mTOR Signaling and the Molecular

Adaptation to Resistance Exercise

SUE C. BODINE
Section of Neurobiology, Physiology & Behavior, University of California, Davis, Davis, CA

ABSTRACT
BODINE, S. C. mTOR Signaling and the Molecular Adaptation to Resistance Exercise. Med. Sci. Sports Exerc., Vol. 38, No. 11,
pp. 1950–1957, 2006. Skeletal muscle size is dynamic and responsive to extracellular signals such as mechanical load, neural activity,
hormones, growth factors, and cytokines. The signaling pathways responsible for regulating cell size in adult skeletal muscle under
growth and atrophy conditions are poorly understood. However, recent evidence suggests a role for the PI3K/Akt/mTOR pathway.
Protein translation is regulated through the phosphorylation of initiation factors that are controlled by signaling pathways downstream
of PI3K/Akt. Recent work in mammals has suggested that activation of Akt/PKB, a Ser-Thr phosphatidylinositol-regulated kinase,
and its downstream targets, glycogen synthase kinase-3 (GSK3) and the mammalian target of rapamycin (mTOR), may be critical
regulators of postnatal cell size in multiple organ systems, including skeletal muscle. This paper will review some of the recent data
that demonstrate the critical role of Akt/mTOR signaling in the regulation of postnatal muscle size, especially under conditions of
increased external loading. Key Words: SKELETAL MUSCLE, HYPERTROPHY, PROTEIN TRANSLATION, PROTEIN
KINASE B/AKT

REGULATION OF PROTEIN TRANSLATION

I
t is well established that chronic loading and resistance
exercise lead to increases in muscle fiber size and tension
Skeletal muscle growth occurs when the balance between
output (2,22,41,45). The cellular and molecular mecha-
protein synthesis and degradation shifts toward protein syn-
nisms responsible for signaling muscle growth in response
thesis. Protein synthesis is known to increase under
to increases in external load are poorly understood. How-
conditions of increased loading and during the recovery
ever, recent studies suggest that activation of the Akt/mTOR
phase after resistance exercise (24,41,52). Increases in
signaling pathways plays a critical role in the regulation of
protein synthesis occur before an increase in RNA and
skeletal muscle size, especially under conditions of altered
DNA levels (52) and can be explained in part by an in-
external loading (5–7,18,33,37). The target of rapamycin
crease in the initiation of mRNA translation, a key
(TOR) is a highly conserved protein kinase that integrates
regulatory step in protein synthesis. The initiation of
signals from nutrients and growth factors to regulate cell
protein translation requires the assembly of a translation-
growth (an increase in cell mass and size) and cell cycle
ally competent ribosome at the AUG start site near the
progression (14,19). Although mTOR is best known as a
5-end of the mRNA (24). Protein translation can be con-
regulator of cell cycle progression and cell proliferation,
trolled at two key steps: 1) modulation of the binding of the
recent data have highlighted its important role in the
initiator methionyl-tRNA (met-tRNAi) to the 40S ribo-
regulation of cell growth (15,34). The purpose of this paper
somal subunit to form the 43S preinitiation complex, and
is to summarize some of the evidence supporting the hy-
2) the binding of the mRNA to the 43S preinitiation com-
pothesis that adaptive muscle growth is dependent on the
plex to form the 48S preinitiation complex. Regulation of
activation of mTOR, a Ser/Thr protein kinase, and its
these translation initiation steps is controlled through the
downstream targets, the ribosomal protein S6 kinase (S6K1/
phosphorylation of initiation factors and protein kinases
p70s6k) and the translational repressor eukaryotic initiation
that are controlled by signaling pathways downstream of
factor 4E-binding protein 1 (4E-BP1/PHAS-I).
phosphatidylinositol 3-kinase (PI3K) (24,40).
Recent work in mammals has suggested that activation of
PKB/Akt, a Ser-Thr phosphatidylinositol-regulated kinase, and
Address for correspondence: Sue C. Bodine, Ph.D., University of its downstream targets, glycogen synthase kinase (GSK3) and
California, Davis, Section of Neurobiology, Physiology & Behavior, 196
the mammalian target of rapamycin (mTOR), may be critical
Briggs Hall, One Shields Avenue, Davis, California 95616; E-mail:
scbodine@ucdavis.edu. regulators of postnatal cell size in multiple organ systems,
Submitted for publication December 2005. including skeletal and cardiac muscle (Fig. 1) (14,24,39). Akt
Accepted for publication May 2006. signaling promotes protein synthesis in a number of ways.
0195-9131/06/3811-1950/0 For example, Akt phosphorylation of GSK3 leads to its
MEDICINE & SCIENCE IN SPORTS & EXERCISEÒ inhibition and an increase in global protein synthesis through
Copyright Ó 2006 by the American College of Sports Medicine an increase in the activity of eurakyotic initiation factor 2B
DOI: 10.1249/01.mss.0000233797.24035.35 (eIF2B) (24). eIF2B is a guanine nucleotide exchange factor
1950

