ARTICLE

Carbohydrate Availability and Training

Adaptation: Effects on Cell Metabolism
John A. Hawley1 and Louise M. Burke 2
1
Health Innovations Research Institute, School of Medical Sciences, RMIT University, Bundoora, Victoria;
and 2Department of Sports Nutrition, Australian Institute of Sport, Belconnen, ACT, Australia

HAWLEY, J.A. and L.M. BURKE. Carbohydrate availability and training adaptation: effects on cell metabolism. Exerc. Sport Sci.
Rev., Vol. 38, No. 4, pp. 152Y160, 2010. Several markers of endurance training adaptation are enhanced to a greater extent when
individuals undertake selected training sessions with low compared with normal muscle glycogen content or with low exogenous
carbohydrate availability. The potential mechanisms underlying the cellular responses arising from such nutrient-exercise interactions
are discussed in the context of promoting training adaptation. Key Words: AMPK, citrate synthase, endurance training, fat
oxidation, muscle glycogen, p38 MAPK, PGC-1

INTRODUCTION mass (BM)), the guidelines now promote a sliding scale of

The relationship between dietary carbohydrate intake, mus-
cle glycogen content, and endurance exercise capacity is well
documented, and it has become widely accepted that a high-
carbohydrate intake before, combined with carbohydrate sup-
plementation during prolonged, submaximal exercise, can
postpone the development of muscular fatigue and enhance
performance (5,11). A common belief arising from this premise
is that a high-carbohydrate intake during training will permit
an athlete to train harder and longer and thus achieve a
superior training response. Accordingly, sport nutritionists and
exercise physiologists consistently have recommended that
athletes who undertake training that is reliant on muscle gly-
cogen as a primary or limiting fuel source consume a diet that
provides high carbohydrate availability (6).
We purposefully highlight the terminology used here, noting
that there has been a subtle and frequently overlooked mod-
ification to the current sports nutrition guidelines regarding
carbohydrate intake in the athlete’s daily diet. Rather than
promoting a standardized ‘‘high-carbohydrate’’ intake for all
athletes (regardless of whether this is expressed as a percentage
of energy intake, total grams, or grams per kilogram of body
Address for correspondence: John A. Hawley, Ph.D., Health Innovations Research
Institute, School of Medical Sciences, RMIT University, PO Box 71, Bundoora, Victoria
3083, Australia (E-mail: john.hawley@rmit.edu.au).
Accepted for publication: April 30, 2010.
Associate Editor: Mark Hargreaves, Ph.D., FACSM
0091-6331/3804/152Y160
Exercise and Sport Sciences Reviews
Copyright * 2010 by the American College of Sports Medicine
intake that is more closely matched to the predicted fuel costs of
the athlete’s training and recovery (6). This recommendation is
underpinned by the rationale that training sessions should be
undertaken with adequate fuel supplies from muscle glycogen
and other carbohydrate-based fuels.
However, this standpoint does not consider the question of
whether it is a lack or a surplus of substrate that triggers and
promotes the training adaptation process. Indeed, the value
of high carbohydrate availability for supporting the demands
of training has been met by some with skepticism. Such a
viewpoint is, no doubt, based on the failure of long-term
studies of trained individuals to show clear evidence of supe-
rior performance outcomes from high-carbohydrate diets
compared with an energy-matched diet low in carbohydrate.
However, recent reviews of this surprisingly sparse litera-
ture have identified that many of the studies may not have
achieved major differences in carbohydrate availability for
training needs, certainly in the context of the contemporary
definition of recommended carbohydrate intake (6).
Changes in an athletes’ dietary intake and training program
that alter the concentration of blood-borne substrates and the
hormonal milieu cause large perturbations in the macro-
nutrient storage profile of skeletal muscle and other insulin-
sensitive tissues. As such, altering nutrient availability can
exert profound effects on both resting energy metabolism and
subsequent fuel utilization patterns during training and/or
competition, as well as the acute regulatory processes under-
lying gene expression (3) and cell signaling (9,32,34). An
intriguing study, albeit in untrained individuals, reported
superior training adaptation and subsequent exercise capac-
ity after 10 wk of training, incorporating alternate sessions

152

Copyright @ 2010 by the American College of Sports Medicine. Unauthorized reproduction of this article is prohibited.
commenced with low carbohydrate availability compared TABLE. Strategies to reduce carbohydrate availability to alter the
with training with high carbohydrate support (15). The arti- molecular responses to endurance-based training sessionsa.
cle (discussed subsequently) coined the term ‘‘train low, com- Exercise-Diet Strategy Main Outcomes
pete high’’ to describe this novel training-nutrient approach.
