OriginalInvestigations

in Mice after Long-Term

Training Effects
Voluntary Exercise
SARA R. DAVIDSON', MARGARET BURNETT 2, and LAURIE HOFFMAN-GOETZ'
'Department of Health Studies and Gerontology and 2Department of Kinesiology, Faculty of Applied Health Studies,
University of Waterloo, Waterloo, Ontario, CANADA

ABSTRACT
DAVIDSON, S. R., M. BURNETT, and L. HOFFMAN-GOETZ. Training Effects in Mice after Long-Term Voluntary Exercise. Med.
Sc. Sports Exerc.,Vol. 38, No. 2, pp. 250-255, 2006. Background: Mice are an important animal model in exercise studies on the

immune system, cancer, and aging. There is limited research about the training effects of long-term voluntary exercise in this species.
Purpose: To describe the training effects in mice given long-term aerobic voluntary exercise. Methods: Female C57BL/6 mice were
randomly assigned to 1) individual cages with in-cage running wheels with 24-h access (WR; N = 31), or 2) individual cages without
running wheels for 16 wk (NR; N = 20). Run-to-exhaustion (RTE) times, 902,A speed at V'02a,A, and citrate synthase (CS),

succinate dehydrogenase (SDH), and phosphofructokinase (PFK) activity in the soleus, plantaris, and red and white gastrocnemius
were assessed. Results: Final body weight and speed at 9O2,ak did not differ by training condition. WR mice had significantly longer
RTE times (P < 0.001) and higher V'O2,A (P < 0.05) compared with NR mice. Higher CS and SDH activities were found in WR

compared with NR mice for soleus (P < 0.01), red gastrocnemius (P < 0.01), and plantaris (P < 0.01) muscles. PFK activity was
higher in WR mice in white gastrocnemius compared with NR mice (P < 0.01). Conclusions: Voluntary running wheel activity for
16 wk in female C57BL/6 mice resulted in longer run times to exhaustion, higher VO2,ak, and higher SDH and CS activities in
oxidative muscles. These findings suggest that wheel running in female C57BL/6 mice: 1) produces a measurable aerobic training
effect and 2) is an effective exercise modality for long-term training studies. Key Words: WHEEL RUNNING, CITRATE
SYNTHASE, SUCCINATE DEHYDROGENASE, PHOSPHOFRUCTOKINASE, V702p,jAK, RUN TO EXHAUSTION

exercise, such as wheel running, tends not to be associated

with activation of the stress response (8) and, therefore, has
been proposed as a training protocol to circumvent this
problem.
A significant limitation in the literature on voluntary
exercise paradigms involving mice is that few studies document physiological and biochemical indices indicative of
training effects. V0 2 uptake, VCO2 production, respiratory
exchange ratio (RER), and citrate synthase activity in
soleus muscle have been reported, but the duration of the
voluntary exercise was relatively short (8 wk) (19). Concurrent changes in performance measures (such as VO2p,ak) and
in skeletal enzyme activity in mice as a result of voluntary
wheel exercise have not been systematically assessed.
The purpose of this study was to describe the performance and biochemical training effects of mice given
access to running wheels for 4 months compared with mice
not given access to running wheels. The physiological.
performance measures included run-to-exhaustion (RTE)
times, VO2peak and speed at 902peak. The skeletal muscle
enzyme measures included citrate synthase (CS), succinate
dehydrogenase (SDH) and phosphofructokinase (PFK)
activity in the soleus, plantaris, and red and white
gastrocnemius. Due to the nature of wheel running as an
aerobic endurance activity, we hypothesized that there
would be longer RTE times, and higher VO 2pe,ak, and CS

ice are frequently used as an animal model to

the effects of exercise on a variety of
physiological processes including immune
L
responses (13), aging (28), and carcinogenesis (30). The
exercise modality used is often in the form of long-term
training and typically involves forced activity such as
treadmill running (4). However, forced activity protocols
activate the hypothalamic-pituitary-adrenal (HPA) axis
and the "stress" response with a concomitant elevation in
plasma corticosterone levels, catecholamines, heart rate
and blood pressure (21). Thus, results of studies employing
forced exercise models may be confounded by chronic
activation of the stress response. In contrast, voluntary

Sstudy

Address for correspondence: Laurie Hoffman-Goetz, PhD, MPH, Department of Health Studies and Gerontology, Faculty of Applied Health
Studies and Gerontology, University of Waterloo, 200 University Avenue
West, Waterloo, Ontario, N2L 3G1 Canada; E-mail: lhgoetz@healthy.
uwaterloo.ca.
Submitted for publication April 2005.
Accepted for publication August 2005.
0195-9131/06/3802-0250/0
MEDICINE & SCIENCE IN SPORTS & EXERCISE,
Copyright 2006 by the American College of Sports Medicine
DOI: 10.1249/01.mss.0000183179.86594.4f

250

and SDH activities in the soleus, red gastrocnemius, and

plantaris in wheel-run mice compared with nonrun mice.