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FIGURE 1—Signaling pathways downstream of insulin and insulin-
like growth factor 1.
that mediates the exchange of GDP for GTP on eukaryotic
initiation factor 2 (eIF2). The active eIF2-GTP binds to met-
tRNAi, which then binds to the 40S ribosomal subunit. In
addition, Akt can phosphorylate mTOR, which, in turn, leads
to phosphorylation of p70s6k and 4E-BP1/PHAS-I (14,19).
Phosphorylation and activation of p70s6k lead to hyper-
phosphorylation of the ribosomal protein S6, which is
associated with enhanced translation of a specific class of
mRNAs that contain a 5-terminal oligopyramidine structure.
Proteins encoded by such mRNAs include the ribosomal
proteins and elongation factors. Phosphorylation of the
translational repressor 4E-BP1 leads to its release from within
an inhibitory complex with the translation initiation factor 4E
(eIF4E). eIF4E is the rate-limiting translation initiation factor
that binds to the cap structure at the 5-end of mRNA
transcripts to initiate cap-dependent translation. Release of
eIF4E from 4E-BP1 permits eIF4E to bind to the scaffolding
protein eIF4G, which then binds to the cap structure of the
mRNA and assembles the eIF4F initiation complex by
recruiting other initiation factors.
mTOR AND THE REGULATION OF
ORGAN GROWTH
The growth of an organ is mediated by increases in both
cell size and cell number. Genetic studies in drosophilas and
mice have clearly demonstrated that the PI3K and TOR
pathways are important regulators of cell and organ growth.
In drosophilas, inactivation of PI3K, chico (insulin-receptor
substrate (IRS) ortholog), PDK1, Akt, TOR, and S6K
decreases organ and cell size, whereas overexpression of
many of these molecules increases organ and cell size
(35,51). Inactivation of PI3K results in a reduction in organ
size through a decrease in both cell size and number, and
inactivation of S6K reduces organ size through a decrease
in cell size only (51). In mice, homozygous deletion of
IRS-1, Akt1, or S6K1 results in a small animal phenotype
mTOR AND THE REGULATION OF MUSCLE SIZE
Copyright @ 2006 by the American College of Sports Medicine. Unauthorized reproduction of this article is prohibited.
(11,29,49). In the S6K1 knockout mice, skeletal muscle
mass is reduced by a decrease in fiber cross-sectional area,
with no decrease in the number of myonuclei per fiber
(34). Furthermore, modulation of PI3K, PDK1, or Akt
expression in the heart affects heart size as a result
of changes in cardiomyocyte size but not cell number
(28,39).
The activation of mTOR-mediated pathways seems to be
critical for normal postnatal growth of mammalian skeletal
muscle, but it is not required for the maintenance of muscle
mass. The phosphorylation state of p70s6k is elevated in
skeletal muscle of early postnatal versus young adult rats
(Fig. 2A). The relative importance of mTOR in the control
of early postnatal muscle growth was demonstrated by
treating growing rat pups with rapamycin, a selective
inhibitor of mTOR. Rapamycin binds to its intracellular
receptor, FKBP12, forming a complex that binds mTOR
with high affinity and blocks its ability to phosphorylate its
downstream targets (19). Delivery of rapamycin (1.5
mgIkgj1Idj1 intraperitoneally) to 2-wk-old rat pups for
11 d resulted in a 30% reduction in body weight in both
male and female pups (Fig. 2B). Interestingly, the effects
of rapamycin on muscle growth were reduced as the rats
developed into young adults. Fourteen days of rapamycin
treatment beginning at 2 wk of age resulted in a 40%
decrease in hindlimb muscle mass, and treatment begin-
ning at 4 wk of age produced only a 20–25% decrease in
muscle mass relative to untreated controls (Fig. 2C). Most
surprising was the observation that rapamycin treatment in
young adult rats (10–12 wk old) produced no change in
muscle mass, suggesting that the maintenance of muscle
mass under steady-state conditions is not dependent on the
activation of mTOR pathways.
It should be noted that much of what is known about
mTOR function has been inferred from experiments
employing rapamycin. Rapamycin-dependent inhibition of
mTOR decreases cell growth through inhibition of protein
translation and ribosome biosynthesis (19). However,
inhibition of mTOR with rapamycin can also decrease the
rate of cell cycle progression and proliferation of mamma-
lian cells in vitro for reasons not currently understood
(15,34). The mechanism by which mTOR coordinates cell
growth and cell cycle progression are currently unknown.
The deletion of S6K1/2 genes in combination with the
expression of a hypophosphorylated form of 4E-BP1
(which would bind eIF4E) is not sufficient to mimic the
effect of rapamycin on cell proliferation (34). Conse-
quently, mTOR must regulate cell proliferation through
effectors other than S6K1/2 and 4E-BP1. It is not known
whether there is a direct effect of rapamycin on the
activation, proliferation, and/or fusion of satellite cells
in vivo in skeletal muscle under growth conditions. Several
studies have demonstrated that during hypertrophic growth
of myotubes in vitro, there is an upregulation of the cell
cycle gene, cyclin D1, and an increase in cyclin
D1jdependent CDK4 activity (20,33). The upregulation
of cell cycle genes, however, was not associated with an
increase in the number of myonuclei per fiber. The
Medicine & Science in Sports & Exercised 1951