It should be emphasized that training with low muscle gly- Chronically low-carbohydrate diet Reduction in muscle carbohydrate
(carbohydrate intake less than availability (endogenous and
cogen content in that study (15) comprised only 50% of the
fuel requirements for training) potentially exogenous sources)
total training load during the intervention period. for all training sessions,
‘‘Train low’’ has now become a catchphrase in athletic depending on degree of fuel
circles, as well as in scientific literature; however, we note mismatch.
that this terminology is used to describe both a range of Chronic low carbohydrate
practices other than the original protocol and as a generic or availability: whole-body effects
‘‘one-size-fits-all’’ theme promoted as a replacement to the including immune system and
era of the ‘‘high-carbohydrate diet’’ in sport. We have wit- central nervous system
nessed firsthand the confusion caused by misunderstood ter- Twice-a-day training Reduction in endogenous and
minology in sports nutrition (6). Accordingly, we encourage (low endogenous carbohydrate exogenous carbohydrate
the concept of low and high carbohydrate availability to be availability for the second session availability for the muscle
promoted. Furthermore, we observe that there are many ways in a day achieved by limiting during the second training
of achieving low carbohydrate availability before, during, and the duration and carbohydrate session
after training sessions that differ in the site of low carbohy- intake in recovery period after
the first session) Acute reduction in carbohydrate
drate availability (i.e., endogenous glycogen vs exogenous availability for immune and
glucose), in the duration of exposure, the number of tissues central nervous systems,
affected (i.e., muscle, liver), and the frequency and timing depending on duration of
of their incorporation into an athlete’s periodized train- carbohydrate restriction and
muscle fuel requirements of
ing program (Table). To understand the importance of these second session
subtleties and to examine the support for ‘‘training high’’
or ‘‘training low,’’ we provide a brief overview of the results Training after an overnight fast Reduction in exogenous
carbohydrate availability for the
of contemporary studies that have determined the effects of
muscle for the specific session
chronically manipulating endogenous (muscle glycogen)
and/or exogenous (blood glucose) carbohydrate availability Potential reduction in endogenous
carbohydrate availability if there
on endurance training adaptation and, where appropriate, per-
is inadequate glycogen restoration
formance outcomes. The potential mechanisms underlying the from previous day’s training
cellular responses that arise from these nutrient-training inter-
actions are discussed in the context of promoting training Acute reduction in carbohydrate
availability for immune and
adaptation. central nervous systems,
depending on duration of
THE TRAINING RESPONSE-ADAPTATION PROCESS carbohydrate restriction and fuel
requirements of the session
From a cellular perspective, (endurance) training adapta- Prolonged training with or without Reduction in exogenous
tion can be viewed as a consequence of the accumulation of an overnight fast and/or carbohydrate sources for the
specific proteins required for sustaining energy metabolism withholding carbohydrate intake muscle for the specific session
during and after exercise. Thus, the training-induced increase during the session
Acute reduction in carbohydrate
in gene expression that allows for subsequent changes in availability for immune and
protein abundance is crucial to the adaptation process (15). central nervous systems,
Although exercise alone is a powerful stimulus for the tran- depending on duration of
scription of multiple ‘‘metabolic’’ genes, nutrition V in par- carbohydrate restriction and fuel
ticular, altered carbohydrate availability (i.e., nutrient exercise requirements of the session
interaction) V also is a potent modulator of this transcrip- Withholding carbohydrate during Could provide adequate fuel
tional response. For example, the rate of translation of post- the first hours of recovery availability for the session but
exercise skeletal muscle interleukin 6 (IL-6) messenger restrict availability for
postexercise signaling activities
ribonucleic acid (mRNA) is reduced by feeding glucose during
exercise, whereas the transcriptional rate of IL-6 from the a
Note that permutations and combinations of these strategies could alter
nuclei of contracting skeletal muscle fibers also is influenced exogenous and endogenous carbohydrate supplies independently or in-
by muscle glycogen content (20). An acute bout of endurance teractively. [Adapted from Burke, L.M. Fuelling strategies to optimise
exercise commenced with low muscle glycogen stores also performance V training high or training low? Scand. J. Sports Med. Sci.
(in press, 2010). Copyright * 2010 John Wiley & Sons, Inc. Used with
results in a greater transcriptional activation of enzymes
permission.]
involved in carbohydrate metabolism (i.e., the adenosine
monophosphateYactivated protein kinase [AMPK], glucose
transporter 4 [GLUT-4], hexokinase, and the pyruvate dehy- information underpins the recent postulate that a ‘‘cycling’’ of
drogenase [PDH] complex) compared with when glyco- muscle substrate stores is required to obtain the optimal adap-
gen is normal or elevated before exercise (27,28,32,33). Such tations to exercise training and provides the impetus for the

Volume 38 Number 4 October 2010

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Carbohydrate Availability and Training 153

Copyright @ 2010 by the American College of Sports Medicine. Unauthorized reproduction of this article is prohibited.
hypothesis that training with low muscle glycogen availability
may enhance training adaptation to a greater extent than
training with normal or elevated glycogen stores (15). Extend-
ing this paradigm, Baar and McGee (4) have proposed that the
classic principles of training incorporating systematic pro-
gressive overload are no longer adequate for optimal perfor-
mance, and based on our increasing knowledge of the role of
nutrition and training, this century-old principle is in need of
revision. Specifically, these workers recommend athletes
deliberately train in a glycogen-depleted state to maximize the
physiological adaptation to endurance exercise. Others (29)
also have noted that training-nutrient ‘‘periodization’’ is nec-
essary to optimize phenotypic adaptation and performance. We
now examine the scientific evidence for the hypothesis that
training undertaken with low carbohydrate availability pro-
motes endurance-training adaptation to a greater extent than
when training undertaken with high carbohydrate availability.
