Pharmacological Investigation of Serial Anxiety Tests in the Mouse: A Pilot Study

Main Article Content

A. M. Umarudeen
M. G. Magaji
O. S. Bello
C. Aminu
M. I. Abdullahi


The use of test batteries has been suggested as a means to enhance throughput, to broaden the spectrum of anxiety parameters detectable and to minimise the numbers of experimental animals required in behavioural studies. This study was undertaken to determine the sensitivity of test batteries each of open field, zero mazes and staircase tests to anxiety behaviours in randomised mouse groups. Mice (n = 8) were exposed to these apparatuses serially in that order, thirty minutes following intraperitoneal administration of distilled water, 0.5 and 1.0 mg/kg diazepam.  Each mouse was allowed to spend 5 minutes to freely explore each test apparatus.  The results showed diazepam-treated mice exhibited significantly (p<0.05) reduced anxiety behaviours compared to the placebo group on most rodent anxiety parameters evaluated. The findings of this study suggest these behavioural test apparatuses, when used in serial combination, are sensitive and reliable to measure murine anxiety-related behaviours and the anxiolytic effects of standard/putative agents.

Evaluation, open-field, elevated zero-maze, staircase, mice

Article Details

How to Cite
Umarudeen, A. M., Magaji, M. G., Bello, O. S., Aminu, C., & Abdullahi, M. I. (2020). Pharmacological Investigation of Serial Anxiety Tests in the Mouse: A Pilot Study. Journal of Advances in Medical and Pharmaceutical Sciences, 22(4), 25-31.
Original Research Article


Craske MG, Stein MB. Anxiety. The Lancet. 2016;388(10063):3048-3059.

Gale C, Herbison GP, Glue P, Coverdale J, Guaiana G. Benzodiazepines for generalised anxiety disorder (GAD). Cochrane Database of Systematic Reviews; 2012.

Bystritsky A, Khalsa SS, Cameron ME, Schiffman J. Current diagnosis and treatment of anxiety disorders. P and T. 2013;38:30–44.

Chioca LR, Ferro MM, Baretta IP, Oliveira SM, Silva CR, Ferreira J, Losso EM, Andreatini R. Anxiolytic-like effect of lavender essential oil inhalation in mice: Participation of serotonergic but not GABAA/benzodiazepine neuro-transmission. Journal of Ethnopharmacology. 2013;147(2): 412-418.

Baxter AJ, Vos T, Scott KM, Ferrari AJ, Whiteford HA. The global burden of anxiety disorders in 2010. Psychological Medicine. 2014;44:2363–2374.

Bandelow B, Michaelis S. Epidemiology of anxiety disorders in the 21st century. Dialogues in Clinical Neuroscience. 2015; 17(3):327-335.

Evans BE, Rittle KE, Bock MG, DiPardo RM, Freidinger RM, Whitter WL, et al. Methods for drug discovery: Development of potent, selective, orally effective cholecystokinin antagonists. Journal of Medicinal Chemistry. 1988;31(12):2235-2246.

Cryan JF, Sweeney FF. The age of anxiety: Role of animal models of anxiolytic action in drug discovery. British Journal of Pharmacology. 2011;164:1129–1161.

Cryan JF, Holmes A. Model organisms: The ascent of mouse: Advances in modelling human depression and anxiety. Nature Reviews Drug Discovery. 2005;4: 775–790.

Ago Y, Takahashi K, Nakamura S, Hashimoto H, Baba A, Matsuda T. Anxiety-like and exploratory behaviours of isolation-reared mice in the staircase test. Journal of Pharmacological Sciences. 2007;104:153–158.

Campos AC, Fogaça MV, Aguiar DC, Guimarães FS. Animal models of anxiety disorders and stress. Revista Brasileira de Psiquiatria. 2013;35(SUPPL. 2):S101-11.

DOI: 10.1590/1516-4446-2013-1139

Crawley JN, Paylor R. A proposed test battery and constellations of specific behavioural paradigms to investigate the behavioural phenotypes of transgenic and knockout mice. Hormones and Behavior. 1997;31:197–211.