FIGURE 2—Postnatal growth is associated with activation of p70s6k
and blocked by rapamycin treatment. A, Western blot of p70s6k in the
medial gastrocnemius of 10- 12-wk-old (adult) and 4-wk-old (4 wk)
rats. The p70s6k gel shift observed during postnatal growth (lane 2) is
reduced in the adult rats (lane 1). Each lane represents 25 Kg of total
protein extracted from a pool of three MG muscles. For each group,
duplicate lanes represent different pools of MG muscles. B, Plot of
body weight in grams (mean T SE ) of female (f) and male (m) 2-wk-
old rat pups during 11 d of treatment with saline (Con/m, Con/f) or
rapamycin (Rap/m, Rap/f). C, Muscle mass, expressed as a percent of
control, of the medial gastrocnemius (MG), lateral gastrocnemius
(LG), soleus (Sol), and tibialis anterior (TA) in rats after 14 d of
rapamycin treatement. Rapamycin was given to 2-wk-old (early,
empty bars) and 4-wk-old (late, solid bars) rat pups. Data are
expressed as mean T SD.
activation of cell cycle regulators during hypertrophy may
be controlling processes other than cell cycle progression,
such as ribosome biosynthesis (33) or the transcription of
specific genes (20).
1952 Official Journal of the American College of Sports Medicine
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mTOR AND THE REGULATION OF ADAPTIVE
MUSCLE GROWTH
Protein kinase B/Akt activity increases in response to
numerous stimuli, including a wide variety of growth factors
and hormones (e.g., insulin-like growth factor-1 (IGF1) and
insulin) (24,43). IGF1 binding to its receptor leads to the
production of phosphatidylinositol 3,4,5-triphosphate,
which leads to the recruitment of Akt from the cytosol to
the membrane, where it becomes activated through the
phosphorylation of Thr 308 and Ser 473 (36). Transgenic
overexpression of IGF-1 from a muscle-specific promoter
results in an enlargement in skeletal muscle size (12,48).
Furthermore, IGF-1 given to C2C12 myotubes in vitro leads
to an increase in cell size because of activation of the PI3k/
Akt/mTOR signaling pathway (43). The IGF-1–induced
growth of myotubes in culture is not the result of an
increase in the number of myonuclei and can be inhibited
by the coadministration of rapamycin (34,43).
To obtain evidence for a role of the Akt/mTOR pathway
in regulating adaptive muscle hypertrophy in adult animals,
the phosphorylation status of key protein kinases and
initiation factors in the Akt/mTOR pathway were examined
in the plantaris muscle of young adult female Sprague–
Dawley rats after functional overload (compensatory
hypertrophy). The plantaris was exposed to a chronic
increase in activity and external loading after bilateral
removal of its functional synergists, the soleus, and medial/
lateral gastrocnemius muscles (6). The amount of Akt and,
more importantly, the phosphorylation state representing
activated Akt, increased throughout the hypertrophy pro-
cess (from day 3 to 14) (Fig. 3A). After 14 d, the total
amount of Akt had increased fourfold more than controls,
and the level of phosphorylated/activated form increased
ninefold in the hypertrophied plantaris. In addition,
phosphorylation of mTOR on Ser 2448 increased twofold
(42). Examination of the downstream targets of mTOR
revealed an increase in the phosphorylation status and
activity of p70S6K, as well as hyperphosphorylation of
PHAS-1/4E-BP1 (Fig. 3B). Hyperphosphorylation of 4E-
BP1 resulted in its release from eIF4E, allowing it to bind
eIF4G, as demonstrated by a decrease in the amount of 4E-
BP1:eIF4E complex and an increase in 4E-BP1:eIF4G
complex (6,42).
These findings indicated that Akt and its downstream
targets were activated in vivo during load-induced muscle
hypertrophy. To determine whether activation of mTOR
was necessary to produce muscle hypertrophy, rats sub-
jected to functional overload were treated with rapamycin.
Daily treatment with rapamycin (1.5 mgIkgj1 intraperito-
neally) did not alter phosphorylation or activity of Akt
itself or its mTOR-independent target GSK-3A, but it did
specifically block the activation of targets known to be
downstream of mTOR (6). Most importantly, rapamycin
treatment led to a 95% reduction in plantaris growth
relative to control at 7 and 14 d of functional overload, as
assessed by both muscle mass (Fig. 4A) and fiber cross-
sectional area (6). These data demonstrate that Akt and its
http://www.acsm-msse.org
 ways that increase translation initiation and/or elongation,
or through the addition of myonuclei. The extent to which
each mechanism is required to produce growth in response
to load is debated, and it may differ depending on the
extent of the increase in cell size. It is well established that
activation, proliferation, and fusion of satellite cells is
critical for growth and repair after injuries that induce
muscle degeneration (10). However, the role of satellite

