Effects of high-intensity intermittent swimming
on glucose transport in rat epitrochlearis muscle
KENTARO KAWANAKA, IZUMI TABATA, AYUMI TANAKA, AND MITSURU HIGUCHI
Division of Health Promotion, National Institute of Health and Nutrition,
Toyama 1-23-1, Shinjuku City, Tokyo 162-8636, Japan
Kawanaka, Kentaro, Izumi Tabata, Ayumi Tanaka,
and Mitsuru Higuchi. Effects of high-intensity intermittent
swimming on glucose transport in rat epitrochlearis muscle.
J. Appl. Physiol. 84(6): 1852–1857, 1998.—Recently (K. Kawa-
naka, I. Tabata, and M. Higuchi. J. Appl. Physiol. 83: 429–433,
1997), we demonstrated that glucose transport activity after
repeated 10-s-long in vitro tetani in rat epitrochlearis (Epi)
muscle was negatively correlated with the postcontraction
muscle glycogen concentration. Therefore, we examined
whether high-intensity intermittent swimming, which de-
pletes muscle glycogen to a lower level than that observed
after ten 10-s-long in vitro tetani, elicits higher glucose
transport than that observed after ten 10-s-long in vitro
tetani, which has been regarded as the exercise-induced
maximal stimulus for glucose transport. In male rats, 2-deoxy-
D-glucose transport rate in Epi muscle after eight bouts of
high-intensity intermittent swimming with a weight equal to
18% of body mass (exercise duration: 20 s, rest duration
between exercise bouts: 40 s) was higher than that observed
after the ten 10-s-long tetani (2.25 6 0.08 vs. 1.02 6 0.16
µmol · ml intracellular water21 · 20 min21). Muscle glycogen
concentration in Epi after eight bouts of high-intensity inter-
mittent swimming was significantly lower than that observed
after ten 10-s-long in vitro tetani (7.6 6 0.5 vs. 14.8 6 1.4
µmol glucose/g muscle). These observations show that the
high-intensity intermittent swimming increases glucose trans-
port in rat Epi to a much higher level than that induced by ten
10-s-long in vitro tetani, which has been regarded as the
exercise-related maximal stimulus for glucose transport.
Furthermore, this finding suggests that the lower muscle
glycogen level after high-intensity intermittent swimming
than after in vitro tetani may play a role, because there was a
significant negative correlation between glucose transport
and muscle glycogen concentration in Epi after high-intensity
swimming and in vitro tetani.
muscle glycogen; 2-deoxy-D-glucose transport; exercise-
induced maximal glucose transport
INSULIN INCREASES THE PERMEABILITY of skeletal muscle
to glucose. Furthermore, it is well known that low- or
moderate-intensity swimming or running has an insu-
linlike effect on glucose transport in skeletal muscle
(14, 15). In the absence of insulin, in vitro twitch or
tetanic contractions induced by electrical stimulation
produce an increase in muscle glucose transport (12,
22, 23). Therefore, it is suggested that there are two
separate pathways for stimulation of glucose transport
in skeletal muscle: one pathway is activated by insulin,
the other by muscle contraction.
Several previous studies reported enhanced insulin-
stimulated glucose uptake after exercise when the
muscle glycogen content was low (5, 7, 28). These
results suggest that muscle glycogen content affects
insulin-stimulated glucose transport in skeletal muscle.
1852 8750-7587/98 $5.00 Copyright r 1998 the American Physiological Society
Furthermore, we found recently that in vitro tetani-
stimulated glucose transport in rat epitrochlearis (Epi)
muscle was negatively correlated with postcontraction
muscle glycogen concentration (17). This result also
suggests that the lower the concentration of muscle
glycogen, the higher the contraction-stimulated glucose
transport rate.
In rat Epi muscle, ten 10-s-long in vitro tetani had a
maximal effect on glucose transport, and more tetanic
contractions did not induce a further increase in glu-
cose transport (11, 17). Furthermore, glucose transport
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after this protocol was elevated to a level as high as that
observed after 120 min of swimming with a weight
equal to 2% of body mass (3). Therefore, activation of
glucose transport after ten 10-s-long in vitro tetani has
been considered as the exercise-induced maximal acti-
vation of glucose transport in Epi. However, our previ-
ous study showed that mean muscle glycogen concentra-
tion was ,15 µmol glucose/g muscle after ten 10-s-long
in vitro tetani, and more tetani did not further decrease
muscle glycogen content (17). Therefore, it is not known
whether a significant negative relationship between
postcontraction muscle glycogen and glucose transport
exists in the lower range of muscle glycogen concentra-
tion (i.e., much less than 15 µmol glucose/g muscle).
