Hindawi Publishing Corporation

ISRN Physiology
Volume 2013, Article ID 371235, 8 pages
http://dx.doi.org/10.1155/2013/371235

Research Article
Reduced Muscle Glycogen Differentially Affects Exercise
Performance and Muscle Fatigue

Jay H. Williams,1 Timothy W. Batts,1 and Simon Lees1, 2

1
Department of Human Nutrition, Foods and Exercise, Virginia Polytechnic Institute & State University, Blacksburg, VA 24060, USA
2
Medical Sciences Division, Northern Ontario School of Medicine, under Bay, ON, Canada P7B 5E1

Correspondence should be addressed to Jay H. Williams; [email protected]

Received 21 August 2012; Accepted 26 September 2012

Academic Editors: J. Cannon and K. Sakuma

Copyright © 2013 Jay H. Williams et al. is is an open access article distributed under the Creative Commons Attribution License,
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

is investigation examined the effects of reduced muscle glycogen on exercise performance and muscle fatigue. Male rats were
assigned to a low glycogen group (LG) that participated in a protocol of exercise and fasting, a high glycogen group (HG) that
exercised but were allowed free access to food, or control group (CON) that did not exercise but were allowed free access to food.
Following the protocol, muscle glycogen content of the LG animals was reduced by 45%. e LG animals also performed 79 and
81% less voluntary treadmill exercise than the HG and CON groups. At exhaustion, the LG group had lower blood glucose than
HG and CON but exhibited no reduction in sarcoplasmic reticulum (SR) function. During 30 min of in situ stimulation, the rates
and magnitudes of muscle fatigue were not signi�cantly different between groups, and fatigue-induced reductions in SR function
were similar between groups. e results indicate that reduced muscle glycogen markedly impairs voluntary exercise performance
but does not appreciably affect isolated muscle function. It is likely that exercise exhaustion due to reduced muscle glycogen is due,
in large part, to hypoglycemia and central fatigue as opposed to peripheral mechanisms.

1. Introduction in intracellular ATP concentration, resulting in an inability

to meet the energy demands of the contraction process. As a
As early as the 19th century a relationship between car- consequence, the muscle would be unable to sustain maximal
bohydrate metabolism and fatigue was proposed. In 1807, force output. Green [7] de�ned this notion as the “energy
Berzelius suggested “the amount of free lactic acid in skeletal crisis” hypothesis. While this remains a widely accepted
muscle is proportional to the extent to which it has been exer- explanation to account for the link between muscle glycogen
cised” [1]. Exhaustive exercise is known to cause to dramatic and fatigue, it has received little experimental support. In fact,
reductions in muscle glycogen content within the active mus- it is oen observed that during prolonged or intense exercise,
cle [2–4]. Also, the onset of exhaustion is dependent on the there is little or no reduction in intracellular ATP despite near
initial muscle glycogen content [2] and dietary manipulation complete depletion of muscle glycogen (see [8]).
of muscle glycogen can increase or decrease exercise duration e process of voluntary muscle contraction consists of
[5]. Based on this, most researchers readily accept the notion a long chain of events beginning with input to the motor
that muscle glycogen depletion is somehow involved in the cortex and ending with force production via actin-myosin
development of fatigue during prolonged exercise. interaction. If any one of the steps in this sequence fails, force
Despite this, more than 40 years of investigation have output will be compromised. In simple terms, fatigue can be
failed to identify a speci�c “cause-and-effect” relationship thought of as “central” or “peripheral” [8]. Central fatigue
between muscle glycogen and fatigue during exercise. New- involves events responsible for 𝛼𝛼-motoneuron activation. In
sholm and Leech [6] proposed that during muscular activity, this case, force output is limited by failure to adequately
the loss of glycogen results in an inability to sustain an activate the 𝛼𝛼-motoneuron pool. Central fatigue can be
adequate glycolytic �ux rate to support ATP regeneration. further described as “supraspinal”, failure at the level of the
Reductions in ATP resynthesis would then cause a decline motor cortex, or “spinal”, involving afferent activation or
2 ISRN Physiology