FIGURE 3—Muscle hypertrophy is associated with activation of the

Akt/mTOR pathway. A, Western blots of native and phosphorylated
Akt in the rat plantaris during compensatory hypertrophy (CH). Each
lane represents 200 Kg of total protein extracted from a pool of three
plantaris muscles after control (Con, lane 1), 3d of CH (lane 2), 7 d of
CH (lane 3), or 14 d of CH (lane 4). For each group, duplicate lanes
represent different pools of plantaris muscles. B, Western blot of
p70s6k in the rat plantaris after 7 and 14 d of CH. The p70s6k gel shift
observed during hypertrophy (CH/-, lanes 2 and 5) was inhibited by
daily injections of rapamycin (CH/rap, lanes 3 and 6). Each lane
represents 25 Kg of total protein extracted from a pool of three
plantaris muscles. C, Western blots of native and phosphorylated Akt
and p70s6k in the medial gastrocnemius (MG) after hindlimb
suspension (HLS) and reloading (Rec). Each lane represents 200 Kg
(Akt) or 25 Kg (p70s6k) of total protein extracted from a pool of three
MG muscles after control (Con, lanes 1), 14 d of HLS (lanes 2), or 14
HLS followed by 7 d of recovery (lanes 3). For each group, duplicate
lanes represent different pools of MG muscles.

