Measurement and Characterization of Energy Intake in the Mouse

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

1 Biovitrum, Stockholm, null, 2 McArthur and Associates GmbH, Basel, null
Publication Name:  Current Protocols in Pharmacology
Unit Number:  Unit 5.40
DOI:  10.1002/0471141755.ph0540s28
Online Posting Date:  April, 2005
GO TO THE FULL TEXT: PDF or HTML at Wiley Online Library

Abstract

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

     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Table of Contents

  • Commentary
  • Literature Cited
  • Figures
  • Tables
     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Materials

Basic Protocol 1:

  Materials
  • 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 1.0.0.0; Coulbourn Instruments) software (updated version “Graphic State Notation” available)
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Figures

  •   FigureFigure 5.40.1 Flow diagram of data collection and analysis of feeding, drinking, and activity events by the WinLinc process control program (Coulbourn Instruments). WinLinc is used to control each eight‐station test system for a user‐specified length of time. It controls the switching on and off of pellet dispensers, light switches, and other components. It also records all feeding, drinking, and activity events in real time and time stamps each event. The files produced by WinLinc can be converted into Excel files for further manipulation of data.
  •   FigureFigure 5.40.2 A Habitest Animal Behavior Test System continuous‐monitoring chamber (Coulbourn Instruments). The entire apparatus is divided into four systems (A‐D) of eight chambers each. Each continuous‐monitoring chamber consists of a single pellet feeder (H14‐23), a combination pellet trough (H14‐01) and optical sensor (H20‐93), a single optical lickometer (H24‐01) attached to a 120‐ml water bottle, and an infrared motion detector sensor (H24‐61) mounted in the ceiling of each chamber. The system uses a Kissileff‐type eatometer (Kissileff, ) consisting of an optical sensor that detects the presence of a 20‐mg food pellet in the trough. Each time the mouse removes a pellet, an electronic signal from the sensor activates the pellet dispenser to deliver a new pellet.
  •   FigureFigure 5.40.3 Hourly interval representation of the number of 20‐mg food pellets taken by a single ob/ob mouse over the course of a 22‐hr recording session from the data set in Figure . The black bar extending from 1‐2 hr to 12‐13 hr indicates the 12‐hr dark period imposed on this animal.
  •   FigureFigure 5.40.4 Cumulative recording of the number of 20‐mg food pellets taken by a single ob/ob mouse over the course of a 22‐hr recording session. The black bar extending from the start of recording (17:00) indicates the 12‐hr dark period imposed on this animal. This is a typical pattern and shows that although the ob/ob mouse eats in clusters or meals during both the dark and light phases, most of the pellets are eaten during the dark phase.
  •   FigureFigure 5.40.5 Frequency distribution of number of meals generated as a function of increasing the minimum interval between pellet deliveries from 0 to 2500 sec during 12 hr of dark phase eating in sixteen ob/ob mice. The point of inflection of this frequency distribution indicates that most concentrated bouts of feeding activity occur at intervals of ≤200 sec. Consequently, this interval of 200 sec has been used as the minimal interval to define the start and end of one meal in this species of mouse under these experimental conditions.
  •   FigureFigure 5.40.6 The effect of varying the number of pellets per meal while maintaining a constant intermeal interval (IMI) of 200 sec on the number of meals generated by the MDAT/MLAN analysis program. These data represent the mean number of meals (± standard error) generated by the 12‐hr dark period feeding activity of a group of 8 ob/ob mice. The assumption is that a full meal represents not a single pellet but five or more pellets. The data indicate that this criterion does not influence the number of meals appreciably. The relationship between meal number and meal size is linear.
  •   FigureFigure 5.40.7 Basal food intake, water intake, and total motor activity of ob/ob ( n = 28; right panel) and their C57BL/6J ( n = 32; left panel) background strain controls during 3 days of continuous monitoring in eatometer chambers (day 1‐2, day 2‐3, and day 3‐4). The mice were handled daily before being placed in the eatometer chambers and allowed to eat undisturbed while their behaviors were continuously monitored for 22 hr each day. The black bar represents the 12 hr of dark from 17:00 to 05:00. Each data point indicates the daily group mean of each strain. Standard errors of the means (∼20%) have been omitted for the sake of clarity.
  •   FigureFigure 5.40.8 Effects of m‐1‐(3‐chlorophenyl)piperazine dihydrochloride (mCPP) on the feeding, water intake, and total motor activity of three groups each of ob/ob mice (right panel) and their C57BL/6J (left panel) background strain controls ( n = 8 per group). The mice were handled daily for 4 days to habituate them to the experimental procedure. At the end of the fourth day (day 4‐5), the mice were administered either saline or 3.0 or 10.0 mg/kg mCPP, p.o., within 1 hr before the start of a 12‐hr dark phase. These 12 hr are represented by the dark bar in each graph. The animals were replaced in the chambers and left undisturbed while their feeding, drinking, and motor activity were recorded for 21 hr.
  •   FigureFigure 5.40.9 Effects of amphetamine on food intake, water intake, and total motor activity in ob/ob mice. The procedure is the same as used for mCPP (Fig. ). At the end of the fourth day (day 4‐5), the mice were administered either saline or 0.5 or 2.0 mg/kg of amphetamine intraperitoneally within 1 hr before the start of a 12‐hr dark phase. These 12 hr are represented by the dark bar in each graph. The animals were replaced in the chambers and left undisturbed while their feeding, drinking, and motor activity were recorded for 21 hr.

