Advertisement

Behavior Genetics

, Volume 43, Issue 1, pp 34–50 | Cite as

What are We Measuring When We Test Strain Differences in Anxiety in Mice?

  • Timothy P. O’Leary
  • Rhian K. Gunn
  • Richard E. Brown
Original Research

Abstract

We examined measures of locomotor and anxiety-like behavior in male and female mice of 15 inbred strains on the elevated-plus maze, light/dark transition box and open field. Strain differences were found on all measures of locomotor activity and anxiety. Strain means for measures of locomotor activity on the three apparatus were significantly correlated, but strain means for commonly used measures of anxiety were not correlated. Principal component analysis revealed a common locomotor activity factor, which accounted for 28.6 % of the variance, but no common anxiety factor. Species-typical behaviors (defecations, stretch-attend postures, grooming) accounted for smaller proportions (<11 %) of the variance. These results plus comparisons with previously published data suggest that the elevated-plus maze, light/dark box and open field measure different facets of anxiety, and that the reliability of genetic differences on anxiety is highly dependent on apparatus, procedural variables and laboratory factors. Locomotor activity, however, is a stable trait that differs across strains and is reliably measured in different apparatus and laboratories. We conclude that anxiety traits of inbred mouse strains are best reflected by species-typical behaviors in each apparatus. These results suggest that new ways of measuring trait anxiety are required in order to determine the neural and genetic correlates of anxiety-like behaviour in mice.

Keywords

Inbred mice Elevated plus maze Light dark box Open field Species-typical behaviors Anxiety Locomotion Strain differences Laboratory effects 

Notes

Acknowledgments

We wish to thank Valerie Bolivar of the Wadsworth Genomics Institute for her donation of BTBR mice, and Nicola Hoffman, Lisa Currie, Vicki Savoie, Martin Williamson, Dirk Luchtman and Amanda Arnold for assistance in behavioral testing. Equipment was purchased with funds from an NSERC equipment grant. This research was funded by an NSERC grant to REB. Generous funds from AstraZeneca R&D Boston were used to defray the cost of mice through the Mouse Phenome Project (The Jackson Laboratory, Bar Harbor, ME).

Supplementary material

10519_2012_9572_MOESM1_ESM.pdf (140 kb)
Supplementary material 1 (PDF 140 kb)