downstream target, mTOR, were phosphorylated/activated

during load-induced adapative muscle hypertrophy. Fur-
ther, the finding that specific inhibition of mTOR with
rapamycin resulted in a nearly complete blockage of
growth suggests that the activation of mTOR and its
targets, p70S6K and PHAS-1/4E-BP1, were necessary for
adaptive hypertrophy.
The relative importance of p70S6K versus PHAS-1/4E-
BP1 in the regulation of muscle fiber growth is unclear.
Manipulation of S6K1, S6K2, 4E-BP1, and eIF4E levels in
myotubes suggests that S6K1 is the primary regulator of
muscle fiber size (34). However, another study (15)
suggests that both S6K1 and 4EBP1/eIF4E can influence
mammalian cell size. Increases in protein synthesis can be
achieved by increasing protein translation through path-
FIGURE 4—Muscle hypertrophy in response to altered mechanical
loading is blocked by rapamycin. A, Mass of the plantaris muscle,
expressed as a percent increase relative to control, after 7 and 14 d of
compensatory hypertrophy with vehicle (CH, empty bars) or rapamy-
cin treatment (CH + rap, solid bars). Data are expressed as mean T SD
(10 rats per group). * Significant difference from CH group (P G 0.05).
B, Muscle mass, expressed as a percent loss relative to control, for the
medial gastrocnemius (MG), lateral gastrocnemius (LG), plantaris
(PL), soleus (SOL), and tibialis anterior (TA). Muscle wet weights
were taken after 14 d of hindlimb suspension (HLS), 14 d of HLS
followed by 7 d of reloading (REC), and 14d HLS followed by 7 d of
reloading plus treatement with rapamycin (REC/RAP). Data are
expressed as mean T SD (10 rats per group). * Significant difference
between REC and REC/RAP (P G 0.05).

mTOR AND THE REGULATION OF MUSCLE SIZE Medicine & Science in Sports & Exercised 1953

