Effect of consecutive repeated sprint and resistance

exercise bouts on acute adaptive responses in human

skeletal muscle
Vernon G. Coffey, Bozena Jemiolo, Johann Edge, Andrew P. Garnham, Scott W.
Trappe and John A. Hawley
Am J Physiol Regul Integr Comp Physiol 297:R1441-R1451, 2009. First published 19 August 2009;
doi:10.1152/ajpregu.00351.2009

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 Am J Physiol Regul Integr Comp Physiol 297: R1441–R1451, 2009.
First published August 19, 2009; doi:10.1152/ajpregu.00351.2009.

Effect of consecutive repeated sprint and resistance exercise bouts on acute

adaptive responses in human skeletal muscle
Vernon G. Coffey,1 Bozena Jemiolo,2 Johann Edge,3,4 Andrew P. Garnham,5 Scott W. Trappe,2 and
John A. Hawley1
1
Exercise Metabolism Group, School of Medical Science, RMIT, Melbourne, Australia; 2Human Performance Laboratory,
Ball State University, Indiana; 3Sport and Exercise Science Division, Institute of Food, Nutrition and Human Health, Massey
University, New Zealand; 4Department of Sport and Exercise, University of Auckland, New Zealand; and 5School of Exercise
and Nutrition Sciences, Deakin University, Melbourne, Australia
Submitted 19 June 2009; accepted in final form 18 August 2009

Coffey VG, Jemiolo B, Edge J, Garnham AP, Trappe SW, Hawley effect of concurrent training. In this regard, results of research
JA. Effect of consecutive repeated sprint and resistance exercise bouts on investigating adaptation and performance changes, when con-
acute adaptive responses in human skeletal muscle. Am J Physiol Regul current training is undertaken, has been equivocal (39, 40).
Integr Comp Physiol 297: R1441–R1451, 2009. First published August While the specific physiological effects of concurrent training
19, 2009; doi:10.1152/ajpregu.00351.2009.—We examined acute mo- in skeletal muscle are far from established, work from our
lecular responses in skeletal muscle to repeated sprint and resistance
laboratory has revealed divergent responses for several molec-

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exercise bouts. Six men [age, 24.7 ⫾ 6.3 yr; body mass, 81.6 ⫾ 7.3
kg; peak oxygen uptake, 47 ⫾ 9.9 ml 䡠 kg⫺1 䡠 min⫺1; one repetition ular pathways when athletes undertake acute concurrent train-
maximum (1-RM) leg extension 92.2 ⫾ 12.5 kg; means ⫾ SD] were ing (15). Indeed, we have shown that the proximity of endur-
randomly assigned to trials consisting of either resistance exercise ance and resistance exercise reduced the extent of the desired
(8 ⫻ 5 leg extension, 80% 1-RM) followed by repeated sprints (10 ⫻ molecular response and did not promote optimal activation of
6 s, 0.75 N 䡠 m torque 䡠 kg⫺1) or vice-versa. Muscle biopsies from pathways to promote training specificity for each exercise
vastus lateralis were obtained at rest, 15 min after each exercise bout, mode.
and following 3-h recovery to determine early signaling and mRNA An integral part of training programs for individuals partic-
responses. There was divergent exercise order-dependent phosphory- ipating in many sporting activities is repeated sprint ability.
lation of p70 S6K (S6K). Specifically, initial resistance exercise However, we currently lack information on the molecular
increased S6K phosphorylation (⬃75% P ⬍ 0.05), but there was no training responses in skeletal muscle that occur in response to
effect when resistance exercise was undertaken after sprints. Exercise this type of activity. Sprint training promotes alterations in ion
decreased IGF-I mRNA following 3-h recovery (⬃50%, P ⫽ 0.06)
regulation and buffering capacity with little or no increase in
independent of order, while muscle RING finger mRNA was elevated
with a moderate exercise order effect (P ⬍ 0.01). When resistance muscle size (4, 21, 49). Moreover, high-intensity sprint exer-
exercise was followed by repeated sprints PGC-1␣ mRNA was cise results in a large ionic disturbance, including the accumu-
increased (REX1-SPR2; P ⫽ 0.02) with a modest distinction between lation of muscle lactate and hydrogen ions (H⫹). Of note is that
exercise orders. Repeated sprints may promote acute interference on sprint exercise has recently been shown to induce a molecular
resistance exercise responses by attenuating translation initiation sig- adaptive profile associated with mitochondrial biogenesis and
naling and exacerbating ubiquitin ligase expression. Indeed, repeated increased capacity for glucose and fatty acid oxidation (23). To
sprints appear to generate the overriding acute exercise-induced re- the best of our knowledge, the molecular interactions of con-
sponse when undertaking concurrent repeated sprint and resistance current resistance and repeated sprint training have not been
exercise. Accordingly, we suggest that sprint-activities are isolated investigated. Indeed, such an approach represents a novel
from resistance training and that adequate recovery time is considered expansion of the conventional concurrent training paradigm.
within periodized training plans that incorporate these divergent Consequently, we currently have no understanding of the
exercise modes.
potential mechanisms that might underlie exercise-induced
repeated sprint ability; resistance training; adaptation adaptation and enhanced/impaired muscle function with con-
current resistance and repeated sprint training. Despite the
potential similarity in the work-to-recovery ratio between
CONCURRENT TRAINING CAN BE defined as the incorporation of two sprint and resistance exercise, the greater rate of force produc-
or more divergent exercise modes in a periodized training tion and inadequate recovery during repeated sprints generates
program. Since the classic work of Hickson (30), studies of large disturbance to ion homeostasis, the accumulation of
concurrent training have attempted to determine the interaction lactate, and altered substrate metabolism during ensuing repe-
of endurance and resistance exercise on subsequent adaptation titions that characterize a distinct exercise overload. (7). Thus,
in skeletal muscle. Given the distinct phenotypes generated in in contrast to the resistance exercise-induced increases in
response to either chronic endurance or resistance training and translation initiation and protein synthesis (65), repeated
the apparent incompatibility of the exercise-induced adaptation sprints might be expected to inhibit anabolic processes. Ac-
(fatigue resistance vs. high-force production), previous work in cordingly, the aim of the current study was to quantify the
this area has endeavoured to elucidate the potential interference acute cellular responses in skeletal muscle when resistance and
sprint training sessions were performed successively and to
Address for reprint requests and other correspondence: V. Coffey, Exercise
examine the effect of alternate exercise order (i.e., an initial
Metabolism Group, School of Medical Science, RMIT Univ., PO Box 71, bout of heavy resistance exercise, then repeated sprint exercise
Bundoora, Victoria 3083, Australia (e-mail: [email protected]). or vice versa). Given the paucity of data regarding the molec-
http://www.ajpregu.org 0363-6119/09 $8.00 Copyright © 2009 the American Physiological Society R1441
R1442 CONCURRENT SPRINTS AND RESISTANCE EXERCISE