References

  1. Andreatini R, Bacellar LF (2000) Animal models: trait or state measure? The test-retest reliability of the elevated plus-maze and behavioral despair. Prog Neuropsychopharmacol Biol Psychiatr 24:549–5604CrossRefGoogle Scholar
  2. Archer J (1977) Sex differences in the emotional behaviour of laboratory mice. Br J Psychiatr 68:125–131Google Scholar
  3. Beuzen A, Belzung C (1995) Link between emotional memory and anxiety states: a study by principal component analysis. Physiol Behav 58:111–118PubMedCrossRefGoogle Scholar
  4. Blanchard DC, Griebel G, Blanchard RJ (2003) The mouse defense test battery: pharmacological and behavioral assays for anxiety and panic. Eur J Pharmacol 463:97–116PubMedCrossRefGoogle Scholar
  5. Blanchard DC, Griebel G, Pobbe R, Blanchard RJ (2011) Risk assessment as an evolved threat detection and analysis process. Neurosci Biobehav Rev 35:991–998PubMedCrossRefGoogle Scholar
  6. Bolivar VJ (2009) Intrasession and intersession habituation in mice: from inbred strain variability to linkage analysis. Neurobiol Learn Mem 92:206–214PubMedCrossRefGoogle Scholar
  7. Bolivar VJ, Caldarone BJ, Reilly AA, Flaherty L (2000) Habituation of activity in an open field: a survey of inbred strains and F1 hybrids. Behav Genet 30:285–293PubMedCrossRefGoogle Scholar
  8. Bothe GWM, Bolivar VJ, Vedder MJ, Geistfeld JG (2005) Behavioural differences among fourteen inbred mouse strains commonly used as disease models. Comp Med 55:326–334PubMedGoogle Scholar
  9. Bourin M, Hascoët M (2003) The mouse light/dark box test. Eur J Pharmacol 463:55–65PubMedCrossRefGoogle Scholar
  10. Bourin M, Petit-Demoulière B, Dhonnchadha BN, Hascoët M (2007) Animal models of anxiety in mice. Fundam Clin Pharmacol 21:567–574PubMedCrossRefGoogle Scholar
  11. Brigman JL, Mathur P, Lu L, Williams RW, Holmes A (2009) Genetic relationship between anxiety-related and fear-related behaviors in BXD recombinant inbred mice. Behav Pharmacol 20:204–209PubMedCrossRefGoogle Scholar
  12. Brown RE, Wong AA (2007) The influence of visual ability on learning and memory in 13 strains of mice. Learn Mem 14:134–144PubMedCrossRefGoogle Scholar
  13. Brown RE, Corey SC, Moore AK (1999) Differences in measures of exploration and fear in MHC-congenic C57BL/6 J and B6-H-2 K mice. Behav Genet 29:263–271CrossRefGoogle Scholar
  14. Brown RE, Gunn RK, Schellinck HM, Wong AA, O’Leary TP (2004) Anxiety, exploratory behavior, and motor activity in 14 inbred strains of mice. MPD:Brown1. Mouse Phenome Database website, The Jackson Laboratory, Bar Harbor, ME. http://phenome.jax.org, Dec, 2012
  15. Calatayud F, Coubard S, Belzung C (2004a) Emotional reactivity in mice may not be inherited but influenced by parents. Physiol Behav 80:465–474PubMedCrossRefGoogle Scholar
  16. Calatayud F, Belzung C, Aubert A (2004b) Ethological validation and the assessment of anxiety-like behaviours: methodological comparison of classical analyses and structural approaches. Behav Processes 67:195–206PubMedCrossRefGoogle Scholar
  17. Carobrez AP, Bertoglio LJ (2005) Ethological and temporal analyses of anxiety-like behavior: the elevated plus-maze model 20 years on. Neurosci Biobehav Rev 29:1193–1205PubMedCrossRefGoogle Scholar
  18. Clément Y, Calatayud F, Belzung C (2002) Genetic basis of anxiety-like behaviour: a review. Brain Res Bull 57:57–71PubMedCrossRefGoogle Scholar
  19. Clenet F, Bouyon E, Hascoet M, Bourin M (2006) Light/dark cycle manipulation influences mice behaviour in the elevated-plus maze. Behav Brain Res 166:140–149PubMedCrossRefGoogle Scholar
