Review

Changes in Akt/mTOR signaling pathway in response to concurrent exercise:

A review

Carl-Erik Dang
Department of Physical Education and Health, rebro University, rebro, Sweden


Dang CE. Changes in mTOR signaling pathway in response
to resistance, endurance and concurrent exercise: A review.

Exercise is known to cause morphological, architectural
(Seynnes, 2007) and biochemical changes in human skeletal
muscle (Zoll, 2002). However, different exercise modes
cause different adaptations in the human body. Resistance
training has proven to cause increases in muscle fiber size
and strength (Seynnes, 2007), whereas endurance exercise
adaptations leads to increases in maximal oxygen uptake,
formation of capillaries and mitochondrial biogenesis (Ingjer,
1979).
A combination of exercise modes are widely used to gain
benefits from both resistance and endurance exercise, known
as concurrent training. Concurrent training is used to induce
adaptations that occur with both endurance and resistance
training. However, there has been a debate whether the
adaptations of these exercises may interfere with one another,
known as the interference effect (Hickson, 1980). In 1980
this phenomenon was investigated by Hickson (1980) who in
fact revealed that strength gains were compromised with 10
weeks of concurrent training. However, maximal oxygen
uptake improvements were not diminished when compared to
endurance training alone. This pioneering work of Hickson
(1980) has led to several investigations of concurrent
training. A more recent study (Silva, 2014) investigated the
effects of concurrent training on leg strength compared to
resistance training alone. Forty-eight physically active
females were randomly divided into four groups with
resistance training combined with continuous running,
interval running, continuous cycle ergometer training or
resistance training only. Interestingly, the study reported that
leg strength was not inferior with 11 weeks of concurrent
training when compared to resistance training alone.
Akt/mTOR signaling pathway. The up regulation of protein
synthesis has been proven to be regulated by the mammalian
target of rapamycin (mTOR). There are quite many different
factors that affect the Akt/mTOR signaling pathway, one of
them being phosphorylation of eukaryotic translation factors
(eukaryotic initiation, elongation and release factors). It has
been demonstrated that leucine administers the activity of
S6K1 by increasing the availability of eIF4E for formation
with active eIF4G and eIF4E complex, all which partakes in
up regulation of protein synthesis (Anthony 2000).
Furthermore, it appears that its possible to inhibit
hypertrophy by blocking phosphorylation of p70
S6K
without
altering the phosphorylation of Akt or mTOR. Conversely,
Anthony et al. (2000) demonstrated an increase in
phosphorylation of downstream targets of mTOR,
specifically 4E-BP1 and S6K1, but with lack of increases in
the rate of protein synthesis. The downstream targets of
mTOR proven to regulate hypertrophy in skeletal muscle are
GSK-3, p70
S6K
and PHAS-1/4E-BP1. Blockage of mTOR
signaling and its downstream targets inhibits hypertrophy
(Bodine, 2001). The mTOR signaling pathway is therefore
crucial in order to achieve skeletal muscle hypertrophy.
Atherton et al (2005) found in mice that low frequency
stimulation (LFS) caused increases in phosphorylated AMPK
and that mTOR and protein synthesis was increased
following high frequency stimulation (HFS). Furthermore,
AMPK has been proven to attenuate phosphorylation of
S6K1, 4E-BP1 and activity of eEF2 signaling responses in
rats (Thomson, 2008). HFS increased phosphorylation of
Akt/TSC2/mTOR signaling, GSK-3 and activated
translation initiation regulators p70
S6K
, 4E-BP1, eIF2B and
elongation factor eEF2. All of which has an important role in
anabolic processes in skeletal muscle. However, some of
these anabolic responses, specifically the downstream targets
of TSC2, were inhibited following LFS. Findings also
suggest that TSC2 mediates anabolic responses to HFS via
Akt/TSC2/mTOR pathway and inhibits protein synthesis
during LFS. Inhibition of inactivity of AMPK occurs with
accumulated high concentrations of AMP (Hawley, 1995)
and inhibits phosphorylation of mTOR and some of its
downstream targets (Bolster, 2002). It has been suggested
that short contractions, similar to high frequency stimulation,
does not accumulate as high concentration of AMP as seen in
longer and continuous stimulation (Atherton, 2005). It has
also been proposed that resistance and endurance exercise
combined attenuates desired molecular adaptations when
compared to each exercise alone (Lundberg, 2012, Apro
2013, Donges 2012).
There appears to be many physiological factors that may
influence and interfere with the adaptations of concurrent
training. The purpose of this brief review is to focus on the
molecular response to concurrent training, more specifically
the Akt/mTOR signaling pathway.