ular response following repeated sprints in skeletal muscle, we Maximal strength. The strength of quadriceps was determined
examined signaling associated with the translational machinery during a series of single repetitions on a plate-loaded leg extension
(Akt-mTOR-p70S6K) and changes in mRNA abundance re- machine until the maximum load lifted was established (1-RM).
lated to anabolic growth [insulin-like growth factor (IGF)], Repetitions were separated by 3-min recovery and were used to
establish the maximum load/weight that could be moved through the
proteasome-dependent protein breakdown [muscle RING fin- full range of motion once, but not a second time. Exercise range of
ger (MuRF), antrogin], and mitochondrial biogenesis/carbohy- motion was 85 degrees, with leg extension endpoint set at ⫺5 degrees
drate metabolism [peroxisome proliferator-activated recep- from full extension.
tor-␥ coactivator-1␣ (PGC-1␣), hexokinase] (14). We also 30-s Wingate. Wingate testing was undertaken on a Lode Excalibur
specifically chose this suite of molecular markers to compare Sport cycle ergometer (Groningen, The Netherlands). Subjects re-
the responses to repeated sprints with our previous work mained seated throughout the protocol that commenced with 60-s
showing concurrent resistance and endurance exercise-induced cycling at 75 W immediately followed by 30-s maximal sprint cycling
changes during the acute postexercise recovery period (15). against a resistance equal to 0.7 N 䡠 m torque⫺1 䡠 kg body mass⫺1.
We hypothesized that the different exercise modes undertaken Diet/Exercise Control
in close proximity would amplify the potential incompatibility
of the acute molecular profile for each form of contractile Before an experimental trial (described below), subjects refrained
activity. from training and other vigorous physical activity for a minimum of
48 h. Subjects were provided with standardized prepacked meals that
METHODS consisted of 3 g CHO/kg body mass, 0.5 g protein/kg body mass, and
0.3 g fat/kg body mass consumed as the final caloric intake the
Subjects evening prior to reporting for an exercise testing session.

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Six male subjects [age, 24.7 ⫾ 6.3 yr; body mass, 81.6 ⫾ 7.3 kg; Experimental Trials
peak oxygen uptake (V̇O2 peak), 47 ⫾ 9.9 ml 䡠 kg⫺1 䡠 min⫺1; leg exten-
sion one repetition maximum (1-RM), 92.2 ⫾ 12.5 kg (means ⫾ SD)] An overview of the study testing protocol is shown in Fig. 1. On the
who had been participating in regular concurrent resistance and morning of an experimental trial, subjects reported to the laboratory
endurance training (⬎3 ⫻ per wk) volunteered for this study. The after an ⬃10-h overnight fast. After resting in the supine position for
experimental procedures and possible risks associated with the study ⬃15 min, local anesthesia (2–3 ml of 1% Xylocaine) was adminis-
were explained to each subject who gave written informed consent tered to the skin, subcutaneous tissue, and fascia of the vastus lateralis
prior to participation. This study was approved by the Human Re- of the subject’s leg in preparation for the series of muscle biopsies. A
search Ethics Committee of the Royal Melbourne Institute of Tech- resting (basal) biopsy was taken using a 5-mm Bergstrom needle
nology, Melbourne, Australia. modified with suction. Approximately 150 mg of muscle was removed
and blotted to remove excess blood; ⬃15–20 mg was then cut and
Study Design placed in RNALater (Ambion, Austin, TX), and the remainder was
immediately frozen in liquid nitrogen. Subjects then completed the
The study employed a randomized cross-over design where each two exercise sessions (described in detail below). Fifteen minutes
subject completed two acute concurrent repeated sprint and resistance after completion of the first exercise bout, a second biopsy was taken.
exercise sessions separated by a 2-wk recovery period, during which Subjects then performed the second exercise bout. After 15 min of
time subjects were instructed to continue their habitual physical recovery from the second exercise session, a third muscle biopsy was
activity. One experimental trial consisted of consecutive resistance taken, and subjects rested in the supine position until a fourth muscle
and then repeated sprint exercise bouts, while the other trial was biopsy was taken following the 3-h recovery period. Each muscle
performed using the reverse exercise order, i.e., repeated sprint then biopsy was taken from a separate site, and all samples were stored at
resistance exercise. ⫺80°C until subsequent analysis. In addition, blood samples (⬃2 ml)
were taken at rest preexercise, 0, 5, 10, and 15 min after the first
Preliminary Testing exercise bout and 0, 5, 10, 15, and 30 min after the second exercise
V̇O2 peak. V̇O2 peak was determined during an incremental test to bout.
exhaustion on a Lode cycle ergometer (Groningen, The Netherlands). Resistance Exercise
The protocol has been described in detail previously (29). In brief,
subjects commenced cycling at a workload equivalent to 2 W/kg for Following a standardized warm-up (2 ⫻ 5 repetitions at 50% and
150 s. Thereafter, the workload was increased by 25 W every 150 s 60% 1-RM, respectively), subjects performed eight sets of five repe-
until volitional fatigue, defined as the inability to maintain a ca- titions at 80% 1-RM. Each set was separated by a 3-min recovery
dence ⬎ 70 rev/min. Throughout the test, subjects breathed through a period during which the subject remained seated on the leg extension
mouthpiece attached to a metabolic cart (Parvomedics, Sandy, Utah) machine. Contractions were performed at a set metronome cadence
that was calibrated before testing using a 3-liter Hans-Rudolph sy- approximately equal to 30 degrees/s and strong verbal encouragement
ringe and gases of known concentration (4% CO2 and 16% O2). was provided during each set.

Fig. 1. Schematic of the experimental trials

incorporating consecutive bouts of diverse ex-
ercise. This study utilized a randomized cross-
over design where subjects completed both
exercise orders [1 repetition maximum (1-
RM)]. The duration of each exercise bout was
⬃25 min vs. ⬃10 min for resistance (REX)
and repeated sprint (SPR) exercise, respec-
tively.