excitability of the 𝛼𝛼-motoneuron pool [9]. On the other hand, Next, a fatigue protocol was applied, which consisted of
peripheral fatigue occurs distal to the spinal cord and usually 333 ms trains of pulses (20 Hz) delivered at a rate of one per
refers to mechanisms at the level of the muscle �ber. second for 30 min. Immediately aer the fatigue protocol,
In the present study, we sought to gain insight into a single titanic contraction was elicited. All contractions
the mechanisms whereby reduced muscle glycogen limits were evoked and sampled via microcomputer then analyzed
exercise performance through central or peripheral fatigue off-line for peak force and peak rate or relaxation. For the
mechanisms. We utilized a rodent model of reduced muscle rested condition, the three contractions were averaged. For
glycogen [10] and examined the effects on exercise perfor- the fatigued condition, the single contraction was used. is
mance (central and peripheral mechanisms) and in situ mus- was because there was some, albeit small recovery of force
cle function (peripheral mechanisms). We �nd that reduced between three postfatigue contractions separated by 1 min.
muscle glycogen severely limits exercise performance but has
little effect on in situ muscle force or fatigue.
2.3. Biochemical and Sarcoplasmic Reticulum (SR) Function
Measurements. Immediately before (rested) and aer (ex-
2. Methods hausted) the exercise performance treadmill bout, animals
were quickly anesthetized with CO2 inhalation, and a blood
2.1. Experimental Model. For all experiments, female Spra- sample was obtained for plasma glucose measurement. ey
gue-Dawley rats were used (200–250 g). All procedures used were then decapitated, and both gastrocnemius muscles were
were reviewed and approved by the Virginia Tech Animal removed. One muscle was prepared for glycogen analysis
Care Committee. e experimental model used in this study and the other prepared for SR function measurements. For
has been described in detail [10]. �rie�y, to reduce muscle comparison with exhausted LG animals, separate groups of
glycogen, a protocol combining exercise and fasting was used. HG and CON animals were sacri�ced aer 35 minutes of
Rats were randomly divided into three groups, high glycogen exercise (35 min). is resulted in three conditions for these
(HG), low glycogen (LG), and control (CON). During the groups: rested, exhausted, and 35 min.
�rst 24 hrs of the protocol, food was removed from the HG For the in situ stimulation experiments, both gastroc-
and LG groups. Aer this initial fast, these animals were nemius muscles were removed (rested and fatigued muscle
placed on a motor-driven treadmill and exercised for 90 min from each animal) and prepared for glycogen or SR function
at a running pace of 21 m/min and 5% grade. e treadmill analysis (separate animals were used for each assay). A blood
was equipped with an electrical grid that provided a mild sample was also obtained for plasma glucose measurement.
external stimulus during exercise. Immediately aer exercise, Animals were then euthanized by anesthesia overdose. In a
animals were returned to their cages. e HG group was separate group of animals, muscles were removed for glyco-
then given free access to standard rodent chow and water gen measurements. ey were immediately frozen in liquid
supplemented with 5% sucrose. e LG group was given free N2 then freeze dried. Freeze-dried samples were weighed
access to water (no sucrose) and fasted for another 24 hrs. e and then homogenized in 5 vol of ice-cold perchloric acid.
CON group was given free access to food and water and did Glycogen content was determined using the glucoamylase
not participate in the exercise protocol. (E.C. 3.2.1.3) method described by Keppler and Decker [14].
Plasma glucose was determined using the glucose oxidase
2.2. Exercise and Muscle Performance. Exercise performance method using a commercially available kit (Stanbio).
was determined using treadmill running to exhaustion. ATP, ADP, and PCr measurements were made using
Animals were exercised at a pace of 21 m/min, 5% grade. high-performance liquid chromatography (HPLC). A por-
Exhaustion was determined when the animals could no tion of each freeze-dried muscle was weighed, minced, and
longer keep pace with the treadmill and failed to right then homogenized in 5 volumes of ice-cold perchloric acid.
themselves when placed on their back. Aer incubating the samples on ice for 20 minutes, the pH
e in situ muscle stimulation was carried out as was neutralized with KOH. Samples were then centrifuged
described by Williams and Ward [11] and Lees et al. [12]. at 1600 xg to sediment proteins and precipitated KCl. HPLC
�rie�y, animals were anesthetized using ketamine (40 mg/kg) analysis was carried out as described [15, 16] using a Waters
and xylazine (8 mg/kg) delivered ip. e le gastrocnemius HPLC system equipped with a diode array detector and a
muscle was isolated; the Achilles tendon was cut at the Supelchem C18 3 𝜇𝜇m reversed phased column (0.46 × 15 cm).
calcaneus and tied via surgical thread, to an isometric For the SR function measurements, dissected muscles
transducer (Harvard Apparatus). Muscle temperature was were immediately placed in ice-cold buffer containing 20 mM
maintained at 37∘ C using a warming lamp, and hydration was HEPES, 0.2% sodium azide, 0.2 mM PMSF, and 1 mM
preserved by bathing muscle with mineral oil and covering EDTA (VirtiShear, 3 × 15 s). Differential centrifugation was
the area with polyvinylidene chloride �lm (Saran). e then used to isolate SR vesicles (8,000–60,000 xg pellet) as
muscle was stimulated through electrodes placed around the described previously [17]. Following centrifugation, the �nal
sciatic nerve. Contractions were evoked via 0.5 msec pulses SR pellets were frozen and stored at −80∘ C until used.
using a voltage that elicited maximal twitch force. Ca2+ uptake was determined as described by Lees and
Aer resting length was determined (i.e., the length that Williams [17]. For the uptake measurements, SR protein
resulted in peak twitch force), three tetanic contractions (25 𝜇𝜇g) was added to 1.5 mL of buffer (100 mM KCl, 20 mM
(333 msec, 100 Hz) were elicited (separated by 1 min) [13]. HEPES, 5 mM MgCl2 , 5 mM KH2 PO4 , 2 mM ATP and
ISRN Physiology 3