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cells in adaptive hypertrophy and regrowth after atrophy is
less clear. During compensatory hypertrophy it has been
postulated that increases in protein synthesis are initially
derived through increases in translational efficiency,
followed later by changes in translational capacity through
the addition of myonuclei. Indeed, increases have been
found in the number of myonuclei/fiber after months of
functional overload (3), and irradiation studies suggest that
satellite cell incorporation is critical for compensatory
hypertrophy of the rat EDL and plantaris muscles (1,44).
However, irradiation results in the loss of all proliferating
cells, including satellite cells, and may interfere with the
activation of protein synthesis pathways (1). For example,
Adams et al. (1) demonstrated that the rat plantaris is
capable of early muscle growth (15 d) after irradiation
and functional overload, whereas the later growth is
blocked, suggesting the need for myonuclei addition (1).
However, in those overloaded plantaris muscles exposed to
irradiation, there was a decrease in p70s6k and 4E-BP1
phosphorylation, relative to overloaded nonirradiated,
which could impact protein synthesis (1).
Functional overload represents a very specific model of
altered use and external loading. To determine whether
activation of Akt/mTOR signaling pathways represented a
general response to increased loading, the response of
muscle to reloading after disuse muscle atrophy was
examined. Fourteen days of hindlimb suspension (HLS)
in young adult female Sprague–Dawley rats resulted in a
15–55% loss of mass in various lower-limb muscles
(Fig. 4B). On release from suspension, the atrophied hind-
limb muscles experience a chronic increase in external
loading and hypertrophy, achieving a 15–20% recovery in
muscle mass after 7 d of reloading (Fig. 4B). The exception
was the tibialis anterior muscle, which showed a 4%
decrease in muscle mass during the first 7 d of recovery.
Hindlimb unloading of the medial gastrocnemius resulted in
a decrease in the phosphorylation state of Akt, as measured
by phospho-specific antibodies (Fig. 3C). Further, there was
a 60% decrease in phosphorylation of mTOR on Ser 2448,
with no change in total protein (42). Consistent with the
changes in Akt and mTOR activation, there was a decrease
in the activation state of p70S6K and an increase in the
amount of 4E-BP1 bound to eIF4E (6). These changes
reverted on reloading of the muscle (Fig. 3C).
As in the functional overload model, treatment with
rapamycin during the reloading period resulted in a signifi-
cant block of muscle growth (Fig. 4B). The inhibition of
growth by rapamycin varied across the muscle types and
was less complete in the HLS/reloading model compared
with the functional overload model. This suggests that
muscle growth after atrophy differs from functional over-
load in that it requires the activation of mTOR and
additional signaling pathways. One possible candidate is
the GSK3/eIF2 pathway, which was inhibited during HLS
and activated during reloading (data not shown). Another
protein that was modified during HLS was elongation factor
2 (eEF2). eEF2 is regulated through the mTOR pathway and
by cellular energy stress status (9) and mediates the
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translocation step of elongation. Phosphorylation of eEF2
at Thr 56 inhibits its activity by preventing binding to the
ribosome. After HLS, eEF2 phosphorylation increased
immediately (day 1) and remained elevated relative to control
for at least 7 d (data not shown). Phosphorylation of eEF2 is
catalyzed by eEF2kinase, the activity of which is also
inhibited by phosphorylation. Recent studies have reported
that eEF2 kinase is phosphorylated by p70s6k and p90RSK1,
suggesting control not only by mTOR pathways but also by
MAP kinase pathways (8,9).
mTOR AND CLENBUTEROL-INDUCED
MUSCLE GROWTH
Muscle growth occurs in response to increases in
mechanical loading and also in response to selective
chemical agents, such as the A2-adrenergic agonist,
clenbuterol (21,30). The mechanism by which clenbuterol
induces muscle growth is unclear, although increases in
protein synthesis have been measured after clenbuterol
treatment (21). The phosphorylation status of p70s6k and
PHAS-1/4E-BP1 is elevated in the rat plantaris (50) and
the medial gastrocnemius (unpublished data) after daily
clenbuterol treatment. Further, a role for mTOR in
mediating the effects of clenbuterol was demonstrated by
the ability of rapamycin to inhibit muscle growth during
clenbuterol treatment. Clenbuterol (3 mgIkgj1 subcutane-
ously) treatment produced a 30–35% increase in the mass
of the medial gastrocnemius and tibialis anterior muscles,
whereas coadministration of clenbuterol and rapamycin
(1.5 mgIkgj1 intraperitoneally) treatment produced no
significant muscle growth (Fig. 5). These data suggest that
clenbuterol-induced muscle growth is mediated through the
activation of mTOR and its downstream targets. The
inhibition of clenbuterol-induced growth by rapamycin is
most likely the result of blocking protein synthesis and
FIGURE 5—Muscle hypertrophy in response to clenbuterol is blocked
by rapamycin. Mass of the medial gastrocnemius muscle, expressed as
a percent increase relative to control, after 14 d of clenbuterol
treatment (Clen, empty bars) or clenbuterol plus rapamycin treatment
(Clen + rapa, solid bars). Data are expressed as mean T SEM (10 rats
per group). * Significant difference from the Clen group (P G 0.05).
http://www.acsm-msse.org
not satellite cell proliferation/fusion, because satellite cell
fusion into myofibers has not been observed during
clenbuterol-induced hypertrophy (31).
The above data demonstrate that the Akt/mTOR path-
way is activated in, and requisite for, muscle hypertrophy.
To demonstrate that activation of Akt/mTOR is sufficient
to induce muscle hypertrophy, constitutively activate Akt
(c.a.Akt) has been overexpressed in skeletal muscle using
two methods: plasmid injection and electroporation of
individual muscles in adult animals (6,37), and transgenic
overexpression (27). Overexpression of c.a.Akt in skeletal