ROLE OF MTOR IN ENDURANCE EXERCISE

Animal studies. As previously mentioned, 3 h low frequency
stimulation done on mice did not induce significant increases
in phosphorylated Akt, mTOR via Ser2448 or its downstream
targets, p70
S6K
, 4-BP1 and eIF2B post stimulation or 3 h
after. However, a slight increase was found in GSK-3. In
contrast, a study (Edgett, 2013) performed on mice found
quite the opposite. 3 h treadmill exercise induced expression
of mRNA translation initiation by significantly increasing
phosphorylated Akt, mTOR, rpS6 and eIF2B. Interestingly,
another study (Souza, 2013) done on rats revealed no
increases directly after exercise or 2 h post-exercise in
phosphorylation of Akt, AMPK, mTOR or p70
S6K1
following
60 minutes of running at 60% of V
max
.

Human skeletal muscle. Mascher et al. (2007) studied the
effects of 1 h cycling exercise on changes in signaling
pathways. Six male subjects performing endurance exercise
less than twice a week participated. Phosphorylated GSK-3

was constantly increased during a time course of 3 h post-
exercise. Interestingly, there was an acute increase of
phosphorylated Akt 1 h and 2 h post-exercise, but not after 3
h. Phosphorylated mTOR revealed an increase directly after,
30 min and 2 h post-exercise, but not after 3 h. mTOR and
Akt were partially greater during different time points, it
suggests that there are alternate pathways in which mTOR
can be activated. In support of this notion, Mascher et al.
(2010) revealed an increase of phosphorylated Akt 3 h post-
exercise, whereas mTOR and p70
S6K
was immediately greater
in the working and resting leg following 60 minutes of one-
legged cycling. Interestingly, despite an acute increase in
mTOR post-exercise, a significant increase in the mTOR
inhibitor AMPK was reported. Moreover, the fractional
protein synthesis was higher in the exercising leg. The
evidence suggests that increases in phosphorylated AMPK
and mTOR can occur concomitantly. Mascher et al. (2006)
did not only prove that endurance exercise induces acute
mRNA translation initiation, but also that gene expression
fluctuates during a time course of 3 h post-exercise. Studies
examining at one time point may therefore miss out on
valuable information concerning changes in gene expression.

ROLE OF MTOR IN RESISTANCE EXERCISE

Changes in mTOR signaling pathway in response to
resistance exercise has been studied well more than changes
that occurs with endurance exercise. Several studies done on
human subjects support the notion that resistance exercise
induces acute increases in mTOR phosphorylation (insert
studies). In addition to this notion, intake of BCAA or whey
protein isolate can further elevate mTOR signaling (Apro,
2010, Farnfield 2009).
Resistance exercise with subjects performing 4 sets of 10
repetitions at 80% of 1RM on leg press for a total of 20
minutes induced increases in phosphorylated mTOR, p70
S6K
,
rpS6 and decrease in eEF2 15 min, 1 h and 2 h post-exercise
(Mascher, 2008). In accordance with previously mentioned
studies (insert), increases in phosphorylated mTOR were
established with absence of elevated phosphorylated Akt after
resistance exercise (Apro, 2010, Mascher 2008). However,
the elevation of phosphorylated mTOR did not differ between
the BCAA and the placebo group (Apro, 2010). Moreover,
phosphorylated p70
S6K
were significantly higher than before
exercise and the placebo group, whereas placebo group did
not elevate p70
S6K
. Increases in the downstream target of
p70
S6K
, rpS6 and eEF2, were found, but with no differences
between groups. Interestingly, Apr et al. (2010) failed to
elevate phosphorylated p70
S6K
while elevations of rpS6 were
reported. Roux et al. (2007) showed that RAS/ERK
(Extracellular signal-regulated kinase) signaling pathway can
promote phosphorylation of rpS6 at Ser
235/236
independently
of the mTOR pathway.
It appears that four sets of 10 repetitions of 80% of 1RM
followed by four sets of 15 repetitions of 65% of 1RM in leg
press with 5 minutes rest between sets did not elevate levels
of phosphorylated AMPK (Apro, 2010). However, a study
with a slightly different exercise protocol (10 sets of 10
repetitions leg extension at 70% of 1RM, 3 minute rest
between sets) reported elevated activity levels of AMPK2,
but not AMPK1, during and 1 h post-exercise (Dreyer,
2006).