250 𝜇𝜇M antipyrylazo III, pH. 7.0, 37∘ C). Uptake was initiated 250
by adding 1.2 𝜇𝜇mol/mg CaCl2 and was allowed to continue
until free [Ca2+ ] in the cuvette declined to a plateau. APIII
∗
absorbance was monitored at 790 nm and 710 nm using 200
a diode array spectrophotometer (Aligent) and converted

Muscle glycogen (mmol/kg)

∗
to free [Ca2+ ]. Peak rate of uptake was determined as the
steepest slope of the free [Ca2+ ] time curves. 150
Ca2+ ATPase activities were determined using a coupled-
enzyme assay [18]. Samples (20 𝜇𝜇g protein) were added to
incubation buffer (25 mM HEPES, 100 mM KCl, 10 mM 100
MgCl2 , 1 mM EGTA, 0.2% NaN2 , 2 𝜇𝜇M A23187, 5 U/mL
lactate dehydrogenase, 7.5 U/mL pyruvate kinase, 3.0 mM
phosphoenolpyruvate, and 0.6 mM NADH, pH 7.0, 37∘ C).
∗†
50 †
NADH absorbance was monitored continuously at 340 nm †
using a diode array spectrophotometer (Aligent). e reac-
tion was started with addition of ATP (5 mM) and basal 0
activity was recorded for 3 minutes. Ca2+ was then added Rested 35 min Exhaustion
(2.0 𝜇𝜇M free [Ca2+ ]), and total activity was recorded for an 8
additional 3 minutes. Ca2+ -stimulated activity was computed
∗
by subtracting basal activity from total activity. 7 ∗

2.4. Statistics. Analyses of variance with Tukey’s post-hoc 6

Plasma glucose (µmol/mL)

exams were used to identify differences between the HG, LG,

and CON groups for all variables recorded. For the stimu- 5 ∗†
∗†
lation experiments, the analyses of variance were adjusted
4
for repeated measures performed on the same animal (rested
and fatigued muscles). Signi�cance was established at the 0.05
3
level of con�dence. †