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 CONCURRENT SPRINTS AND RESISTANCE EXERCISE R1443
Repeated Sprint Exercise Signaling Technology (Danvers, MA). Antiphospho-AMPK␣thr172 was
raised against AMPK-␣ peptide (KDGEFLRpTSCGAPNY) as described
Subjects undertook 10 ⫻ 6-s maximal sprint efforts at a constant previously (13) and anti-phospho-p70 S6Kthr389 was from Millipore (cat.
load. Subjects performed 10 maximal effort sprint cycling repetitions no. 04 –392; Temecula, CA). Monoclonal anti-␣-tubulin control protein
of 6-s duration against 0.75 N 䡠 m torque⫺1 䡠 kg⫺1 body mass on a Lode antibody (cat. no. T6074) was from Sigma-Aldrich (St Louis, MO).
Excalibur Sport cycle ergometer. The initial sprint repetition was Total RNA extraction and RNA quality check. Total RNA (23.1 ⫾
preceded by 60 s cycling at 75 W, and subjects were instructed to 1.28 mg of tissue) was extracted in TRI reagent (Molecular Research
initiate each sprint at 80 rpm and remain seated throughout the sprint Center, Cincinnati, OH). The quality and integrity (RIN of 8.56 ⫾
exercise. Each sprint repetition was separated by 49-s passive rest 0.18 SE) of extracted RNA (0.28 ␮g/ mg of tissue) was evaluated
followed by a 5-s rolling start for each subsequent sprint (i.e., each using an RNA 6000 Nano LabChip kit on an Agilent 2100
sprint started at 1-min intervals). Bioanalyzer.
Analytical Procedures RT and real-time PCR. Oligo(dT) primed first-strand cDNA was
synthesized (150 ng total RNA) using SuperScript II RT (Invitrogen,
Blood/muscle lactate and pH/H⫹. Whole blood was immediately Carlsbad, CA). Quantification of mRNA (in duplicate) was performed
analyzed for lactate concentration using an automated glucose/lactate in a 72-well Rotor-Gene 3000 Centrifugal Real-Time Cycler (Corbett
analyzer (model YSI 2300; Yellow Springs, Ohio). Approximately 25 Research, Mortlake, NSW, Australia). The current study was limited
mg of frozen muscle was freeze dried and blood/connective tissue was in that, to the best of our knowledge, no previous work has determined
removed from the powdered muscle tissue. For determining muscle the suitability or otherwise of selected housekeeping genes following
pH, samples (⬃3 mg) were homogenized on ice for 2 min in a solution repeated sprint exercise. We selected GAPDH based on a validation
containing sodium fluoride (10 mM NaF) at a dilution of 30 mg dry study from our laboratory that has shown GAPDH to be most stable
muscle/ml of homogenizing solution. The muscle homogenate was in all human skeletal muscle fiber types following acute exercise (36,

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then placed in a circulating water bath at 37°C for 5 min prior to and 66). All primers used in this study were mRNA specific (on different
during the measurement of pH. The pH measurements were made exons and over an intron) and were designed for SYBR Green chemistry
with a microelectrode (model MI-415; Microelectrodes, Bedford, NH) using Vector NTI Advance 9 software (Invitrogen) [Table 1; sequences
connected to a pH meter (model SA 520; Orion Research, Cambridge, published elsewhere: PGC-1␣ (27), antrogin-1, muscle RING finger,
MA). For determining muscle lactate, samples (⬃4 mg) were enzy- forkhead box O 3a (Foxo3a) (43), and myogenic differentiation factor
matically assayed by using the spectrophotometry methods of Harris (MyoD) (56)]. A melting curve analysis was generated to validate that
et al. (28). only one product was present. The details regarding RT and PCR
Western blot analysis. Muscle samples were homogenized in buffer reaction parameters have been reported previously (43, 56).
containing 50 mM Tris䡠HCL, pH 7.5, 1 mM EDTA, 1 mM EGTA, 10% Relative qPCR data analysis. Relative gene expression analysis
glycerol, 1% Triton X-100, 50 mM NaF, 5 mM Na pyrophosphate, 1 mM comparing expression of a gene-of-interest in relation to a reference-
DTT, 10 ␮g/ml trypsin inhibitor, 2 ␮g/ml aprotinin, 1 mM benzamidine, gene, based on the distinct cycle (Ct) differences, the ⌬Ct model (42),
and 1 mM PMSF. After determination of protein concentration (Pierce, was used in this study as described in detail previously (56, 57, 68).
Rockford, IL), lysate was resuspended in Laemmli sample buffer, sepa-
rated by SDS-PAGE, and transferred to polyvinylidine fluoride mem- Statistical Analysis
branes blocked with 5% nonfat milk, washed with TBST (10 mM
Tris䡠HCl, 100 mM NaCl, 0.02% Tween 20) and incubated with primary We calculated sample size using the methods of Hopkins (32), and
antibody (1:1,000) overnight at 4°C. Membranes were incubated with we have previously reported statistical significance for related phys-
secondary antibody (1:2,000), and proteins were detected via chemilu- iological variables in cross-over studies employing six to eight sub-
minescence (Amersham Biosciences, Buckinghamshire, UK; Pierce Bio- jects (15–17, 52). All data were analyzed by two-way repeated-
technology, Rockford, IL) and quantified by densitometry. Sample (50 measures ANOVA (2-factor: time ⫻ exercise order) with Student-
␮g) time points for each subject were run on the same gel and quantified Newman-Kuels post hoc analysis. Statistical significance was
relative to ␣-tubulin protein abundance. Polyclonal antiphospho-mam- established when P ⬍ 0.05 (SigmaStat for windows Version 3.11).
malian target of rapamycin (mTOR)ser2448 (cat. no. 2971); and monoclo- Lacking any information on the smallest substantial changes in acute
nal antiphospho-Aktser473 (cat. no. 9271), -tuberin (TSC2)thr1462 (cat. no. responses to divergent exercise, we performed additional post hoc
3617), -S6 ribosomal proteinser235/6 (cat. no. 4856) were from Cell analysis in which each individual’s data were back transformed

Table 1. Summary of mRNA primers for genes of interest used in PCR reactions
Name Sequence GeneBank No. Amplicon Size, bp Annealing Temperature,°C mRNA Region, bp

IGF-1
Sense 5⬘-CAGGGGCTTTTATTTCAACAAGCCCA-3⬘ NM_000618 127 59 419–545
Antisense 5⬘-TGCGCAATACATCTCCAGCCTCCTTA-3⬘
MGF (IGF-IEc)
Sense 5⬘-CAGAAGTATCAGCCCCCATCTACCAACAA-3⬘ NM_001111283 91 59 616–706
Antisense 5⬘-TGCACTCCCTCTACTTGCGTTCTTCAA-3⬘
TFAM
Sense 5⬘-GGAAAACCAAAAAGACCTCGTTCAGCTT-3⬘ NM_003201 86 59 589–674
Antisense 5⬘-TTTTCCTGCGGTGAATCACCCTT-3⬘
HK II
Sense 5⬘-CAGCAGAACAGCCTGGACGAGAGCAT-3⬘ MN_000189 126 60 3745–3870
Antisense 5⬘-GTCAAACTCCTCTCGCCGGTGGAT-3⬘
CS
Sense 5⬘-CATGGACTGGCAAATCAGGAAGTGCTT-3⬘ MN_004077 144 58 1091–1220
Antisense 5⬘-CTGGAACAACCCGTCCTGAGTTGAG-3⬘
Sense, forward primer; anti-sense, reverse primer; IGF-I, insulin-like growth factor 1; MGF, mechano-growth factor; TFAM, mitochondrial transcription factor
A; HK II, hexokinase II; CS, citrate synthase.

