Reliability, robustness, and reproducibility in mouse behavioral phenotyping: a cross-laboratory study

Abstract

Establishing standard operating procedures (SOPs) as tools for the analysis of behavioral phenotypes is fundamental to mouse functional genomics. It is essential that the tests designed provide reliable measures of the process under investigation but most importantly that these are reproducible across both time and laboratories. For this reason, we devised and tested a set of SOPs to investigate mouse behavior. Five research centers were involved across France, Germany, Italy, and the UK in this study, as part of the EUMORPHIA program. All the procedures underwent a cross-validation experimental study to investigate the robustness of the designed protocols. Four inbred reference strains (C57BL/6J, C3HeB/FeJ, BALB/cByJ, 129S2/SvPas), reflecting their use as common background strains in mutagenesis programs, were analyzed to validate these tests. We demonstrate that the operating procedures employed, which includes open field, SHIRPA, grip-strength, rotarod, Y-maze, prepulse inhibition of acoustic startle response, and tail flick tests, generated reproducible results between laboratories for a number of the test output parameters. However, we also identified several uncontrolled variables that constitute confounding factors in behavioral phenotyping. The EUMORPHIA SOPs described here are an important start-point for the ongoing development of increasingly robust phenotyping platforms and their application in large-scale, multicentre mouse phenotyping programs.

Unpredictable chronic stress decreases inhibitory avoidance learning in Tuebingen Long-Fin zebrafish (Danio rerio Hamilton): stronger effects in the resting phase than in the active phase

“…Reproducibility of results within and between laboratories is crucial for advancing the field of behaviour and genetics (Crabbe et al, 1999;Mandillo et al, 2008;Wahlsten et al, 2006). The zebrafish is a relatively new model in genetics and brain-behaviour research and differences between strains of zebrafish (Egan et al, 2009;Vignet et al, 2013;Wahlsten et al, 2006) as well as rearing conditions within and between laboratories (Mahabir et al, 2013;Parker et al, 2012;Pavlidis et al, 2013) may affect the outcome of experiments.…”

Absence of TI-VAMP/Vamp7 Leads to Increased Anxiety in Mice

“…All experiments were performed in accordance with the European Communities Council Directive of 24 November 1986. Modified SHIRPA, rotarod test, grip test, tail flick, hot plate, open field test, tail suspension, auditory startle reflex reactivity and prepulse inhibition, Y-maze spontaneous alternation task (Meziane et al, 2007;Mandillo et al, 2008), string test (Andrieu et al, 2006), shock threshold test, and Pavlovian fear conditioning (Lamprianou et al, 2006) were previously described. Noninvasive blood pressure was performed in conscious animals by tail cuff sphygmomanometry as described previously (Koutnikova et al, 2009).…”

Retinoic Acid Deficiency Impairs the Vestibular Function

“…The test was performed as described previously as part of the SHIRPA (acronym of SmithKline Beecham, Harwell, Imperial College, Royal London Hospital, phenotype assessment) analysis (Mandillo et al, 2008). Briefly, mice were placed in a narrow, 3-cm-wide clear Plexiglas cylinder.…”