2
3. Results
1
3.1. Treadmill Exercise. Animals in the LG group (35.21 ±
6.72 min) exercised for signi�cantly less time than did those
0
in the HG (166.84 ± 23.21 min) and CON (182.40 ± 17.90) Rested 35 min Exhaustion
groups (𝑃𝑃 𝑃 𝑃𝑃𝑃, 𝑛𝑛 𝑛 𝑛 for each group, mean ± SEM). Means
for the later two groups were not signi�cantly different. LG
Muscle glycogen and plasma glucose levels in rested and HG
exercise animals are shown in Figure 1. At rest, e LG ani- CON
mals displayed reduced levels of both glycogen and glucose. F 1: Muscle glycogen (top) and plasma glucose (bottom)
Compared to the HG animals, the LG animals’ glycogen was measured at rest and during exercise. Muscle glycogen values are
reduced by 45.0%. At exhaustion (∼35 minutes of exercise), expressed as glucosyl units per g dry mass. ∗ 𝑃𝑃 𝑃 𝑃𝑃𝑃 versus LG,
these animals showed marked glycogen depletion and hypo- †
𝑃𝑃 𝑃 𝑃𝑃𝑃 versus rested. 𝑛𝑛 𝑛 𝑛 for each group and time point.
glycemia. Glycogen was reduced by 85.0% compared to rest
and plasma glucose dropped to nearly 2 𝜇𝜇mol/mL. On the
other hand, the HG and CON animals showed no signi�cant As shown previously [10], Both SR Ca2+ uptake and Ca2+ -
changes in either parameter aer 35 minutes of exercise. At stimulated ATPase activity were increased in the LG animals
exhaustion, these animals did experience signi�cant declines by about 25% compared to the other groups (Table 2).
in both glycogen and glucose (compared to rest), but �nal At exhaustion, these animals did not display any exercise-
glycogen and glucose levels were higher than the LG group. induced reductions in SR function. Aer 35 minutes of
Also, plasma glucose in the HG and LG groups remained exercise, there were also no changes in SR Ca2+ uptake or
above 4.0 umol/mL at exhaustion. ATPase activity in the HG or CON animals. At exhaustion,
Intramuscular ATP, ADP, and PCr levels in rested and however, these latter groups showed 30–38% reductions in
exercised muscles are shown in Table 1. Under all three con- both parameters.
ditions, there were no signi�cant exercise-induced changes in
ATP levels but signi�cant increases in ADP and signi�cant 3.2. In Situ Stimulation. Force responses to electrical stim-
decrements in PCr. However, values were not signi�cantly ulation are shown in Figure 2. All three groups showed a
different between conditions in either rested or exhausted staircase response during the initial minute. is staircase
animals. response was signi�cantly smaller in the LG group than
4 ISRN Physiology

T 1: Muscle metabolites in rested and exercised animals. Values are expressed as 𝜇𝜇mol/g dry mass.

LG group HG group CON group

Rested 22.12 ± 1.80 25.34 ± 2.03 25.46 ± 1.98
ATP 35 min — 24.92 ± 2.20 25.65 ± 1.39
Exhausted 19.36 ± 2.45 20.56 ± 3.11 22.38 ± 2.06

Rested 3.12 ± 0.41 2.98 ± 0.56 3.20 ± 0.69

ADP 35 min — 4.94 ± 1.02† 5.22 ± 1.34†
Exhausted 5.36 ± 1.34† 5.06 ± 0.93† 5.27 ± 2.03†

Rested 78.33 ± 4.22 81.24 ± 3.22 80.44 ± 5.10

PCr 35 min — 19.34 ± 4.00† 20.67 ± 4.68†
Exhausted 17.56 ± 4.10† 18.21 ± 3.29† 22.64 ± 4.06†
†
𝑃𝑃 𝑃 𝑃𝑃𝑃 versus rested. 𝑛𝑛 𝑛 𝑛 for each group and time point.

T 2: Effects of treadmill exercise on SR function. Values are expressed per mg of protein.