  20. Cook MN, Williams RW, Flaherty L (2001) Anxiety-related behaviors in the elevated zero-maze are affected by genetic factors and retinal degeneration. Behav Neurosci 115:468–476PubMedCrossRefGoogle Scholar
  21. Cook MN, Bolivar VJ, McFadyen MP, Flaherty L (2002) Behavioral differences among 129 substrains: implications for knockout and transgenic mice. Behav Neurosci 116:600–611PubMedCrossRefGoogle Scholar
  22. Costall B, Jones BJ, Kelly ME, Naylor RJ, Tomkins DM (1989) Exploration of mice in a black and white test box: validation as a model of anxiety. Pharmacol Biochem Behav 32:777–785PubMedCrossRefGoogle Scholar
  23. Crabbe JC, Wahlsten D, Dudek BC (1999) Genetics of mouse behavior: interactions with laboratory environment. Science 284:1670–1672PubMedCrossRefGoogle Scholar
  24. Cryan JF, Holmes A (2005) The ascent of mouse: advances in modeling human depression and anxiety. Nat Rev Drug Discov 4:775–790PubMedCrossRefGoogle Scholar
  25. Cryan JF, Sweeney FF (2011) The age of anxiety: role of animal models of anxiolytic action in drug discovery. Brit J Pharmacol 164:1129–1161CrossRefGoogle Scholar
  26. Dawson GR, Tricklebank MD (1995) Use of the elevated-plus maze in the search for novel anxiolytic agents. Trends Pharmacol Sci 16:33–36PubMedCrossRefGoogle Scholar
  27. DeFries JC, Hegmann JP, Weir MW (1966) Open-field behavior in mice: evidence for major gene effect mediated by the visual system. Science 154:1577–1579PubMedCrossRefGoogle Scholar
  28. Depino AM, Gross C (2007) Simultaneous assessment of autonomic function and anxiety-related behavior in BALB/c and C57BL/6 mice. Behav Brain Res 177:254–260PubMedCrossRefGoogle Scholar
  29. Durant C, Christmas D, Nutt D (2010) The pharmacology of anxiety. Curr Top Behav Neurosci 2:303–330PubMedCrossRefGoogle Scholar
  30. Estanislau C (2012) Cues to the usefulness of grooming behavior in the evaluation of anxiety in the elevated plus-maze. Psychol Neurosci 5:105–112CrossRefGoogle Scholar
  31. Flint J (2003) Analysis of quantitative trait loci that influence animal behavior. J Neurobiol 54:46–77PubMedCrossRefGoogle Scholar
  32. Fonio E, Benjamin Y, Golani I (2009) Freedom of movement and the stability of its unfolding in free exploration of mice. Proc Nat Acad Sci USA 106:21335–21340PubMedCrossRefGoogle Scholar
  33. Fraser LM, Brown RE, Hussin A, Fontana M, Whittaker A, O’Leary TP, Lederle L, Holmes A, Ramos A (2010) Measuring anxiety- and locomotion-related behaviours in mice: a new way of using old tests. Psychopharm 211:99–112CrossRefGoogle Scholar
  34. Geronikaki A, Babaev E, Dearden J, Dehaen W, Filimonov D, Galaeva I, Krajneva V, Lagunin A, Macaev F, Molodavkin G, Poroikov V, Pogrebnoi S, Saloutin V, Stepanchikova A, Stingaci E, Tkach N, Vlad L, Voronina T (2004) Design, synthesis, computational and biological evaluation of new anxiolytics. Bioorg Med Chem 12:6559–6568PubMedCrossRefGoogle Scholar
  35. Griebel G, Belzung C, Misslin R, Vogel E (1993) The free-exploratory paradigm: an effective method for measuring neophobic behaviour in mice and testing potential neophobia-reducing drugs. Behav Pharmacol 4:637–644PubMedCrossRefGoogle Scholar
  36. Griebel G, Blanchard DC, Blanchard RJ (1996) Evidence that the behaviors in the mouse defense test battery relate to different emotional states: a factor analytic study. Physiol Behav 60:1255–1260PubMedCrossRefGoogle Scholar
  37. Griebel G, Belzung C, Perrault G, Sanger DJ (2000) Differences in anxiety-related behaviours and in sensitivity to diazepam in inbred and outbred strains of mice. Psychopharm 148:164–170CrossRefGoogle Scholar