Therefore, we examined glucose transport in Epi
after high-intensity intermittent exercise that has been
used as a tool to deplete muscle glycogen (4). For this
purpose, we adopted intermittent swimming with a
weight equivalent to 18% of body mass. After this
protocol, muscle glycogen concentration decreased to
much less than 15 µmol glucose/g muscle, and glucose
transport was further increased to a level that was
much higher than that observed after ten 10-s-long in
vitro tetani.
METHODS
Male Sprague-Dawley rats (Crea Japan, Tokyo) with body
weights of 90–110 g were used for this study. Ethical approval
for this work was obtained from the Committees on Animal
Care at the National Institute of Health and Nutrition, Japan.
Treatment of animals and swimming program. All animals
were housed in rooms lighted from 7 AM to 7 PM and were
maintained with ad libitum feeding on standard chow and
water. Room temperature was maintained at 20–22°C. Food
was restricted to 8 g on the evening of the last day before the
experiment.
Some of the rats swam in a barrel (4 rats/barrel) filled to a
depth of 50 cm, with water maintained at 35°C. A weight
equal to 3% of body mass was tied to the body of the rat
(low-intensity continuous swimming). These rats swam for
10, 30, or 120 min. At the end of 120 min of low-intensity
continuous swimming, the blood lactate concentration was
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 GLUCOSE TRANSPORT AFTER HIGH-INTENSITY INTERMITTENT SWIMMING
1.8 6 0.2 (SE) mM (n 5 7), which was not significantly
different from the resting level of 1.3 6 0.2 mM (n 5 7).
Some other rats swam with a weight equal to 18% of body
mass for 20 s at a rate of 1 swim/min (high-intensity
intermittent swimming). The rats repeated this swimming
protocol 1, 3, or 8 times. At the end of eight bouts of
high-intensity intermittent swimming, blood lactate concen-
trations were significantly elevated from the resting level of
1.3 6 0.2 mM (n 5 7) to 10.4 6 0.3 mM (n 5 7). When the rats
swam to exhaustion with a weight equal to 18% of body mass,
the mean exhaustion time was 63 6 2 s (n 5 8).
Muscle preparation. Rested and exercised rats were anes-
thetized with an intraperitoneal injection of pentobarbital
sodium, 5 mg/100 g body wt, and the Epi muscles were
dissected out to measure glucose transport in vitro. Some
muscles from high-intensity intermittently swimming rats
were incubated at 35°C for 2 h in 3 ml of oxygenated
Krebs-Henseleit bicarbonate buffer (KHB) containing 8 mM
Na pyruvate and 24 mM mannitol. The gas phase in the
flasks was 95% O2 –5% CO2. The Epi muscle is a small, thin
muscle weighing ,25 mg in rats weighing 90–110 g. It is
made up predominantly (,85%) of fast-twitch fibers (21).
Electrical stimulation. For electrical stimulation, the distal
end of the muscle was attached to a vertical Lucite rod
containing two platinum electrodes. The proximal end was
clipped to a jeweler’s chain that connected to an isometric
force transducer (TB-653t, Nihon Kohden, Tokyo, Japan), and
resting tension was adjusted to 0.5 g. The mounted muscle
was immersed in 100 ml KHB containing 32 mM mannitol
and 8 mM glucose. The muscles were continuously oxygen-
ated with 95% O2 –5% CO2 at 35°C. For activation of glucose
transport by tetanic contractions, the muscles were stimu-
lated with supramaximal square-wave pulses of 0.2-ms dura-
tion with an electrical stimulator (SEN-7203, Nihon Kohden).
Tetanic contractions were produced by stimulating the muscle
at 100 Hz for 10 s. Tetanic contractions were repeated 1, 5, 10,
15, and 20 times at a rate of 1 contraction/min.
Insulin treatment. The muscles to be used for the measure-
ment of insulin-stimulated glucose transport activity were
placed in 3 ml of oxygenated KHB containing 8 mM Na
pyruvate, 24 mM mannitol, 0.1% radioimmunoassay-grade
bovine serum albumin, with 2 mU/ml insulin (this concentra-
tion of insulin stimulates glucose transport to a maximal
level). The gas phase in the flasks was 95% O2 –5% CO2 at
35°C.