LG group HG group CON group

Rested 1.25 ± 0.19 1.03 ± 0.12∗ 0.97 ± 0.19∗
Ca2+ Uptake rate (𝜇𝜇mol/mg/min) 35 min — 1.13 ± 0.10 1.08 ± 0.11
Exhausted 1.18 ± 0.13 0.64 ± 0.09∗† 0.68 ± 0.08∗†

Rested 3.16 ± 0.25 2.55 ± 0.39∗ 2.67 ± 0.29∗

2+
Ca -Stim ATPase (𝜇𝜇mol/mg/min) 35 min — 2.85 ± 0.28 2.56 ± 0.33
Exhausted 3.14 ± 0.21 1.76 ± 0.12∗† 1.77 ± 0.19∗†
∗
𝑃𝑃 𝑃 𝑃𝑃𝑃 versus LG, † 𝑃𝑃 𝑃 𝑃𝑃𝑃 versus rested. 𝑛𝑛 𝑛 𝑛 for each group and time point.

the other groups (Table 2). is was followed by a steady 1.6
decline in force during the following 90 sec. From minutes 3 1.4
to 30, force showed a much slower steady reduction in force.
To characterize the rate and magnitude of fatigue for each 1.2
animal, these forces were �t by nonlinear regression to an
Force (P/Pi )

1
equation of the form: 𝑃𝑃𝑃𝑃𝑃𝑜𝑜 = 𝑎𝑎 𝑎 𝑎𝑎−𝑘𝑘𝑘𝑘𝑘 + 𝑏𝑏, where −𝑘𝑘 is the 0.8
rate constant for force loss or fatigue and 𝑏𝑏 is the �nal force.
0.6
Neither of these variables was signi�cantly different between
the three groups. at is neither the rate nor magnitude of 0.4
fatigue during stimulation differed between groups. 0.2
Tetanic forces, recorded before stimulation, were not
signi�cantly different between groups (Table 3). Aer stim- 0
0 1 2 3 4 5 30
ulation, titanic forces were reduced by 46–50% but were
Time (min)
not signi�cantly different between groups. Relaxation rate
in rested muscles was signi�cantly higher in the LG group LG
(Table 2). In the fatigued muscles, these rates were decreased HG
compared to the rested condition in all three groups but were CON
not signi�cantly different between groups. F 2: Forces generated during constant 30 Hz stimulation.
Muscle glycogen recorded in the rested and stimulated Values are means for each group. Error bars are omitted for clarity.
limbs is shown in Figure 3. As can be seen, the rested limb 𝑛𝑛 𝑛 𝑛𝑛 for each group.
of the LG animals had signi�cantly lower glycogen content
than those of the other two groups. In the stimulated limb,
glycogen was lower than the rested limb in all three groups.
However, the stimulated limb of the LG animals contained stimulation-induced changes in ATP levels in any of the three
less glycogen than that of the HG and CON groups. Also, groups of animals. However, signi�cant increases in ADP and
plasma glucose at the end of stimulation was signi�cantly signi�cant decrements in PCr occurred in all three. Values
lower in the LG group than in the HG and CON animals were not signi�cantly different between conditions in either
(5.03 ± 0.52, 5.92 ± 0.61 and 6.01 ± 0.58 𝜇𝜇mol/mL, resp.). rested or fatigued muscles.
Table 4 shows intramuscular ATP, ADP, and PCr levels Both Ca2+ uptake and ATPase activity recorded in the
in rested and stimulated muscles. ere were no signi�cant rested limb of the LG animals were greater than values of the
ISRN Physiology 5

T 3: Effects of fatiguing stimulation of tetanic force and fatigue.

LG group HG group CON group

Staircase (𝑃𝑃/𝑃𝑃𝑖𝑖 ) 1.25 ± 0.07 1.40 ± 0.08∗ 1.42 ± 0.07∗
Rate constant (/𝑠𝑠) 1.54 ± 0.09 1.39 ± 0.09 1.47 ± 0.07
Final force (𝑃𝑃/𝑃𝑃𝑖𝑖 ) 0.29 ± 0.03 0.34 ± 0.03 0.32 ± 0.04

Rested 36.98 ± 3.42 33.15 ± 3.62 35.22 ± 4.14

Tetanic force (mN/gm)
Fatigued 18.33 ± 2.11† 17.53 ± 1.77† 19.02 ± 2.06†

Rested 61.9 ± 2.0 55.0 ± 2.5∗ 51.3 ± 2.1∗

Relaxation rate (mN/ms)
Fatigued 21.7 ± 2.7† 20.3 ± 3.1† 19.8 ± 2.6†
𝑃𝑃𝑖𝑖 : initial force. Tetanic forces are expressed per gram of dry muscle mass. ∗ 𝑃𝑃 𝑃 𝑃𝑃𝑃 versus LG, † 𝑃𝑃 𝑃 𝑃𝑃𝑃 versus rest. 𝑛𝑛 𝑛 𝑛𝑛 for each group and condition (rest
or fatigue).

