Tests for Anxiety‐Related Behavior in Mice

Sabine M. Hölter1, Jan Einicke1, Bettina Sperling1, Annemarie Zimprich1, Lillian Garrett1, Helmut Fuchs2, Valerie Gailus‐Durner2, Martin Hrabé de Angelis3, Wolfgang Wurst4

1 German Mouse Clinic, Helmholtz Zentrum München, Munich, 2 Institute of Experimental Genetics, Helmholtz Zentrum München, German Research Centre for Environmental Health, Munich, 3 German Centre for Diabetes Research (DZD), Neuherberg, 4 Munich Cluster for Systems Neurology (SyNergy), Munich
Publication Name:  Current Protocols in Mouse Biology
Unit Number:   
DOI:  10.1002/9780470942390.mo150010
Online Posting Date:  December, 2015
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Abstract

Phenotyping of inbred mouse strains and genetically modified mouse models for characteristics related to neuropsychiatric diseases includes assessing their anxiety‐related behavior. A variety of tests have been developed to measure anxiety in laboratory rodents and these tests have been placed under scrutiny over the years concerning their validity. Here we describe the most widely used tests for anxiety in mice. The protocols we present are established methods used in the German Mouse Clinic (GMC), with which alterations in anxiety could successfully be discovered in mouse mutants. Moreover, since baseline anxiety levels in mice are easily influenced by a great variety of disturbances, we carefully outline the critical parameters that need to be considered. © 2015 by John Wiley & Sons, Inc.

Keywords: open field; light/dark box; elevated plus‐maze; social interaction; anxiety

     
 
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Table of Contents

  • Introduction
  • Basic Protocol 1: Open Field Test to Measure Anxiety in Mice
  • Basic Protocol 2: Light/Dark Box Test to Measure Anxiety in Mice
  • Basic Protocol 3: Elevated Plus‐Maze Test to Measure Anxiety in Mice
  • Basic Protocol 4: Social Interaction Test to Measure Social Anxiety in Mice
  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1: Open Field Test to Measure Anxiety in Mice

  Materials
  • Mice (e.g., Charles River)
  • Disinfectant solution (e.g., Pursept‐A Xpress, Merz Hygiene GmbH)
  • Sound‐attenuated testing room with adjustable light source
  • Plexiglas Open Field arenas (TSE systems; dimensions: 45 × 45 × 40 cm), elevated off the ground and positioned in an infrared beam‐break sensor frame connected with a beam‐break tracking interface and software (e.g., Actimot system, TSE systems)
  • Example worksheet.xlsx: worksheet containing list of mouse ID and cage number, genotype, sex, file number, date, weight (see http://www.currentprotocols.com/protocol/mo150010)
  • Light (lux) meter

Basic Protocol 2: Light/Dark Box Test to Measure Anxiety in Mice

  Materials
  • Mice (e.g., Charles River)
  • Disinfectant solution (e.g., Pursept‐A Xpress, Merz Hygiene GmbH)
  • Sound‐attenuated testing room with adjustable light source
  • Plexiglas light/dark box testing chamber (TSE systems; dimensions: dark box 14 × 22.6 × 26 cm; light box 26.1 × 22.6 × 26 cm; tunnel 4.5 × 5.6 × 13 cm; see Fig.  ), elevated off the ground and positioned in an infrared beam‐break sensor frame connected with a beam‐break tracking interface and software (e.g., Actimot system, TSE systems)
  • Example worksheet.xlsx: worksheet containing list of mouse ID and cage number, genotype, sex, file number, date, weight (see http://www.currentprotocols.com/protocol/mo150010)
  • Lux meter

Basic Protocol 3: Elevated Plus‐Maze Test to Measure Anxiety in Mice

  Materials
  • Mice (e.g., Charles River)
  • Disinfectant solution (e.g., Pursept‐A Xpress, Merz Hygiene GmbH)
  • Sound‐attenuated testing room with adjustable light source
  • Elevated Plus‐Maze (EPM; see Fig.  )
    • EPM floor illuminated by infrared light; dimensions of arena: 66 × 6 cm; height of wall of enclosed arms: 15 cm, open arms are surrounded by a small rim of 0.5 cm height; EPM elevated ∼70 cm above the ground
    • Small chamber (1 × 1 × 3.6 m) to house EPM apparatus
  • Lamps (two; for illumination of open arms of EPM from above, which illuminate the ends of the open arms each directly with 300 lux, resulting in an indirect illumination of enclosed arms with ∼35 lux).
  • Infrared‐sensitive camera (position above EPM)
  • Video‐tracking software (e.g., Ethovision Software, Noldus Information Technology)
  • Example worksheet.xlsx: worksheet containing list of mouse ID and cage number, genotype, sex, file number, date, weight (see http://www.currentprotocols.com/protocol/mo150010)
  • Lux meter

Basic Protocol 4: Social Interaction Test to Measure Social Anxiety in Mice

  Materials
  • Mice (e.g., Charles River)
  • Disinfectant solution (e.g., Pursept‐A Xpres, Merz Hygiene GmbH)
  • Sound‐attenuated testing room with adjustable light source
  • Psion workabout (handheld computer) with pocket observer program (Noldus Information Technology)
  • Fresh cages without litter as test cages
  • Timer
  • Example worksheet.xlsx: worksheet containing list of mouse ID and cage number, genotype, sex, file number, date, weight (see http://www.currentprotocols.com/protocol/mo150010)
  • Lux meter
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Figures

Videos

Literature Cited

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Supplementary Material