MUSCLE GLYCOGEN AVAILABILITY AND
TRAINING ADAPTATION
The first modern-day investigation of the effects of reducing
endogenous carbohydrate availability on training adaptation
and performance was undertaken by Hansen and colleagues
(15). They studied seven previously untrained male subjects
who completed a training program of leg-knee extensor exer-
cise 5 dIwkj1 for 10 wk. The subjects’ legs were trained
according to a different schedule, but the total amount of
work undertaken by each leg was the same: one leg was trained
twice a day, every second day in which the second training
session was commenced with low glycogen content (LOW),
whereas the contralateral leg trained daily under conditions
of high glycogen availability (HIGH). On the first day of
each 5-day training cycle, both legs trained simultaneously for
1 h at 75% of (one leg) maximal power output. After 2 h of
recovery, during which subjects refrained from carbohydrate
intake, the LOW leg trained again for a further 1 h at 75% of
single-leg peak power output. On the second day, only the
HIGH leg trained. Muscle biopsies were taken from both legs
before and after 5- and 10-wk of training. Submaximal and
maximal exercise testing was performed before and after
training. Resting muscle glycogen content before training was
similar for both groups, but was increased only in LOW after
training (P G 0.05). There was a training-induced increase in
the maximal activity of citrate synthase in both legs (P G 0.05),
with the magnitude of increase being significantly greater
in LOW than in HIGH (P G 0.05). Exercise performance
(measured as the time to exhaustion at 90% of posttraining
maximal power output) was similar for both legs before training
(5.0 T 0.7 vs 5.6 T 1.2 min for LOW and HIGH, respectively).
Noticeably, the magnitude of increase in posttraining exercise
time to exhaustion was twice as great for LOW as HIGH
(19.7 T 2.4 vs 11.9 T 1.3 min; P G 0.05). These results clearly
demonstrate that adaptation and endurance performance are
augmented by lack of substrate (i.e., muscle glycogen) avail-
ability, at least for previously untrained subjects undergoing a
short-term training intervention (15).
To investigate whether well-trained individuals might at-
tain the same benefit as untrained, less fit individuals who un-
dertake a training regimen with lowered glycogen availability,
154 Exercise and Sport Sciences Reviews
Copyright @ 2010 by the American College of Sports Medicine. Unauthorized reproduction of this article is prohibited.
we recruited male cyclists or triathletes who had a history
(93 yr) of endurance training and who were riding 300 to
500 kmIwkj1 in the months before study participation (35).
The athletes were divided into two groups (matched for age,
peak oxygen uptake [V̇O2peak], and training history) and un-
dertook supervised laboratory training sessions during a 3-wk
intervention. The control group (HIGH) trained 6 dIwkj1
with 1 rest day (day 7), alternating between 100-min steady-
state aerobic training (AT; È70% V̇O2peak) on the first day
and high-intensity interval training (HIT; 8  5-min work
bouts at maximal effort with 1-min recovery in between work
bouts at È100 W) the next day. The AT and HIT session
were deliberately chosen, as these workouts deplete È50% of
resting muscle glycogen stores in the fed state in well-nourished,
trained subjects (J.A. Hawley, unpublished observations, 2008).
The experimental group (LOW) trained twice per day, every
second day, performing the AT in the morning to decrease
muscle glycogen content, followed by 2 h of rest without car-
bohydrate intake, and then HIT. During the time between
these two training sessions, subjects rested in the laboratory and
were given ad libitum access to water. Accordingly, HIGH
completed all HIT sessions at a time when muscle glycogen
levels were restored, whereas LOW commenced this interval
set when muscle glycogen stores were depleted by È50% of
resting values.
The novel findings from the study of Yeo et al. (35) were
that in skeletal muscle of already well-trained individuals,
resting glycogen content, the maximal activity of citrate
synthase, the content of the electron transport chain com-
ponent cytochrome-oxidase subunit IV (COX-IV), and rates
of whole-body fat oxidation during submaximal exercise were
all enhanced to a greater extent by training twice every sec-
ond day (LOW) compared with training daily (HIGH) after
the 3-wk intervention (P G 0.05). Although power output
during a 60-min time-trial significantly improved by È11%
(P G 0.05) after both training regimens, there was no differ-
ence between HIGH and LOW. A notable observation was
that self-selected maximal power output was significantly
lower (P G 0.05) for the first six interval training sessions for
athletes who commenced these workouts with low muscle
glycogen content (i.e., the first 2 wk of the training program),
but by the third week of the study, there were no differences
in average power output whether subjects commenced the
workouts with low or normal glycogen stores (Figure).
Thus, despite a compromised (i.e., lower power output)
training capacity (Figure), the twice-every-second-day regi-
men elicited a comparable increase in endurance performance
to that attained after training every day. Yeo et al. (35) pro-
posed that for an athlete unable to train daily but who can
perform two workouts in close proximity, with the second
session performed under conditions of low starting muscle
glycogen, this nutrient-exercise protocol may offer a time-
efficient method of maintaining training adaptations and
performance.