METHODS
Experimental Animals
Female C57BL/6 mice, age 4-6 wk and weighing 16.9 d
0.2 g, were obtained from Harlan Sprague Dawley
(Indianapolis, IN), acclimated to our vivarium for 1 wk,
and then randomly assigned to one of two groups: 1)
individual cages (29.5 x 18.5 x 12.5 cm) with in-cage
running wheels allowing 24 h access (WR; N = 31) and
with supplemental toys for enrichment (i.e., plastic tubes,
nesting materials) or 2) individual cages without running
wheels (NR; N = 20) and with supplemental toys. The mice
were housed for 16 wk in a humidity (65%) and
temperature (21 l1QC) controlled room, maintained on a
12-h reversed light-dark cycle and provided with tap water
and food (Laboratory Rodent Chow, PMI Foods, Richmond, IN) ad libitum. The guidelines for experimental
procedures established by the Canadian Council on Animal
Care were followed and all protocols with live animals
were approved by the university animal care committee.

Wheel Running Activity

Activity of mice on the running wheels (23.0 cm in
diameter) was monitored by a magnetic switch affixed to
each wheel, which recorded the number of completed
revolutions; data were captured by an automated computer
monitoring system and software (Vital View Application
software, Mini Mittei Company, Sunriver, OR). Physical
activity was recorded continuously as wheel revolutions
per 15-min interval, converted to kilometers and summed
by week for analysis. Free open-field locomotor activity of
mice within cages was not measured. Exercise performance measures and skeletal muscle enzyme activities
were obtained for a randomly selected subset of mice from
each of the WR (N = 10) and NR (N = 10) groups.

Performance Measures
Run to exhaustion (RTE). Two weeks before sacrifice, mice were acclimated once to run on a small rodent
treadmill (Omni-max metabolic small rodent treadmill,
Omni Tech Electronics, C61umbus, OH) (5-min warm-up
followed by 15-20 min at 28 m,min-1). One day after
acclimation to the treadmill, mice were run during the dark
cycle until they reached volitional exhaustion using a
modification of the protocol of Campisi et al. (2). In brief,
for the first 5 min, the speed of the treadmill was gradually
increased to 28 m'min- 1 , and thereafter, the mice ran at
28 m-min-1 until exhaustion. To encourage the mice to
run, an electric shock grid at the base of the treadmill was
activated to deliver a 0.2-mA pulse. This delivered an
uncomfortable shock but did not injure or harm the mouse.
Volitional fatigue or exhaustion was defined as the refusal
to run after 10 consecutive tail shocks.
VOLUNTARY TRAINING INMICE

