Measurement and Characterization of Energy Intake in the Mouse

Jan Svartengren1, Ali‐Reza Modiri1, Robert A. McArthur2

1 Biovitrum, Stockholm, 2 McArthur and Associates GmbH, Basel
Publication Name:  Current Protocols in Pharmacology
Unit Number:  Unit 5.40
DOI:  10.1002/0471141755.ph0540s28
Online Posting Date:  April, 2005
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Because of the dramatic increase in obesity and related conditions, such as type 2 diabetes, efforts have intensified to develop medications to assist in losing weight or in minimizing weight gain. To this end, methods that allow for the continuous monitoring of metabolically relevant functions in laboratory animals have been developed to help identify novel anorectic and thermogenic agents. Described in this unit is an in vivo procedure for simultaneous recording of feeding, drinking, and motor activity in mice. Data obtained using reference compounds are presented to illustrate how results are calculated, including the minimum effective dose and the dose producing a half‐maximal effect (ED50), as well as the time of onset and duration of action. Information derived from this procedure reveals the specificity of an anorectic effect, which, when combined with parameters of meal patterns, allows for inferences to be made about the effects of test compounds on satiety and hunger.

Keywords: amphetamine; appetite; drinking; eatometer; energy homeostasis; feeding; hunger; intermeal interval; mCPP; obesity; satiety

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Table of Contents

  • Commentary
  • Literature Cited
  • Figures
  • Tables
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Basic Protocol 1:

  • 8‐ to 9‐week‐old male C57BL/6J or C57BL/6JBom‐Lepob (ob/ob; Taconic) mice weighing approximately 25 and 45 g, respectively, upon arrival to the animal care facility
  • Nesting material, such as Nestlets (Ancare) or Happi‐Mat (Scanbur)
  • Standard laboratory mouse chow (e.g., Diet R34; Lactamin)
  • 20‐mg dust‐free precision pellets (Bio‐Serv)
  • Test compound (preferably dissolved in water or saline, acidified to no lower than pH 5 if needed to improve solubility) and appropriate vehicle
  • Appropriate reference compound dissolved in 0.9% (w/v) NaCl on the day of treatment, such as:
    • m‐1‐(3‐Chlorophenyl)piperazine dihydrochloride (mCPP; Sigma)
    • D‐Amphetamine sulfate (Sigma)
    • (±)‐Fenfluramine hydrochloride (Sigma)
  • Macrolon mouse cages (Scanbur)
  • Monitoring chamber (Habitest Animal Behavior Test System; Coulbourn Instruments; Fig. ) including appropriate computer with WinLinc (Version; Coulbourn Instruments) software (updated version “Graphic State Notation” available)
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Literature Cited

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   Pelleymounter, M.A., Kant, R.H., and Aravich, P. 1999. Models for environmentally inducing eating disorders: Dietary hyperphagia and anorexia nervosa. In Current Protocols in Pharmacology (S.J. Enna, M. Williams, J.F. Barrett, J.W. Ferkany, T. Kenakin, and R.D. Porsolt, eds.) pp. 5.39.1‐5.39.15. John Wiley & Sons, Hoboken, N.J.
   Petersen, S. and McCarthy, J.C. 1981. Correlated changes in feeding behavior on selection for large and small body size in mice. Behav. Genet. 11:57‐64.
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Key References
   Alberts, P., Johansson, B.G., and McArthur, R.A. 2005. Measurement and characterization of energy expenditure as a tool in the development of drugs for metabolic diseases. In Current Protocols in Pharmacology (S.J. Enna, M. Williams, J.F. Barrett, J.W. Ferkany, T. Kenakin, and R.D. Porsolt, eds.) pp. 5.39.1‐5.39.15. John Wiley & Sons, Hoboken, N.J.
  Detailed protocol on measuring energy expenditure in mice as a fundamental step in development of antiobesity drugs.
   Blundell and Latham, 1982. See above.
  Theoretical discussion of the process of continuous monitoring of feeding behavior and the microstructure of feeding techniques used in drug discovery of antiobesity drugs.
   Clifton, 2000. See above.
  Theoretical discussion of the determination of meal criteria, use of meal parameters to elucidate seritonergic and dopaminergic mechanisms and their neural substrates.
   Halford and Blundell, 1998. See above.
  Detailed protocol on behavioral satiety sequence (BSS) for identifying behavioral specificity of appetite suppressants used in development of antiobesity drugs.
   Panksepp, 1978. See above.
  Review of meal patterns, data reduction, and the theoretical implications for these techniques when studying feeding behavior and drug‐induced changes.
   Pelleymounter et al., 1999. See above.
  Detailed protocol on developing rodent models of obesity and the modification of continuous‐monitoring techniques to measure intake from nonpellet diets as a fundamental step in development of antiobesity drugs.
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