Videos

Literature Cited

   Anliker, J. and Mayer, J. 1956. An operant conditioning technique for studying feeding‐fasting patterns in normal and obese mice. J. Appl. Physiol. 8:667‐670.
   Bailey, C.J. and Flatt, P.R. 1986. Anorectic effect of fenfluramine, cholecystokinin and neurotensin in genetically obese (ob/ob) mice. Comp. Biochem. Physiol. A 84:451‐454.
   Bickerdike, M.J. 2003. 5‐HT2C receptor agonists as potential drugs for the treatment of obesity. Curr. Top. Med. Chem. 3:885‐897.
   Blundell, J.E. and Latham, C.J. 1979. Serotonergic influences on food intake: Effect of 5‐hydroxytryptophan on parameters of feeding behavior in deprived and free‐feeding rats. Pharmacol. Biochem. Behav. 11:431‐437.
   Blundell, J.E. and Latham, C.J. 1982. Behavioral pharmacology of feeding. In Drugs and Appetite (T. Silverstone, ed.) pp. 41‐80. Academic Press, London.
   Blundell, J.E., Latham, C.J., and Leshem, M.B. 1976. Differences between the anorexic actions of amphetamine and fenfluramine—possible effects on hunger and satiety. J. Pharmacy Pharmacol. 28:471‐477.
   Bray, G.A. 2001. Drug treatment of obesity. Rev. Endocr. Metab. Disord. 2:403‐418.
   Brockmann, G.A. and Bevova, M.R. 2002. Using mouse models to dissect the genetics of obesity. Trends Genet. 18:367‐376.
   Clapham, J.C. 2004. Treating obesity: Pharmacology of energy expenditure. Curr. Drug Targets 5:309‐323.
   Clifton, P.G. 1987. Analysis of feeding and drinking patterns. In Methods for the Study of Feeding and Drinking (F.M. Toates and N.R. Rowland, eds.) pp. 19‐35. Elsevier Science Publishers B.V., Amsterdam.
   Clifton, P.G. 2000. Meal patterning in rodents: Psychopharmacological and neuroanatomical studies. Neurosci. Biobehav. Rev. 24:213‐222.
   Clifton, P.G., Lee, M.D., and Dourish, C.T. 2000. Similarities in the action of Ro 60‐0175, a 5‐HT2C receptor agonist and D‐fenfluramine on feeding patterns in the rat. Psychopharmacology 152:256‐267.
   Cox, J.E. and Powley, T.L. 1977. Development of obesity in diabetic mice pair‐fed with lean siblings. J. Comp. Physiol. Psychol. 91:347‐ 358.
   Craddock, D. 1976. Anorectic drugs: Use in general practice. Drugs 11:378‐393.
   Dantzer, R., Bluthe, R.M., Gheusi, G., Cremona, S., Laye, S., Parnet, P., and Kelley, K.W. 1998. Molecular basis of sickness behavior. Ann. N.Y. Acad. Sci. 856:132‐138.
   Fox, E.A., Phillips, R.J, Baronowsky, E.A., Byerly, M.S., Jones, S., and Powley, T.L. 2001. Neurotrophin‐4 deficient mice have a loss of vagal intraganglionic mechanoreceptors from the small intestine and a disruption of short‐term satiety. J. Neurosci. 21:8602‐8615.