Using an identical protocol to that of Yeo et al. (35),
Hulston et al. (19) also showed that power output was com-
promised when trained cyclists commenced HIT sessions with
low versus normal glycogen stores. In addition, they reported
that tracer-derived measures of fat oxidation during sub-
maximal cycling were greater after low-glycogen training
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Figure. Training intensity (expressed as a percentage of peak sustained
power output (PPO)) sustained during high-intensity interval training (HIT)
sessions undertaken on three occasions per wk during a 3-wk intervention
period. Each HIT session consisted of eight repetitions of 5-min work bouts
separated by 1 min of active recovery (100 W). The 5-min work bouts were
performed at the subject’s maximal self-selected power output. (See text for
further details of nutrient-exercise manipulation.) Values are reported as
mean T standard error. *Significantly different between train high (HIGH)
and train low (LOW). (Reprinted from Yeo WK, Paton CD, Garnham AP,
Burke LM, Carey AL, and Hawley JA. Skeletal muscle adaptation and per-
formance responses to once a day versus twice every second day endurance
training regimens. J. Appl. Physiol. 2008;105:1462Y70. Copyright * 2008
The American Physiological Society. Used with permission.)
(26 T 2 compared with 34 T 2 KmolIkgj1Iminj1; P G 0.01),
with the majority of this training-induced increase being
derived from muscle triacylglycerol oxidation (from 16 T 1
to 23 T 1 KmolIkgj1Iminj1; P G 0.05). Commencing se-
lected training sessions with low muscle glycogen levels
also increased the protein content of A-hydroxyacyl-CoA-
dehydrogenase (A-HAD; P G 0.01), but in agreement with
the findings of Yeo et al. (35), these metabolic changes failed
to improve cycling time-trial performance. Taken collec-
tively, the results from these studies (15,19,35) clearly dem-
onstrate that, independent of prior training status, short-term
(3Y10 wk) interventions in which approximately 50% of the
number of sessions are commenced with low muscle glycogen
levels promote training adaptations (i.e., increases the activ-
ities of enzymes involved in energy metabolism and mito-
chondrial biogenesis, increases rates of whole-body and
muscle-derived triacylglycerol oxidation) to a greater extent
than when all workouts are undertaken with normal or ele-
vated glycogen stores. However, despite creating conditions
that should, in theory, enhance exercise capacity, the effects
of this train-low strategy on a range of performance measures
are equivocal (discussed subsequently).
EXOGENOUS GLUCOSE AVAILABILITY AND
TRAINING ADAPTATION
Another strategy to alter carbohydrate availability is to
alter the exogenous supply of glucose. Glucose supplementa-
tion during exercise inhibits whole-body fat oxidation by
suppressing plasma free fatty acid (FFA) levels while con-
comitantly reducing the entry of long-chain fatty acids into
the mitochondrion, an effect that persists for several hours
Volume 38 Number 4 October 2010
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Copyright @ 2010 by the American College of Sports Medicine. Unauthorized reproduction of this article is prohibited.
after ingestion. Glucose ingestion also has been reported to
attenuate the activation of the AMPK during exercise in
some (2), but not all (21), studies. If AMPK activation is
reduced by increasing glucose availability, then a chronic
downregulation of the typical exercise-induced rise in AMPK
may attenuate the training response-adaptation process. This
is because AMPK activation has a putative role in promoting
metabolic and mitochondrial enzyme content in skeletal
muscle (17,18).
Akerstrom et al. (1) studied the effects of altered exoge-
nous glucose availability in healthy males during a 10-wk
program of leg-knee extensor training. Subjects trained one
leg while ingesting a glucose solution (6% weight-volume for
an intake of 0.7 g carbohydrateIkgj1 BMIhj1), and ingested
a placebo when training the other leg. Training consisted of
2 h of submaximal ‘‘kicking,’’ and each leg was trained on
alternate days. Although there were training-induced in-
creases in the maximal activities of both oxidative and li-
polytic enzymes (citrate synthase and A-HAD), tracer-derived
measures of palmitate turnover, and exercise capacity in both
legs, the magnitude of improvement was similar, independent
of exogenous carbohydrate availability.
De Bock et al. (13) also have investigated whether mus-
cle adaptation is affected by the nutritional status during
training sessions. They recruited moderately active males who
performed 6 wk of training (3 dIwkj1 for 1Y2 h at 75% of
V̇O2peak) during which workouts were commenced in either a
fasted state or 90 min after a carbohydrate-rich breakfast and
additional carbohydrate supplementation (1 gIkgj1 BMIhj1)
throughout the exercise bout. In agreement with the results
of Akerstrom et al. (1), a variety of metabolic markers (in-
cluding succinate dehydrogenase (SDH) activity, GLUT-4,
and hexokinase II content) were increased by a similar extent
with or without carbohydrate supplementation. Despite a sig-
nificant increase in fatty acidYbinding protein after ‘‘fasted’’
training (P G 0.05), rates of fat oxidation during submaximal
exercise were not altered by either training intervention. The
results from these studies (1,13) suggest that the major adap-
tations to endurance training are not augmented by reduced
exogenous carbohydrate availability.
Contrasting results were reported by Nybo and colleagues
(25), who determined the effects of 8-wk endurance training
in previously untrained males who were allocated into either
a group that consumed a sweetened placebo during workouts
(low carbohydrate availability) or a cohort who received a 10%
carbohydrate solution (high carbohydrate availability). They
found that undertaking training without exogenous carbohy-
drate support produced greater enhancement of the increases
in resting muscle glycogen, GLUT-4, and A-HAD. Yet despite
these metabolic changes, there was an unclear effect on time-
trial performance undertaken after 2 h of submaximal cycling,
even when this performance session was undertaken without
carbohydrate intake. Both intervention groups achieved similar
benefits in fat loss, increases in aerobic capacity, loss of intra-
myocellular lipid, and improved blood lipid profile, whereas
only the carbohydrate-supported training group achieved an
increase in lean BM (25). These results suggest that in pre-
viously unconditioned subjects, there may be an impact of
altering the exogenous glucose supply during training sessions
on selected muscular adaptations, but these are without a
Carbohydrate Availability and Training 155
functional transfer to the many of benefits of training on health
and performance parameters.
