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Exercise Performance Tests in Mice

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  • Abstract
  • Table of Contents
  • Materials
  • Figures
  • Literature Cited

Abstract

 

Maximal exercise performance is a multifactorial process in which the cardiovascular component, the innervation of the musculature, and the contractile and metabolic properties of skeletal muscle all play key roles. Here, protocols are provided for assessment of maximal running capacity of mice on a treadmill, with a combination of short high?intensity paradigms primarily intended to test for maximal power and cardiovascular function, and longer low?intensity paradigms to assess endurance and oxidative metabolism in skeletal muscle. The coupling of treadmill running to indirect calorimetry, to correlate performance measurements to maximal oxygen consumption, is also described. Curr. Protoc. Mouse Biol. 1:141?154. © 2011 by John Wiley & Sons, Inc.

Keywords: exercise; running; treadmill; VO2max; endurance power

     
 
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Table of Contents

  • Introduction
  • Basic Protocol 1: Measurement of Forced Exercise Performance on a Treadmill
  • Basic Protocol 2: VO2max Determination with a Treadmill Coupled to Indirect Calorimetry
  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1: Measurement of Forced Exercise Performance on a Treadmill

  Materials
  • Appropriate mouse strain (e.g., C57BL/6J) and housing facility
  • Spray bottle of distilled water
  • Mild cleaning agent for cleaning the belt, e.g., 0.75% lysoform
  • Treadmill: see, e.g., Fig. ; the ideal set‐up should include:
    • Revolving belt with adjustable speed (0 to 100 cm/sec)
    • Adjustable slope for up‐ and downhill running (+20°/−20°)
    • Independent lanes with covered tops adapted to the size of mice (∼100 mm width and 450 mm length)
    • A platform at the rear of the belt where the animal can escape when exhausted or in case of a major issue; this platform should, however, be equipped with a system to generate aversive stimulation that forces animals to run during the test (e.g., electrical stimulation grid with adjustable intensity from 0 to 2 mA and automatic detection of each stimulation received)
    • Real‐time control of belt speed, slope, time spent running, distance traveled, and number of aversive stimulations received
    • A number of lanes adapted to the number of animals to assess: commercial systems have been developed, e.g., by Panlab (http://www.panlab.com/), Columbus Instruments (http://www.colinst.com/), and TSE (http://www.tse‐systems.com/)
NOTE: It is recommended but not essential to use a treadmill that communicates with a computer and allows the experimenter to control and record the treadmill parameters (velocity, distance traveled, aversive stimulations per min/cumulative number of aversive stimulations).NOTE: To ensure optimal test results, a few sessions of familiarization with the setup are required a few days preceding the actual treadmill test. For C57BL6/J mice, 1 to 2 sessions is typically sufficient (Fig. ), but this number should be adapted to the particular strain of mice under consideration.

Basic Protocol 2: VO2max Determination with a Treadmill Coupled to Indirect Calorimetry

  Materials
  • Appropriate mouse strain (e.g., C57BL/6J) and housing facility
  • Spray bottle of distilled water
  • Mild cleaning agent for cleaning the belt, e.g., 0.75% lysoform
  • A metabolic treadmill (see, e.g., Fig. ) consisting of a chamber tightly closed at both ends by removable walls (approximate volume, 2 liters); the chamber encompasses:
    • A revolving belt with adjustable speed (0 to 120 cm/sec) adapted to the size of the mice (approximately 50 mm width and 26 cm length)
    • A system at the rear of the belt to encourage animals to run during the test (e.g., electrical stimulation grid with adjustable intensity)
    • An inlet port connected to a pump and an air‐flow controller to ensure chamber ventilation with ambient air at a constant flow
    • An outlet port connected to an air sampler delivering the air inside the chamber to the gas analyzer at regular intervals
    • A fan at the front of the belt to ensure circulation of the air over the animal
    • Ability to adjust the slope of the entire setup in 5° increments from −10° to +25°: commercial systems have been developed, e.g., by Columbus Instruments (http://www.colinst.com/) and TSE (http://www.tse‐systems.com/)
  • An open‐circuit calorimeter (indirect calorimetry) (e.g., Oxymax, Columbus Instruments, http://www.colinst.com/) to monitor changes in gas concentration (O 2 and CO 2 ) in the air of the chamber; ambient air is used as a reference after calibration of the system with a calibration gas (20.5% O 2 /0.5% CO 2 with remainder N 2 )
  • Computer and software communicating with the open‐circuit calorimeter to define the settings of the experiment (air flow, sampling time, etc.) and to display measurements in real time (VO 2 , VCO 2 , and RER) throughout the experiment; ideally, the same software should allow to control and record the treadmill parameters
  • Calibration gas (high‐purity grade): exactly 20.5% O 2 and 0.5% CO 2 with remainder N 2 (custom prepared by gas supplier)
  • Scale to monitor body weight of the mouse
  • Timer
NOTE: To ensure optimal test results, a few sessions of familiarization to the setup are required every day preceding the actual treadmill test. For C57BL6/J mice 1 to 2 sessions is typically sufficient, but this number should be adapted to the particular strain of mice under consideration.
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Figures