200
∗
muscle glycogen affects blood glucose and contributes to the
180 development of central fatigue rather than peripheral fatigue.
∗
160
It should be pointed out that the in situ stimulation
Muscle glycogen (mmol/kg)

protocol used here did not completely mimic the activation

140
pattern of the gastrocnemius muscle during treadmill exer-
120 cise. For the pace and incline used here, it is estimated that
100 the gastrocnemius muscle was activated at 2–2.5 Hz [19, 20].
80
∗† ∗†
Also the duration of activation was estimated at 200–225 ms
or a 50% duty cycle [19, 20]. For the repetitive stimulation
60 ∗
protocol, contractions were evoked at 1 Hz with a duty cycle
40 of 33%. Also, activation of the gastrocnemius is submaximal
20 during treadmill exercise (at least during the early stages
0 of exercise), whereas with nerve stimulation it is likely
Rested Fatigue that all motor units were activated. Lastly, stimuli during
each evoked contraction were delivered at 20 Hz to avoid
LG possible high-frequency fatigue. As for treadmill running,
HG it is difficult to determine an average motor unit activation
CON frequency for a mixed muscle such as the gastrocnemius
F 3: Muscle glycogen rested muscles and muscle subjected to during activity. However, 20 Hz is well within the range of
electrical stimulation for 30 min. ∗ 𝑃𝑃 𝑃 𝑃𝑃𝑃 versus LG, † 𝑃𝑃 𝑃 𝑃𝑃𝑃 versus �ring frequencies of other muscles performing submaximal
rest. 𝑛𝑛 𝑛 𝑛 for each group and condition. contractions [21]. us, there were important differences
in the muscle activation pattern between treadmill exercise
and in situ stimulation. Despite these, it seems reasonable
to suggest that the differences between the two muscle
activation protocols were small enough such that they did not
HG and CON animals (Table 5). However, all three groups substantially contribute to the differential effects of reduced
demonstrated signi�cant reductions in SR function following muscle glycogen on treadmill running performance and in
electrical stimulation. Ca2+ uptake was reduced by 30–35%, situ fatigue.
and ATPase activity was lowered by 30–32% by stimulation. e peak staircase response was signi�cantly less in the
LG animals. is is likely due to the effects of reduced muscle
glycogen on SR function and relaxation rate. As shown earlier
4. Discussion by Batts et al. [10] reducing muscle glycogen through diet
Reducing muscle glycogen by approximately 45% through a and exercise leads to increased SR Ca2+ uptake rate and
protocol of exercise and fasting [10] resulted in a dramatic ATPase activity. Lees and Williams [17] also found that
reduction in treadmill exercise performance. However, in situ extraction of glycogen from the SR had the same effect on SR
muscle force and fatigue were not affected. Also, reduced Ca2+ pump function. Increasing the rate of SR Ca2+ uptake
muscle glycogen resulted in greater hypoglycemia following should accelerate relaxation and −𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑 (as shown here).
both treadmill exercise and in situ stimulation. us, when is, in turn would lead to a diminished staircase response
the central nervous system (CNS) is required for muscle during repetitive stimulation. us it is likely that the reduced
activity (e.g., treadmill exercise), glycogen depletion and staircase response was a direct effect of reduced glycogen on
hypoglycemia markedly impair performance. But, when the the SR Ca2+ ATPase.
CNS is bypassed by in situ stimulation, the effects are min- We did not assess in situ muscle function immediately
imal. Taken together, this supports the notion that reduced aer treadmill exercise. us it is not clear to what extent
6 ISRN Physiology

T 4: Muscle metabolites in muscles and muscle subjected to stimulation. Values are expressed as 𝜇𝜇mol/g dry mass.