Recently, we determined the chronic effects of undertaking
daily endurance training with either high or low carbohydrate
availability during workouts (10). During a 28-d intervention
period, 16 endurance-trained subjects were all fed a standard
diet consisting of 5 gIkgj1 BM. Eight subjects were randomly
allocated to a high-carbohydrate-intake group (HICHO) and
consumed a carbohydrate nutritional supplement (a 10% glu-
cose solution that provided an additional 25 kJIkgj1 BM
of carbohydrate for every hour of training). The other eight
subjects (LOCHO) were fed a placebo during training and
ingested energy-matched, fat- and protein-rich snacks after
training sessions. There were no clear effects of the dietary
intervention on resting muscle glycogen or GLUT-4 protein
content. However, the maximal activity of citrate synthase
increased to a greater extent in LOCHO than HICHO
(P G 0.05), whereas tracer-derived estimates of exogenous
glucose oxidation were increased only in HICHO (14% vs 1%;
P G 0.05). Cycling performance (a 7-kJIkgj1 time-trial lasting
approximately 30 min undertaken after 100 min of steady-
state submaximal cycling and performed after the intake of
a carbohydrate-rich meal) was improved to a similar extent
in both groups after the diet-training intervention, regard-
less of whether the bout was undertaken with or without
carbohydrate intake during the bout. These results suggest
that, although there were some differences in the training
adaptations arising from altering carbohydrate availability
during training sessions, these did not transfer into clear
performance differences under the specific conditions of the
cycling trials (10).
INTERACTION OF MUSCLE GLYCOGEN AND GLUCOSE
AVAILABILITY AND TRAINING ADAPTATION
To date, the only investigation to systematically deter-
mine the interactive effects of low muscle glycogen content
combined with altered exogenous carbohydrate availability
on training adaptation and performance was performed by
Morton and colleagues (24). They studied three groups of
recreationally active males who completed 6 wk of high-
intensity, intermittent run training (a total of 24 train-
ing sessions each consisting of 15 min of running at a speed
corresponding to È90% of V̇O2peak, 15 min of running at
25%Y50% of V̇O2peak, and 20 min of running at È70% of
V̇O2peak). In this study, two groups trained twice a day, two
sessions per wk (one session in the morning, the other in
the afternoon), whereas the third group trained once per day,
4 d/wk. This design ensured each subject completed the same
amount of training, but that subjects in groups 1 and 2 per-
formed the second exercise session with a 35% to 45% reduced
glycogen level. To allow for the determination of the effects of
exogenous glucose supplementation, subjects in group 1 con-
sumed a 6.4% glucose solution (GLU) immediately before
and throughout every second training session (LOW+GLU),
whereas subjects in group 2 consumed an identical volume of
placebo (PLA) (LOW+PLA). A control group commenced
each training session with normal glycogen stores and did not
consume any beverages throughout the sessions. Muscle biop-
156 Exercise and Sport Sciences Reviews
Copyright @ 2010 by the American College of Sports Medicine. Unauthorized reproduction of this article is prohibited.
sies from the vastus lateralis and gastrocnemius were taken
before and after the training intervention.
In contrast to the findings of Hansen et al. (15), performance
(determined as intermittent run time to exhaustion) was sim-
ilar in all three groups (22%Y24% increase; P G 0.001). The
training-induced increase in V̇O2peak also was similar between
groups (8%Y10%; P G 0.001). Several training-related proteins,
including COX-IV and the peroxisome proliferatorYactivated
receptor F coactivator (PGC-1), were significantly increased
after training (P G 0.05), but there were no differences in the
magnitude of change between groups. In contrast, the training-
induced increase in the maximal activity of SDH was greater
in LOW+PLA than the other conditions (P G 0.05). Morton
and coworkers (24) concluded that ‘‘training under conditions
of reduced carbohydrate availability from both endogenous
and exogenous sources provides an enhanced stimulus for
inducing oxidative enzyme adaptations of skeletal muscle,
although this does not translate to improved performance
during high-intensity exercise.’’
POTENTIAL MECHANISMS UNDERLYING
AUGMENTED TRAINING ADAPTATION WITH LOW
CARBOHYDRATE AVAILABILITY
To date, the balance of evidence demonstrates that com-
mencing a portion of short-term endurance-based training
programs in the face of low carbohydrate availability pro-
motes training adaptation (i.e., mitochondrial biogenesis) to
a greater extent than when subjects undertake similar training
regimens with high carbohydrate availability. Certainly, in all
of the studies reviewed, there is no evidence of impaired
adaptation or even a decrement to any performance outcome
with low carbohydrate availability. So what are some of the
potential mechanisms that underlie this amplified adaptive
process when both previously untrained subjects and well-
trained athletes deliberately commence selected training ses-
sions with low endogenous and/or exogenous carbohydrate
availability?