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R1444 CONCURRENT SPRINTS AND RESISTANCE EXERCISE

Fig. 2. Blood lactate at rest preexercise, immediately and at 5-min intervals during 15-min postexercise recovery following each individual exercise bout (Ex
1 and 2) incorporating REX (8 ⫻ 5 leg extensions at 80% 1-RM) and SPR (10 ⫻ 6 s maximal effort) in alternate exercise orders. Results are group means ⫾
SD. Significant difference (P ⬍ 0.05) vs. *rest, †SPR2 vs. REX1, #REX2 vs. REX1, and §REX2 vs. SPR1.

against the mean of their corresponding resting preexercise values. contrast, blood lactate concentration was significantly elevated

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For further details of the statistical methods employed for magnitude- above rest at every time point throughout the 15-min recovery
based inferences the reader is referred to Hopkins et al. (33). Briefly, when resistance exercise was undertaken after repeated sprints
log-transformed delta values between data time points were directly (REX2 ⬃3– 4.7 mmol/l, P ⬍ 0.05; Fig. 2). In addition, pos-
compared and converted to Cohen effect sizes (ES) to determine the
magnitude of change for each exercise bout and alternate exercise
texercise blood lactate concentration was consistently in-
order (31). We chose a default confidence interval of 90% to calculate creased above rest following repeated sprints regardless of
ES via a spreadsheet, making the same assumptions about sampling exercise order (P ⬍ 0.05; Fig. 2).
distributions that statistical packages use to derive P values (33). We There were significant main effects among the levels of time
interpreted the magnitude of the ES by using conventional threshold for muscle lactate concentration (Table 2). Repeated sprint
values of 0.2 as the smallest ES, 0.5 as a moderate ES, and 0.8 as a exercise significantly increased muscle lactate above resting
large ES (18, 33). Data are expressed as arbitrary units ⫾ SD. levels regardless of exercise order (P ⬍ 0.05), and lactate
RESULTS
concentration was higher following SPR1 compared with
REX2 (P ⬍ 0.05). Moreover, muscle lactate was only signif-
Repeated Sprint Performance icantly elevated above rest following resistance exercise when
preceded by repeated sprints (rest vs. REX2, P ⬍ 0.05; Table
There was a significant main effect for mean power output 2). There were also significant main effects among the levels of
within the different sprint repetition bouts (P ⬍ 0.01). More- time for both pH (rest vs. REX, rest vs. SPR, P ⬍ 0.05) and H⫹
over, mean power declined ⬃11% (sprint 1–10) when repeated (rest vs. SPR, P ⬍ 0.05; Table 2), but there were no significant
sprints were undertaken after resistance exercise (REX1-SPR2, interactions. The decrease in pH and increase in H⫹ was
965 ⫾ 140 W vs. 860 ⫾ 106 W) and ⬃19% when sprints greatest when sprints were undertaken prior to resistance ex-
preceded resistance exercise (SPR1-REX2, 990 ⫾ 194 W vs. ercise compared with sprints performed subsequent to resis-
804 ⫾ 76 W). However, mean power generated during the tance exercise (Table 2).
entire set of 10 ⫻ 6 s sprints was similar regardless of exercise
order, with a significant difference between exercise orders
only within SPR4 (P ⬍ 0.05). Signaling Responses

Blood/Muscle Lactate and pH/H⫹ Main effects for Aktser473 phosphorylation verged on signif-
icance for the different levels of time (P ⫽ 0.055). The initial
Changes in postexercise blood lactate concentration resulted bout of exercise had little effect on phosphorylation of Akt
in differences for time and exercise order, and significant regardless of contraction mode (Fig. 3A). However, increased
time ⫻ exercise order interactions (P ⬍ 0.01; Fig. 2). Specif- Akt phosphorylation 15 min after resistance exercise was
ically, when resistance exercise preceded repeated sprints, undertaken, subsequent to repeated sprints (SPR1-REX2), re-
blood lactate levels were only elevated above rest immediately sulted in a large disparity compared with initial resistance
after REX1 (0 min, ⬃2.4 ⫾ 1.5 mmol/l, P ⬍ 0.05; Fig. 2). In exercise (REX2 vs. REX1, ⬃180%; ES ⬎ 1.0; Fig. 3A).

Table 2. Muscle lactate (La) and H⫹ content, and pH measured at rest and 15 min postexercise following each exercise
bout in the alternate exercise orders
Rest REX1 SPR2 Rest SPR1 REX2

La, mmol/kg dry wt 9.2⫾4 15.8⫾4 28.2⫾16* 8.3⫾4.6 42.5⫾19*† 25.5⫾21*

pH 7.06⫾0.09 6.98⫾0.13 6.95⫾0.20 7.06⫾0.09 6.92⫾0.15* 6.99⫾0.12
H⫹, mmol/kg dry wt 89⫾18 109⫾36 123⫾63 88⫾16 125⫾40 105⫾27
Values are means ⫾ SD. REX, resistance exercise; SPR, repeated sprints. Significantly different (P ⬍ 0.05) vs. *rest, vs. †REX2.

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 CONCURRENT SPRINTS AND RESISTANCE EXERCISE R1445

Fig. 3. Phosphorylated Aktser473 (A), tuber-

inthr1462 (TSC2; B), mammalian target of
rapamycinser2448 (mTOR; C), p70 S6Kthr389
(D), S6 ribosomal proteinser235/6 (p-rpS6; E),
and adenosine monophosphate-activated ki-
nase ␣thr172 (AMPK; F) relative to ␣-tubulin

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at rest preexercise, 15 min after each indi-
vidual exercise bout (Ex 1 and 2), and 3 h
after cessation of training. Subjects com-
pleted 2 experimental trials incorporating
consecutive REX (8 ⫻ 5 leg extensions at
80% 1-RM) and SPR (10 ⫻ 6 s maximal
effort) bouts performed in alternate order.
Results are group means ⫾ SD, and data are
log-transformed values as arbitrary units.
Significant difference (P ⬍ 0.05) vs. *rest,
†SPR2, (†)REX2, and ‡3 h.

