Long-term exposure to daily ethanol injections in DBA/2J and Swiss mice: Lessons for the interpretation of ethanol sensitization

Autoři: Vincent Didone aff001;  Théo van Ingelgom aff001;  Ezio Tirelli aff001;  Etienne Quertemont aff001
Působiště autorů: Psychology and Neuroscience of Cognition Research Unit (PsyNCog), , University of Liège, Liège, Belgium aff001
Vyšlo v časopise: PLoS ONE 14(11)
Kategorie: Research Article
doi: 10.1371/journal.pone.0214696


Most mice ethanol sensitization studies focused on neurobiology at the expense of its behavioral characterization. Furthermore, relatively short ethanol exposures (10 to 20 injections) were used in these studies. The first aim of the present study is to better characterize the development and expression of ethanol sensitization after an extended exposure of 45 daily injections. In some previous studies, mice were classified as “respondent” and “resistant” to ethanol sensitization. The second aim of the present study is to test the long-term reliability of such categorizations and the consequences of their use on the interpretation of the ethanol sensitization results. Swiss and DBA/2J female mice received 45 consecutive daily ethanol administrations (respectively 2.5 and 2.0 g/kg) and their locomotor activity was daily recorded to test the development of ethanol sensitization. At the end of the procedure, a challenge test assessed the inter-group ethanol sensitization.The results of the present study show that ethanol sensitization continues to develop beyond 20 days to reach maximal levels after about 25 injections in DBA/2J mice and 40 injections in Swiss mice, although the core phase of the development of ethanol sensitization occurred in both strains during the first 20 days. Remarkably, ethanol sensitization after such a long daily ethanol treatment resulted in both an upward shift of the magnitude of ethanol stimulant effects and a prolongation of these effects in time (up to 30 minutes). Mice classified as “resistant to ethanol sensitization” according to previous studies developed very significant levels of ethanol sensitization when tested after 45 ethanol injections and are best described as showing a delayed development of ethanol sensitization. Furthermore, mice classified as respondent or resistant to ethanol sensitization also differ in their acute response to ethanol, such that it is difficult to ascertain whether these classifications are specifically related to the sensitization process.

Klíčová slova:

Analysis of variance – Animal behavior – Behavioral addiction – Biological locomotion – Conditioned response – Data acquisition – Ethanol – Mice


1. Kalivas PW, Stewart J. Dopamine transmission in the initiation and expression of drug- and stress-induced sensitization of motor activity. Brain Res Brain Res Rev. 1991;16: 223–244. doi: 10.1016/0165-0173(91)90007-u 1665095

2. Robinson TE, Berridge KC. The neural basis of drug craving: an incentive-sensitization theory of addiction. Brain Res Brain Res Rev. 1993;18: 247–291. doi: 10.1016/0165-0173(93)90013-p 8401595

3. Quoilin C, Didone V, Tirelli E, Quertemont E. Chronic ethanol exposure during adolescence alters the behavioral responsiveness to ethanol in adult mice. Behav Brain Res. 2012;229: 1–9. doi: 10.1016/j.bbr.2011.12.039 22227505

4. Didone V, Quoilin C, Tirelli E, Quertemont E. Parametric analysis of the development and expression of ethanol-induced behavioral sensitization in female Swiss mice: effects of dose, injection schedule, and test context. Psychopharmacology (Berl). 2008;201: 249–260. doi: 10.1007/s00213-008-1266-9 18685830

5. Hunt WA, Lands WE. A role for behavioral sensitization in uncontrolled ethanol intake. Alcohol Fayettev N. 1992;9: 327–328.

6. Vanderschuren LJMJ, Pierce RC. Sensitization processes in drug addiction. Curr Top Behav Neurosci. 2010;3: 179–195. doi: 10.1007/7854_2009_21 21161753

7. Newlin DB, Thomson JB. Chronic tolerance and sensitization to alcohol in sons of alcoholics: II. Replication and reanalysis. Exp Clin Psychopharmacol. 1999;7: 234–243. doi: 10.1037//1064-1297.7.3.234 10472511

8. Newlin DB, Thomson JB. Alcohol challenge with sons of alcoholics: a critical review and analysis. Psychol Bull. 1990;108: 383–402. doi: 10.1037/0033-2909.108.3.383 2270234

9. Didone V, Quoilin C, Dieupart J, Tirelli E, Quertemont E. Differential effects of context on psychomotor sensitization to ethanol and cocaine. Behav Pharmacol. 2016;27: 173–181. doi: 10.1097/FBP.0000000000000161 26164714