LG group HG group CON group

Rested 26.45 ± 2.61 25.84 ± 3.02 27.19 ± 3.12
ATP
Fatigued 21.55 ± 3.56 22.32 ± 4.20 23.10 ± 3.09

Rested 2.95 ± 0.55 2.79 ± 0.65 3.06 ± 0.96

ADP
Fatigued 5.64 ± 1.44† 5.57 ± 1.00† 5.63 ± 1.56†

Rested 82.11 ± 4.33 80.26 ± 4.02 80.65 ± 4.65

PCr
Fatigued 29.32 ± 4.32† 27.64 ± 4.03† 27.99 ± 3.96†
†
𝑃𝑃 𝑃 𝑃𝑃𝑃 versus rested. 𝑛𝑛 𝑛 𝑛 for each group and time point.

T 5: Effects of fatiguing stimulation on SR function.

LG group HG group CON group

Rested 1.23 ± 0.18 0.91 ± 0.15∗ 1.02 ± 0.16∗
Ca2+ uptake rate (𝜇𝜇mol/mg/min)
Fatigued 0.79 ± 0.10† 0.59 ± 0.12† 0.62 ± 0.12†

Rested 2.95 ± 0.32 2.13 ± 0.31∗ 2.22 ± 0.25∗

Ca2+ -stimulated ATPase (𝜇𝜇mol/mg/min)
Fatigued 0.91 ± 0.19† 0.64 ± 0.17∗† 0.72 ± 0.19†
∗
𝑃𝑃 𝑃 𝑃𝑃𝑃 versus LG, † 𝑃𝑃 𝑃 𝑃𝑃𝑃 versus Rested. 𝑛𝑛 𝑛 𝑛 for each group and condition.

the LG animals were experiencing muscle fatigue at the point Others have reported a link between muscle glyco-
of exhaustion. However, we did measure SR function. A gen, excitation-contraction coupling, and SR function using
number of groups have shown that muscle fatigue following dietary manipulation or chemical extraction [10, 17, 25–
stimulation is clearly associated with depressions in SR Ca2+ 27]. ese studies suggest that the loss of glycogen during
uptake and ATPase activity (see [22, 23]). Also, in well-feed exercise may affect force during fatigue via direct actions
animals, exhaustion aer treadmill exercise is associated with on SR Ca2+ uptake and release. In addition, a recent study
reductions in SR function. In the exhausted state, the LG by Ørtenblad et al. [28] used glycogen-depleting exercise
animals showed no change in SR function. Likewise, the HG coupled with carbohydrate supplementation and found a
and CON groups showed no change in the SR function at close relationship between low muscle glycogen levels and
approximately the same exercise duration (35 min). However, reduced SR Ca2+ release. ey found that depressed SR
both the HG and CON groups showed marked depressions in function returned to normal four hours postexercise when
SR Ca2+ pump function at exhaustion. Following stimulation carbohydrate was provided. Without supplementation, both
where force was reduced by nearly 70%, all three groups glycogen and SR Ca2+ release remained depressed. However,
showed reduced SR Ca2+ function. While SR Ca2+ uptake Ørtenblad et al. [28] did not measure muscle function or
and ATPase activity not necessarily indicated fatigue, these exercise performance. us it is not clear to what extent
results are consistent with the idea that the LG animals likely the muscle had recovered and it is not clear if the same
experienced little muscle fatigue at the point of exhaustion, at relationship between glycogen and SR function would hold
least not to the extent as the HG and CON groups. in a rested muscle that is deplete of glycogen. In the present
It should be pointed out that Karelis et al. [24] showed study, the muscle function was allowed to full recovery for
that glucose infusion during in situ stimulation attenuated 24 hours (as indicated by in situ force measurements) aer
the extent of fatigue and partially restored force in fatigued exercise. is allowed for the relationship between glycogen
muscle, possibly by maintaining Na/K pump function. is and SR function to be examined in the absence of fatigue.
suggests that blood glucose may have a direct effect on muscle In this case, a reduction in glycogen resulted in increased SR
function during fatiguing exercise. It also raises the possibility Ca2+ uptake and ATPase activity. Given this, it is possible that
that the hypoglycemia experienced by the LG animals during a more complex relationship exists between muscle glycogen
exercise may have induced some degree of peripheral fatigue and SR function at rest and during recovery from exercise.
during treadmill exercise. However, in the Karelis et al. [24] When viewed as a whole, the results of this investigation
study, plasma glucose levels were very high during infusion, suggest that reduced muscle glycogen during exercise can
nearly twice the initial and control levels and, in the control lead to the development of hypoglycemia and central fatigue.
condition, there was no decline in blood glucose. Further, Mosso [29] was one of the �rst to suggest that �mental
the slight hypoglycemia experience by the LG animals during fatigue” contributes to muscular performance. e idea that
stimulation (present study) was not associated with alter- central fatigue contributes to force loss during exercise and
ations in force or fatigue. While it is possible that plasma that it may be linked to hypoglycemia is not novel (e.g.,
glucose may directly affect muscle force during exercise, more [7, 30–32]). Glucose oxidation provides a major energy
evidence is needed to support such a notion. source for the brain [33]. Since the brain maintains very
ISRN Physiology 7

