Advertisement

Genetics of Ethanol-Related Behaviors

  • Cynthia A. Dlugos
Protocol
Part of the Neuromethods book series (NM, volume 52)

Abstract

Alcoholism is a disorder that affects human beings during every stage of the lifespan. Many animal models have been developed to study alcoholism, including those used to assess alcohol preference, the effects of alcohol withdrawal, and the development of tolerance. Knowledge gained from multiple studies on twins has supported a strong genetic basis for predisposition to alcohol. Our laboratory has chosen to investigate the use of the zebrafish, a vertebrate with an accessible and 75% sequenced genome as a possible model for ethanol research. We have used a simple, noninvasive evaluation of swimming behavior in which we measured the distance between each fish and its nearest neighbor to gage the response of the central nervous system to pharmacologically relevant doses of acute and chronic ethanol. In the acute studies, we have shown that WT (wild type) zebrafish show a dose dependent increase in nearest neighbor distance. Conversely, another strain, the LFS (long-fin striped) zebrafish demonstrated a biphasic response to acute alcohol exposure in that change from baseline was larger at the 0.5 than at the 1.0% (v/v) ethanol concentration. A third strain, the BLF (blue longfin) zebrafish, showed no apparent response to acute alcohol exposure. Subsequent studies showed that behavioral response to ethanol in BLF zebrafish was age dependent, as nearest neighbor distance was increased in juvenile but not in adult fish. Investigations using chronic ethanol exposure in zebrafish also support differential strain sensitivity to ethanol and the capacity to develop tolerance. Ethanol-induced alterations in gender were also investigated. Gender does not appear to be a factor in acute sensitivity to ethanol. Chronic ethanol treatment demonstrated that female WT zebrafish are preferentially affected compared to males of the WT strain. The results of chronic studies suggest that the zebrafish may be a useful model for dissecting the rather complex differential effects of ethanol on gender. Taken together, these studies demonstrate with a simple noninvasive behavioral test that zebrafish of three strains demonstrate differential sensitivity to ethanol and suggest that zebrafish are useful models in sorting out the genetic factors concerning the mechanisms of ethanol’s actions.

Key words

Alcoholism alcohol withdrawal tolerance ethanol genetic differences genetic long fin striped blue longfin nearest neighbor distance strain sensitivity chronic treatment 

Notes

Acknowledgments

I would like to thank Dr. Richard Rabin, my collaborator, for his help throughout these studies and in preparation of this manuscript. I would also like to acknowledge the participation of Dr. Shereene Brown in the gender studies.