Schmitt U, Hiemke C. Combination of open field and elevated plus-maze: A suitable test battery to assess strain as well as treatment differences in rat behaviour. Progress in Neuro-Psychopharmacology and Biological Psychiatry. 1998;22(7): 1197-1215.

Ramos A, Pereira E, Martins GC, Wehrmeister TD, Izídio GS. Integrating the open field, elevated plus maze and light/dark box to assess different types of emotional behaviours in one single trial. Behavioural Brain Research. 2008;193: 277–288.

Sartori SB, Whittle N, Hetzenauer A, Singewald N. Magnesium deficiency induces anxiety and HPA axis dysregulation: Modulation by therapeutic drug treatment. In Neuropharmacology. 2012;62(1):304–312.

Holmes A, Yang RJ, Crawley JN. Evaluation of an anxiety-related phenotype in galanin overexpressing transgenic mice. In the Journal of Molecular Neuroscience. 2002;18(1-2):151-165.

Qiu ZK, Zhang LM, Zhao N, Chen HX, Zhang YZ, Liu YQ, Mi TY, Zhou WW, Li Y, Yang RF, Xu JP, Li YF. Repeated administration of AC-5216, a ligand for the 18kDa translocator protein, improves behavioural deficits in a mouse model of post-traumatic stress disorder. Progress in Neuro-Psychopharmacology and Biological Psychiatry. 2013;45:40–46.

Gray VC, Hughes RN. Drug-, dose- and sex-dependent effects of chronic fluoxetine, reboxetine and venlafaxine on open-field behaviour and spatial memory in rats. Behavioural Brain Research. 2015; 281:43–54.

Singer ML, Oreschak K, Rhinehart Z, Robison BD. Anxiolytic effects of fluoxetine and nicotine exposure on exploratory behaviour in zebrafish. Peer J. 2016;4:e2352.

Ramos A. Animal models of anxiety: do I need multiple tests? Trends in Pharmacological Sciences. 2008;29:493–498.

Seibenhener ML, Wooten MC. Use of the Open Field Maze to Measure Locomotor and Anxiety-like Behavior in Mice. Journal of Visualized Experiments. 2015;96: e52434.

Pellow S, Chopin P, File SE, Briley M. Journal of Neuroscience Methods. 1985; 14(3):149-167.

Lister RG. The use of a plus-maze to measure anxiety in the mouse. Psychopharmacology. 1987;92(2):180-185.

Singh K, Bishnoi M, Kulkarni SK. Elevated Zero-maze: A paradigm to evaluate anti-anxiety effects of drugs. Methods and Findings in Experimental and Clinical Pharmacology. 2007;29(5):343-8.

Braun AA, Skelton MR, Vorhees CV, Williams MT. Comparison of the elevated plus and elevated zero mazes in treated and untreated male Sprague-Dawley rats: Effects of anxiolytic and anxiogenic agents. Pharmacology Biochemistry and Behavior. 2011;97:406–415.

Simiand J, Keane PE, Morre M. The staircase test in mice: A simple and efficient procedure for the primary screening of anxiolytic agents. Psychopharmacology. 1984;84:48–53.

Mönnikes H, Schmidt BG, Taché Y. Psychological stress-induced accelerated colonic transit in rats involves hypothalamic corticotropin-releasing factor. Gastroenterology. 1883;104:716–723.

Babygirija R, Yoshimoto S, Gribovskaja-Rupp I, Bülbül M, Ludwig K, Takahashi T. Social interaction attenuates stress responses following chronic stress in maternally separated rats. Brain Research. 1469:2012:54–62.

Kim TE, Jung JH, Yoon S, Kim SJ. Anti-anxiety effect of methanol extract of Pericarpium zanthoxyli using a strychnine-sensitive glycine receptor model. Tropical Journal of Pharmaceutical Research. 2016;15:951–957.

Kulkarni SK, Singh K, Bishnoi M. Involvement of adenosinergic receptors in anxiety related behaviours. Indian Journal of Experimental Biology. 2007;45:439–443.