  38. Grubb SC, Maddatu TP, Bult CJ, Bogue MA (2009) Mouse phenome database. Nucleic Acids Res 35:643–649Google Scholar
  39. Hall C (1934) Emotional behavior in the rat: i. defecation and urination as measures of individual differences in emotionality. J Comp Psychol 18:385–403CrossRefGoogle Scholar
  40. Haller J, Alicki M (2012) Current animal models of anixiety, anxiety disorders, and anxiolytic drugs. Curr Opin Psychiatry 1:59–64CrossRefGoogle Scholar
  41. Henderson ND, Turri MG, DeFries JC, Flint J (2004) QTL analysis of multiple behavioral measures of anxiety in mice. Behav Genet 34:267–293PubMedCrossRefGoogle Scholar
  42. Hohoff C (2009) Anxiety in mice and men: a comparison. J Neural Transm 116:679–687PubMedCrossRefGoogle Scholar
  43. Hussin AT, Fraser LM, Ramos A, Brown RE (2012) The effect of chlordiazepoxide on measures of activity and anxiety in Swiss-Webster mice in the triple test. Neuropharm 63:883–889CrossRefGoogle Scholar
  44. Izídio GS, Lopes DM, Spricigo L Jr, Ramos A (2005) Common variations in the pretest environment influence genotypic comparisons in models of anxiety. Genes Brain Behav 4:412–419PubMedCrossRefGoogle Scholar
  45. Jacobson LH, Cryan JF (2010) Genetic approaches to modeling anxiety in animals. Curr Top Behav Neurosci 2:161–201PubMedCrossRefGoogle Scholar
  46. Jain A, Dvorkin A, Fonio E, Golani I, Gross CT (2012) Validation of the dimensionality emergence assay for the measurement of innate anxiety in laboratory mice. Eur Neuropsychopharm 22:153–163CrossRefGoogle Scholar
  47. Kalueff AV, Tuohimaa P (2005a) Contrasting grooming phenotypes in three mouse strains markedly different in anxiety and activity (129S1, BALB/c and NMRI). Behav Brain Res 160:1–10PubMedCrossRefGoogle Scholar
  48. Kalueff AV, Tuohimaa P (2005b) The grooming analysis algorithm discriminates between different levels of anxiety in rats: potential utility for neurobehavioural stress research. J Neurosci Methods 143:169–177PubMedCrossRefGoogle Scholar
  49. Kessler RC (2007) The global burden of anxiety and mood disorders: putting ESEMeD findings into perspective. J Clin Psychiatr 68:10–19CrossRefGoogle Scholar
  50. Kessler RC, Ruscio AM, Shear K, Wittchen HU (2010) Epidemiology of anxiety disorders. Curr Top Behav Neurosci 2:21–35PubMedCrossRefGoogle Scholar
  51. Krömer SA, Keßler MS, Milfay D, Birg IN, Bunck M, Czibere L, Panhuysen M, Pütz B, Deussing JM, Holsboer F, Landgraf R, Turck CW (2005) Identification of glyoxalase-I as a protein marker in a mouse model of extremes in trait anxiety. J Neurosci 25:4375–4384PubMedCrossRefGoogle Scholar
  52. Lad HV, Liu L, Paya-Cano JL, Parsons MJ, Kember R, Fernandes C, Schalkwyk LC (2010) Behavioural battery testing: evaluation and behavioural outcomes in 8 inbred mouse strains. Physiol Behav 99:301–316PubMedCrossRefGoogle Scholar
  53. Lister RG (1987) The use of a plus-maze to measure anxiety in the mouse. Psychopharmacology 92:180–185PubMedGoogle Scholar
  54. Lister RG (1990) Ethologically-based animal models of anxiety disorders. Pharmacol Ther 46:321–340PubMedCrossRefGoogle Scholar
  55. Logue SF, Owen EH, Rasmussen DL, Wehner JM (1997) Assessment of locomotor activity, acoustic and tactile startle, and prepulse inhibition of startle in inbred mouse strains and F1 hybrids: implications of genetic background for single gene and quantitative trait loci analyses. Neuroscience 80:1075–1086PubMedCrossRefGoogle Scholar
  56. Mandillo S, Tucci V, Hölter SM, Meziane H, Al Banchaabouchi M, Kallnik M, Lad HV, Nolan PM, Ougazzal A-M, Coghill EL, Gale K, Golini E, Jacquot S, Krezel W, Parker A, Riet F, Schneider I, Marazziti D, Auwerx J, Brown SDM, Chambon P, Rosenthal N, Tocchini-Valentini G, Wurst W (2008) Reliability, robustness, and reproducibility in mouse behavioral phenotyping: a cross-laboratory study. Physiol Genomics 34:243–255PubMedCrossRefGoogle Scholar