References

  1. 1.
    National Institute on Alcohol Abuse and Alcoholism (US), National Institutes of Health (US) and United States. Department of Health and Human Services. (2006) National Epidemiologic Survey on Alcohol and Related Conditions. Rockville, MD, US Department of Health and Human Services, National Institutes of Health, National Institute on Alcohol Abuse and Alcoholism.Google Scholar
  2. 2.
    Stratton, K. R., Howe, C. J. & Battaglia, F. C., Institute of Medicine (US). Division of Biobehavioral Sciences and Mental Disorders. Committee to Whom It May Concern: It May Concern: Study Fetal Alcohol Syndrome & National Institute on Alcohol Abuse and Alcoholism (US) (1996) Fetal Alcohol Syndrome: Diagnosis, Epidemiology, Prevention, and Treatment. Summary. Washington, DC, National Academy Press.Google Scholar
  3. 3.
    National Institute on Alcohol Abuse and Alcoholism (US) (2008) Alcohol Research: A Lifespan Perspective. Rockville, MD, US Department of Health and Human Services, National Institutes of Health, National Institute on Alcohol Abuse and Alcoholism.Google Scholar
  4. 4.
    Heath, A., Kk, B., Paf, M., Dinwiddie, S., Ws, S., Bierut, L., Statham, D., Dunne, M., Jb, W. & Martin, N. (1997) Genetic and environmental contributions to alcohol dependence risk in a national twin sample: consistency of findings in women and men. Psychol Med 27, 1381–1396.PubMedCrossRefGoogle Scholar
  5. 5.
    Heath, A. C. & Martin, N. G. (1994) Genetic influences on alcohol consumption patterns and problem drinking: results from the Australian NH&MRC twin panel follow-up survey. Ann N Y Acad Sci 708, 72–85.PubMedCrossRefGoogle Scholar
  6. 6.
    Kendler, K. S., Neale, M. C., Heath, A. C., Kessler, R. C. & Eaves, L. J. (1994) A twin-family study of alcoholism in women. Am J Psychiatry 151, 707–715.PubMedGoogle Scholar
  7. 7.
    Prescott, C. A. & Kendler, K. S. (1999) Genetic and environmental contributions to alcohol abuse and dependence in a population-based. Am J Psychiatry 156, 7p.Google Scholar
  8. 8.
    Goist, K. C., Jr. & Sutker, P. B. (1985) Acute alcohol intoxication and body composition in women and men. Pharmacol Biochem Behav 22, 811–814.PubMedCrossRefGoogle Scholar
  9. 9.
    Jones, B. & Jones, M. (1976) Women and Alcohol. Intoxication, metabolism, and the menstrual cycle. In Greenblatt, M. & Ma, S. (Eds.) Alcoholism Problems in Women and Children. New York, NY, Grune & Sttratton.Google Scholar
  10. 10.
    Ashley, M. J., Olin, J. S., Le Riche, W. H., Kornaczewski, A., Schmidt, W. & Rankin, J. G. (1977) Morbidity in alcoholics. Evidence for accelerated development of physical disease in women. Arch Int Med 137, 883–887.CrossRefGoogle Scholar
  11. 11.
    Blume, S. B. (1986) Women and alcohol. A review. JAMA 256, 1467–1470.PubMedCrossRefGoogle Scholar
  12. 12.
    Colantoni, A., Idilman, R., de Maria, N., La Paglia, N., Belmonte, J., Wezeman, F., Emanuele, N., van Thiel, D. H., Kovacs, E. J. & Emanuele, M. A. (2003) Hepatic apoptosis and proliferation in male and female rats fed alcohol: role of cytokines. Alcohol Clin Exp Res 27, 1184–1189.PubMedCrossRefGoogle Scholar
  13. 13.
    Dlugos, C. A. (2006b) Smooth endoplasmic reticulum dilation and degeneration in Purkinje neuron dendrites of aging ethanol-fed female rats. Cerebellum 5, 155–162.PubMedCrossRefGoogle Scholar
  14. 14.
    Klintsova, A. Y., Cowell, R. M., Swain, R. A., Napper, R. M., Goodlett, C. R. & Greenough, W. T. (1998) Therapeutic effects of complex motor training on motor performance deficits induced by neonatal binge-like alcohol exposure in rats. I. Behavioral results. Brain Res 800, 48–61.PubMedCrossRefGoogle Scholar
  15. 15.
    Hashimoto, J. G. & Wiren, K. M. (2008) Neurotoxic consequences of chronic alcohol withdrawal: expression profiling reveals importance of gender over withdrawal severity. Neuropsychopharmacology 33, 1084–1096.PubMedCrossRefGoogle Scholar
  16. 16.