muscles results in an increase in muscle fiber size
(6,27,37). Further, the effects of c.a.Akt can be blocked
by the concurrent administration of rapamycin (6,37),
demonstrating that the growth of muscle fibers in response
to c.a.Akt is mediated through the activation of mTOR and
its downstream targets.
mTOR AND RESISTANCE EXERCISE
Finally, are the effects of resistance exercise mediated
through the Akt/mTOR pathway? The first evidence that
mTOR and its downstream targets could have a role in
mediating the effects of resistance exercise came from a
study by Barr and Esser (5). In this study, p70s6k
phosphorylation was shown to increase in the tibialis
anterior and extensor digitorum longus muscles at 3 and
6 h after an acute bout of lengthening contractions
produced by high-frequency neural stimulation. More
interesting was the observation that there was a direct
correlation (r = 0.998) between the increase in p70s6k
measured at 6 h after an acute exercise bout and the percent
change in muscle mass measured after 6 wk of training (5).
These data suggested that p70s6k activation was involved in
the adaptive growth response to chronic resistance exer-
cise. However, no studies to date have examined the
activation of Akt and its downstream targets during chronic
resistance exercise training in vivo.
An increase in the phosphorylation of Akt, mTOR, and
GSK3 is detected in isolated rat muscles immediately after
high-frequency stimulation (4). Further, activation of the
translational regulators p70s6k, 4E-BP1, eIF2B, and eEF2 is
observed at 3 h poststimulation (4). The best in vivo support
for a role of mTOR signaling in resistance exercise,
however, comes from the work of Kubica et al. (25,26). In
these studies, male Sprague–Dawley rats were trained to
perform repetitive ankle extensions with a 0.6-g weighted
vest. In this model, protein synthesis and polysomal aggre-
gation were shown to significantly increase in the medial
gastrocnemius at 16 h postexercise. The status of ribosomal
assembly can be assessed from the separation of muscle
extracts on a sucrose gradient where polysomes migrate to
the denser fractions and monosomes remain in the less
dense fractions. A shift in the distribution of ribosomes from
monosomes to polysomes is interpreted as an increase in
protein translation. The increase in protein synthesis and
polysomal aggregation observed after the acute resistance
exercise was completely prevented with the administration
mTOR AND THE REGULATION OF MUSCLE SIZE
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of rapamycin 2 h before the exercise. At 16 h postexercise, a
small but significant increase in p70s6k and 4E-BP1
phosphorylation relative to sedentary controls was mea-
sured. Interestingly, although there was an increase in S6
phosphorylation, there was no detectable change in the
amount of 4E-BP1 bound to eIF4E or eIF4G. Of particular
interest was the finding that the increase in eIF2BD protein
expression after resistance exercise was rapamycin depen-
dent, suggesting that mTOR activation was responsible for
the early increase in eIF2BD translation.
These data and others (5,18,32,38) provide strong support
for the hypothesis that mTOR signaling is a critical mediator
of muscle growth after resistance training. Although the
majority of studies have examined acute exercise from
muscles that have experienced lengthening contractions,
muscle hypertrophy can occur in response to isometric and
shortening contractions as demonstrated by Adams et al. (2).
The extent to which mTOR and its downstream targets are
activated under all types of muscle contractions remains to
be determined. Additional pathways may also be important
in signaling protein synthesis after both acute and chronic
resistance exercise. For example, activation of MAPK
(17,39) and GSK3 (46) pathways have been measured after
mechanical loading, which could enhance protein synthesis.
CONCLUSIONS
The present literature supports a role for the activation of
Akt/mTOR signaling pathways in regulating muscle
growth through an increase in protein synthesis. However,
the mechanisms regulating muscle fiber size are complex,
and the relative contribution of changes in translation
efficiency and/or addition of myonuclei to adaptive hyper-
trophy require further study. The use of rapamycin has
greatly increased our understanding of the role of mTOR
in regulating muscle growth. However, the generation of
mice with muscle-specific conditional deletions and over-
expression of mTOR, S6K1, 4E-BP1, and eIF4E would
assist in defining the specific function of mTOR and its
downstream targets in muscle growth under varying
conditions. The relative contribution of mTOR pathways
in regulating load-induced fiber growth in normal versus
atrophied muscles is unclear and needs further investiga-
tion. For example, whereas activation of mTOR and its
downstream pathways seem to be critical for muscle
growth after functional overload (6), ‘‘regrowth’’ of muscle
after atrophy may rely on mTOR as well as additional
pathways (6,16). Cell growth (an increase in cell size or
mass) is tightly coupled to nutrient availability, growth
factors, and the energy status of the cell (13,23). mTOR is
capable of integrating signals from all of these inputs in the
regulation of protein translation and cell growth. The extent
to which any or all of these signals are involved in regulating
the activity of mTOR during mechanical loading requires
further investigation. It is interesting to note that Akt
activation occurs after low-intensity, long-duration contrac-
tions, which are typically associated with endurance-type
training, with no activation of mTOR and its downstream
Medicine & Science in Sports & Exercised 1955
targets (47). The upstream effectors responsible for the
selective activation of Akt and mTOR signaling after
increases in activity and load are unknown. Potential
upstream effectors include growth factors, insulin, integ-
rins, G protein–coupled receptors, and amino acids. Our
knowledge of the mechanisms that regulate muscle fiber
size has grown considerably in recent years. However,
additional research is needed to identify the critical signal-
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