, Gulliver SB, Gordon A, Abrams DB (1999) Naltrexone’s effect on cue-elicited craving among alcoholics in treatment. Alcohol Clin Exp Res 23:1386–1394PubMedGoogle Scholar
  37. Neisewander JL, Baker DA, Fuchs RA, Tran-Nguyen LTL, Palmer A, Marshall J (2000) Fos protein expression and cocaine-seeking behavior in rats after exposure to a cocaine self-administration environment. J Neurosci 20:798–805PubMedGoogle Scholar
  38. Nisell M, Nomikos GG, Chergui K, Grillner P, Svensson TH (1997) Chronic nicotine enhances basal and nicotine-induced Fos immunoreactivity preferentially in the medial prefrontal cortex. Neuropsychopharmacology 17:151–161CrossRefPubMedGoogle Scholar
  39. Paxinos G, Watson C (1986) The rat brain in stereotaxic coordinates. Academic Press, San DiegoGoogle Scholar
  40. Piasecki J, Koros E, Dyr W, Kostowski W, Danysz W, Bienkowski P (1998) Ethanol-reinforced behavior in the rat: effects of uncompetitive NMDA receptor antagonist, memantine. Eur J Pharmacol 354:135–143CrossRefPubMedGoogle Scholar
  41. Rodriguez de Fonseca F, Navarro M (1998) Role of the limbic system in dependence on drugs. Ann Med 30:397–405PubMedGoogle Scholar
  42. Rohsenow DJ, Monti PM, Abrams DB, Rubonis AV, Niaura VS, Colby SM, Wunshel SM, Abrams DB (1994) Cue-reactivity as a predictor of drinking among male alcoholics. J Consult Clin Psychol 62:620–626CrossRefPubMedGoogle Scholar
  43. Rolls ET (1997) Taste and olfactory processing in the brain and its relation to the control of eating. Crit Rev Neurobiol 11:263–287PubMedGoogle Scholar
  44. Rolls ET (2000) The orbitofrontal cortex and reward. Cereb Cortex 10:284–294CrossRefPubMedGoogle Scholar
  45. Rosenkranz JA, Grace AA (2002) Cellular mechanisms of infralimbic and prelimbic prefrontal cortical inhibition and dopaminergic modulation of basolateral amygdala neurons in vivo. J Neurosci 22:324–337PubMedGoogle Scholar
  46. Ryabinin AE, Bachtell RK, Freeman P, Risinger FO (2001) ITF expression in mouse brain during acquisition of alcohol self-administration. Brain Res 890:192–195CrossRefPubMedGoogle Scholar
  47. Samson HH (1986) Initiation of ethanol reinforcement using a sucrose-substitution procedure in food- and water-sated rats. Alcohol Clin Exp Res 10:436–442PubMedGoogle Scholar
  48. Schneider F, Habel U, Wagner M, Franke P, Salloum M, Shah NJ, Toni L, Sulzbach C, Honig K, Maier W, Gaebel W, Zilles K (2001) Subcortical correlates of craving in recently abstinent alcoholic patients. Am J Psychiatry 158:1075–1083CrossRefPubMedGoogle Scholar
  49. Schroeder BE, Holahan MR, Landry CF, Kelley AE (2000) Morphine-associated environmental cues elicit conditioned gene expression. Synapse 37:146–158PubMedGoogle Scholar
  50. Schroeder BE, Binzak MJ, Kelley AE (2001) A common profile of prefrontal cortical activation following exposure to nicotine- or chocolate-associated contextual cues. Neuroscience 105:535–545CrossRefPubMedGoogle Scholar
  51. Shaham Y, Erb S, Stewart J (2000) Stress-induced relapse to heroin and cocaine seeking in rats: a review. Brain Res Rev 33:13–33PubMedGoogle Scholar
  52. Shalev U, Morales M, Hope BT, Yap J, Shaham Y (2001) Time-dependent changes in extinction behavior and stress-induced reinstatement of drug-seeking following withdrawal from heroin in rats. Psychopharmacology 156:98–107Google Scholar