Measurement of glucose transport activity and glycogen
concentration. Glucose transport activity was measured by
using the glucose analog 2-deoxy-D-glucose (2-DG) and the
procedure of Young et al. (27). After exercise, electrical
stimulation, or the incubation with insulin, the muscles were
transfered to flasks containing 3 ml of KHB with 40 mM
mannitol and were incubated with shaking for 15 min at 29°C
to remove glucose. In the case of insulin stimulation, 2 mU/ml
of insulin were added to the 3 ml of KHB. The muscles were
then incubated for 20 min at 29°C in 2 ml of KHB containing
Table 1. Effects of high-intensity intermittent swimming on glycogen concentration and glucose transport activity
in rat epitrochlearis muscle
Basal (n 5 14)
Muscle glycogen, µmol glucose units/g wet wt 26.2 6 1.4
2-Deoxy-D-glucose transport, µmol · ml21 · 20 min21 0.34 6 0.02
Values are means 6 SE, n 5 no. of muscles. Rats swam for 20 s at a rate of 1 bout in 1 min with a weight equal to 18% body wt for 1, 3, and 8
bouts. Immediately after swimming, muscles were excised and analyzed. * Significantly different from basal, P , 0.05. † Significantly different
from 1 bout of swimming, P , 0.05. ‡ Significantly different from 3 bouts of swimming, P , 0.05.
1853
1 mM 2-deoxy-(1,2-3H)glucose (1.5 mCi/mmol) and 39 mM
(U-14C)mannitol (8 µCi/mmol) (Moravek Biochemicals). In
the case of insulin stimulation, 2 mU/ml of insulin were also
added to 2 ml of KHB. The gas phase in the flasks during both
the rinse and incubation periods was 95% O2 –5% CO2.
Hansen et al. (10) reported that the accumulation of 2-DG
remains linear until the total intracellular 2-DG-6-phosphate
concentration exceeds 30 mM. Therefore, under our experi-
mental conditions, 2-DG uptake accurately reflects glucose
transport activity. After incubation, the muscles were blotted
briefly on filter paper dampened with incubation medium,
trimmed, and frozen in liquid N2. The extracellular space and
intracellular 2-DG concentrations were determined as de-
scribed previously (27). The samples were weighed and
homogenized in 0.3 M perchloric acid, and an aliquot of the
homogenate was stored for later determination of glycogen
concentration. Glycogen concentrations were determined by
enzymatic methods according to Lowry and Passonneau (18)
after acid hydrolysis. The remainder of the homogenate was
centrifuged at 1,000 g. Aliquots of the muscle extracts and of
the incubation media were placed in scintillation vials contain-
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ing 4 ml of Aquasol-2 (NEN) and were counted in a liquid
scintillation counter with channels preset for simultaneous
3H and 14C counting. The amount of each isotope present in
the samples was determined, and this information was used
to calculate the extracellular space and the intracellular
concentration of 2-DG. The intracellular water content of the
muscles was calculated by subtracting the measured extracel-
lular space water from the total muscle water. Glucose
transport activity is expressed as micromoles of 2-DG per
milliliter intracellular water per 20 minutes.
Statistics. All values are means 6 SE. Statistical compari-
sons of the groups were made by one-way analysis of vari-
ance, and each group was compared with Fisher’s paired least
significant difference analysis. The correlation analysis was
done with Pearson’s test. Statistical significance was defined
as P , 0.05.
RESULTS
Effects of high-intensity intermittent swimming on
muscle glucose transport and glycogen in rat Epi muscle.
When rats swam with a weight equal to 18% of body
mass, muscle glycogen concentration in Epi decreased
as the number of bouts of swimming was increased
(Table 1). Eight bouts of swimming reduced muscle
glycogen concentration to a level of 7.6 6 0.5 µmol
glucose/g muscle (Table 1). 2-DG transport also in-
creased as the number of bouts of swimming was
increased (Table 1).
Effects of in vitro tetanic contractions on muscle
glucose transport and glycogen in rat Epi muscle.