VO2peak, One week before sacrifice, mice were placed

in the metabolic treadmill over a 10-min period to measure
resting V02. Thereafter, mice were run on the treadmill
with the speed increasing by 2 m'min-1 every 90 s (22)
until exhaustion was reached. Volitional exhaustion was
defined as above at which time VO2peak was measured.
Muscle isolation. Mice were sacrificed by sodium
pentobarbital (0.6-0.8 cm3 per mouse, i.p.) overdose.
Muscle samples from WR and NR mice were obtained
for soleus (SOL), plantaris (PLANT), and red (RG) and
white gastrocnemius muscle (WG), which were isolated,
flash frozen in liquid nitrogen, and stored at -90'C until
assessed for enzyme activity.
Muscle enzyme activity. All muscles were cut into
5- to 10-mg segments, homogenized in homogenizing buffer
(glycerol (50%), sodium phosphate buffer (20 mM),
2-mercaptoethanol (5 mM), ethylendiaminetetraacetic acid
(EDTA, 0.5 mM), BSA (10%)) to yield a 50:1 dilution and
assayed for succinate dehydrogenase (SDH), citrate
synthase (CS), and phosphofructokinase (PFK) activities.
An aliquot of each muscle homogenate was sonicated (with
a 3-mm tip, 2 s on, 5 s off, for a total of 20 s at 60 Hz; Vibra
Cell, Sonics and Materials, Danbury, CT) and subsequently
analyzed for PFK and SDH activity on the same day (3). The
remaining homogenate was stored at -90'C for CS and
protein analysis.
SDH. Muscle homogenates were diluted in diluting
medium (imidazole-HCl (20 mM), BSA (0.02%)) to yield
a 1:150 dilution. Twenty-five microliters of the diluted
samples were aliquoted and placed in a 37C water bath for
10 min, then 100 pL of SDH reagent (phosphate buffer
(0.03 M), sodium succinate (115 mM), phosphate
(0.0015%), and BSA (10%)) were added. Thirty minutes
later, 200 pL of 1 N NaOH were added, the samples were
removed from the water bath,, and 100 pL of
bromobenzene were added, followed by centrifugation for
3 min at 15,000 'x g. Twenty-five microliters of supematant were aliquoted, 1 mL of dilute reagent (hydrazine
(100 mM) EDTA (4 mM), and NAD (0.4 mM)) was added,
and the fluorescence of NAD' was measured
fluorometrically. Ten microliters of dilute MDH-fumarase
reagent (hydrazine, EDTA, NAD, fumarase (700 U.mL-1),
malate dehydrogenase (6000 U'mL-')) were added, and
the samples were incubated in the dark for 2 h at room
temperature (RT) and read fluorometrically.
PFK. Muscle homogenates were diluted in PFK diluting
medium (Tris-HCl (50 mM), K2 HPO 4 (10 mM), 2mercaptoethanol (5 mM), EDTA (0.5 mM), BSA
(0.02%)) to yield a 1:15,000 dilution. Five microliters of
the diluted sample were aliquoted and 100 pL of reagent 1
(Tris-HCl (50 mM), ATP (1 mM), fructose-6-phosphate
(1 mM), K2HPO 4 (10 mM), MgCL 2 (2 mM), NADH

(200 pM), 2-mercaptoethanol (1 mM), BSA (0.05%),

aldolase (0.09 U'mL-'), glycerol-3-phosphate dehydrogenase/triosphosphate isomerase (GDH:TIM)) were added.
After 1 h at RT, this reaction was stopped by adding 10 pL
of 0.75 N HCI and left for 10 min at RT. One milliliter
of 6 N NaOH (10 mM imidazole) was then added,
Medicine &Science in Sports &Exercises

251

the samples were incubated for 20 min at 60'C, and the

fluorescence of NAD' was measured fluorometrically.
CS. Muscle homogenates were diluted to yield a 1:2500
dilution. Ten microliters of the diluted sample were
aliquoted, 100 pL of reagent 1 (Tris-HCl (50 nM), acetyl
CoA (0.4 mM), oxaloacetate (0.5 mM) and BSA (0.25%))
added, and samples incubated at RT for I h. Ten microliters of 0.75 N NaOH were added, the samples incubated
at RT for 10 min, I mL of reagent 2 was added (Tris-HC1
(100 mM), ZnC12 (100 pM), BSA (0.01%), NADH
(30 pM), citrate lyase (0.003 U-mL-'), malate dehydrogenase (3 U'mL-')) and incubated at RT for 20 min. Sixty
microliters of 1 N HCl were added, the samples were
incubated at RT for 10 min, and 100-pL aliquots of the
solution were added to 1 mL 6 N NaOH (10 mM
Imidazole). Samples were incubated at 60'C for 20 min,
and the fluorescence of NAD' was measured fluorometrically.
Protein concentration. The concentration of protein
in muscle homogenates was determined using Lowry
method (18). All enzymes are expressed as moles per hour
per kilogram of protein.
Statistical analysis. Data were analyzed using SPSS
v12.0 software, and values are reported as group means
SEM. Running wheel data were analyzed with a repeated
measures ANOVA, and all other dependent variables were
analyzed with a one-way ANOVA (for data that were
normally distributed) or with a Mann-Whitney U test (for
data that were not normally distributed). A threshold of
significance of a = 0.05 was accepted as being different
from chance alone. Unless otherwise indicated, there were
10 mice in each of the WR and NR groups used for
analysis of performance measures and muscle enzyme
activities.
RESULTS
Body weight. There were no significant differences
in final body weight between WR (24.6 0.7 g) and NR
(26.2 0.9 g) mice at sacrifice.
Average running distance. Figure 1 shows the
average running distances of female C57BL/6 mice for
16 wk. Mice ran on average 2.21 0.49 km'd-1 for a total
of 364.72 1 28.83 km over the 16-wk period. At week 1,
average running distance was 14.0 2.0 km. Running
distance varied significantly over time (F1,3o = 4.37,
P < 0.05) and each weekly time point was significantly
higher than week 1 (all P values < 0.05).
Exercise performance measures. There was a significant difference in RTE times between NR and WR
groups (Mann-Whitney U statistic = 0.00, P < 0.001), with
WR mice having almost double the time to exhaustion
during treadmill challenge. Initial analysis of VO2peak
between NR and WR groups indicated no difference
between groups. This lack of difference was the result of
one animal in the NR group with a V'02peak of
91 mL-kg-'.min- 1, which was significantly higher than
values for WR mice and resulted in a large standard
252

