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.
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References
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 24:549–5604
Archer J (1977) Sex differences in the emotional behaviour of laboratory mice. Br J Psychiatr 68:125–131
Beuzen A, Belzung C (1995) Link between emotional memory and anxiety states: a study by principal component analysis. Physiol Behav 58:111–118
Blanchard DC, Griebel G, Blanchard RJ (2003) The mouse defense test battery: pharmacological and behavioral assays for anxiety and panic. Eur J Pharmacol 463:97–116
Blanchard DC, Griebel G, Pobbe R, Blanchard RJ (2011) Risk assessment as an evolved threat detection and analysis process. Neurosci Biobehav Rev 35:991–998
Bolivar VJ (2009) Intrasession and intersession habituation in mice: from inbred strain variability to linkage analysis. Neurobiol Learn Mem 92:206–214
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 30:285–293
Bothe GWM, Bolivar VJ, Vedder MJ, Geistfeld JG (2005) Behavioural differences among fourteen inbred mouse strains commonly used as disease models. Comp Med 55:326–334
Bourin M, Hascoët M (2003) The mouse light/dark box test. Eur J Pharmacol 463:55–65
Bourin M, Petit-Demoulière B, Dhonnchadha BN, Hascoët M (2007) Animal models of anxiety in mice. Fundam Clin Pharmacol 21:567–574
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 20:204–209
Brown RE, Wong AA (2007) The influence of visual ability on learning and memory in 13 strains of mice. Learn Mem 14:134–144
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 29:263–271
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
Calatayud F, Coubard S, Belzung C (2004a) Emotional reactivity in mice may not be inherited but influenced by parents. Physiol Behav 80:465–474
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 67:195–206
Carobrez AP, Bertoglio LJ (2005) Ethological and temporal analyses of anxiety-like behavior: the elevated plus-maze model 20 years on. Neurosci Biobehav Rev 29:1193–1205
Clément Y, Calatayud F, Belzung C (2002) Genetic basis of anxiety-like behaviour: a review. Brain Res Bull 57:57–71
Clenet F, Bouyon E, Hascoet M, Bourin M (2006) Light/dark cycle manipulation influences mice behaviour in the elevated-plus maze. Behav Brain Res 166:140–149
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 115:468–476
Cook MN, Bolivar VJ, McFadyen MP, Flaherty L (2002) Behavioral differences among 129 substrains: implications for knockout and transgenic mice. Behav Neurosci 116:600–611
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 32:777–785
Crabbe JC, Wahlsten D, Dudek BC (1999) Genetics of mouse behavior: interactions with laboratory environment. Science 284:1670–1672
Cryan JF, Holmes A (2005) The ascent of mouse: advances in modeling human depression and anxiety. Nat Rev Drug Discov 4:775–790
Cryan JF, Sweeney FF (2011) The age of anxiety: role of animal models of anxiolytic action in drug discovery. Brit J Pharmacol 164:1129–1161
Dawson GR, Tricklebank MD (1995) Use of the elevated-plus maze in the search for novel anxiolytic agents. Trends Pharmacol Sci 16:33–36
DeFries JC, Hegmann JP, Weir MW (1966) Open-field behavior in mice: evidence for major gene effect mediated by the visual system. Science 154:1577–1579
Depino AM, Gross C (2007) Simultaneous assessment of autonomic function and anxiety-related behavior in BALB/c and C57BL/6 mice. Behav Brain Res 177:254–260
Durant C, Christmas D, Nutt D (2010) The pharmacology of anxiety. Curr Top Behav Neurosci 2:303–330
Estanislau C (2012) Cues to the usefulness of grooming behavior in the evaluation of anxiety in the elevated plus-maze. Psychol Neurosci 5:105–112
Flint J (2003) Analysis of quantitative trait loci that influence animal behavior. J Neurobiol 54:46–77
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 106:21335–21340
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 211:99–112
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 12:6559–6568
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 4:637–644
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 60:1255–1260
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 148:164–170
Grubb SC, Maddatu TP, Bult CJ, Bogue MA (2009) Mouse phenome database. Nucleic Acids Res 35:643–649
Hall C (1934) Emotional behavior in the rat: i. defecation and urination as measures of individual differences in emotionality. J Comp Psychol 18:385–403
Haller J, Alicki M (2012) Current animal models of anixiety, anxiety disorders, and anxiolytic drugs. Curr Opin Psychiatry 1:59–64
Henderson ND, Turri MG, DeFries JC, Flint J (2004) QTL analysis of multiple behavioral measures of anxiety in mice. Behav Genet 34:267–293