Phosphorylation of Akt was largely unaffected 15 min after TSC2 and mTOR phosphorylation abated and returned to
each bout of repeated sprints, and phosphorylation state re- resting levels following 3-h recovery from the consecutive
turned to resting levels 3 h following cessation of contractile exercise bouts.
activity, regardless of exercise order. Diverse contractile activity and alternating exercise order
Changes in TSC2thr1462 phosphorylation in response to re- induced divergent p70 S6Kthr389 phosphorylation responses
sistance and repeated sprint exercise were modest (Fig. 3B). (P ⬍ 0.05; Fig. 3D). Specifically, an initial bout of resistance
There were minor changes to TSC2 phosphorylation following exercise generated an increase in S6K phosphorylation that
an initial bout of contractile activity regardless of mode, while was different from all other time points within the REX1-SPR2
a moderate increase was observed with the successive exercise exercise order (P ⬍ 0.05; Fig. 3D). Moreover, repeated sprints
bout (REX2 vs. REX1, ⬃28%; ES, 0.5; SPR2 vs. SPR1, did not promote phosphorylation of S6K, and there was no
⬃56%; ES, 0.8; Fig. 3B). There was a significant main effect effect when resistance exercise was undertaken after sprints,
for mTORser2448 phosphorylation 15 min following resistance resulting in a significant order effect within resistance exercise
exercise (P ⬍ 0.05), while elevated phosphorylation above rest bouts (REX1 vs. REX2, ⬃74%; ES ⬎ 1.0, P ⬍ 0.05). There
after repeated sprints approached significance (P ⫽ 0.056; Fig. was also disparity in ⌬S6K phosphorylation from rest follow-
3C), but there were no significant interactions. Of note, there ing the 3-h recovery period, with elevated phosphorylation
were corresponding mTOR phosphorylation responses to con- observed when resistance exercise preceded repeated sprints
tractile activity regardless of exercise mode (Fig. 3C). Elevated (REX1-SPR2, ⬃56%; ES ⬎ 1.0; Fig. 3D). Changes in ribo-
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R1446 CONCURRENT SPRINTS AND RESISTANCE EXERCISE

somal protein (rp) S6ser235/6 phosphorylation resulted in a

significant main effect within the different levels of time (P ⬍
0.05; Fig. 3E). The enhanced phosphorylation of rpS6 15 min
after initial resistance exercise was significantly different from
rest and other postexercise time points (REX1-SPR2, P ⬍
0.05; Fig. 3E). Of note, prior repeated sprints did not prevent
increased rpS6 phosphorylation with subsequent resistance
exercise, but the magnitude of elevation from rest was moder-
ately lower (REX1 vs. REX2, ⬃50%; ES, 0.66). After 3-h
recovery, rpS6 phosphorylation abated but remained higher
above rest following the resistance exercise-repeated sprints
exercise order (REX1-SPR2, ⬃69%; ES ⬎ 1.0; Fig. 3E).
Changes in AMPKthr172 phosphorylation resulted in a sig-
nificant main effect within the different levels of time (P ⬍
0.05; Fig. 3F). There were similar increases in AMPK phos-
phorylation 15 min after an initial bout of contractile activity
regardless of exercise mode. Moreover, the increase in phos-
phorylation of AMPK was sustained following the subsequent
exercise bouts with no disparity in the magnitude of change

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from rest (Fig. 3F). Following 3-h recovery, AMPK phosphor-
ylation was moderately higher above rest when repeated sprints
preceded resistance exercise (SPR1-REX2, ⬃22%; ES, 0.53;
Fig. 3F).

mRNA Responses
IGF-I-mechano-growth factor-MyoD. Due to limitations in
tissue sample size, the mRNA analysis was restricted to com-
parisons at rest and 3 h after cessation of contractile activity.
Comparable decreases in IGF-I mRNA from rest following 3-h
recovery from exercise generated a large magnitude of effect
(ES ⬃1.0) that verged on significance (⬃50%, P ⫽ 0.06; Fig. 4A).
However, there was little difference in the magnitude of effect
between the alternate exercise orders (⫺16%; ES, 0.2). Simi-
larly, there was a moderate decrease in mechano-growth factor
(MGF) mRNA abundance 3 h postexercise that was not sig-
nificantly different from rest (⬃50%; ES, ⬃0.7, P ⫽ 0.08),
with negligible disparity in the exercise order effect (⫺28%;
ES, 0.26; Fig. 4B). The elevation in MyoD mRNA 3 h after the
cessation of contractile activity was large (ES ⬎ 1.0) but failed
to reach significance and was similar regardless of exercise
order (⬃300%, P ⫽ 0.06; Fig. 4C).
Atrogin-MuRF-Foxo3a. Variation in individual responses
resulted in little change in atrogin and Foxo3a (Foxo3a not Fig. 4. Insulin-like growth factor 1 (IGF-I; A), mechano-growth factor (MGF;
shown) mRNA abundance following the consecutive exercise B), and myogenic differentiation factor (MyoD; C) mRNA abundance at rest
bouts, with little discrepancy in the effect of the different preexercise and 3 h after cessation of training. Subjects completed 2 experi-
exercise orders (ES ⬍ 0.2; Fig. 5A). In contrast, the mRNA mental trials incorporating consecutive REX (8 ⫻ 5 leg extensions at 80%
1-RM) and SPR (10 ⫻ 6 s maximal effort) bouts performed in alternate order.
abundance of MuRF was significantly elevated above rest Results are individual responses and group means in arbitrary units.
following 3-h recovery from resistance exercise undertaken
prior to repeated sprints, and also after sprints were performed
before resistance exercise (ES ⬎ 1.0, P ⬍ 0.01; Fig. 5B). The effect (SPR1-REX2, ES ⬎ 1.0, P ⫽ 0.09), while the resistance
magnitude of increase in MuRF mRNA abundance was mod- exercise followed by repeated sprints exercise order was sig-
erately exacerbated when repeated sprints were undertaken nificantly different from rest (REX1-SPR2, ES ⬎ 1.0, P ⫽
subsequent to resistance exercise (REX1-SPR2 vs. SPR1- 0.02). There was a moderate distinction in the relative increase
REX2, ⬃25%; ES, 0.75; Fig. 5B). above resting levels with augmented PGC-1␣ mRNA when
PGC-1␣-hexokinase II-mitochondrial transcription fac- repeated sprints followed resistance exercise (REX1-SPR2,
tor A-citrate synthase. There was a significant main effect for ⬃35%; ES, 0.46; Fig. 6A). Hexokinase II mRNA abundance
the levels of time for PGC-1␣ mRNA in response to the was enhanced in response to contractile activity and was
combined exercise bouts (P ⫽ 0.04; Fig. 6A). The increased significantly elevated above rest after both exercise orders
abundance of PGC-1␣ mRNA, when repeated sprints preceded (ES ⬎ 1.0, P ⬍ 0.05; Fig. 6B). The magnitude of increase in
resistance exercise, failed to reach significance despite a large hexokinase II mRNA 3 h postexercise was moderately higher
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 CONCURRENT SPRINTS AND RESISTANCE EXERCISE R1447
robust increases in the mRNA abundance of PGC-1␣ and
hexokinase with only a modest exercise order distinction,
highlighting the capacity of high-intensity sprint exercise to
stimulate acute mitochondrial and metabolic adaptive re-
sponses.
There were minimal differences in the decline of mean
power despite a greater decrement with initial repeated sprints,
likely due to higher initial power outputs and accompanying
metabolic perturbations when undertaking sprints first (i.e.,
without any metabolic activity or residual fatigue from prior
contractile activity) generating a slightly greater fatigue index
(8). Moreover, during the 15-min recovery period following
each repeated sprint bout, blood lactate concentration was
similar (Fig. 2). However, the effect of elevated blood lactate
levels following initial repeated sprint exercise resulted in
higher concentrations after subsequent REX2. This finding
demonstrates that circulating blood lactate remained elevated
during resistance exercise undertaken after sprints (Fig. 2).
Muscle lactate was only significantly elevated above rest fol-