10. Botia B, Legastelois R, Alaux-Cantin S, Naassila M. Expression of ethanol-induced behavioral sensitization is associated with alteration of chromatin remodeling in mice. PloS One. 2012;7: e47527. doi: 10.1371/journal.pone.0047527 23110077

11. Camarini R, Pautassi RM, Méndez M, Quadros IM, Souza-Formigoni ML, Boerngen-Lacerda R. Behavioral and neurochemical studies in distinct animal models of ethanol’s motivational effects. Curr Drug Abuse Rev. 2010;3: 205–221. 21208169

12. de Araujo NP, Fukushiro DF, Grassl C, Hipólide DC, Souza-Formigoni MLO, Tufik S, et al. Ethanol-induced behavioral sensitization is associated with dopamine receptor changes in the mouse olfactory tubercle. Physiol Behav. 2009;96: 12–17. doi: 10.1016/j.physbeh.2008.07.029 18761028

13. Legastelois R, Botia B, Naassila M. Blockade of ethanol-induced behavioral sensitization by sodium butyrate: descriptive analysis of gene regulations in the striatum. Alcohol Clin Exp Res. 2013;37: 1143–1153. doi: 10.1111/acer.12088 23488934

14. Quadros IMH, Hipólide DC, Frussa-Filho R, De Lucca EM, Nobrega JN, Souza-Formigoni MLO. Resistance to ethanol sensitization is associated with increased NMDA receptor binding in specific brain areas. Eur J Pharmacol. 2002;442: 55–61. doi: 10.1016/s0014-2999(02)01503-0 12020682

15. Quadros IMH, Nobrega JN, Hipólide DC, de Lucca EM, Souza-Formigoni MLO. Differential propensity to ethanol sensitization is not associated with altered binding to D1 receptors or dopamine transporters in mouse brain. Addict Biol. 2002;7: 291–299. doi: 10.1080/13556210220139505 12126488

16. Quadros IMH, Nobrega JN, Hipolide DC, Souza-Formigoni MLO. Increased brain dopamine D4-like binding after chronic ethanol is not associated with behavioral sensitization in mice. Alcohol Fayettev N. 2005;37: 99–104. doi: 10.1016/j.alcohol.2005.12.001 16584973

17. Quadros IMH, Souza-Formigoni MLO, Fornari RV, Nobrega JN, Oliveira MGM. Is behavioral sensitization to ethanol associated with contextual conditioning in mice? Behav Pharmacol. 2003;14: 129–136. doi: 10.1097/01.fbp.0000063262.43827.57 12658073

18. Quoilin C, Didone V, Tirelli E, Quertemont E. Developmental differences in ethanol-induced sensitization using postweanling, adolescent, and adult Swiss mice. Psychopharmacology (Berl). 2012;219: 1165–1177. doi: 10.1007/s00213-011-2453-7 21881875

19. Coune F, Silvestre de Ferron B, González-Marín MC, Antol J, Naassila M, Pierrefiche O. Resistance to ethanol sensitization is associated with a loss of synaptic plasticity in the hippocampus. Synap N Y N. 2017;71. doi: 10.1002/syn.21899 26898905

20. Legastelois R, Botia B, Coune F, Jeanblanc J, Naassila M. Deciphering the relationship between vulnerability to ethanol-induced behavioral sensitization and ethanol consumption in outbred mice. Addict Biol. 2014;19: 210–224. doi: 10.1111/adb.12104 24164956

21. Botia B, Legastelois R, Houchi H, Naassila M. Basal anxiety negatively correlates with vulnerability to ethanol-induced behavioral sensitization in DBA/2J mice: modulation by diazepam. Alcohol Clin Exp Res. 2015;39: 45–54. doi: 10.1111/acer.12595 25623405

22. Masur J, Boerngen R. The excitatory component of ethanol in mice: a chronic study. Pharmacol Biochem Behav. 1980;13: 777–780. doi: 10.1016/0091-3057(80)90206-3 7208545

23. Masur J, Oliveira de Souza ML, Zwicker AP. The excitatory effect of ethanol: absence in rats, no tolerance and increased sensitivity in mice. Pharmacol Biochem Behav. 1986;24: 1225–1228. doi: 10.1016/0091-3057(86)90175-9 3725828

24. Brabant C, Quertemont E, Tirelli E. Influence of the dose and the number of drug-context pairings on the magnitude and the long-lasting retention of cocaine-induced conditioned place preference in C57BL/6J mice. Psychopharmacology (Berl). 2005;180: 33–40. doi: 10.1007/s00213-004-2138-6 15682299