Muscle glycogen concentration in Epi decreased with
increasing numbers of in vitro tetani (Table 2). After
Bouts of 20-s Swimming With Weight 5 18% body wt
1 (n 5 9) 3 (n 5 11) 8 (n 5 18)
22.3 6 2.0* 12.9 6 1.3*† 7.6 6 0.5*†‡
0.55 6 0.05 1.36 6 0.11*† 2.25 6 0.08*†‡
1854 GLUCOSE TRANSPORT AFTER HIGH-INTENSITY INTERMITTENT SWIMMING

Table 2. Effects of in vitro tetanic contractions on glycogen concentration and glucose transport activity in rat
epitrochlearis muscle
No. of Tetani

Basal (n 5 6) 1 (n 5 5) 5 (n 5 5) 10 (n 5 6) 15 (n 5 7) 20 (n 5 7)

Muscle glycogen, µmol glucose units/g wet wt 25.9 6 2.3 22.8 6 1.6 18.1 6 3.6* 14.8 6 1.4*† 14.9 6 1.0*† 13.7 6 1.0*†
2-Deoxy-D-glucose transport, µmol · ml21 · 20 min21 0.33 6 0.05 0.58 6 0.10 0.93 6 0.06*† 1.02 6 0.16*† 1.05 6 0.12*† 1.14 6 0.10*†
Values are means 6 SE; n 5 no. of muscles. Muscles were excised and stimulated at 100 Hz for 10 s at a rate of 1 contraction/min for 1, 5, 10,
15, and 20 min. * Significantly different from basal, P , 0.05. † Significantly different from 1 tetani. P , 0.05.

five tetani, muscle glycogen concentration was de-
creased, compared with the basal level (P , 0.01), but
more tetani did not significantly induce any further
decrease in muscle glycogen (Table 2). Twenty tetani
reduced muscle glycogen concentration to a level of
13.7 6 1.0 µmol glucose/g muscle (Table 2). 2-DG
transport also increased with increasing numbers of in
vitro tetani (Table 2). Although 2-DG transport was
significantly increased after five tetani compared with
2.83 2 0.11x, r 5 20.95, P , 0.01). When the data after
high-intensity intermittent swimming alone were plot-
ted, 2-DG transport rate was negatively correlated with
postexercise muscle glycogen level ( y 5 3.06 2 0.12x,
r 5 20.99, P , 0.01).
Effects of high-intensity intermittent swimming, insu-
lin, and combined stimuli on muscle glucose transport
and glycogen in rat Epi muscle. Incubation of Epi
muscle with 2 mU/ml of insulin increased 2-DG trans-

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the basal level (P , 0.01), more tetani did not result in
further increase in 2-DG transport (Table 2).
Effects of low-intensity continuous swimming on
muscle glucose transport and glycogen in rat Epi muscle.
When rats swam with a weight equal to 3% of body
mass, muscle glycogen concentration in Epi decreased
as the duration of swimming was increased (Table 3).
2-DG transport also increased as the duration of swim-
ming was prolonged to 120 min (Table 3).
Effects of high-intensity intermittent swimming, in
vitro tetani, and combined stimuli on muscle glucose
transport and glycogen in rat Epi. Fifteen tetanic
contractions, which provide a maximal effect of in vitro
tetani on 2-DG transport, resulted in a 3.2-fold increase
in 2-DG transport rate above the basal level (Table 4).
Eight bouts of high-intensity swimming induced a
7.3-fold increase in the rate of 2-DG transport above the
basal level, which was significantly greater than that
induced by 15 tetani alone (P , 0.01; Table 4). We also
examined the effects of swimming followed by 15 tetani
in vitro on 2-DG transport. The combined effects of
swimming and subsequent in vitro tetani on 2-DG
transport were not significantly greater than the effect
of swimming alone (Table 4).
Figure 1 shows the relationship between the mean
2-DG transport rate and the mean muscle glycogen
level at the end of swimming and in vitro tetani. When
all the data after in vitro tetani, high-intensity intermit-
tent swimming, and low-intensity continuous swim-
ming were plotted, 2-DG transport rate was negatively
correlated with muscle glycogen concentration ( y 5
port from a basal value of 0.33 6 0.04 µmol · ml intracel-
lular water21 · 20 min21 (n 5 8) to 1.34 6 0.07 µmol · ml
intracellular water21 · 20 min21 (n 5 5). High-intensity
intermittent swimming followed by 15 tetani in vitro
increased 2-DG transport to 2.32 6 0.13 µmol · ml
intracellular water21 · 20 min21 (n 5 5). Furthermore,
swimming followed by in vitro tetani and subsequent
insulin (2 mU/ml) incubation increased 2-DG transport
to 3.41 6 0.29 µmol · ml intracellular water21 · 20 min21
(n 5 7). Thus the combined effects of insulin and swim-
ming followed by in vitro tetani were additive.