Official Journal of the American College of Sports Medicine

aj
U
In
01 S

01
0
cJ
cC

3530"

*
*

25201510.

5.
1 2 3 4 5 6 7 8 9 1011 1213141516
Week

FIGURE 1-Average running wheel distance for 16 wk for female

C57BL/6 mice. *Average running wheel distance is significantly
different from week 1 (P < 0.05). A total of 31 mice were given access
to in-cage running wheels (details in text).

deviation for the NR group. When this outlier was

removed, there was a significant difference in V'O2peak

(-10%) in WR mice compared with NR mice

(17
1, 17 = 4.417, P < 0.05), but no significant difference in
the speed at which VO2p,ak was elicited (Table 1).
Skeletal muscle enzyme activity. Table 2 shows
the skeletal muscle enzyme activity from SOL, RG, WG,
and PLANT of female C57BL/6 mice. For SOL, there was
significantly greater activity of CS (F1,18 = 5.051, P < 0.05)
(-18%) and SDH (F1 ,18 = 8.836, P < 0.01) (-23%) in the
SOL from WR compared with NR mice. In contrast, PFK
activity did not differ by training group. For RG, there was
significantly greater CS (F1,18 = 4.994, P < 0.05) (-26%)

and SDH (F1 ,18 = 16.107, P < 0.01) (-61%) activity in the
RG from WR relative to NR mice. As with the SOL, no
difference in PFK activity was found by training group for
RG. For PLANT, WR mice had higher CS (-16%)
(Fi,i 8 = 13.074, P < 0.01) and SDH (Mann-Whitney U
Statistic = 1, P < 0.01) activity (-56%) in the PLANT
muscle compared with NR mice. No significant differences
were found for PFK activity between the two groups. For
WG, no significant difference was found in CS activity in
the WG by training group. However, the activity of SDH,
an oxidative enzyme' (F1,18 = 16.107, P < 0.01) and of

PFK, a glycolytic enzyme (Mann-Whitney U Statistic = 3,

P < 0.01) differed significantly as a function of training
group, with activity higher in WR (-87%) compared with
NR mice.

DISCUSSION
The main findings of this paper are that beginning at
ages 5-7 wk, female mice that were given 16 wk of access
to in-cage running wheels had physiological performance
and skeletal muscle enzyme changes suggestive of a
training effect. These changes included longer treadmill
run times to exhaustion, and higher 'VO2p,Ak, oxidative
enzyme activity in the SOL, RG and PLANT, and
glycolytic enzyme activity in the WG. This study is unique
in its comprehensive characterization of skeletal muscle
biochemistry and performance characteristics for mice
given long-term voluntary training. Nonetheless, the data
are specific to adult (6 months old) female C57BL/6 mice,
http://www.acsm-msse.org

TABLE 1. Physiological performance measures for female C57B3J6 mice given 16 wk

of in-cage running wheel access.
Speed at V02pak
(m-min')

Run to Exhaustion
(min)

1
(mL.kgi--min- )

56.20 t 1.97

74.44 - 1.76t

36.4 + 1.7

104.00 : 6.98*

81.27 - 2.701

38.8 t 1.7

Nonrunning wheel

V021pe0k

(N= 10)
Running wheel
(N = 10)
*

Significant at P< 0.001 by nonparametric Mann-Whitney U test.