  57. McFadyen MP, Kusek G, Bolivar VJ, Flaherty L (2003) Differences among eight strains of mice in motor ability and motor learning on a rotorod. Genes Brain Behav 2:214–219PubMedCrossRefGoogle Scholar
  58. McIlwain KL, Merriweather MY, Yuva-Paylor LA, Paylor R (2001) The use of behavioral test batteries: effects of training history. Physiol Behav 73:705–717PubMedCrossRefGoogle Scholar
  59. Miller BH, Schulz LE, Gulati A, Su AI, Pletcher MT (2010) Phenotypic characterization of a genetically diverse panel of mice for behavioural despair and anxiety. PLoS ONE 10:e14458CrossRefGoogle Scholar
  60. Milner LC, Crabbe JC (2008) Three murine anxiety models: results from multiple inbred strain comparisons. Genes Brain Behav 7:496–505PubMedCrossRefGoogle Scholar
  61. Nadel L (1968) Dorsal and ventral hippocampal lesions and behavior. Physiol Behav 3:891–900CrossRefGoogle Scholar
  62. Nagy ZM, Forrest EJ (1970) Open-field behavior of C3H mice: effect of size and illumination of field. Psychon Sci 20:19–21Google Scholar
  63. Nagy ZM, Glaser HD (1970) Open-field behavior of C57BL/6 J mice: effect of illumination, age, and number of test days. Psychon Sci 19:143–145Google Scholar
  64. Nagy ZM, Holm M (1970) Open-field behavior of C3H mice: effect of early handling, field illumination, and age at testing. Psychon Sci 19:273–275Google Scholar
  65. Owen DR, Rupprecht R, Nutt DJ (2012) Stratified medicine in psychiatry: a worrying example or new opportunity in the treatment of anxiety? J Psychopharm. doi: 10.1177/0269881112443746
  66. Palanza P (2001) Animal models of anxiety and depression: how are females different? Neurosci Biobehav Rev 25:219–233PubMedCrossRefGoogle Scholar
  67. Paulus MP, Dulawa SC, Ralph RJ, Geyer MA (1999) Behavioral organization is independent of locomotor activity in 129 and C57 mouse strains. Brain Res 835:27–36PubMedCrossRefGoogle Scholar
  68. Pletcher MT (2007) Open field and tail suspension tests in males of 32 inbred strains of mice. In: MPD Pletcher (ed) Mouse Phenome Database website, The Jackson Laboratory, Bar Harbor, Maine USA. http://phenome.jax.org, Mar, 2012
  69. Podhorna J, Brown RE (2002) Strain differences in activity and emotionality do not account for differences in learning and memory performance between C57BL/6 and DBA/2 mice. Genes Brain Behav 1:96–110PubMedCrossRefGoogle Scholar
  70. Post AM, Weyers P, Holzer P, Painsipp E, Pauli P, Wultsch T, Reif A, Lesh K-P (2011) Gene-environment interaction influences anxiety-like behaviour in ethologically based mouse models. Behav Brain Res 218:99–105PubMedCrossRefGoogle Scholar
  71. Ramos A (2008) Animal models of anxiety: do I need multiple tests? Trends Pharmacol Sci 29:493–498PubMedCrossRefGoogle Scholar
  72. Ramos A, Pereira E, Martins GC, Wehrmeister TD, Izídio GS (2008) Integrating the open field, elevated plus maze and light/dark box to assess different types of emotional behaviors in one single trial. Behav Brain Res 193:277–288PubMedCrossRefGoogle Scholar
  73. Rodgers RJ, Cole JC (1993) Influence of social isolation, gender, strain and prior novelty on plus-maze behaviour in mice. Physiol Behav 54:729–736PubMedCrossRefGoogle Scholar
  74. Rodgers RJ, Cao B-J, Dalvi A, Holmes A (1997) Animal models of anxiety: an ethological perspective. Braz J Med Biol Res 30:289–304PubMedCrossRefGoogle Scholar