There are several putative exercise and/or nutrient-induced
signaling pathways that promote mitochondrial biogenesis in
skeletal muscle. The initial breakthrough in understanding
how repeated contractions (i.e., exercise training) promoted
mitochondrial biogenesis was the discovery of the tran-
scription factors that regulate expression of the nuclear genes
that encode mitochondrial proteins. These include nuclear-
respiratory factor 1 (NRF-1) and NRF-2, which bind to the
promoters and activate transcription of the genes that encode
mitochondrial respiratory chain proteins. The second break-
through was the discovery of an inducible coactivator, PGC-
1>, which docks on and activates these transcription factors
and thus activates and regulates the coordinated expression of
mitochondrial proteins encoded in the nuclear and mito-
chondrial genomes (17).
Much interest has focused on elucidating a possible role for
the AMPK in promoting many of the contractile-induced
adaptations in skeletal muscle, including mitochondrial bio-
genesis. Given the role of the AMPK in regulating cellular
energy metabolism, this is perhaps not surprising: during
exercise, the perturbations in cellular energy balance lead to
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AMPK-induced activation of several metabolic and catabolic
pathways that restore energy equilibrium (i.e., match ATP
supply to ATP demand). To explore a potential role for the
AMPK in training adaptation, we recently investigated acute
skeletal muscle signaling responses to a single bout of HIT
commenced with low or normal muscle glycogen stores in
endurance-trained cyclists/triathletes (34). Six athletes per-
formed a 100-min ride at È70% V̇O2peak (AT) on day 1 and
HIT (8  5-min work bouts at maximal self-selected effort
with 1-min rest) 24 h later (HIGH), whereas another six sub-
jects (matched for fitness and training history) performed AT
on day 1, then, 1 to 2 h later, the HIT session (LOW). Muscle
biopsies were taken before and after both AT and HIT. AMPK
phosphorylation increased significantly in both cohorts after
HIT (P G 0.05), independent of starting muscle glycogen sta-
tus, but the magnitude of increase was greater in LOW
than HIGH (P G 0.05). A possible explanation for the finding
of a higher AMPK activation in the face of low muscle glyco-
gen availability is evidence that glycogen binding to the gly-
cogen-binding domain on the AMPK A subunit allosterically
inhibits AMPK activity and phosphorylation by upstream
kinases (23). McBride et al. (23) recently reported that
AMPK is inhibited by glycogen, particularly preparations
with high branching content. Moreover, they also demon-
strated that this inhibition of AMPK activation by carbohy-
drates was largely dependent on the glycogen-binding domain
being abolished by mutation of residues required for carbo-
hydrate binding. Collectively, these results strongly suggest
that glycogen is a potent regulator of AMPK activity through
its association with the glycogen-binding domain on the
AMPK A subunit.
Another nutrient-sensitive signaling molecule potentially
involved in the altered skeletal muscle adaptive response after
training under conditions of restricted carbohydrate avail-
ability is the p38 mitogen-activated protein kinase (MAPK).
The p38 MAPK phosphorylates and activates PGC-1> and
also increases PGC-1> expression by phosphorylating the
activating transcription factor 2, which increases PGC-1
protein expression by binding to and activating the CREB site
on the PGC-1> promoter. Exercise results in rapid activation
of p38 MAPK, which mediates both the activation and
increased expression of PGC-1>. To investigate the role of
altered carbohydrate availability on the p38 MAPK response
in muscle, Cochran et al. (9) had untrained subjects perform
two training sessions the same day (a morning and afternoon
session both consisting of 5  4 min cycling at È90% of
maximal heart rate) separated by 3 h of passive recovery
during which subjects ingested either a high-carbohydrate
drink or placebo. Biopsies of the vastus lateralis revealed an
exercise-induced increase in the phosphorylation of p38
MAPK (È4-fold; P G 0.05) with a return to baseline levels
before the second training bout, regardless of nutritional
manipulation. However, after the second training session p38
MAPK phosphorylation was higher after the placebo trial
compared with when carbohydrate availability was increased
(P G 0.05). Further support for the contention that chronic
elevation of p38 MAPK signaling may play a role in pro-
moting the greater response-adaptation reported after training
with low carbohydrate availability comes from the data of
Morton et al. (24). They showed that when individuals in-
Volume 38 Number 4 October 2010
c c
Copyright @ 2010 by the American College of Sports Medicine. Unauthorized reproduction of this article is prohibited.
creased carbohydrate availability during the second of twice-
daily training sessions for 6 wk, the increase in SDH activity
was blunted compared with when subjects were carbohydrate
restricted between training sessions (25).
Low Carbohydrate or Increased Fat Availability?
A major problem for the basic scientist when trying to
unravel potential mechanism(s) underlying the benefit to
training adaptation with reduced carbohydrate availability is
the fact that carbohydrate restriction has reciprocal and pro-
nounced effects on lipid availability, (i.e., increased circulat-
ing FFA concentrations and/or elevated muscle triacylglycerol
levels). Evidence linking the increased p38 MAPK response
to low carbohydrate rather than high FFA availability comes
from the study of Watt et al. (31). These workers showed that
p38 MAPK phosphorylation levels were increased during
prolonged (3 h) cycling exercise in humans when circulating
FFA levels were artificially suppressed by administration of
nicotinic acid. Results from animal studies, however, show
that prolonged (4 wk) elevation of FFA promotes mito-
chondrial biogenesis and the capacity to oxidize fatty acids to
a greater extent than chow-fed animals (14).