    White, A. M., Bae, J. G., Truesdale, M. C., Ahmad, S., Wilson, W. A. & Swartzwelder, H. S. (2002) Chronic-intermittent ethanol exposure during adolescence prevents normal developmental changes in sensitivity to ethanol-induced motor impairments. Alcohol Clin Exp Res 26, 960–968.PubMedCrossRefGoogle Scholar
  17. 17.
    White, A. M., Ghia, A. J., Levin, E. D. & Swartzwelder, H. S. (2000) Binge pattern ethanol exposure in adolescent and adult rats: differential impact on subsequent responsiveness to ethanol. Alcohol Clin Exp Res 24, 1251–1256.PubMedCrossRefGoogle Scholar
  18. 18.
    Breslow, R. A., Faden, V. B. & Smothers, B. (2003) Alcohol consumption by elderly Americans. J Stud Alcohol 64, 884–892.PubMedGoogle Scholar
  19. 19.
    Lovinger, D. M. & Crabbe, J. C. (2005) Laboratory models of alcoholism: treatment target identification and insight into mechanisms. Nat Neurosci 8, 1471–1480.PubMedCrossRefGoogle Scholar
  20. 20.
    Tabakoff, B. & Hoffman, P. L. (2000) Animal models in alcohol research. Alcohol Res Health J Natl Inst Alcohol Abuse Alcohol 24, 77–84.Google Scholar
  21. 21.
    Lithgow, G. J. & Andersen, J. K. (2005) Models of oxidative stress in the biology of aging. Drug Discov Today Dis Models 2, 273–277.CrossRefGoogle Scholar
  22. 22.
    Scholz, H., Franz, M. & Heberlein, U. (2005) The hangover gene defines a stress pathway required for ethanol tolerance development. Nature 436, 845–847.PubMedCrossRefGoogle Scholar
  23. 23.
    Wolf, F. W. & Heberlein, U. (2003) Invertebrate models of drug abuse. J Neurobiol 54, 161–178.PubMedCrossRefGoogle Scholar
  24. 24.
    Davies, A. G., Pierce-Shimomura, J. T., Kim, H., Vanhoven, M. K., Thiele, T. R., Bonci, A., Bargmann, C. I. & McIntire, S. L. (2003) A central role of the BK potassium channel in behavioral responses to ethanol in C. elegans. Cell 115, 655–666.PubMedCrossRefGoogle Scholar
  25. 25.
    Corl, A. B., Rodan, A. R. & Heberlein, U. (2005) Insulin signaling in the nervous system regulates ethanol intoxication in Drosophila melanogaster. Nat Neurosci 8, 18–19.PubMedCrossRefGoogle Scholar
  26. 26.
    Holley, S. A., Geisler, R. & Nusslein-Volhard, C. (2000) Control of her1 expression during zebrafish somitogenesis by a Delta-dependent oscillator and an independent wave-front activity. Genes Dev 14, 1678–1690.Google Scholar
  27. 27.
    Fishman, M. C. (2001) Genomics. Zebrafish – the canonical vertebrate. Science 294, 1290–1291.PubMedCrossRefGoogle Scholar
  28. 28.
    Nüsslein-Volhard, C. & Dahm, R. (2002) Zebrafish: A Practical Approach. Oxford, Oxford University Press.Google Scholar
  29. 29.
    Rubinstein, A. L. (2003) Zebrafish: from disease modeling to drug discovery. Curr Opin Drug Discov Dev 6, 218–223.Google Scholar
  30. 30.
    Boehmler, W., Carr, T., Thisse, C., Thisse, B., Canfield, V. A. & Levenson, R. (2007) D4 dopamine receptor genes of zebrafish and effects of the antipsychotic clozapine on larval swimming behaviour. Genes Brain Behav 6, 155–166.PubMedCrossRefGoogle Scholar
  31. 31.
    Giacomini, N. J., Rose, B., Kobayashi, K. & Guo, S. (2006) Antipsychotics produce locomotor impairment in larval zebrafish. Neurotoxicol Teratol 28, 245–250.PubMedCrossRefGoogle Scholar
  32. 32.
    Parker, B. & Connaughton, V. P. (2007) Effects of nicotine on growth and development in larval zebrafish. Zebrafish 4, 59–68.PubMedCrossRefGoogle Scholar
  33. 33.
    Langheinrich, U., Hennen, E., Stott, G. & Vacun, G. (2002) Zebrafish as a model organism for the identification and characterization of drugs and genes affecting p53 signaling. Curr Biol 12, 2023–2028.PubMedCrossRefGoogle Scholar
  34. 34.
    Darland, T. & Dowling, J. E. (2001) Behavioral screening for cocaine sensitivity in mutagenized zebrafish. Proc Natl Acad Sci USA 98, 11691–11696.PubMedCrossRefGoogle Scholar
  35. 35.
    Farber, S. A., Pack, M., Ho, S. Y., Johnson, I. D., Wagner, D. S., Dosch, R., Mullins, M. C., Hendrickson, H. S., Hendrickson, E. K. & Halpern, M. E. (2001) Genetic analysis of digestive physiology using fluorescent phospholipid reporters. Science 292, 1385–1388.PubMedCrossRefGoogle Scholar