DISCUSSION
In the present study, the high-intensity intermittent
swimming increased glucose transport in rat Epi to a
much higher level than that induced by repeated
10-s-long in vitro tetani alone (Table 4). A previous
study (3) reported that ten 10-s-long in vitro tetanic
contractions increased glucose transport to a level as
high as that achieved after four bouts of 30-min swim-
ming with a weight equal to 2% of body mass. There-
fore, glucose transport after ten 10-s-long in vitro tetani
has been considered as the exercise-induced maximal
activation of glucose transport in Epi. However, the
present study showed that high-intensity intermittent
swimming raises glucose transport in rat Epi to a
higher level than do ten 10-s-long in vitro tetani.
We do not know whether other in vitro contraction- or
exercise-related protocols increase glucose transport to
a higher level than that observed after ten 10-s-long in

Table 3. Effects of low-intensity continuous swimming on glycogen concentration and glucose transport activity
in rat epitrochlearis muscle
Duration of Swimming With Weight 5 3% Body Wt

Basal (n 5 5) 10 min (n 5 4) 30 min (n 5 6) 120 min (n 5 5)

Muscle glycogen, µmol glucose units/g wet wt 26.8 6 2.8 17.0 6 1.4* 13.5 6 1.2* 9.9 6 0.8*†
2-Deoxy-D-glucose transport, µmol · ml21 · 20 min21 0.38 6 0.02 0.78 6 0.09* 1.23 6 0.14*† 1.67 6 0.15*†‡
Values are means 6 SE; n 5 no. of muscles. Immediately after swimming, muscles were excised and analyzed. * Significantly different from
basal, P , 0.05. † Significantly different from 10-min swimming, P , 0.05. ‡ Significantly different from 30-min swimming, P , 0.05.
 GLUCOSE TRANSPORT AFTER HIGH-INTENSITY INTERMITTENT SWIMMING
Table 4. Interactions among effects of in vitro tetanic contractions and high-intensity swimming on glycogen
concentration and glucose transport activity in rat epitrochlearis muscle
Basal
(n 5 8)
Muscle glycogen, µmol glucose units/g wet wt 26.7 6 1.8
2-Deoxy-D-glucose transport, µmol · ml21 · 20 min21 0.33 6 0.04
Values are means 6 SE; n 5 no. of muscles. Swimming 1 Tetani, rats swam with weight 5 18% body wt for 20 s at a rate of 1 swim/min for 8
min, then muscles were excised and stimulated at 100 Hz to contract for 10 s at a rate of 1 contraction/min for 15 min. * Significantly different
from basal, P , 0.05. † Significantly different from 15 tetani, P , 0.05.
vitro tetani. Therefore, it is not, so far, plausible to
conclude that this high-intensity intermittent swim-
ming protocol induces the exercise-related maximal
stimulus for glucose transport. However, our finding
showed that the high-intensity intermittent swimming
protocol increases glucose transport to a higher level
than that induced by ten 10-s-long in vitro tetani,
which was regarded as the exercise-related maximal
stimulus for glucose transport in Epi.
We previously showed that glucose transport in Epi
after repeated in vitro tetani was negatively correlated
with postcontraction muscle glycogen content (17). In
the present study, we also demonstrated a significant
negative correlation between glucose transport and
muscle glycogen content at the end of high-intensity
intermittent swimming (Fig. 1). Furthermore, the data
of muscle glycogen and glucose transport after in vitro
tetani fell on the same regression line obtained from
the data after high-intensity intermittent swimming
(Fig. 1). Therefore, these results suggest that glucose
transport in Epi is influenced by glycogen content
regardless of the mode of contraction (in vitro tetani or
swimming). In the present study, high-intensity inter-
mittent swimming reduced muscle glycogen concentra-
tion in Epi to a level of 7.6 µmol glucose/g muscle, which
has never been observed after repeated 10-s-long in
vitro tetani (Table 1). Therefore, the higher glucose
Fig. 1. Relationship between mean value of postexercise glycogen
concentration and mean rate of 2-deoxy-D-glucose (2-DG) transport
in rat epitrochlearis muscle after 1 (h), 5 (M), 10 (°), 15 (b), and 20
(j) in vitro tetanic contractions; 10 (S), 30 (u ), and 120 (Q) min
of low-intensity continuous swimming; 1 (X), 3 (a), and 8 (d) bouts
of high-intensity intermittent swimming; and 8 bouts of high-
intensity swimming following 15 in vitro tetani (B). Data are shown
in Tables 1, 2, 3, and 4. Values are means 6 SE.