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lowing resistance exercise when preceded by repeated sprints
(Table 2). Differences in muscle H⫹ content were less clear,
but the largest changes were observed following an initial bout
of repeated sprints (Table 2). It may be reasonable to conclude
that resistance exercise performed after the repeated sprint

Fig. 5. Atrogin (A) and muscle-ring finger (MuRF; B) mRNA abundance at

rest preexercise and 3 h after cessation of training. Subjects completed 2
experimental trials incorporating consecutive REX (8 ⫻ 5 leg extensions at
80% 1-RM) and SPR (10 ⫻ 6 s maximal effort) bouts performed in alternate
order. Results are individual responses and group means in arbitrary units.
*Significant difference (P ⬍ 0.05) vs. rest.

when repeated sprints were undertaken before resistance exer-

cise (SPR1-REX2, ⬃46%; ES, 0.4; Fig. 6B). There was dis-
parity in individual mitochondrial transcription factor A and
citrate synthase mRNA responses (data not shown) culminat-
ing in minor changes in mean mRNA abundance from rest
following the 3-h recovery period.
DISCUSSION

Individual training sessions provide the overload stimulus

that generates specific adaptive events that ultimately alter
cellular characteristics for enhanced performance capacity in
skeletal muscle. We have recently established adaptive re-
sponses for several cellular and molecular pathways when
athletes undertake consecutive acute resistance and endurance
(concurrent) training (15). In the present study, we have taken
a novel approach to the concurrent training paradigm by
combining resistance exercise with repeated sprint perfor-
mance to examine the acute specificity of training adaptation
and exercise-order effects in skeletal muscle. Specifically, we
show attenuated S6K and rpS6 phosphorylation following Fig. 6. Peroxisome proliferator-activated receptor-␥ coactivator-1␣ (PGC-1␣;
repeated sprints in an exercise order-dependent manner. More- A) and hexokinase II (HKII; B) mRNA abundance at rest preexercise and 3 h
over, repeated sprints also appeared to diminish the anabolic after cessation of training. Subjects completed two experimental trials incor-
porating consecutive REX (8 ⫻ 5 leg extensions at 80% 1-RM) and SPR (10 ⫻
response to resistance exercise highlighted by a decrease in 6 s maximal effort) bouts performed in alternate order. Results are individual
IGF-I mRNA and concomitant increase in MuRF mRNA responses and group means in arbitrary units. *Significant difference (P ⬍
abundance following 3-h recovery. In addition, we observed 0.05) vs. rest.

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R1448 CONCURRENT SPRINTS AND RESISTANCE EXERCISE

repetitions was undertaken in the presence of greater intramus- cher et al. (45) have also examined Akt-mTOR signaling after
cular and whole body metabolic acidosis compared with the 60 min of cycling and observed increased mTOR phosphory-
initial resistance exercise bout. These findings provide support lation 0 and 30 min postexercise, a time when AMPK activity
for the notion that the different exercise modes generate diver- is also elevated. In addition, work in a transgenic animal model
gent stimuli that promote dissimilar signaling activity and gene by McGee et al. (47) provides support for the role of AMPK in
expression. Thus, the close proximity to resistance exercise and promoting, rather than inhibiting, skeletal muscle growth in-
metabolic changes as a consequence of the repeated sprints is dependent of its role in metabolism. Clearly, much work
significant given its potential to create an incompatible envi- remains to elucidate the interactions and subsequent physio-
ronment for resistance exercise-induced acute responses within logical function of these signaling proteins in human skeletal
the muscle milieu. muscle.
Akt has been characterized as a critical junction for multiple A novel finding of the present study was the divergence in
cell processes including glucose transport and hypertrophy in S6KThr389 and rpS6Ser235/6 phosphorylation following the di-
skeletal muscle (10, 64). In the present study, changes in verse exercise modes. The S6K and rpS6 proteins are proposed
Akt-TSC2-mTOR and AMPK phosphorylation and any subse- to regulate translational efficiency of mRNAs with a 5⬘-termi-
quent exercise-order effect were modest. Following 15-min nal oligopyrimidine tract (59), although a causative relation-
recovery from the initial exercise bout there was little diver- ship remains contentious (59), and increased phosphorylation
gence in Aktser473 phosphorylation between exercise modes. In of S6K and rpS6 is associated with the adaptive response to
contrast, when resistance exercise was undertaken after sprints, resistance exercise (17, 20, 22, 24). The mechanisms regulating
we observed elevated phosphorylation of Akt 15 min postex- these proteins are complex, and Akt and mitogen-activated