N= nine mice.
Significant at P< 0.05 by one-way ANOVA.

and additional studies will be needed to determine whether

performance and skeletal muscle enzyme changes occur in
other strains, in both genders, and at different ages.
During the 16 wk of the study, WR female mice
accumulated an average daily running distance of 2.21 +
0.49 km'd-', a weekly distance of 22.8 1.8 km'wk-1, and
a total running distance of 364.72 28.83 km. These
running distances are similar to those found by Mehl et al.
(20) involving female C57BL/6 mice, but lower than those
found by Valentinuzzi et al. (28) involving male C57BL/6.
Wheel running distance is variable and dependent on many
factors including gender, strain, and age (6). The results
presented here indicate that the maximal average running
distance in female C57BL/6 mice occurred at week 12.
Others have reported peak running distance for mice and
rats to occur at weeks 3-4 (9,25), week 5 (23), week 6
(17), and week 8 (1,29). The peak and plateau'in distances
on running wheels also reflect strain and gender contributions. Finally, there were no significant differences in final
body weights between the WR and NR mice. This result
was not unexpected, because female rodents increase their
food intake in proportion to their energy expenditure (26),
whereas males decrease their food intake (11).
WR mice had approximately 10% higher VO2peak values

than NR mice indicating that wheel running is able to

invoke a change in aerobic function. Increases in VO2peak
after wheel running have been observed after 8 (17), 10
(29), and 12 wk (23) of voluntary wheel exposure in
rodents. Similar increases in VO2pak (-14%) have been
observed after 12 wk of treadmill training (16). Nonetheless, the actual VO2pek values for mice in the present study
were lower than those reported by Fueger et al. (7). The
reasons for this difference are not known. Because mice in
the two studies differed by substrain (6J vs 6), backcrossing (Ukko I backcrossed onto C57BL/6J for 5
generations vs no backcrossing across strains), age (4 vs
6 months), and gender (mixed vs females only), V02pak

may have differed as a result; it is also possible that in the

present study mice were running at submaximal rather than
maximal exertion, which would result in lower VO2p,ak.
WR mice ran nearly twice as long in the RTE as NR mice,
providing further evidence that wheel running invokes
changes to aerobic capacity. Other studies report significantly longer RTE times in rats following 8 wk of wheel
access (2,17,23). Similarly, 13 wk of treadmill training
significantly increased RTE times (5).
CS and SDH are widely used as metabolic markers
for muscle oxidative capacity (9,29), and PFK and
hexokinase for muscle glycolytic capacity (9,10). The
SOL, RG, PLANT, and WG were selected for evaluation
because of their involvement in wheel locomotion and
differences in fiber type and metabolic capacity: SOL
(Type I, oxidative), RG (Type HI, oxidative), PLANT (Type
II, oxidative, glycolytic), and WG (Type II, glycolytic).
Significantly increased CS activity in the WR group was
found in the SOL (18%), RG (26%), and PLANT (16%)
muscles. SDH activity was also higher in the WR group for
the SOL (23%), RG (38%), and PLANT (56%) muscles.
Increased activity of these oxidative enzymes is consistent
with the hypothesis that voluntary wheel running is
an endurance exercise, and leads to an upregulation of
oxidative enzyme activity in oxidative muscle. Our results
agree with previous studies in rats showing, increased CS
activity in SOL between 4 and 8 wk (9,15) and'PLANT
between 2 and 12 wk of voluntary wheel exposure
(9,10,25). Increased SDH activity in rat SOL (29) and rat
red vastus lateralis (Type II oxidative, glycolytic) (1) occur
after 4 wk of voluntary wheel exposure, results similar to
enzymatic changes occurring with treadmill training
(5,26,27).
No significant increase in CS activity was found in WG
of WR mice that were given 16 wk of running. This was
expected because WG is predominantly a, glycolytic
muscle, and no increase in rat CS activity for other
glycolytic muscles (e.g.,^white head of tricep brachii) have
been reported after voluntary wheel exposure (25).
In contrast, Podolin et al. (24) found no significant
differences in rat CS activity-in the soleus following 8 wk
of wheel exposure. Bagby et al. (1) also reported no
significant change in SDH in rat soleus after 7 or 16 wk of
wheel exposure. Differences in wheel running speed may
contribute to these divergent results, because 'higher speeds
require muscles with a higher percentage of Type HI fibers
than Type I fibers (1).

TABLE 2. Skeletal muscle enzyme activities in female C57BL/6 mice given 16 wk of in-cage running wheels.
Skeletal Muscle'

SSoleus
Enzyme Activliy'

Training Status

(Type 1,
Oxidative)

NR
WR
NR
WR
NR

12.98 t 0.75
15.31 0.71
9.05 0.53
11.15:t 0.461
3.36 0.15

WR

4.36 : 0.58

Citrate synthase
Succinate dehydrogenase
Pyruvate fructose kinase

Red Gastracnemius '

(Type II, Oxidative)
11.13
13.98 :
7.50 :
12.10
8.07 :
9.40

0.68
1.08*
0.414
1.07
0.44

0.56

Plantarls (Type 11,

Oxidative, Glycolytic)

White Gastrocnernlus
(Type II, Glycolytlc)

10.42 0.29
12.09 - 0.36
9.97 0.31
15.58 0.881
11.99 0.32

5.96,
6.5
4.69
7.57
.9.42

.12.25 0.56

17.64 1.49

0.11
0.26
0.61
0.38
0.69

a Enzyme activity expressed as moles per hour per kilogram of protein.