1855
15 Tetani 8 Bouts of Swimming 1 Tetani
(n 5 5) 18% Swimming (n 5 7) (n 5 5)
13.7 6 1.0* 8.3 6 0.7*† 6.9 6 0.5*†
1.07 6 0.12* 2.40 6 0.07*† 2.32 6 0.13*†
transport induced by high-intensity intermittent swim-
ming seems to be related to a lower muscle glycogen
content after swimming than that observed after re-
peated 10-s-long in vitro tetani. However, a causal
mechanism explaining the negative correlation be-
tween muscle glycogen and glucose transport after
exercise is not known. Some novel experimental ap-
proach seems necessary to explain the high glucose
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transport activity after high-intensity intermittent
swimming.
In the present study, 120-min-long, low-intensity
swimming reduced muscle glycogen concentration to a
lower level and activated glucose transport to a greater
extent than did ten 10-s-long in vitro tetani (Tables 2
and 3), whereas a previous study (3) reported that
low-intensity swimming with the same duration acti-
vated glucose transport to the same extent as did ten
10-s-long in vitro tetani. Apparently, the results ob-
tained from the two studies are in conflict. Because the
weight attached to the swimming rat (3% of body weight)
in the present study was heavier than that used in the
previous study (2% of body weight), the total muscle
work (probably substrate consumption) of the Epi during
swimming in the present study is thought to be greater
than that in the previous study. Therefore, it is possible
that muscle glycogen content in the present study was
lower than that in the previous study. Consequently, we
believe that this apparent difference can also be explained
by our hypothesis that the lower the glycogen content
in muscle, the higher the glucose transport activity.
In a previous study by Gulve et al. (9), when the Epi
was allowed to recover for 3 h after prolonged low-
intensity swimming in incubation medium without
glucose, glucose transport returned to basal levels. In
this study, during the recovery period, muscle glycogen
in Epi was not resynthesized, because the incubation
medium did not contain glucose. Furthermore, when
Epi was allowed to recover in incubation medium
without glucose for 2 h after high-intensity intermit-
tent swimming, we observed that glucose transport
returned to near-basal levels while muscle glycogen
content was as low as that observed immediately after
the swim (unpublished observations). Therefore, this
suggests that a reduction of muscle glycogen itself does
not initiate the biochemical reactions that lead to
activation of glucose transport. In addition to reduced
muscle glycogen content, other factor(s) may be neces-
sary for activating glucose transport. The transient
increase in the cytoplasmic Ca21 during muscle contrac-
tion may be a possible factor. Previous studies (1, 13,
1856 GLUCOSE TRANSPORT AFTER HIGH-INTENSITY INTERMITTENT SWIMMING
24, 26) suggest that an increase in cytoplasmic Ca21
that activates phosphorylase kinase and induces a
burst of glycogenolysis activates the process that re-
sults in increased glucose transport. An increase in
cytoplasmic Ca21 may increase glucose transport
through activation of a Ca21-activated enzyme, in coop-
eration with a reduction in muscle glycogen.
Muscle contraction stimulates glucose transport by
translocating glucose transporter GLUT-4-containing
vesicles from an intracellular pool to the plasma mem-
brane (6, 8, 19). An increase in GLUT-4 at the plasma
membrane is considered to be due both to stimulation of
translocation from an intracellular pool to the plasma
membrane and to a decrease in the rate of endocytosis
(16, 25). In terms of explaining the data obtained in the
present investigation, we raise the hypothesis that
translocation of GLUT-4 vesicles is interfered with by
muscle glycogen. Coderre et al. (2) reported that ,30%
of the GLUT-4 in rat skeletal muscle coprecipitated
with glycogen and that transporters could be released
from the glycogen particles by amylase digestion. There-
fore, it may be possible that the more muscle glycogen
is broken down, the more GLUT-4 is freed from glyco-
gen, resulting in an increased intracellular pool of free
GLUT-4 available in response to muscle contraction.