  36. 36.
    Bilotta, J., Barnett, J. A., Hancock, L. & Saszik, S. (2004) Ethanol exposure alters zebrafish development: a novel model of fetal alcohol syndrome. Neurotoxicol Teratol 26, 737–743.PubMedCrossRefGoogle Scholar
  37. 37.
    Bilotta, J., Saszik, S., Givin, C. M., Hardesty, H. R. & Sutherland, S. E. (2002) Effects of embryonic exposure to ethanol on zebrafish visual function. Neurotoxicol Teratol 24, 759–766.PubMedCrossRefGoogle Scholar
  38. 38.
    Blader, P. & Strahle, U. (1998) Ethanol impairs migration of the prechordal plate in the zebrafish embryo. Dev Biol 201, 185–201.PubMedCrossRefGoogle Scholar
  39. 39.
    Carvan, M. J., 3rd, Loucks, E., Weber, D. N. & Williams, F. E. (2004) Ethanol effects on the developing zebrafish: neurobehavior and skeletal morphogenesis. Neurotoxicol Teratol 26, 757–768.PubMedCrossRefGoogle Scholar
  40. 40.
    Dlugos, C. & Rabin, R. (2007) Ocular deficits associated with alcohol exposure during zebrafish development. J Comp Neurol 502, 497–506.PubMedCrossRefGoogle Scholar
  41. 41.
    Gerlai, R., Lahav, M., Guo, S. & Rosenthal, A. (2000) Drinks like a fish: zebra fish Danio rerio as a behavior genetic model to study alcohol effects. Pharmacol Biochem Behav 67, 773–782.PubMedCrossRefGoogle Scholar
  42. 42.
    Lockwood, B., Bjerke, S., Kobayashi, K. & Guo, S. (2004) Acute effects of alcohol on larval zebrafish: a genetic system for large-scale screening. Pharmacol Biochem Behav 77, 647–654.PubMedCrossRefGoogle Scholar
  43. 43.
    Ryback, R. S. (1970) The use of fish, especially goldfish, in alcohol research. Quart J Stud Alcohol 31, 162–166.PubMedGoogle Scholar
  44. 44.
    Ryback, R., Percarpio, B. & Vitale, J. (1969) Equilibration and metabolism of ethanol in the goldfish. Nature 222, 1068–1070.PubMedCrossRefGoogle Scholar
  45. 45.
    Dlugos, C. A. & Rabin, R. A. (2003) Ethanol effects on three strains of zebrafish: model system for genetic investigations. Pharmacol Biochem Behav 74, 471–480.PubMedCrossRefGoogle Scholar
  46. 46.
    Scobie, S. R. & Bliss, D. K. (1974) Ethyl alcohol: relationships to memory for aversive learning in goldfish (Carassius auratus). J Comp Physiol Psychol 86, 867–874.PubMedCrossRefGoogle Scholar
  47. 47.
    Rayes, A. E., Ryback, R. S. & Ingle, D. J. (1968) The effect of alcohol on aggression in Betta splendens. Commun Behav Biol 2, 141–146.Google Scholar
  48. 48.
    Peeke, H. V., Peeke, S. C., Avis, H. H. & Ellman, G. (1975) Alcohol, habituation and the patterning of aggressive responses in a cichlid fish. Pharmacol Biochem Behav 3, 1031–1036.PubMedCrossRefGoogle Scholar
  49. 49.
    Gerlai, R., Lee, V. & Blaser, R. (2006) Effects of acute and chronic ethanol exposure on the behavior of adult zebrafish Danio rerio. Pharmacol Biochem Behav 85, 752–761.PubMedCrossRefGoogle Scholar
  50. 50.
    Gerlai, R., Ahmad, F. & Prajapati, S. (2008) Differences in acute alcohol-induced behavioral responses among zebrafish populations. Alcohol Clin Exp Res 32, 1763–1773.PubMedCrossRefGoogle Scholar
  51. 51.
    Ruhl, N. & McRobert, S. P. (2005) The effect of sex and shoal size on shoaling behaviour in Danio rerio. J Fish Biol 67, 1318–1326.CrossRefGoogle Scholar
  52. 52.
    Damodaran, S., Dlugos, C. A., Wood, T. D. & Rabin, R. A. (2006) Effects of chronic ethanol administration on brain protein levels: a proteomic investigation using 2-D DIGE system. Eur J Pharmacol 547, 75–82.PubMedCrossRefGoogle Scholar
  53. 53.
    Chester, J. A., Blose, A. M. & Froehlich, J. C. (2004) Acoustic startle reactivity during acute alcohol withdrawal in rats that differ in genetic predisposition toward alcohol drinking: effect of stimulus characteristics. Alcohol Clin Exp Res 28, 677–687.PubMedCrossRefGoogle Scholar
  54. 54.
    Grillon, C., Sinha, R., Ameli, R. & O’malley, S. S. (2000) Effects of alcohol on baseline startle and prepulse inhibition in young men at risk for alcoholism and/or anxiety disorders. J Stud Alcohol 61, 46–54.PubMedGoogle Scholar