25. Brabant C, Tambour S, Quertemont E, Ferrara A, Tirelli E. Do excitatory and inhibitory conditioning processes underlie psychomotor sensitization to amphetamine? An analysis using simple and multiple regressions. Behav Brain Res. 2011;221: 227–236. doi: 10.1016/j.bbr.2011.02.040 21377496

26. Meyer PJ, Phillips TJ. Bivalent effects of MK-801 on ethanol-induced sensitization do not parallel its effects on ethanol-induced tolerance. Behav Neurosci. 2003;117: 641–649. doi: 10.1037/0735-7044.117.3.641 12802892

27. Abrahao KP, Quadros IMH, Souza-Formigoni MLO. Individual differences to repeated ethanol administration may predict locomotor response to other drugs, and vice versa. Behav Brain Res. 2009;197: 404–410. doi: 10.1016/j.bbr.2008.10.009 18984011

28. Abrahao KP, Ariwodola OJ, Butler TR, Rau AR, Skelly MJ, Carter E, et al. Locomotor sensitization to ethanol impairs NMDA receptor-dependent synaptic plasticity in the nucleus accumbens and increases ethanol self-administration. J Neurosci Off J Soc Neurosci. 2013;33: 4834–4842. doi: 10.1523/JNEUROSCI.5839-11.2013 23486954

29. Abrahao KP, Quadros IMH, Andrade ALM, Souza-Formigoni MLO. Accumbal dopamine D2 receptor function is associated with individual variability in ethanol behavioral sensitization. Neuropharmacology. 2012;62: 882–889. doi: 10.1016/j.neuropharm.2011.09.017 21964435

30. Ferreira SE, Abrahao KP, Souza-Formigoni MLO. Expression of behavioral sensitization to ethanol is increased by energy drink administration. Pharmacol Biochem Behav. 2013;110: 245–248. doi: 10.1016/j.pbb.2013.07.014 23911904

31. Nona CN, Guirguis S, Nobrega JN. Susceptibility to ethanol sensitization is differentially associated with changes in pCREB, trkB and BDNF mRNA expression in the mouse brain. Behav Brain Res. 2013;242: 25–33. doi: 10.1016/j.bbr.2012.12.035 23291223

32. Nona CN, Li R, Nobrega JN. Altered NMDA receptor subunit gene expression in brains of mice showing high vs. low sensitization to ethanol. Behav Brain Res. 2014;260: 58–66. doi: 10.1016/j.bbr.2013.11.037 24315834

33. Nona CN, Bermejo MK, Ramsey AJ, Nobrega JN. Changes in dendritic spine density in the nucleus accumbens do not underlie ethanol sensitization. Synap N Y N. 2015;69: 607–610. doi: 10.1002/syn.21862 26340045

34. Nona CN, Lam M, Nobrega JN. Localized brain differences in Arc expression between mice showing low vs. high propensity to ethanol sensitization. Pharmacol Biochem Behav. 2016;142: 15–22. doi: 10.1016/j.pbb.2015.12.006 26708208

35. Nona CN, Nobrega JN. A role for nucleus accumbens glutamate in the expression but not the induction of behavioural sensitization to ethanol. Behav Brain Res. 2018;336: 269–281. doi: 10.1016/j.bbr.2017.09.024 28919158

36. Abrahao KP, Souza-Formigoni MLO. Behavioral sensitization to ethanol results in cross-sensitization to MK-801 but not to NMDA administered intra-accumbens. Behav Brain Res. 2012;235: 218–224. doi: 10.1016/j.bbr.2012.07.034 22871257

37. Abrahao KP, Quadros IM, Souza-Formigoni MLO. Morphine attenuates the expression of sensitization to ethanol, but opioid antagonists do not. Neuroscience. 2008;156: 857–864. doi: 10.1016/j.neuroscience.2008.08.012 18804151

38. Abrahao KP, Goeldner FO, Souza-Formigoni MLO. Individual differences in ethanol locomotor sensitization are associated with dopamine D1 receptor intra-cellular signaling of DARPP-32 in the nucleus accumbens. PloS One. 2014;9: e98296. doi: 10.1371/journal.pone.0098296 24919054

39. Pildervasser JVN, Abrahao KP, Souza-Formigoni MLO. Distinct behavioral phenotypes in ethanol-induced place preference are associated with different extinction and reinstatement but not behavioral sensitization responses. Front Behav Neurosci. 2014;8: 267. doi: 10.3389/fnbeh.2014.00267 25152719

40. Souza-Formigoni ML, De Lucca EM, Hipólide DC, Enns SC, Oliveira MG, Nobrega JN. Sensitization to ethanol’s stimulant effect is associated with region-specific increases in brain D2 receptor binding. Psychopharmacology (Berl). 1999;146: 262–267.

