Behavior Genetics

, Volume 36, Issue 4, pp 536–552 | Cite as

Effects of Genetic and Procedural Variation on Measurement of Alcohol Sensitivity in Mouse Inbred Strains

  • John C. Crabbe
  • Pamela Metten
  • Igor Ponomarev
  • Carol A. Prescott
  • Douglas Wahlsten


Mice from eight inbred strains were studied for their acute sensitivity to ethanol as indexed by the degree of hypothermia (HT), indexed as the reduction from pre-injection baseline of their body temperature. Two weeks later, mice were tested for their loss of righting reflex (LRR) after a higher dose of ethanol. The LRR was tested using the “classical” method of watching for recovery in animals placed on their backs in a V-shaped trough and recording duration of LRR. In a separate test, naive animals of the same strains were tested for HT repeatedly to assess the development of rapid (RTOL) and chronic tolerance (CTOL). We have recently developed a new method for testing LRR that leads to a substantial increase in the sensitivity of the test. Strains also have been found to differ in the new LRR test, as well as in the development of acute functional tolerance (AFT) to this response. In addition, our laboratory has periodically published strain difference data on the older versions of the HT and LRR responses. The earlier tests used some of the exact substrains tested currently, while for some strains, different substrains (usually, Nih versus Jax) were tested. We examined correlations of strain means to see whether patterns of strain differences were stable across time and across different test variants assessing the same behavioral construct. HT strain sensitivity scores were generally highly correlated across a 10–23 years period and test variants. The CTOL to HT was well-correlated across studies, and was also genetically similar to RTOL. The AFT, however, was related to neither RTOL nor CTOL, although this may be because different phenotypic end points were compared. The LRR data, which included a variant of the classical test, were not as stable. Measures of LRR onset were reasonably well correlated, as were those taken at recovery (e.g., duration). However, the two types of measures of LRR sensitivity to ethanol appear to be tapping traits that differ genetically. Also, the pattern of genetic correlation between HT and LRR initially reported in 1983 was not seen in current and contemporaneous studies. In certain instances, substrain seems to matter little, while in others, substrains differed a great deal. These data are generally encouraging about the stability of genetic differences.


Acute tolerance chronic tolerance ethanol hypothermia inbred mouse strains loss of righting reflex pharmacogenetics 



These studies and the preparation of this manuscript were supported by NIH grants AA19760, AA12714, AA13519, NSERC grant 45825, the Mouse Phenome Project, and the Department of Veterans Affairs. Thanks to Christina J. Cotnam, Andy J. Cameron, Chia-Hua Yu, Jason P. Schlumbohm, Katie L. Mordarski, and Karyn L. Best for expert technical assistance with Experiments 1–3, and many former students and research assistants for help with the earlier work.


