ORIGINAL INVESTIGATION Measuring anxiety- and locomotion-related behaviours in mice: a new way of using old tests Leanne M. Fraser & Richard E. Brown & Ahmed Hussin & Mara Fontana & Ashley Whittaker & Timothy P. O’Leary & Lauren Lederle & Andrew Holmes & André Ramos Received: 26 December 2009 / Accepted: 19 April 2010 / Published online: 8 May 2010 # Springer-Verlag 2010 Abstract Rationale Batteries of tests that are thought to measure different aspects of anxiety-related behaviour are used to characterise mice after genetic or pharmacological manip- ulation. However, because of the potentially confounding effects of repeated testing and natural intra-individual variations in behaviour over time, subjecting mice to a succession of tests is not ideal. Objectives The aim of this study was to investigate, in mice, the utility of an integrated apparatus that combines three classical tests of anxiety, the open field, elevated plus maze (EPM) and light/dark box. Methods Mice from four different strains (CD-1, BALB/cJ, DBA/2J, C57BL/6J) were used in a series of five experi- ments where their behaviour was observed for 15 min in the integrated apparatus. Responses to anxiety-modulating drugs and 2-day repeated testing were evaluated. Results CD-1 mice explored the apparatus thoroughly, providing measures from all areas throughout the entire testing session. Factor analysis showed that measures of locomotion and anxiety-related behaviour were dissociable. BALB/cJ, DBA/2J and C57BL/6J showed markedly differ- ent behavioural profiles, largely consistent with previous studies examining individual tests. Avoidance of aversive environments did not increase with repeated testing. In CD- 1 mice, the anxiolytics diazepam and alprazolam (4 and 2 mg/kg, respectively) increased the approach towards the EPM open arms. Alprazolam also had sedative effects, whereas the anxiogenic pentylenetetrazole had no effects. Conclusions These findings suggest that the triple test is sensitive to genetic/pharmacological influences on anxiety and locomotion and that, by providing quasi-simultaneous measures from three different apparatuses, it may represent an alternative to the use of test batteries. Keywords Anxiety . Behavioural test . Mice . Open field . Elevated plus maze . Light/dark box . Strain differences . Phenotyping Introduction Numerous animal species, including humans, display defensive behaviours (e.g. escape, avoidance, risk assess- ment) as an adaptive response to potentially threatening situations. Whereas momentary defensive reactions that ensure survival can be associated with a normal state of anxiety, the exaggerated or inappropriately prolonged expression of these behaviours is thought to lead to the so-called anxiety-related disorders (Rodgers and Johnson 1995; Bakshi and Kalin 2002; Cryan and Holmes 2005). The study of defensive behaviours (hereafter referred to as L. M. Fraser was funded by NSERC, Canada and A. Ramos had a fellowship from CAPES, Brazil. L. M. Fraser : R. E. Brown : A. Hussin : M. Fontana : A. Whittaker : T. P. O’Leary Psychology Department and Neuroscience Institute, Dalhousie University, Halifax, Nova Scotia, Canada B3H 4J1 L. Lederle : A. Holmes Section on Behavioral Science and Genetics, Laboratory for Integrative Neuroscience, National Institute on Alcoholism and Alcohol Abuse, NIH, 5625 Fishers Lane Room 2N09, Rockville, MD 20852-9411, USA A. Ramos (*) Laboratório de Genética do Comportamento, Departamento de Biologia Celular, Embriologia e Genética, Universidade Federal de Santa Catarina, 88.040-900 Florianópolis, Santa Catarina, Brazil e-mail: andre@ccb.ufsc.br Psychopharmacology (2010) 211:99–112 DOI 10.1007/s00213-010-1873-0