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ercise, resulting in a disparity in the magnitude of effect from protein kinase-mediated signaling pathways may converge at
rest (ES ⬎ 1.0; Fig. 3A). To the best of our knowledge, this is rpS6 (58). Nonetheless, we (15) and others (5, 44) have also
the first study to examine the insulin/IGF signaling pathway in shown increased S6K/rpS6 phosphorylation after a single bout
skeletal muscle following short, repeated sprint activity. Ac- of endurance exercise in human skeletal muscle. In contrast,
cordingly, we present new data indicating that repeated sprints Gibala et al. (23) reported no change in S6K phosphorylation
do not appear to promote Akt-TSC2-mediated kinase activity. following multiple 30 s sprints. We provide new data indicat-
In support of this observation, Gibala et al. (23) have reported ing that short, repeated sprint activity (10 ⫻ 6 s) does not
a decrease in Akt phosphorylation immediately (0 min) after promote translation initiation signaling in the early recovery
4 ⫻ 30-s all-out cycling bouts, each separated by 4-min period, suggesting that the overriding acute response necessi-
recovery. However, it should be noted that the acute postex- tates enhanced ion transport and buffering capacity (21, 49).
ercise cell-signaling time course, following both resistance and The suppression of S6K/rpS6 phosphorylation when compar-
sprint exercise is currently unknown. Whether the disparity in ing resistance exercise undertaken after repeated sprints with
Akt phosphorylation after a subsequent bout of resistance initial resistance exercise is intriguing. In previous work from
exercise in the present study represents the cumulative effect of our laboratory we failed to see a significant exercise order
the combined exercise bouts or is related to the time course of effect and observed increased S6K and rpS6 phosphorylation
activation, remains to be established. with consecutive endurance and resistance exercise bouts (15).
mTOR is a key regulatory protein for cell processes includ- Of note, metabolic acidosis has been shown to decrease protein
ing translation and protein synthesis, and mitochondrial func- synthesis in skeletal muscle of rodents and humans (11, 38).
tion (6, 19, 60). In the present study, there was a moderate Thus, the elevated muscle lactate and low pH immediately
increase in mTORser2448 phosphorylation from rest following prior to resistance exercise when undertaken after the initial
the initial exercise bouts that was sustained with subsequent repeated sprint bout implicates local acidosis as an inhibitor of
exercise, regardless of mode and order (Fig. 3C). The putative resistance exercise-induced S6K and rpS6 phosphorylation
evidence for an Akt-independent mechanism capable of regu- (Table 2). Collectively, these results provide support for the
lating mTOR phosphorylation in response to mechanical putative capacity of an initial bout of repeated sprints to
stretch (34, 35) may provide some rationale for the modest attenuate subsequent translation initiation up to ⬃3 h postex-
increases (main effects) in exercise-induced phosphorylation of ercise, due at least in part to changes in the ionic state of the
mTOR without concomitant changes in Akt phosphorylation. muscle milieu.
We also observed similar changes in AMPKThr172 phosphory- We also examined mRNA responses of select genes associ-
lation independent of exercise order (Fig. 3F). Given the ated with hypertrophy, atrophy, and metabolism in skeletal
potential for high-intensity contraction during repeated sprints muscle (14) to provide an adaptive profile generated by the
to induce rapid changes in energy turnover and alter the consecutive bouts of diverse exercise. Changes in mRNA
ATP-to-AMP ratio in skeletal muscle, a significant increase in abundance for genes of interest implicated in regulation of
AMPK activation might be expected. Indeed, Chen et al. (12) hypertrophy and atrophy did not reveal any obvious exercise
and Gibala et al. (23) have shown elevated AMPK activity order effect but the dual exercise bouts appeared to inhibit the
immediately after 30-s sprints. The results of the present study IGF-I mRNA response and promote mRNA abundance of
indicate that the 15-min recovery time point may not have MuRF 3 h after cessation of contractile activity (Figs. 4 and 5).
coincided with peak AMPK activity following the divergent A potential limitation of the present study was that muscle
contractile activity. Although few previous studies have mea- samples for equivalent 3-h postexercise recovery time points
sured both AMPK and mTOR phosphorylation/activity in vivo for each individual exercise bout were not taken. Nonetheless,
human skeletal muscle, Dreyer et al. (20) have previously our findings represent an adaptation “snapshot” 3 h after the
shown a significant increase in both AMPK activity and mTOR cessation of contractile activity that characterizes the cumula-
phosphorylation following a bout of resistance training. Mas- tive effect of the combined divergent stimuli.
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 CONCURRENT SPRINTS AND RESISTANCE EXERCISE R1449
IGF-I is a major growth factor that induces anabolic effects increase in markers of exercise-induced inflammation and may
in skeletal muscle (3, 51, 62) and increased IGF-I expression is actually accentuate the muscle damage response following
closely associated with muscle hypertrophy (1, 2). While heavy resistance exercise. However, there is a paucity of data
recent work by Spangenburg et al. (63) offers putative evi- on the effect of repeated sprints on mRNA expression and
dence for IGF-I independent muscle hypertrophy, elevated acute concurrent training interactions, and more work is
mRNA abundance of IGF-I variants (IGF-IEa and MGF) needed to elucidate the effect on anabolic and catabolic adap-
corresponds with increases in muscle cross-sectional area fol- tation responses in skeletal muscle.
lowing chronic exercise training (53). Few acute exercise PGC-1␣ has been described as a master controller of mito-
studies have examined IGF-I mRNA and a range of IGF-I chondrial biogenesis, and increases in PGC-1␣ mRNA have
responses have been observed between 2– 4 h recovery follow- been clearly evident following an acute bout of aerobic endur-
ing resistance exercise (26, 48, 55). We have previously shown ance exercise (16, 46, 50, 54). We have previously shown that
that following concurrent endurance and resistance exercise, an acute bout of concurrent endurance and resistance exercise
IGF-IEa and MGF mRNA abundance was only attenuated incorporating 30 min of cycling (70% V̇O2 peak) is insufficient
when cycling immediately preceded resistance exercise (15). to induce a robust increase in PGC-1␣ mRNA abundance,
In contrast, the results of the present study revealed a decrease regardless of exercise order. In contrast, the results of the
in IGF-I mRNA 3 h after the consecutive repeated sprint and present study reveal a significant increase in PGC-1␣ mRNA
resistance exercise bouts that was similar regardless of exercise despite the proximity of the divergent exercise bouts and short
order (Fig. 4A and B). Thus, we suggest that repeated sprints cumulative duration of the sprint activity (60 s). Gibala et al.
may have a greater negative effect on growth factor mRNA (23) previously showed only a small dose of intense exercise