  1. Allan A. M., and Harris R. A. (1991). Neurochemical studies of genetic differences in alcohol action. In: Crabbe J. C. and Harris R. A. (eds), The Genetic Basis for Alcohol and Drug Actions. New York, Plenum, pp. 105–152Google Scholar
  2. Barbosa A. D., and Morato G. S. (2000). Effect of epipregnanolone and pregnenolone sulfate on chronic tolerance to ethanol. Pharmacol. Biochem. Behav. 67:459–464PubMedCrossRefGoogle Scholar
  3. Barbosa A. D., and Morato G. S. (2001). Influence of neurosteroids on the development of rapid tolerance to ethanol in mice. Eur. J. Pharmacol. 431:179–188PubMedCrossRefGoogle Scholar
  4. Belknap J. K., Crabbe J. C., and Young E. R. (1993). Voluntary consumption of ethanol in 15 inbred mouse strains. Psychopharmacology 112:503–510PubMedCrossRefGoogle Scholar
  5. Bennett B., Beeson M., Gordon L., Carosone-Link A. P., and Johnson T.E. (2002). Genetic dissection of quantitative trait loci specifying sedative/hypnotic sensitivity to ethanol: mapping with interval-specific congenic recombinant lines. Alcohol. Clin. Exp. Res. 26:1615–1624PubMedGoogle Scholar
  6. Crabbe J. C. (1983). Sensitivity to ethanol in inbred mice: genotypic correlations among several behavioral responses. Behav. Neurosci. 97(2):280–289CrossRefGoogle Scholar
  7. Crabbe J. C. (1994). Tolerance to ethanol hypothermia in HOT and COLD mice. Alcohol. Clin. Exp. Res. 18:42–46PubMedCrossRefGoogle Scholar
  8. Crabbe J. C., Belknap J. K., Mitchell S. R., and Crawshaw L. I. (1994a). Quantitative trait loci mapping of genes that influence the sensitivity and tolerance to ethanol-induced hypothermia in BXD recombinant inbred mice. J. Pharmacol. Exp. Ther. 269:184–192Google Scholar
  9. Crabbe J. C., Cotnam C. J., Cameron A. J., Schlumbohm J. P., Rhodes J. S., Metten P., and Wahlsten D. (2003a). Strain differences in three measures of ethanol intoxication in mice, the screen, dowel and grip strength tests. Genes Brain Behav. 2:201–213CrossRefGoogle Scholar
  10. Crabbe J. C., Feller D. J., and Dorow J. S. (1989). Sensitivity and tolerance to ethanol-induced hypothermia in genetically selected mice. J. Pharmacol. Exp. Ther. 249(2):456–461Google Scholar
  11. Crabbe J. C., Gallaher E. S., Phillips T. J., and Belknap J. K. (1994b). Genetic determinants of sensitivity to ethanol in inbred mice. Behav. Neurosci. 108:186–195CrossRefGoogle Scholar
  12. Crabbe J. C., Janowsky J. S., Young E. R., Kosobud A., Stack J., and Rigter H. (1982). Tolerance to ethanol hypothermia in inbred mice: genotypic correlations with behavioral responses. Alcohol. Clin. Exp. Res. 6(4):446–458CrossRefGoogle Scholar
  13. Crabbe J. C., Kosobud A., Tam B. R., Young E. R., and Deutsch C. M. (1987). Genetic selection of mouse lines sensitive (cold) and resistant (hot) to acute ethanol hypothermia. Alcohol Drug Res. 7:163–174PubMedGoogle Scholar
  14. Crabbe J. C., Metten P., Cameron A. J., and Wahlsten D. (2005). An analysis of the genetics of alcohol intoxication in inbred mice. Neurosci. Biobehav. Rev. 28:785–802PubMedCrossRefGoogle Scholar
  15. Crabbe J. C., Metten P., Yu C.-H., Schlumbohm J. P., Cameron A. J. and Wahlsten D. (2003b). Genotypic differences in ethanol sensitivity in two tests of motor incoordination. J. Appl. Physiol. 95:1338–1351Google Scholar
  16. Crabbe J. C., Phillips T. J., Gallaher E. J., Crawshaw L. I. and Mitchell S. R. (1996). Common genetic determinants of the ataxic and hypothermic effects of ethanol in BXD/Ty recombinant inbred mice: genetic correlations and quantitative trait loci. J. Pharmacol. Exp. Ther. 277:624–632PubMedGoogle Scholar
  17. Crabbe J. C., Rigter H., Uijlen J., and Strijbos C. (1979). Rapid development of tolerance to the hypothermic effect of ethanol in mice. J. Pharmacol. Exp. Ther. 208(1):128–133Google Scholar
  18. Crabbe J. C., Wahlsten D., and Dudek B. C. (1999). Genetics of mouse behavior: interactions with laboratory environment. Science 284:1670–1672PubMedCrossRefGoogle Scholar
  19. Crawshaw L. I., O’Connor, C. S., Crabbe J. C., and Hayteas D. L. (1994). Temperature regulation in mice during withdrawal from ethanol dependence. Am. J. Physiol. 267:R929-R934PubMedGoogle Scholar
  20. Dixon W. J., and Mood A. M. (1948). A method for obtaining and analyzing sensitivity data. J. Am. Stat. Assoc. 43:109–126CrossRefGoogle Scholar