  55. 55.
    Westerfield, M. (2000) The Zebrafish Book. 4th ed. Eugene, Oregon, University of Oregon Press.Google Scholar
  56. 56.
    Eaton, R. & Hackett, J. (1984) The role of the Mauthner cell in fast-starts involving escape in teleost fishes. In Eaton, R. (Ed.) Neural Mechanisms of Startle Behavior. New York, NY, Plenum Press.Google Scholar
  57. 57.
    Tresnake, L. (1981) The long-finned zebra Danio. Trop Fish Hobby 29, 43–56.Google Scholar
  58. 58.
    Itzkowitz, M. & Iovine, M. K. (2007) Single gene mutations causing exaggerated fins also cause non-genetic changes in the display behavior of male zebrafish. Behaviour 144, 787–795.CrossRefGoogle Scholar
  59. 59.
    Kim, S. N., Rhee, J. H., Song, Y. H., Park, D. Y., Hwang, M., Lee, S. L., Kim, J. E., Gim, B. S., Yoon, J. H., Kim, Y. J. & Kim-Ha, J. (2005) Age-dependent changes of gene expression in the Drosophila head. Neurobiol Aging 26, 1083–1091.PubMedCrossRefGoogle Scholar
  60. 60.
    Rico, E. P., Rosemberg, D. B., Dias, R. D., Bogo, M. R. & Bonan, C. D. (2007) Ethanol alters acetylcholinesterase activity and gene expression in zebrafish brain. Toxicol Lett 174, 25–30.PubMedCrossRefGoogle Scholar
  61. 61.
    Dlugos, C. A., Brown, S. & Rabin, R. A. (2010) Gender differences in ethanol-induced behavioral sensitivity in zebrafish. Alcohol.Google Scholar
  62. 62.
    Orger, M. B., Gahtan, E., Muto, A., Page-McCaw, P., Smear, M. C. & Baier, H. (2004) Behavioral screening assays in zebrafish. Methods Cell Biol 77, 53–68.PubMedCrossRefGoogle Scholar
  63. 63.
    Glowa, J. & Hansen, C. (1993) Differences in response to an acoustic startle stimulus among forty-six rat strains. Behav Genet 24, 79–84.CrossRefGoogle Scholar
  64. 64.
    American Psychiatric Association and American Psychiatric association. Task Force on DSM-IV. (1994) Diagnostic and Statistical Manual of Mental Disorders: DSM-IV. Washington, DC, American Psychiatric Association.Google Scholar
  65. 65.
    Tabakoff, B., Cornell, N. & Hoffman, P. L. (1986) Alcohol tolerance. Ann Emerg Med 15, 1005–1012.PubMedCrossRefGoogle Scholar
  66. 66.
    Tabakoff, B. & Hoffman, P. L. (1988) Tolerance and the etiology of alcoholism: hypothesis and mechanism. Alcohol Clin Exp Res 12, 184–186.PubMedCrossRefGoogle Scholar
  67. 67.
    Park, B., Jeong, S. K., Lee, W. S., Seong, J. K. & Paik, Y. K. (2004) A simple pattern classification method for alcohol-responsive proteins that are differentially expressed in mouse brain. Proteomics 4, 3369–3375.PubMedCrossRefGoogle Scholar
  68. 68.
    Lewohl, J. M., van Dyk, D. D., Craft, G. E., Innes, D. J., Mayfield, R. D., Cobon, G., Harris, R. A. & Dodd, P. R. (2004) The application of proteomics to the human alcoholic brain. Ann N Y Acad Sci 1025, 14–26.PubMedCrossRefGoogle Scholar
  69. 69.
    Le, A. D. & Kiianmaa, K. (1988) Characteristics of ethanol tolerance in alcohol drinking (AA) and alcohol avoiding (ANA) Rats. Psychopharmacology 94, 479–483.PubMedCrossRefGoogle Scholar
  70. 70.
    Dlugos, C. A. (2006a) Ethanol-related smooth endoplasmic reticulum dilation in Purkinje dendrites of aging rats. [Erratum appears in Alcohol Clin Exp Res. 2006 Jun;30(6):1091]. Alcohol Clin Exp Res 30, 883–891.PubMedCrossRefGoogle Scholar
  71. 71.
    Dlugos, C. A. (2008) Ethanol-related increases in degenerating bodies in the Purkinje neuron dendrites of aging rats. Brain Res 1221, 98–107.PubMedCrossRefGoogle Scholar
  72. 72.
    Harper, C. & Kril, J. (1989) Patterns of neuronal loss in the cerebral cortex in chronic alcoholic patients. J Neurol Sci 92, 81–89.PubMedCrossRefGoogle Scholar
  73. 73.
    Kril, J. J. & Halliday, G. M. (1999) Brain shrinkage in alcoholics: a decade on and what have we learned? Prog Neurobiol 58, 381–387.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Cynthia A. Dlugos
    • 1
  1. 1.Department of Pathology and Anatomical Sciences, School of Medicine and Biomedical SciencesUniversity of Buffalo/State University of New YorkBuffaloUSA

Personalised recommendations