Models of Anxiety: Stress‐Induced Hyperthermia (SIH) in Singly Housed Mice

Lucianne Groenink1, Christiaan Vinkers1, Ruud van Oorschot1, Berend Olivier2

1 Utrecht Institute for Pharmaceutical Sciences and Rudolf Magnus Institute of Neuroscience, Utrecht, The Netherlands, 2 Yale University School of Medicine, New Haven, Connecticut
Publication Name:  Current Protocols in Pharmacology
Unit Number:  Unit 5.16
DOI:  10.1002/0471141755.ph0516s45
Online Posting Date:  June, 2009
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Described in this unit is the stress‐induced hyperthermia (SIH) test in mice in a single‐housed format. This protocol has proven reliable in detecting the anxiolytic properties of test compounds. In this test, SIH is quantified in singly housed mice using a rectal temperature measurement as the stressor. Rectal temperature is measured twice at a 10‐min interval. Due to the stress experienced during the first temperature measurement, the temperature of the second measurement (T2) is ∼0.8° to 1.5°C higher than that of the first (T1). This difference in temperature (ΔT = T2T1) is defined as the SIH response. The SIH response is reduced by different classes of anxiolytics. The SIH test is simple and robust, it does not require training of animals, and test compound effects on motor behavior, feeding, and nociception do not affect test outcome. Furthermore, it is one of few anxiety tests that focuses on the physiological component of anxiety. Curr. Protoc. Pharmacol. 45:5.16.1‐5.16.12. © 2009 by John Wiley & Sons, Inc.

Keywords: stress; hyperthermia; body temperature; mouse; anxiety

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Table of Contents

  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
  • Tables
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Basic Protocol 1:

  • Male mice of any strain (e.g., NMRI mice, Charles River) weighing 18 to 20 g upon arrival, 10 to 12 per dose group
  • Rodent food
  • Test compound solution (see recipe)
  • Silicon oil, peanut oil, or K‐Y jelly, at room temperature
  • 27 × 16 × 12.5 ×–cm cages, for group housing
  • 12 × 18 × 13–cm cages with sawdust, for individual housing
  • Balances with accuracy of 0.5 g (e.g., Mettler PG5000)
  • 1‐ml syringes, sterile, nontoxic, and pyrogen free
  • 25‐G × 5/8‐in. (16‐mm) needles, sterile
  • Digital thermometer, NiCr‐NiAl thermocoupled with accuracy of 0.1°C (e.g., Keithley 871A)
  • Statistical analysis software (e.g., SAS, SPSS) for data analysis
NOTE: Male mice are preferred over female mice, as the SIH test is best validated in male mice. Furthermore, female mice do show SIH, but the response is considerably smaller and the test may be influenced by their estrous cycle.
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Literature Cited

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Key References
   Borsini et al., 1989. See above.
  First description of the SIH phenomenon in mice.
   Zethof et al., 1994. See above.
  Detailed description of methodology and parameters.
   VanderHeyden et al., 1997. See above.
  Detailed description of methodology and parameters singly housed version of SIH.
   Spooren et al., 2002. See above.
  Pharmacology characterization of the singly housed SIH paradigm.
   Olivier et al., 2003. See above.
  Extensive pharmacology in the singly housed SIH paradigm.
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