  53. Sinclair JD (2001) Evidence about the use of naltrexone and for different ways of using it in the treatment of alcoholism. Alcohol Alcohol 36:2–10CrossRefPubMedGoogle Scholar
  54. Su HS, Bentivoglio M (1990) Thalamic midline cell populations projecting to the nucleus accumbens, amygdala, and hippocampus in the rat. J Comp Neurol 297:582–593PubMedGoogle Scholar
  55. Thiele TE, van Dijk G, Bernstein IL (1997) Ethanol-induced c-Fos expression in rat lines selected for low and high alcohol consumption. Brain Res 756:278–282CrossRefPubMedGoogle Scholar
  56. Topple AN, Hunt GE, McGregor IS (1998) Possible neural substrates of beer-craving in rats. Neurosci Lett 252:99–102CrossRefPubMedGoogle Scholar
  57. Tran-Nguyen LTL, Fuchs RA, Coffey GP, Baker DA, O’Dell LA, Neisewander JL (1998) Time-dependent changes in cocaine-seeking behavior and extracellular dopamine levels in the amygdala during cocaine withdrawal. Neuropsychopharmacology 19:48–59CrossRefPubMedGoogle Scholar
  58. Tran-Nguyen LTL, Bellew JG, Grote KA, Neisewander JL (2001) Serotonin depletion attenuates cocaine seeking but enhances sucrose seeking and the effects of cocaine priming on reinstatement of cocaine seeking in rats. Psychopharmacology 157:340–348Google Scholar
  59. Vertes RP (2002) Analysis of projections from the medial prefrontal cortex to the thalamus in the rat, with emphasis on nucleus reuniens. J Comp Neurol 442:163–187Google Scholar
  60. Volkow ND, Fowler JS (2000) Addiction, a disease of compulsion and drive: involvement of the orbitofrontal cortex. Cereb Cortex 10:318–325PubMedGoogle Scholar
  61. Wang G-J, Volkow ND, Fowler JS, Cervany P, Hitzemann RJ, Pappas NR, Wong CT, Felder Ch (1999) Regional brain metabolic activation during craving elicited by recall of previous drug experiences. Life Sci 64:775–784PubMedGoogle Scholar
  62. Weitemier AZ, Woerner A, Backstrom P, Hyytia P, Ryabinin AE (2001) Expression of c-Fos in Alko Alcohol rats responding for ethanol in an operant paradigm. Alcohol Clin Exp Res 25:704–710Google Scholar
  63. Wexler BE, Gottschalk CH, Fulbright RK, Prohovnik I, Lacadie CM, Rounsaville BJ, Gore JC (2001) Functional magnetic resonance imaging of cocaine craving. Am J Psychiatry 158:86–95PubMedGoogle Scholar
  64. Wu JC, Bell K, Najafi A, Widmark C, Keator D, Tang Ch, Klein E, Bunney BG, Fallon J, Bunney WE (1997) Decreasing striatal 6-FDOPA uptake with increasing duration of cocaine withdrawal. Neuropsychopharmacology 17:402–409CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • Krzysztof Wedzony
    • 1
  • Eliza Koros
    • 2
  • Anna Czyrak
    • 1
  • Agnieszka Chocyk
    • 1
  • Klaudia Czepiel
    • 1
  • Katarzyna Fijal
    • 1
  • Marzena Mackowiak
    • 1
  • Artur Rogowski
    • 2
  • Wojciech Kostowski
    • 2
    • 3
  • Przemyslaw Bienkowski
    • 2
    Email author
  1. 1.Department of Pharmacology, Institute of PharmacologyPolish Academy of SciencesKrakowPoland
  2. 2.Department of PharmacologyInstitute of Psychiatry and NeurologyWarsawPoland
  3. 3.Department of Experimental and Clinical PharmacologyWarsaw Medical AcademyWarsawPoland

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