  •   Figure Figure 1. Example of a commercial 5‐lane treadmill setup for mice (Panlab). The treadmill is a dual system for mice and rats, but is shown in a mouse configuration including dedicated stimulation pads and lane adapters.
    View Image
  •   Figure Figure 2. Total number of aversive stimulations received during daily sessions of familiarization to the setup. Ten wild‐type C57BL/6J mice were subjected to daily sessions of familiarization to the treadmill for 3 days. Each session consisted of a 5‐min run at 15 cm/sec using electrical stimulation at 0.2 mA as aversion. The total number of electrical stimulations over 5 min was recorded for each mouse and plotted.
    View Image
  •   Figure Figure 3. Example of a commercial metabolic treadmill setup for mice (Columbus Instruments).
    View Image
  •   Figure Figure 4. Example of graphical user interface displaying real‐time measurement of VO2 and RER (Columbus Instruments).
    View Image
  •   Figure Figure 5. Individual oxygen consumption and respiratory exchange ratio (RER) during maximal oxygen consumption (VO2 max) protocol. 24‐week‐old male C57BL/6J mice were subjected to a VO2 max protocol with +10° treadmill angle. Left panel: Oxygen consumption (VO2 ) at each belt speed ( n = 8). Right panel: Respiratory exchange ratio at each belt speed ( n = 8).
    View Image
  •   Figure Figure 6. Cumulative number of aversive stimulations received as a function of time in power and endurance performance tests. 18 wild‐type C57BL/6J mice were subjected to a power test and an endurance test as described, using electrical stimulation at 0.2 mA as aversion and a stimulation rate exceeding 20 stimulations per min as termination criteria. The cumulative number of electrical stimulations was recorded during the entire test every minute and plotted as a function of time for every individual mouse.
    View Image
  •   Figure Figure 7. Distance traveled, running time, and maximal speed of wild‐type C57BL6/J mice in power and endurance performance tests. The data from the tests described in Fig. 6 were extracted for each individual mouse and plotted as mean ± SEM.
    View Image
  •   Figure Figure 8. VO2 max and running distance of sedentary and trained lean and diet‐induced obese mice. 8 week old male C57BL/6J mice were fed with a standard chow diet (CD) or a high‐fat diet (HFD) (60% kcal fat) for 16 weeks ( n = 8 per group). These “sedentary” mice were subjected to a VO2 max measurement with a +10° angle for CD‐fed mice and 0° angle for HFD‐fed mice. Upon completion of this experiment, mice were maintained on these diets and allowed free access to a running wheel for 10 days. After this period, a second VO2 max protocol was performed with these “trained” animals under the same condition (+10° angle for CD‐fed mice and 0° angle for HFD‐fed mice). Data are represented as mean ± SEM and * represents a statistical significant difference (p<0.05) using a 1‐way ANOVA followed by a Bonferroni test.
    View Image

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Literature Cited

Literature Cited
   Bernstein, D. 2003. Exercise assessment of transgenic models of human cardiovascular disease. Physiol. Genomics 13:217‐226.
   Billat, V.L., Mouisel, E., Roblot, N., and Melki, J. 2005. Inter‐ and intrastrain variation in mouse critical running speed. J. Appl. Physiol. 98:1258‐1263.
   Booth, F.W., Laye, M.J., and Spangenburg, E.E. 2010. Gold standards for scientists who are conducting animal‐based exercise studies. J. Appl. Physiol. 108:219‐221.
   Dawson, C.A. and Horvath, S.M. 1970. Swimming in small laboratory animals. Med. Sci. Sports 2:51‐78.
   Kemi, O.J., Loennechen, J.P., Wisloff, U., and Ellingsen, O. 2002. Intensity‐controlled treadmill running in mice: Cardiac and skeletal muscle hypertrophy. J. Appl. Physiol. 93:1301‐1309.
   Konhilas, J.P., Maass, A.H., Luckey, S.W., Stauffer, B.L., Olson, E.N., and Leinwand, L.A. 2004. Sex modifies exercise and cardiac adaptation in mice. Am. J. Physiol Heart Circ. Physiol. 287:H2768‐H2776.
   Kregel, K.C., Allen, D.L., Booth, F.W., Fleshner, M., Henriksen, E.J., Musch, T.I., O'Leary, D.S., Parks, C.M., Poole, D.C., Ra'anan, A.W., Sheriff, D.D., Sturek, M.S., and Toth, L.A. 2006. Resource Book for the Design of Animal Exercise Protocols. American Physiological Society, Bethesda, Md.
   Lerman, I., Harrison, B.C., Freeman, K., Hewett, T.E., Allen, D.L., Robbins, J., and Leinwand, L.A. 2002. Genetic variability in forced and voluntary endurance exercise performance in seven inbred mouse strains. J. Appl. Physiol. 92:2245‐2255.
   Lightfoot, J.T., Turner, M.J., Debate, K.A., and Kleeberger, S.R. 2001. Interstrain variation in murine aerobic capacity. Med. Sci. Sports Exerc. 33:2053‐2057.
   Peronnet, F. and Aguilaniu, B. 2006. Lactic acid buffering, nonmetabolic CO2 and exercise hyperventilation: A critical reappraisal. Respir. Physiol. Neurobiol. 150:4‐18.
   Savaglio, S. and Carbone, V. 2000. Scaling in athletic world records. Nature 404:244.
   Thomas, C., Marcaletti, S., and Feige, J.N. 2011. Assessment of spontaneous locomotor and running activity in mice. Curr. Protoc. Mouse Biol. 1:185‐198.
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