  21. Draski L. J. and Deitrich R. A. (1996). Initial effects of ethanol on the central nervous system. In: Deitrich R. A. and Erwin V. G. (eds), Pharmacological Effects of Ethanol on the Nervous System. Boca Raton FL, CRC Press, pp. 227–250Google Scholar
  22. Erwin V. G., and Deitrich R. A. (1996). Genetic selection and characterization of mouse lines for acute functional tolerance to ethanol. J. Pharmacol. Exp. Ther. 279:1310–1317PubMedGoogle Scholar
  23. Erwin V. G., Radcliffe R. A., and Jones B. C. (1992). Chronic ethanol consumption produces genotype-dependent tolerance to ethanol in LS/Ibg and SS/Ibg mice. Pharmacol. Biochem. Behav. 41:275–281PubMedCrossRefGoogle Scholar
  24. Fuller J. L. (1964). Measurement of alcohol preference in genetic experiments. J. Comp. Physiol. Psych. 57:85–88CrossRefGoogle Scholar
  25. Grieve S. J., and Littleton J. M. (1979). Age and strain differences in the rate of development of functional tolerance to ethanol by mice. J. Pharm. Pharmacol. 31:696–700PubMedGoogle Scholar
  26. Grubb S. C., Churchill G. A., and Bogue M. A. (2004) A collaborative database of inbred mouse strain characteristics. Bioinformatics 20:2857–2859PubMedCrossRefGoogle Scholar
  27. Hegmann J. P., and Possidente B. (1981). Estimating genetic correlations from inbred strains. Behav. Genet. 11:103–114PubMedCrossRefGoogle Scholar
  28. Kalant H. (1996). Current state of knowledge about the mechanisms of alcohol tolerance. Addict. Biol. 1:133–141PubMedCrossRefGoogle Scholar
  29. Kalant H., and Lê A. D. (1984). Effects of ethanol on thermoregulation. Pharmacol. Ther. 23:313–364CrossRefGoogle Scholar
  30. Kalant H., LeBlanc A. E., and Gibbins R. J. (1971). Tolerance to, and dependence on, some non-opiate psychotropic drugs. Pharmacol. Rev. 23:135–191PubMedGoogle Scholar
  31. Keir W. J., and Deitrich R. A. (1990). Development of central nervous system sensitivity to ethanol and pentobarbital in short- and long-sleep mice. J. Pharmacol. Exp. Ther. 254(3):831–835Google Scholar
  32. Khanna J. M., Kalant H., Shah G., and Weiner J. (1991). Rapid tolerance as an index of chronic tolerance. Pharmacol. Biochem. Behav. 38:427–432PubMedCrossRefGoogle Scholar
  33. Khanna J. M., Kalant H., Weiner J., Chau A., and Shah G. (1992). Ketamine retards chronic but not acute tolerance to ethanol. Pharmacol. Biochem. Behav. 42:347–350PubMedCrossRefGoogle Scholar
  34. Khanna J. M., Morato G. S., Chau A., Shah G., and Kalant H. (1994). Effect of NMDA antagonists on rapid and chronic tolerance to ethanol: Importance of intoxicated practice. Pharmacol. Biochem. Behav. 48:755–763PubMedCrossRefGoogle Scholar
  35. Khanna J. M., Morato G. S., and Kalant H. (2002). Effect of NMDA antagonists, an NMDA agonist, and serotonin depletion on acute tolerance to ethanol. Pharmacol. Biochem. Behav. 72:291–298PubMedCrossRefGoogle Scholar
  36. Mackay T. F. (2001). The genetic architecture of quantitative traits. Annu. Rev. Genet. 35:303–339PubMedCrossRefGoogle Scholar
  37. Markel P. D., Bennett B., Beeson M., Gordon L., and Johnson T. E. (1997). Confirmation of quantitative trait loci for ethanol sensitivity in long- sleep and short-sleep mice. Genome Res. 7:92–99PubMedCrossRefGoogle Scholar
  38. Markel P. D., DeFries J. C., and Johnson T. E. (1995). Ethanol-induced anesthesia in inbred strains on long-sleep and short-sleep mice: A genetic analysis of repeated measures using censored data. Behav. Genet. 25(1):67–73PubMedCrossRefGoogle Scholar
  39. McClearn G. E. and Kakihana R. (1981). Selective breeding for ethanol sensitivity: short-sleep and long-sleep mice. In: McClearn G. E., Deitrich R. A. and Erwin V. G. (eds), Development of Animal Models as Pharmacogenetic Tools. Rockville MD, USDHHS:NIAAA, pp. 147–159Google Scholar
  40. McClearn G. E., and Rodgers D. A. (1959). Differences in alcohol preference among inbred strains of mice. Q. J. Stud. Alcohol 20:691–695Google Scholar
  41. Mellanby E. (1919). Alcohol: its absorption into and disappearance from the blood under different conditions. London, Medical Research CommitteeGoogle Scholar
  42. Metten P., Best K. L., Cameron A. J., Saultz A. B., Zuraw J. M., Yu C.-H., Wahlsten D. and Crabbe J. C. (2004). Observer-rated ataxia: rating scales for assessment of genetic differences in ethanol-induced intoxication in mice. J. Appl. Physiol. 97:360–368PubMedCrossRefGoogle Scholar