* Significant

(P< 0.05); 'significant (P< 0.01). Atotal of 10 mice were included ineach group (NR and WR) for muscle enzyme measures.

VOLUNTARY TRAINING IN MICE

Medicine &Science in Sports & Exercise

253

Unexpectedly, we found an increase of SDH activity in

the WG. However, increased oxidative enzymes (CS) in
other glycolytic muscles, such as the epitrochlearis, have
been observed after 2-3 wk of wheel exposure in rats (12).
The physiological reasons for increased oxidative enzyme
activity (SDH or CS) in muscle that is primarily Type 11,
glycolytic are not clear from our data. However, there is
some evidence for fiber type change in rat extensor
digitorum longus from Type fI glycolytic to Type Bl
oxidative and Type I in female rats given 45 d of voluntary
exercise (15).
PFK activity in WG was nearly twice as high in the WR
compared with the NR group; however, no significant
difference in PFK activity occurred in the SOL, RG, or
PLANT muscles of WR mice. Our finding of increased
PFK activity in WG, the finding by Kriketos et al. (15) of
increased hexokinase (another glycolytic enzyme) in the
extensor digitorum longus, and by Hokama et al. (12) of
increased hexokinase in epitrochlearis suggest that voluntary running involves both glycolytic and oxidative
muscles. No significant difference between WR and NR
mice were found for PFK activity in the SOL, RG, and the
PLANT muscles. Increases in hexokinase activity have
been found in the SOL after 4 but not 8 wk of voluntary
exercise and in the PLANT after 4 and 8 wk of voluntary
exercise (9). The activity of hexokinase in the SOL may be
upregulated shortly after the initiation of exercise, but this
effect is not persistent. This may also explain why no
significant increases in PFK activity in the SOL were
found after 16 wk in the mice. However, serial measurements of PFK in SOL over the training period would be
necessary to confirm whether up- and then downregulation
of this enzyme occurred.
The observed increases in skeletal muscle oxidative
enzyme activity and performance measures in WR mice
suggest that long-term exposure to running wheels is a
form of aerobic endurance training. However, an alternate
explanation is that mice without running wheels are
deconditioned rather than WR mice being trained animals

(23). Mice are naturally active animals, and removing their

access to activity by using standard caging without wheels
may make them artificially inactive. However, it is
common practice in exercise studies involving rodents to
include a sedentary control group. Further research investigating differences in performance measures and skeletal
enzyme activity between mice allowed free-range locomotion, voluntary wheel access, or treadmill training is
required to clarify the differences in these parameters as a
result of the different types of activity. Detraining has been
found to quickly decrease exercise-induced improvements
in cardiac adaptations (14); therefore, removing physical
activity can result in lower parameter values compared
with animals with regular access to exercise. Nonetheless,
one strength of voluntary wheel running is that this exercise
mode is more representative of human physical activity,
which involves voluntary rather than forced activity.
In conclusion, access to voluntary running wheels for

16 wk in adult female C57BL/6 mice resulted in longer run

times to exhaustion and higher V02peak, oxidative enzyme
activity in oxidative muscles, and glycolytic enzyme

activity in glycolytic muscle. Taken together, our results

suggest that 4 months of voluntary wheel running in this
mouse strain produces an aerobic training effect. The magnitude of this aerobic training effect needs to be evaluated
against other exercise modalities, such as treadmill running. Nonetheless, a recent study shows that training
elicited by voluntary wheel running may be equivalent to
treadmill training: CS activity in gastrocnemius was higher
in wheel-trained ApcMinI+ male mice than treadmill-trained
male mice, and both exercise modes resulted in higher CS
activity than untrained controls (20). Given that wheel

running is a voluntary activity that does not activate the

HPA axis and the stress response, it constitutes an effective

exercise modality for long-term studies in mice.

This research was supported by a grant from the NSERC
Canada to LH-G. The authors thank J. Guan and E. Bombardier
for technical assistance.

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TITLE: Training Effects in Mice after Long-Term Voluntary

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