EFFECTS OF CONCURRENT TRAINING ON MTOR
SIGNALING PATHWAY

Coffey (2009) studied the effect of exercise order on gene
expression. Subjects with more than one year experience in
aerobic and resistance training underwent two trials
consisting of cycling before resistance training, and a reverse
order. Biopsies were taken 15 minutes following each
exercise and 3 h post-exercise. Phosphorylation of AMPK
was higher if cycling was performed subsequently to
resistance exercise (RE). There was a slight increase 3 h post-
exercise in the mTOR inhibitor, TSC2, if cycling was
performed after RE. Interestingly, p70
S6K
and rpS6 were
acutely elevated following the first exercise irrespectively of
exercise mode, with no different between the groups.
Collectively, the study indicates that cycling preceding RE
attenuates anabolic response, whereas RE following cycling
may worsen the inflammatory response and protein
degradation.

Donges et al. (2012) compared the effects RE, endurance and
concurrent training on molecular adaptations on sedentary
subjects. The RE exercise protocol consisted of eight sets of
eight repetitions leg-extensions at 70% of 1RM, whereas
endurance exercise consisted of 40 min ergometer cycling at
55% of peak aerobic power output and concurrent trial did
50% of the work of RE and endurance trial. Measurements of
gene expression were done 1 h and 4 h post-exercise.
Interestingly, none of the trials induced increases in
phosphorylated mTOR, but RE and concurrent training
revealed increases in Akt 1 h post-exercise. In addition,
concurrent and resistance exercise induced an equally high
amount of myofibrillar protein fractional synthesis rate
(FSR), whereas aerobic exercise remained unchanged.
Moreover, mitochondrial FSR was increased with all three
trials, with no difference between trials. Both concurrent and
endurance exercise failed to elevate phosphorylation of rpS6,
but the RE trial received a ten-fold increase 1 h post-exercise.
Surprisingly, this extreme increase did not elevate FSR more
so than the concurrent trial, with concurrent performing half
of the RE work. No changes in AMPK were reported in any
trial, which may be due to subjects getting fed immediately
after exercise. It appears that these particular exercise
protocols (Concurrent and RE) can both induce increases in
equivalent rate of myofibrillar protein synthesis in sedentary
subjects in fed state. It should also be noted that transcription
factors MyoD and MyoG mRNA were only elevated with
RE.

Modest increases in mTOR and p70
S6K
phosphorylation were
reported in subjects performing RE 6 h after endurance
exercise when compared to RE alone (Tommy, 2012).
Subjects had standardized meals before aerobic and
resistance exercise. The RE protocol consisted of leg press
and leg-extensions, two sets of seven repetitions for each
exercise. One-legged cycling was executed 6 h prior to RE.
Biopsies were taken prior to RE, 15 min and 3 h post-
exercise. Acute molecular changes subsequently to endurance
trial were not measured. RE alone failed to elevate mTOR,
p70
S6K
, rpS6 and eEF2 phosphorylation, which may indicate
that the RE protocol was not optimal to induce molecular
changes or that elevations occurred between 15 min and 3 h
post-exercise. Moreover, the purpose of one-legged cycling

was not to maximize cardiovascular stress, but to provide an
aerobic stimulus.

A study comparing endurance and RE to RE alone reported
an increase in phosphorylated Akt with RE and endurance
exercise when compared to rest values and RE trial alone, but
increases in mTOR, p70
S6K1
and eEF2 phosphorylation with
both exercise trials, but no difference between the trials
(Apro, 2013). Interestingly, both trials revealed a reduction of
AMPK 3 h post-exercise. Collectively, the findings indicate
that RE with subsequent endurance training does not impair
gene expression when compared to RE alone in moderately
trained subjects. Moreover, the authors suggested, based on
the results, that prior activation of mTORC1 inhibits elevated
activity of AMPK.



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