  75. Roy-Byrne PP, Davidson KW, Kessler RC, Asmundson GJG, Goodwin RD, Kubzansky L, Lydiard RB, Massie MJ, Katon W, Laden SK, Stein MB (2008) Anxiety disorders and comorbid illness. Gen Hosp Psychiatr 30:208–225CrossRefGoogle Scholar
  76. Sartori SB, Landgraf R, Singewald N (2011) The clinical implications of mouse models of enhanced anxiety. Future Neurol 6:531–571PubMedCrossRefGoogle Scholar
  77. Schalkwyk LC, Fernandes C, Kember R, Lad HV, Liu L, Parsons MJ, Paya-Cano JL (2010) Battery of behavioral testing in males of 8 inbred mouse strains. In: MPD Schalkwyk (ed) Mouse Phenome Database website, The Jackson Laboratory, Bar Harbor, Maine USA. http://phenome.jax.org, Mar, 2012
  78. Schellinck HM, Cyr D, Brown RE (2010) How many ways can mouse behavioral experiments go wrong? Confounding variables in mouse models of neurodegenerative diseases and how to control them. Adv Study Behav 41:255–365CrossRefGoogle Scholar
  79. Schüle C, Eser D, Baghai TC, Nothdurfter C, Kessler JS, Rupprecht R (2011) Neuroactive steroids in affective disorders: target for novel antidepressant or anxiolytic drugs? Neuroscience 191:55–77PubMedCrossRefGoogle Scholar
  80. Spruijt BM, Van Hoof JARAM, Gispen WH (1992) Ethology and neurobiology of grooming behaviour. Physiol Rev 72:825–852PubMedGoogle Scholar
  81. Tasan RO, Bukovac A, Peterschmitt YN, Sartori SB, Landgraf R, Singewald N, Sperk G (2011) Altered GABA transmission in a mouse model of increased trait anxiety. Neuroscience 183:71–80PubMedCrossRefGoogle Scholar
  82. Trullas R, Skolnick P (1993) Differences in fear motivated behaviors among inbred mouse strains. Psychopharm 111:323–331CrossRefGoogle Scholar
  83. Turri MG, Datta SR, DeFries J, Henderson ND, Flint J (2001) QTL analysis identifies multiple behavioral dimensions in ethological tests of anxiety in laboratory mice. Curr Biol 11:725–734PubMedCrossRefGoogle Scholar
  84. Van Bogaert MJV, Groenink L, Oosting RS, Westphal KGC, van der Gugten J, Olivier B (2006) Mouse strain differences in autonomic responses to stress. Genes Brain Behav 5:139–149PubMedCrossRefGoogle Scholar
  85. Võikar V, Kõks S, Vasar E, Rauvala H (2001) Strain and gender differences in the behavior of mouse lines commonly used in transgenic studies. Physio Behav 72:271–281CrossRefGoogle Scholar
  86. Wahlsten D, Crabbe JC (2003) Survey of motor activity, behavior, and forebrain morphology in 21 inbred strains of mice across two laboratories. In: MPD Wahlsten (ed) Mouse Phenome Database website, The Jackson Laboratory, Bar Harbor, Maine USA. http://phenome.jax.org, Mar, 2012
  87. Wahlsten D, Bachmanov A, Finn DA, Crabbe JC (2006) Stability of inbred mouse strain differences in behavior and brain size between laboratories and across decades. Proc Natl Acad Sci USA 103:16364–16369PubMedCrossRefGoogle Scholar
  88. Walsh RN, Cummins RA (1976) The open-field test: a citical review. Psych Bull 83:482–504CrossRefGoogle Scholar
  89. Wehrmeister TD, Izídio GS, Pereira E, Izídio G, Ramos A (2010) Absence of repeated-trial tolerance to the anxiolytic-like effects of chlordiazepoxide in the rat triple test. Pharmacol Biochem Behav 97:301–309PubMedCrossRefGoogle Scholar
  90. Wiltshire T, Tarantino LM (2010) Measurement of exploratory behavior and activity in 38 inbred strains of mice. In: MPD Wiltshire (ed) Mouse Phenome Database website, The Jackson Laboratory, Bar Harbor, Maine USA. http://phenome.jax.org, Mar, 2012

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Timothy P. O’Leary
    • 1
  • Rhian K. Gunn
    • 1
  • Richard E. Brown
    • 1
  1. 1.Department of Psychology and NeuroscienceDalhousie UniversityHalifaxCanada

Personalised recommendations