Hohoff C (2009) Anxiety in mice and men: a comparison. J Neural Transm 116:679–687
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 63:883–889
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 4:412–419
Jacobson LH, Cryan JF (2010) Genetic approaches to modeling anxiety in animals. Curr Top Behav Neurosci 2:161–201
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 22:153–163
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 160:1–10
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 143:169–177
Kessler RC (2007) The global burden of anxiety and mood disorders: putting ESEMeD findings into perspective. J Clin Psychiatr 68:10–19
Kessler RC, Ruscio AM, Shear K, Wittchen HU (2010) Epidemiology of anxiety disorders. Curr Top Behav Neurosci 2:21–35
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 25:4375–4384
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 99:301–316
Lister RG (1987) The use of a plus-maze to measure anxiety in the mouse. Psychopharmacology 92:180–185
Lister RG (1990) Ethologically-based animal models of anxiety disorders. Pharmacol Ther 46:321–340
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 80:1075–1086
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 34:243–255
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 2:214–219
McIlwain KL, Merriweather MY, Yuva-Paylor LA, Paylor R (2001) The use of behavioral test batteries: effects of training history. Physiol Behav 73:705–717
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:e14458
Milner LC, Crabbe JC (2008) Three murine anxiety models: results from multiple inbred strain comparisons. Genes Brain Behav 7:496–505
Nadel L (1968) Dorsal and ventral hippocampal lesions and behavior. Physiol Behav 3:891–900
Nagy ZM, Forrest EJ (1970) Open-field behavior of C3H mice: effect of size and illumination of field. Psychon Sci 20:19–21
Nagy ZM, Glaser HD (1970) Open-field behavior of C57BL/6 J mice: effect of illumination, age, and number of test days. Psychon Sci 19:143–145
Nagy ZM, Holm M (1970) Open-field behavior of C3H mice: effect of early handling, field illumination, and age at testing. Psychon Sci 19:273–275
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
Palanza P (2001) Animal models of anxiety and depression: how are females different? Neurosci Biobehav Rev 25:219–233
Paulus MP, Dulawa SC, Ralph RJ, Geyer MA (1999) Behavioral organization is independent of locomotor activity in 129 and C57 mouse strains. Brain Res 835:27–36
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
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 1:96–110
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 218:99–105
Ramos A (2008) Animal models of anxiety: do I need multiple tests? Trends Pharmacol Sci 29:493–498
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 193:277–288
Rodgers RJ, Cole JC (1993) Influence of social isolation, gender, strain and prior novelty on plus-maze behaviour in mice. Physiol Behav 54:729–736
Rodgers RJ, Cao B-J, Dalvi A, Holmes A (1997) Animal models of anxiety: an ethological perspective. Braz J Med Biol Res 30:289–304
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 30:208–225
Sartori SB, Landgraf R, Singewald N (2011) The clinical implications of mouse models of enhanced anxiety. Future Neurol 6:531–571
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
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 41:255–365
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 191:55–77
Spruijt BM, Van Hoof JARAM, Gispen WH (1992) Ethology and neurobiology of grooming behaviour. Physiol Rev 72:825–852
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 183:71–80
Trullas R, Skolnick P (1993) Differences in fear motivated behaviors among inbred mouse strains. Psychopharm 111:323–331
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 11:725–734
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 5:139–149
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 72:271–281
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
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 103:16364–16369
Walsh RN, Cummins RA (1976) The open-field test: a citical review. Psych Bull 83:482–504
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 97:301–309
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
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).
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O’Leary, T.P., Gunn, R.K. & Brown, R.E. What are We Measuring When We Test Strain Differences in Anxiety in Mice?. Behav Genet 43, 34–50 (2013). https://doi.org/10.1007/s10519-012-9572-8
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DOI: https://doi.org/10.1007/s10519-012-9572-8