limited supplies of endogenous glycogen, plasma glucose [4] L. Hermansen, E. Hultman, and B. Saltin, “Muscle glycogen
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[33]. In fact, cerebral glucose uptake declines when arterial ica, vol. 71, no. 2, pp. 129–139, 1967.
concentration drops below 3.6 mM [34]. Coyle et al.[31] [5] J. Bergström, L. Hermansen, E. Hultman, and B. Saltin, “Diet,
showed that carbohydrate feeding during prolonged exercise muscle glycogen and physical performance,” Acta Physiologica
delayed the development of fatigue, but only in subjects that Scandinavica, vol. 71, no. 2, pp. 140–150, 1967.
experienced hypoglycemia. Nybo et al. [33, 35] found that [6] E. A. Newsholm and A. R. Leech, Biochemistry for the Medical
exercise-induced hypoglycemia reduced both voluntary force Sciences, John Wiley & Sons, New York, NY, USA, 1983.
and the level of muscle activation. In addition, cerebral blood [7] H. J. Green, “How important is endogenous muscle glycogen to
�ow, brain glucose, and oxygen uptakes were reduced as well. fatigue in prolonged exercise?” Canadian Journal of Physiology
ese changes were prevented when glucose supplementa- and Pharmacology, vol. 69, no. 2, pp. 290–297, 1991.
tion was used and hypoglycemia was avoided. us, there [8] R. H. Fitts, “Substrate supply and energy metabolism during
is a strong case that muscle glycogen depletion is linked to brief high intensity exercise: importance in limiting perfor-
voluntary exhaustion via hypoglycemia and central fatigue. mance,” in Perspectives in Exercise Science and Sports Medicine,
It is also possible that muscle glycogen depletion and D. R. Lamb and C. V. Gisol�, Eds., vol. 5 of Energy Metabolism
hypoglycemia during exercise result in brain glycogen deple- in Exercise and Sport, pp. 53–106, Wm. C. Brown, Dubuque,
tion. Matsui et al. [36] found that prolonged exercise resulted Iowa, USA, 1992.
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ey argued that low blood glucose triggered increased cle fatigue,” Physiological Reviews, vol. 81, no. 4, pp. 1725–1789,
astrocytic glycogenolysis leading to reduced brain glycogen 2001.
and possible central fatigue. Unfortunately, they did not [10] T. W. Batts, S. J. Lees, and J. H. Williams, “Combined effects of
measure the recovery of brain glycogen following exercise. exercise and fasting on skeletal muscle glycogen and sarcoplas-
us, it is not clear if our LG animals began their exercise mic reticulum function,” Basic and Applied Myology, vol. 19, pp.
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the early hypoglycemia experienced by the LG group leads to [11] J. H. Williams and C. W. Ward, “Changes in skeletal muscle sar-
reductions in brain glycogen, central fatigue, and diminished coplasmic reticulum function and force production following
exercise performance. However, more information on poten- myocardial infarction in rats,” Experimental Physiology, vol. 83,
no. 1, pp. 85–94, 1998.
tial links between changes in muscle glycogen and central
nervous system metabolism during prolonged exercise and [12] S. J. Lees, P. D. Franks, E. E. Spangenburg, and J. H. Williams,
recovery is needed. “Glycogen and glycogen phosphorylase associated with sar-
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that spinal mechanisms may contribute to central fatigue dur-
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