   Gannon, K.S., Smith, J.C., Henderson, R., and Hendrick, P. 1992. A system for studying the microstructure of ingestive behavior in mice. Physiol. Behav. 51:515‐521.
   Halford, J.C.G. and Blundell, J.E. 1998. Direct and continuous behavioral analysis for the diagnosis of drug action on feeding. In Current Protocols in Neuroscience (J.N. Crawley, C.R. Gerfen, M.A. Rogawski, D.R. Sibley, P. Skolnick, and S. Wray, eds.) pp. 8.6C.1‐8.6C.11. John Wiley & Sons, Hoboken, N.J.
   Halford, J.C.G., Wanninayake, S.C., and Blundell, J.E. 1998. Behavioral satiety sequence (BSS) for the diagnosis of drug action on food intake. Pharmacol. Biochem. Behav. 61:159‐168.
   Halford, J.C.G., Cooper, G.D., and Dovey, T.M. 2004. The pharmacology of human appetite expression. Curr. Drug Targets 5:221‐240.
   Harrold, J.A. 2004. Hypothalamic control of energy balance. Curr. Drug Targets 5:207‐219.
   Ho, A. and Chin, A. 1988. Circadian feeding and drinking patterns of genetically obese mice fed solid chow diet. Physiol. Behav. 43:651‐656.
   Inui, A. 2000. Transgenic approach to the study of body weight regulation. Pharmacol. Rev. 52:35‐61.
   Kissileff, H.R. 1970. Free feeding in normal and “recovered lateral” rats monitored by a pellet‐detecting eatometer. Physiol. Behav. 5:163‐173.
   Ladenheim, E.E., Hampton, L.L., Whitney, A.C., White, W.O., Battey, J.F., and Moran, T.H. 2002. Disruptions in feeding and body weight control in gastrin‐releasing peptide receptor deficient mice. J. Endocrinol. 174:273‐281.
   Meguid, M.M., Kawashima, Y., Campos, A.C.L., Gelling, P.D., Hill, T.W., Chen, T.‐Y., Yang, Z.‐J., Hitch, D.C., Hammond, W.G., and Mueller, W.J. 1990. Automated computerized rat eatometer: Description and application. Physiol. Behav. 48:759‐763.
   Panksepp, J. 1978. Analysis of feeding patterns: Data reduction and theoretical considerations. In Hunger Models: Quantitative Theory of Feeding Control (D. Booth, ed.) pp. 143‐166. Academic Press, London.
   Panksepp, J. and Ritter, M. 1975. Mathematical analysis of energy regulatory patterns of normal and diabetic rats. J. Comp. Physiol. Psychol. 89:1019‐1028.
   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.
   Silverstone, T. 1992. Appetite suppressants. A review. Drugs 43:820‐836.
   Strohmayer, A.J. and Smith, G.P. 1987. The meal pattern of genetically obese (ob/ob) mice. Appetite 8:111‐123.
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.
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library