Finally, it is important to note that carbohydrate availability
is not the only variable manipulated in the investigations
reviewed herein. Many of the studies used different training
modes (cycling vs running vs one-leg kicking), a different
number of training sessions, and variable intervention periods.
It is quite possible that some of the results may not be directly
attributable to differences in carbohydrate availability per se
but rather to the effects of the exercise training protocol itself
(i.e., differences in recovery time between workouts, training
once a day vs twice every second day).
UNANSWERED QUESTIONS AND DIRECTIONS
FOR FUTURE RESEARCH
Cell Signaling Versus Functional Outcomes
A common finding from many of the studies reviewed is the
mismatch between the changes in cellular ‘‘mechanistic’’
variables (typically reported as increases in the phosphor-
ylation status of signaling molecules and/or increases in the
expression of genes and proteins involved in mitochondrial
biogenesis) and whole-body functional outcomes (changes in
training capacity or measures of performance). Several themes
can be proposed to help explain this disconnect, and each
warrants further investigation. First, there may be no direct
relationship between performance and some of the training-
induced changes in selected cellular events that are typically
measured; the functions achieved by upregulating various
muscle proteins may be permissive in promoting the capacity
for exercise but are not quantitatively correlated, or indeed
rate limiting for athletic performance. Muscle function is only
one factor in determining performance, which involves the
integration of whole-body systems including the role of the
central nervous system, in determining pacing strategies and
perceptions of fatigue or effort. A second rationale is that we
currently lack the appropriate tools to accurately measure
Carbohydrate Availability and Training 157
exercise/sports performance, in particular, the ability to detect
small changes that are worthwhile to a competitive athlete to
change the outcomes of real-world events. Within sports sci-
ence, there is much discussion of the challenges of measuring
performance using valid and reliable protocols and of using
different statistical analyses based on magnitude-based infer-
ences to examine the likelihood that changes or differences in
performance are meaningful. In some instances, the technical
variability of various enzymatic assays and/or gene measure-
ments exceeds the small biological changes that manifest as
improvements in performance.
The third possibility is that some train-low strategies have
negative effects on parameters related to an athlete’s health or
performance that either acutely or over the long term coun-
teract the positive effects achieved on isolated muscle char-
acteristics. Acute impairment might directly result because of
the complex interactions between pathways of substrate uti-
lization; as systems to upregulate one pathway occur, there
may be a reciprocal downregulation of others. For example, in
previous work using another dietary periodization strategy
(‘‘fat adaptation’’) in well-trained athletes, we found that 5-d
exposure to a high-fat diet while undertaking a strenuous
training regimen produced a robust enhancement of fat oxi-
dation during submaximal exercise, even when carbohydrate
availability was restored before, or maximized during, exercise
(7). However, we were not able to detect benefits from this
strategy across a range of endurance exercise protocols (7).
In fact, further work clearly demonstrated a reduction in both
the calculated rate of muscle glycogenolysis and the activity
of the rate-limiting enzyme in carbohydrate metabolism,
the PDH complex (30). This impairment to carbohydrate
metabolism would be expected to reduce high-intensity
exercise performance. This is indeed the case. Havemann
et al. (16) investigated the effect of a high-fat diet followed
by 1 d of carbohydrate loading on substrate utilization and
performance during a 100-km cycling time-trial. The 100-km
time-trial incorporated 1-km high-intensity sprints performed
at an intensity of È90% of maximal aerobic power and longer,
4-km work bouts performed at È80% of aerobic power.
Although there was no difference in overall endurance
performance (i.e., the time taken to complete 100 km) or
the 4-km work bouts, sprint performance after fat adaptation
was significantly reduced (P G 0.05).
An indirect outcome of dietary periodization may be a
change in the training stimulus; a common finding when
training sessions are undertaken with low carbohydrate avail-
ability is that subjects frequently chose a lower workload or
intensity because they perceive the effort to be higher, at least
in their initial exposure to training low (35). This outcome
would seem counterintuitive for the preparation of competitive
athletes, where high-intensity workouts and the generation of
high-power outputs are a critical component of a periodized
training program. Interference with such sessions is likely to
impair other adaptations to training (i.e., muscle fiber recruit-
ment). Training with low carbohydrate availability also is
likely to be associated with reduced immune function and ex-
pose the athlete to an increased risk of illness and/or injury.
Finally, it simply may be the case that current studies have
not been sophisticated enough to integrate various com-
binations and permutations of train-low strategies into the
158 Exercise and Sport Sciences Reviews
Copyright @ 2010 by the American College of Sports Medicine. Unauthorized reproduction of this article is prohibited.
periodized training programs of highly trained athletes. The
preparation of elite athletes involves a range of training
activities with various goals (29). It may be that training low
needs to be carefully integrated into parts of this complex
system to allow a performance benefit to be achieved in
concert with the measurable cellular changes. It also should
be considered whether highly trained athletes have a differ-
ent response or require a different stimulus to untrained or
even moderately trained individuals. It has recently been re-
ported that the mitochondrial content and oxidative capacity
of skeletal muscle are key determinants of the activation of
signaling proteins important to muscle plasticity (22). The
attenuation of kinase phosphorylation in muscle with high
mitochondrial content suggests that these proteins may re-
quire a greater stimulus input for activation to propagate these
cues downstream to evoke phenotypic adaptations.