41. MacCallum RC, Zhang S, Preacher KJ, Rucker DD. On the practice of dichotomization of quantitative variables. Psychol Methods. 2002;7: 19–40. doi: 10.1037/1082-989x.7.1.19 11928888

42. Royston P, Altman DG, Sauerbrei W. Dichotomizing continuous predictors in multiple regression: a bad idea. Stat Med. 2006;25: 127–141. doi: 10.1002/sim.2331 16217841

43. Altman DG, Royston P. The cost of dichotomising continuous variables. BMJ. 2006;332: 1080. doi: 10.1136/bmj.332.7549.1080 16675816

44. Irwin JR, McClelland GH. Negative Consequences of Dichotomizing Continuous Predictor Variables. J Mark Res. 2003;40: 366–371. doi: 10.1509/jmkr.40.3.366.19237

45. Sershen H, Hashim A, Vadasz C. Strain and sex differences in repeated ethanol treatment-induced motor activity in quasi-congenic mice. Genes Brain Behav. 2002;1: 156–165. doi: 10.1034/j.1601-183X.2002.10303.x 12884971

46. Camarini R, Pautassi RM. Behavioral sensitization to ethanol: Neural basis and factors that influence its acquisition and expression. Brain Res Bull. 2016;125: 53–78. doi: 10.1016/j.brainresbull.2016.04.006 27093941

47. Post RM, Weiss SR, Pert A, Fontana D. Conditioned components of cocaine sensitization. Clin Neuropharmacol. 1992;15 Suppl 1 Pt A: 650A–651A.

48. Ceyhan M, Kayir H, Uzbay IT. Investigation of the effects of tianeptine and fluoxetine on pentylenetetrazole-induced seizures in rats. J Psychiatr Res. 2005;39: 191–196. doi: 10.1016/j.jpsychires.2004.06.002 15589568

49. Procópio-Souza R, Fukushiro DF, Trombin TF, Wuo-Silva R, Zanlorenci LHF, Lima AJO, et al. Effects of group exposure on single injection-induced behavioral sensitization to drugs of abuse in mice. Drug Alcohol Depend. 2011;118: 349–359. doi: 10.1016/j.drugalcdep.2011.04.017 21596493

50. Durcan MJ, Lister RG. Time course of ethanol’s effects on locomotor activity, exploration and anxiety in mice. Psychopharmacology (Berl). 1988;96: 67–72.

51. Smoothy R, Berry MS. Time course of the locomotor stimulant and depressant effects of a single low dose of ethanol in mice. Psychopharmacology (Berl). 1985;85: 57–61.

52. Lewis MJ, June HL. Neurobehavioral studies of ethanol reward and activation. Alcohol Fayettev N. 1990;7: 213–219.

53. Araujo NP, Fukushiro DF, Cunha JLS, Levin R, Chinen CC, Carvalho RC, et al. Drug-induced home cage conspecifics’ behavior can potentiate behavioral sensitization in mice. Pharmacol Biochem Behav. 2006;84: 142–147. doi: 10.1016/j.pbb.2006.04.019 16753204

54. Faria RR, Lima Rueda AV, Sayuri C, Soares SL, Malta MB, Carrara-Nascimento PF, et al. Environmental modulation of ethanol-induced locomotor activity: Correlation with neuronal activity in distinct brain regions of adolescent and adult Swiss mice. Brain Res. 2008;1239: 127–140. doi: 10.1016/j.brainres.2008.08.056 18789904

55. Itzhak Y, Anderson KL. Ethanol-induced behavioral sensitization in adolescent and adult mice: role of the nNOS gene. Alcohol Clin Exp Res. 2008;32: 1839–1848. doi: 10.1111/j.1530-0277.2008.00766.x 18652592

56. Itzhak Y, Martin JL. Blockade of alcohol-induced locomotor sensitization and conditioned place preference in DBA mice by 7-nitroindazole. Brain Res. 2000;858: 402–407. doi: 10.1016/s0006-8993(00)01940-5 10708693

Článek vyšel v časopise


2019 Číslo 11