Thus less muscle glycogen binding to intracellular
GLUT-4 vesicles may facilitate GLUT-4 vesicles to
undergo translocation in response to muscle contrac-
tion. We speculate that another factor (for example,
Ca21), other than a reduction in muscle glycogen, may
trigger the translocation of GLUT-4 vesicles, and that
the amount of muscle glycogen binding to GLUT-4
vesicles may control the degree of GLUT-4 vesicle
translocation.
Previous studies in which rat Epi muscle was used
indicated that the maximal effects of insulin and in
vitro tetani on glucose transport in Epi were additive
(11, 22). Therefore, these previous studies suggest the
hypothesis that muscle contractile activity and insulin
increase glucose transporter through two different
mechanisms. In the present study, although the effect
of high-intensity intermittent swimming on glucose
transport was much greater than the effect of in vitro
tetani, the combined effects of swimming and insulin on
glucose transport were also additive (see RESULTS).
Therefore, the present study supports the hypothesis
that there are two separate mechanisms of glucose
transport activation in skeletal muscle.
During exercise, muscle glycogen is catabolized to
lactate or CO2 for the resynthesis of ATP. When rats
swam with a weight equal to 18% of body mass, the
blood lactate concentrations were significantly elevated
from resting levels of 1.3 6 0.2 mM (n 5 7) to 10.4 6 0.3
mM (n 5 7), and the mean exhaustion time was 63 s
(see METHODS). Because the increase in blood lactate
concentration roughly reflects production of lactate in
active skeletal muscle, large amounts of glycogen in
active skeletal muscle are considered to be catabolized
to lactate during the high-intensity swimming. Further-
more, Medbo and Tabata (20) demonstrated that most
of the glycogen broken down during 60 s of exhaustive
exercise was catabolized to lactate. On the other hand,
rats with a weight equal to 3% of body mass could swim
for .120 min. During such a low-intensity exercise, the
blood lactate concentration did not increase above
resting levels. Therefore, consumed glycogen during
low-intensity swimming was assumed to be metabo-
lized not to lactate but to CO2. However, the data of
muscle glycogen and glucose transport after low-
intensity continuous swimming fell on the same regres-
sion line obtained from the data after high-intensity
swimming (Fig. 1). Therefore, we suggest that the effect
of exercise on glucose transport is mediated by a
process associated with a decrease in muscle glycogen
content, regardless of the end product from glycogen
breakdown.
In summary, high-intensity intermittent swimming
increases glucose transport in rat Epi to a higher level
than that induced by ten 10-s-long in vitro tetani,
which has been regarded as the maximal exercise-
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related stimulus for glucose transport. This result may
be explained by the lower muscle glycogen level after
high-intensity intermittent swimming than after in
vitro tetani, because there was a significant negative
correlation between glucose transport and muscle glyco-
gen concentration in Epi after the high-intensity swim-
ming and the in vitro tetani.
The authors thank Dr. John O. Holloszy (Washington Univ. School
of Medicine, St. Louis, MO) for his help in editing the English
manuscript.
This work was supported by the research grants from the Research
Development Corporation of Japan (to K. Kawanaka); Grants-in-Aid
for Scientific Research 09480017 and 08680158 from the Ministry of
Education, Science, Sports, and Culture of Japan (to I. Tabata); and
the research grants from the Human Science Foundation (to M.
Higuchi). K. Kawanaka was supported by the National Institute
Postdoctoral Fellowship, Japan.
Present address of K. Kawanaka: Section of Applied Physiology,
Dept. of Internal Medicine, Washington Univ. School of Medicine,
Campus Box 8113, 660 S. Euclid Ave., St. Louis, MO 63110-1093.
Present address of A. Tanaka: Fujisawa City Health and Medical
Center, Ohba 5527-1, Fujisawa City, Kanagawa 251, Japan.
Address for reprint requests: I. Tabata, Division of Health Promo-
tion, National Institute of Health and Nutrition, Toyama 1-23-1,
Shinjuku City, Tokyo 162-8636, Japan.
Received 11 June 1997; accepted in final form 13 February 1998.
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