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abundance compared with an acute endurance exercise bout, was sufficient to elevate PGC-1␣ mRNA abundance. Likewise,
and this effect is mediated by proximity per se rather than our data highlight the capacity for repeated bouts of supra-
exercise order when combined with resistance exercise. Gene maximal exercise to enhance acute PGC-1␣ mRNA expression
expression and resultant synthesis of new functional protein is despite prior resistance exercise that may promote increased
regulated at numerous molecular “check-points”. While strong PGC-1␣ protein content. Indeed, this unexpected revelation
correlations between variance in mRNA and protein levels was achieved utilizing exercise of half the duration (60 s) to
have been observed, the possibility exists that within the that of Gibala et al. (23). In addition, there was a modest
complexity of adaptation in skeletal muscle changes in mRNA disparity in the exercise order effect on PGC-1␣ mRNA,
and protein may not always equate (41). However, more work indicating that resistance exercise undertaken after repeated
is needed to clarify the acute effect of exercise-induced con- sprints may induce mild attenuation of the mitochondrial adap-
traction and concurrent training on IGF-I mRNA responses. tive response, but such an effect remains to be clearly estab-
We also observed similar mRNA changes in the MyoD in an lished. Hexokinase mRNA abundance was increased 3 h after
exercise order-independent manner (Fig. 4C). Yang et al. (67) completing the alternate exercise bouts with only a modest
have shown corresponding increases in skeletal muscle MyoD order effect, demonstrating a comparable metabolic adaptive
mRNA abundance following both endurance and resistance response of pathways regulating energy provision. Taken to-
exercise. Likewise, previous work from our laboratory shows gether, these results strongly suggest repeated sprints are ca-
that the cumulative effect of endurance and resistance exercise pable of stimulating an initial acute response that may ulti-
generates similar increases in MyoD mRNA abundance (15). mately enhance skeletal muscle oxidative and metabolic ca-
Therefore, enhanced MyoD mRNA expression appears to be pacities when undertaken concurrently with resistance exercise
induced by a variety of contractile activity suggesting a mini- regardless of exercise order.
mal role in the specificity of training adaptation.
There were contrasting effects of the acute repeated sprint and Perspectives and Significance
resistance exercise bouts on the mRNA abundance of mediators of
skeletal muscle inflammation and proteolytic responses in skeletal This is the first study to examine the potential interference or
muscle (37, 61). Specifically, the cumulative effect of exercise additive effect of acute concurrent training incorporating re-
induced a pronounced increase in MuRF mRNA, while varia- peated sprint activity and resistance exercise in skeletal mus-
tion in the individual response resulted in no alteration in mean cle. We chose to employ successive exercise bouts with short
atrogin mRNA abundance (Fig. 5, A and B). There was only a recovery based on anecdotal reports from coaches of elite
moderate disparity in the increase above rest in MuRF mRNA athletes regarding selected training regimens in a number of
with the alternate exercise orders (SPR1-REX2 vs. REX1- sports and also for the potential for time-efficient adaptation
SPR2; ES, 0.75). Atrogin and MuRF1 are E3 ubiquitin ligases with acute concurrent training sessions for health and fitness. A
implicated in skeletal muscle proteolysis via covalent tagging novel finding of the present study is that repeated sprints
of specific proteins with ubiquitin for their subsequent degra- appear to promote greater acute interference on markers of
dation by the 26S proteasome (9, 25). We have seen a similar resistance exercise adaptation than previously observed with
discrepancy in an exercise order-dependent manner which endurance exercise (15). Specifically, we observed an attenu-
showed an increased magnitude in the MuRF mRNA response ated phosphorylation of kinases proximal to translation initia-
when an endurance exercise bout was performed following tion and growth factor mRNA abundance. These acute re-
resistance exercise (15). Work by Louis et al. (43) indicates sponses may represent a temporary suppression of molecular
that MuRF mRNA abundance may be elevated 1– 4 h after 30 processes that promote net synthesis or maintenance of muscle
min (75% VO2 max⬎) of running and three sets of 10 repetition mass. Moreover, we also show elevated mRNA for a primary
(70% 1-RM) resistance exercise. Thus, the immediacy of marker of inflammation/proteolysis that would exacerbate the
repeated sprints in the present study did not attenuate the apparent negative effect on muscle mass, and these effects
AJP-Regul Integr Comp Physiol • VOL 297 • NOVEMBER 2009 • www.ajpregu.org
R1450 CONCURRENT SPRINTS AND RESISTANCE EXERCISE

were largely evident, regardless of the order in which the signaling in human skeletal muscle. Am J Physiol Endocrinol Metab 286:
consecutive exercise bouts were undertaken. An additional E737–E743, 2004.
14. Coffey VG, Hawley JA. The molecular bases of training adaptation.
finding of our study was that repeated sprints may provide a Sports Med 37: 737–763, 2007.
concentrated stimulus for mitochondrial biogenesis and high- 15. Coffey VG, Pilegaard H, Garnham AP, O’Brien BJ, Hawley JA.
lights the potential for short, intense exercise to promote Consecutive bouts of diverse contractile activity alter acute responses in
aerobic adaptation responses. Collectively, our results indicate human skeletal muscle. J Appl Physiol 106: 1187–1197, 2009.
16. Coffey VG, Shield A, Canny BJ, Carey KA, Cameron-Smith D,
repeated sprint activity generates the prevailing exercise-in-
Hawley JA. Interaction of contractile activity and training history on
duced adaptive response and subsequently alters the acute mRNA abundance in skeletal muscle from trained athletes. Am J Physiol
adaptive profile in skeletal muscle when undertaking concur- Endocrinol Metab 290: E849 –E855, 2006.
rent repeated sprint and resistance exercise. Accordingly, we 17. Coffey VG, Zhong Z, Shield A, Canny BJ, Chibalin AV, Zierath JR,
suggest that repeated sprint activities are isolated from resis- Hawley JA. Early signaling responses to divergent exercise stimuli in
skeletal muscle from well-trained humans. FASEB J 20: 190 –192, 2005.
tance training and that adequate recovery time is considered 18. Cohen J. Statistical Power Analysis for the Behavioral Sciences. Mah-
within periodized training plans that incorporate these diver- wah, NJ: Lawrence Erlbaum, 1988.
gent exercise modes. 19. Cunningham J, Rodgers J, Arlow D, Vazquez F, Mootha V, Puig-
server P. mTOR controls mitochondrial oxidative function through a
ACKNOWLEDGMENTS YY1-PGC-1␣ transcriptional complex. Nat Cell Biol 450: 736 –740, 2007.
20. Dreyer HC, Fujita S, Cadenas JG, Chinkes DL, Volpi E, Rasmussen
The authors wish to thank Heather Coffey, Ken Ballhause, Donato Rivas, BB. Resistance exercise increases AMPK activity and reduces 4E-BP1
and Jennifer Henderson for technical assistance with experimental trials. We phosphorylation and protein synthesis in human skeletal muscle. J Physiol
are particularly indebted to Michelle McGrath for technical assistance during 576: 613– 624, 2006.
analysis (Sport and Exercise Science Division, Institute of Food, Nutrition and

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21. Edge J, Bishop D, Hill-Haas S, Dawson B, Goodman C. Comparison of
Human Health, Massey University, New Zealand) and also acknowledge Will muscle buffer capacity and repeated-sprint ability of untrained, endurance
Hopkins (Auckland University of Technology, Auckland, New Zealand) for trained and team sport athletes. Eur J Appl Physiol 96: 225–234, 2006.
assistance with statistical analysis. 22. Eliasson J, Elfegoun T, Nilsson J, Kohnke R, Ekblom B, Blomstrand
E. Maximal lengthening contractions increase p70 S6 kinase phosphory-
GRANT lation in human skeletal muscle in the absence of nutritional supply. Am J
This study was funded by a grant from the Australian Sports Commission. Physiol Endocrinol Metab 291: E1197–E1205, 2006.
23. Gibala MJ, McGee SL, Garnham AP, Howlett KF, Snow RJ, Har-
greaves M. Brief intense interval exercise activates AMPK and p38
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