Impact of adrenaline and metabolic stress on exercise-induced intracellular signaling and PGC-1α mRNA response in human skeletal muscle

Nina Brandt, Thomas P Gunnarsson, Morten Hostrup, Jonas Tybirk, Lars Nybo, Henriette Pilegaard, Jens Bangsbo, Nina Brandt, Thomas P Gunnarsson, Morten Hostrup, Jonas Tybirk, Lars Nybo, Henriette Pilegaard, Jens Bangsbo

Abstract

This study tested the hypothesis that elevated plasma adrenaline or metabolic stress enhances exercise-induced PGC-1α mRNA and intracellular signaling in human muscle. Trained (VO2-max: 53.8 ± 1.8 mL min(-1) kg(-1)) male subjects completed four different exercise protocols (work load of the legs was matched): C - cycling at 171 ± 6 W for 60 min (control); A - cycling at 171 ± 6 W for 60 min, with addition of intermittent arm exercise (98 ± 4 W). DS - cycling at 171 ± 6 W interspersed by 30 sec sprints (513 ± 19 W) every 10 min (distributed sprints); and CS - cycling at 171 ± 6 W for 40 min followed by 20 min of six 30 sec sprints (clustered sprints). Sprints were followed by 3:24 min:sec at 111 ± 4 W. A biopsy was obtained from m. vastus lateralis at rest and immediately, and 2 and 5 h after exercise. Muscle PGC-1α mRNA content was elevated (P < 0.05) three- to sixfold 2 h after exercise relative to rest in C, A, and DS, with no differences between protocols. AMPK and p38 phosphorylation was higher (P < 0.05) immediately after exercise than at rest in all protocols, and 1.3- to 2-fold higher (P < 0.05) in CS than in the other protocols. CREB phosphorylation was higher (P < 0.05) 2 and 5 h after exercise than at rest in all protocols, and higher (P < 0.05) in DS than CS 2 h after exercise. This suggests that neither plasma adrenaline nor muscle metabolic stress determines the magnitude of PGC-1α mRNA response in human muscle. Furthermore, higher exercise-induced changes in AMPK, p38, and CREB phosphorylation are not associated with differences in the PGC-1α mRNA response.

Keywords: Exercise; PGC‐1α mRNA; human muscle biopsies; intracellular signaling; metabolic stress.

© 2016 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.

Figures

Figure 1
Figure 1
Schematic presentation of the four experimental days consisting of 60 min of cycling at an average intensity corresponding to 60% of VO 2‐max (171 ± 6 W) as (A) continuous cycling (C), (B) continuous cycling with intermittent arm exercise (A), (C) continuous cycling interspersed by six 30 sec sprints (513 ± 19 W) and cycling at 111 ± 4 W for 3:24 min:sec every 10 min (DS), and (D) 40 min of cycling followed by 20 min with six 30 sec sprints (513 ± 19 W) interspersed by 3:24 min:sec at 111 ± 4 W (CS). On experimental days, muscle biopsies were taken at rest as well as immediately, and 120 and 300 min after exercise. Blood samples were taken at rest, during (23, 39, and 60 min) and after (3, 10, 120, and 300 min) exercise.
Figure 2
Figure 2
Venous plasma adrenaline concentrations before (Rest; black bars), during (23 min: dark gray bars; 39 min: light gray bars; 60 min; white bars) 60 min of cycling at 171 ± 6 W on average as continuous control cycling (C), continuous cycling with arm exercise (A), continuous cycling interspersed by six 30 sec sprints (513 ± 19 W) and cycling at 111 ± 4 W for 3:24 min:sec every 10 min (DS), and 40 min of control cycling followed by 20 min with six 30 sec sprints (513 ± 19 W) interspersed by 3:24 min:sec at 111 ± 4 W (CS). Values (n = 10) are presented as mean ± SE. *Different (P < 0.05) from rest. aDifferent (P < 0.05) from C. bDifferent (P < 0.05) from A.
Figure 3
Figure 3
Muscle glycogen levels (mmol∙kg dw−1) before (rest; black bars), immediately (0 h; dark gray bars), 2 h (light gray bars), and 5 h (white bars) after 60 min of cycling (171 ± 6 W) as continuous control cycling (C), continuous cycling with arm exercise (A), continuous cycling interspersed by six 30 sec sprints (513 ± 19 W) and cycling at 111 ± 4 W for 3:24 min:sec every 10 min (DS), and 40 min of control cycling followed by 20 min with six 30 sec sprints (513 ± 19 W) interspersed by 3:24 min:sec at 111 ± 4 W (CS). Values (n = 10) are presented as mean±SE. *Different (P < 0.05) from rest. aDifferent (P < 0.05) from C. bDifferent (P < 0.05) from A.
Figure 4
Figure 4
(A) Peroxisome proliferator‐activated receptor‐ϒ coactivator‐1α (PGC‐1α), (B) vascular endothelial growth factor (VEGF), and (C) cytochrome c (Cyt C) mRNA content normalized to cyclophilin A mRNA content in vastus lateralis before (rest; black bars), immediately (0 h; dark gray bars), 2 h (light gray bars), and 5 h (white bars) after 60 min of cycling (171 ± 6 W) as continuous control cycling (C), continuous cycling with arm exercise (A), continuous cycling interspersed by six 30 sec sprints (513 ± 19 W) and cycling at 111 ± 4 W for 3:24 min:sec every 10 min (DS), and 40 min of control cycling followed by 20 min with six 30 sec sprints (513 ± 19 W) interspersed by 3:24 min:sec at 111 ± 4 W (CS). Values are presented as mean±SE; n = 8. *Different (P < 0.05) from rest.
Figure 5
Figure 5
(A) AMP‐activated protein kinase (AMPK)Thr172 phosphorylation normalized to AMPKα2 protein, (B) acetyl‐CoA carboxylase (ACC)Ser79 phosphorylation, (C) p38 mitogen‐activated protein kinase (p38)Thr180/Tyr182 phosphorylation normalized to p38 protein content, and (D) cAMP‐response element‐binding protein (CREB)Ser133 phosphorylation normalized to total CREB protein content before (rest; black bars), immediately (0 h; dark gray bars), 2 h (light gray bars), and 5 h (white bars) after 60 min of cycling (171 ± 6 W) as continuous control cycling (C), continuous cycling with arm exercise (A), continuous cycling interspersed by six 30 sec sprints (513 ± 19 W) and cycling at 111 ± 4 W for 3:24 min:sec every 10 min (DS), and 40 min of control cycling followed by 20 min with six 30 s sprints (513 ± 19 W) interspersed by 3:24 min:s at 111 ± 4 W (CS). Protein levels are given in arbitrary units (AU). Values are presented as mean±SE; n = 10. *Different (P < 0.05) from rest. aDifferent (P < 0.05) from C. bDifferent (P < 0.05) from A. dDifferent (P < 0.05) from CS. eDifferent (P < 0.05) from all other protocols.

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