Train Low: How Far and for How Long Do You Have
to Go?
An aspect that is unclear from the present literature is the
degree of glycogen depletion or restricted carbohydrate
availability that is needed to potentiate the effect of the
training stimulus on outcomes such as mitochondrial bio-
genesis or the length of time periodic low-glycogen training
needs to be undertaken to demonstrate functional changes to
training and/or performance outcomes (e.g., weeks to months;
training macrocycles). To answer such questions, a complex
series of studies would need to be undertaken that would
systematically ‘‘titrate’’ levels of carbohydrate availability and
determine subsequent cellular and performance response after
a standardized training regimen. Unfortunately, few of the
present studies have measured actual muscle glycogen content
before and after training in the train-low or control con-
ditions; some have simply assumed that restricted intake of
carbohydrate and/or an abbreviated recovery period between
training sessions will deliver depleted muscle glycogen stores
for subsequent sessions. Investigations to date (15,35) have
used a limited number of total training sessions during the
study duration (18Y45 sessions) when determining both
muscle adaptation and functional performance outcomes.
However, many elite endurance athletes undertake more than
450 total training sessions per year, of which È25% to 30%
would be classified as difficult or hard (T. Stellingwerff,
e-mail, April 15, 2010). It is clearly impractical to extrapolate
the effect of short-term, laboratory-supervised training studies
to an entire year of periodized training and competition.
Therefore, train low currently must be considered somewhat
of a blunt tool.
Perhaps more importantly, we know surprisingly little about
glycogen utilization during the training sessions typically
undertaken by competitive athletes, or how their current real-
world training and dietary practices interact to determine car-
bohydrate availability for various workouts. Indeed, although
sports nutrition guidelines encourage practices to promote car-
bohydrate availability for training, particularly key sessions
involving high-intensity workouts, it is likely that athletes
already undertake some of their sessions with reduced carbo-
hydrate availability, both deliberately and unintentionally.
Some athletes have already adopted specific train-low prac-
tices because of the present and previous interests in this
www.acsm-essr.org
strategy; however, athletes also may restrict carbohydrate in-
take below training requirements as part of the reduced energy
or carbohydrate-modified diets designed to achieve lower BM
or fat levels. Inadvertent causes of training with lowered car-
bohydrate availability include poor nutrition knowledge and
the practical challenges associated with consuming substantial
amounts of carbohydrate before early morning training sessions
or during workouts in which there is restricted access to food
or fluid supplies. Fuel requirements during periods of high-
volume training, with two to three sessions per day, may simply
exceed maximal glycogen storage capacity, which is limited by
both time and carbohydrate intake. Before train-low strategies
can be recommended, it seems important to investigate what
occurs in the sports world and whether some athletes have
already developed successful protocols via trial and error or the
art of coaching. For example, African distance runners, who
often undertake two to three training sessions per day, perform
much of their training in the fasted state (T. Stellingwerff and
H. Stellingwerff, personal communication, 2010) albeit against
a background diet that is higher in carbohydrate (in grams per
kilogram and percentage of energy) than reported intakes from
other free-living athletes (26).
SUMMARY AND CONCLUSIONS
This review has summarized the effects of manipulating
carbohydrate availability on endurance training adaptation.
Current evidence supports the hypothesis that commencing a
portion of short-term (3Y10 wk), endurance-based training
programs in the face of low muscle glycogen content and/or
low exogenous carbohydrate availability promotes training
adaptation (i.e., mitochondrial biogenesis) to a greater extent
than when subjects undertake a similar training regimen with
normal or elevated glycogen levels. Although several putative
cell signaling pathways have been implicated in this nutrient-
exercise adaptation process (i.e., the AMPK and p38 MAPK),
further work is required to determine the precise mechanisms
promoting the amplified endurance training adaptation when
individuals commence selected training sessions with low
carbohydrate availability. There are several studies of the
acute effects of commencing resistance-based exercise in the
face of low muscle glycogen stores (8,12); however, the results
from these investigations suggest that such a practice may
have a negative impact on cellular growth and adaptation.
Indeed, low muscle glycogen content has variable effects on
transcription of select metabolic and myogenic genes at rest,
with any differences in basal transcription being completely
abolished after a single bout of heavy resistance training. For
now, it seems prudent to suggest that competitive athletes
may wish to manipulate carbohydrate availability before,
during, or after selected training sessions that form part of a
long-term periodized training-nutrition plan to promote
metabolic training adaptations that should, in theory, pro-
mote endurance-based performances.
Acknowledgments
The authors thank Dr. Trent Stellingwerff for critical input into this article.
Work from the authors’ laboratory on training-nutrient interactions was
supported, in part, by research grants from GlaxoSmithKline (U.K.) and the
Volume 38 Number 4 October 2010
c c
Copyright @ 2010 by the American College of Sports Medicine. Unauthorized reproduction of this article is prohibited.
Australian Sports Commission to J.A.H. and from the Australian Sports
Commission and Nestlé (Australia) to L.M.B.
The authors have no conflicts of interest relevant to the contents of this
article.
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