  43. Metten P. and Crabbe J. C. (2005). Alcohol withdrawal severity in inbred mouse (Mus musculus) strains. Behav. Neurosci. 119:911–925.PubMedCrossRefGoogle Scholar
  44. Owens J. C., Bennett B., and Johnson T. E. (2002). Possible pleiotropic effects of genes specifying sedative/hypnotic sensitivity to ethanol on other alcohol-related traits. Alcohol. Clin. Exp. Res. 26:1461–1467PubMedGoogle Scholar
  45. Phillips T. J. (1997). Behavior genetics of drug sensitization. Crit. Rev. Neurobiol. 11:21–33PubMedGoogle Scholar
  46. Phillips T. J. and Crabbe J. C. (1991). Behavioral studies of genetic differences in alcohol action. In: Crabbe J. C., Harris R.A. (eds), The Genetic Basis of Alcohol and Drug Actions. New York, Plenum Press, pp. 25–104Google Scholar
  47. Ponomarev I., and Crabbe J. C. (2004). Characterization of acute functional tolerance to the hypnotic effects of ethanol in mice. Alcohol. Clin. Exp. Res. 28:991–997PubMedCrossRefGoogle Scholar
  48. Ponomarev I., and Crabbe J. C. (2002). A novel method to assess initial sensitivity and acute functional tolerance to hypnotic effects of ethanol. J. Pharmacol. Exp. Ther. 302:257–263PubMedCrossRefGoogle Scholar
  49. Preacher K. J., and MacCallum R. C. (2002). Exploratory factor analysis in behavior genetics research: factor recovery with small sample sizes. Behav. Genet. 32:153–161PubMedCrossRefGoogle Scholar
  50. Rodgers D. A. (1972). Factors underlying differences in alcohol preference in inbred strains of mice. In: Kissin B., Begleiter H. (eds), The Biology of Alcoholism. New York, Plenum, pp. 107–130Google Scholar
  51. Rustay N. R., Boehm S. L., Schafer G. L., Browman K. E., Erwin V. G. and Crabbe J. C. (2001). Sensitivity and tolerance to ethanol-induced incoordination and hypothermia in HAFT and LAFT mice. Pharmacol. Biochem. Behav. 70:167–174PubMedCrossRefGoogle Scholar
  52. Rustay N. R., and Crabbe J. C. (2004). Genetic analysis of rapid tolerance to ethanol’s incoordinating effects in mice: Inbred strains and artificial selection. Behav. Genet. 34:441–451PubMedCrossRefGoogle Scholar
  53. Rustay N. R., Wahlsten D., and Crabbe J. C. (2003). Assessment of genetic susceptibility to ethanol intoxication in mice. Proc. Nat. Acad. Sci. USA 100:2917–2922PubMedCrossRefGoogle Scholar
  54. Schuckit M. A. (1994). Low level of response to alcohol as a predictor of future alcoholism. Am. J. Psychiat. 151:184–189PubMedGoogle Scholar
  55. Schuckit M. A., Smith T. L., and Kalmijn J. (2004). The search for genes contributing to the low level of response to alcohol: patterns of findings across studies. Alcohol. Clin. Exp. Res. 28:1449–1458PubMedCrossRefGoogle Scholar
  56. Sokal R. R., and Rohlf F. J. (1981). Biometry. San Francisco, FreemanGoogle Scholar
  57. Wahlsten D., Metten P., and Crabbe J. C. (2003a). A rating scale for wildness and ease of handling laboratory mice: results for 21 inbred strains tested in two laboratories. Genes Brain Behav. 2:71–79CrossRefGoogle Scholar
  58. Wahlsten D., Metten P., Phillips T. J., Boehm S. L., II, Burkhart-Kasch S., Dorow J., Doerksen S., Downing C., Fogarty J., Rodd-Henricks K., Hen R., McKinnon C. S., Merrill C. M., Nolte C., Schalomon M., Schlumbohm J. P., Sibert J. R., Wenger C. D., Dudek B. C., Crabbe J. C. (2003b). Different data from different labs: lessons from studies of gene-environment interaction. J. Neurobiol. 54:283–311CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • John C. Crabbe
    • 1
    • 5
  • Pamela Metten
    • 1
  • Igor Ponomarev
    • 2
  • Carol A. Prescott
    • 3
  • Douglas Wahlsten
    • 4
  1. 1.Portland Alcohol Research Center, Department of Behavioral NeuroscienceOregon Health & Science University, and VA Medical Center (R&D 12)PortlandUSA
  2. 2.Waggoner Center for Alcohol and Addiction ResearchThe University of Texas at AustinAustinUSA
  3. 3.Department of PsychologyUniversity of Southern CaliforniaLos AngelesUSA
  4. 4.Great Lakes Institute for Environmental Research, Department of Biological SciencesUniversity of WindsorWindsorCanada
  5. 5. VA Medical Center (R&D 12)PortlandUSA

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