A novel voluntary weightlifting model in mice promotes muscle adaptation and insulin sensitivity with simultaneous enhancement of autophagy and mTOR pathway.

Our understanding of the molecular mechanisms underlying adaptations to resistance exercise remains elusive despite the significant biological and clinical relevance. We developed a novel voluntary mouse weightlifting model, which elicits squat-like activities against adjustable load during feeding, to investigate the resistance exercise-induced contractile and metabolic adaptations. RNAseq analysis revealed that a single bout of weightlifting induced significant transcriptome responses of genes that function in posttranslational modification, metabolism, and muscle differentiation in recruited skeletal muscles, which were confirmed by increased expression of fibroblast growth factor-inducible 14 (Fn14), Down syndrome critical region 1 (Dscr1) and Nuclear receptor subfamily 4, group A, member 3 (Nr4a3) genes. Long-term (8 weeks) voluntary weightlifting training resulted in significantly increases of muscle mass, protein synthesis (puromycin incorporation in SUnSET assay) and mTOR pathway protein expression (raptor, 4e-bp-1, and p70S6K proteins) along with enhanced muscle power (specific torque and contraction speed), but not endurance capacity, mitochondrial biogenesis, and fiber type transformation. Importantly, weightlifting training profound improved whole-body glucose clearance and skeletal muscle insulin sensitivity along with enhanced autophagy (increased LC3 and LC3-II/I ratio, and decreased p62/Sqstm1). These data suggest that resistance training in mice promotes muscle adaptation and insulin sensitivity with simultaneous enhancement of autophagy and mTOR pathway.

Effects of exercise in a cold environment on transcriptional control of PGC-1α.

Peroxisome proliferator-activated receptor-α coactivator-1α (PGC-1α) mRNA is increased with both exercise and exposure to cold temperature. However, transcriptional control has yet to be examined during exercise in the cold. Additionally, the need for environmental cold exposure after exercise may not be a practical recovery modality. The purpose of this study was to determine mitochondrial-related gene expression and transcriptional control of PGC-1α following exercise in a cold compared with room temperature environment. Eleven recreationally trained males completed two 1-h cycling bouts in a cold (7°C) or room temperature (20°C) environment, followed by 3 h of supine recovery in standard room conditions. Muscle biopsies were taken from the vastus lateralis preexercise, postexercise, and after a 3-h recovery. Gene expression and transcription factor binding to the PGC-1α promoter were analyzed. PGC-1α mRNA increased from preexercise to 3 h of recovery, but there was no difference between trials. Estrogen-related receptor-α (ERRα), myocyte enhancer factor-2 (MEF2A), and nuclear respiratory factor-1 (NRF-1) mRNA were lower in cold than at room temperature. Forkhead box class-O (FOXO1) and cAMP response element-binding protein (CREB) binding to the PGC-1α promoter were increased postexercise and at 3 h of recovery. MEF2A binding increased postexercise, and activating transcription factor 2 (ATF2) binding increased at 3 h of recovery. These data indicate no difference in PGC-1α mRNA or transcriptional control after exercise in cold versus room temperature and 3 h of recovery. However, the observed reductions in the mRNA of select transcription factors downstream of PGC-1α indicate a potential influence of exercise in the cold on the transcriptional response related to mitochondrial biogenesis.

Independent effects of acute normobaric hypoxia and hypobaric hypoxia on human physiology.

The purpose of this study was to examine the effects of acute normobaric (NH, decreased FiO2) and hypobaric (HH, 4200 m ascent) hypoxia exposures compared to sea level (normobaric normoxia, NN). Tissue oxygenation, cardiovascular, and body fluid variables measured during rest and a 3-min step-test following 90-min exposures (NH, HH, NN). Muscle oxygenated hemoglobin (O2Hb) decreased, and muscle deoxygenated hemoglobin (HHb) increased environmentally independent from rest to exercise (p < 0.001). During exercise, brain O2Hb was lower at HH compared to NN (p = 0.007), trending similarly with NH (p = 0.066), but no difference between NN and NH (p = 0.158). During exercise, HR at NH (141 ± 4 beats·min-1) and HH (141 ± 3 beats·min-1) were higher than NN (127 ± 44 beats·min-1, p = 0.002), but not each other (p = 0.208). During exercise, stroke volume at HH (109.6 ± 4.1 mL·beat-1) was higher than NH (97.8 ± 3.3 mL·beat-1) and NN (99.8 ± 3.9 mL·beat-1, p ≤ 0.010) with no difference between NH and NN (p = 0.481). During exercise, cardiac output at NH (13.8 ± 0.6 L) and HH (15.5 ± 0.7 L) were higher than NN (12.6 ± 0.5 L, p ≤ 0.006) with HH also higher than NH (p = 0.001). During acute hypoxic stimuli, skeletal muscle maintains oxygenation whereas the brain does not. These differences may be mediated by environmentally specific cardiovascular compensation. Thus, caution is advised when equating NH and HH.

Skeletal muscle cold shock and heat shock protein mRNA response to aerobic exercise in different environmental temperatures.

The response of cold shock proteins to exercise and environmental temperature in human skeletal muscle is not known. The purpose of this study was to determine the early mRNA response of human stress proteins to endurance exercise and environmental temperatures. Seven recreationally trained males cycled for 1 hour at 60% VO2peak in 7°C, 20°C, and 33°C with biopsies taken pre- and 3 hours post-exercise. Gene expression for heat shock and cold shock proteins were analyzed using qRT-PCR on muscle biopsy samples from the vastus lateralis. RBM3 mRNA was reduced 1.43 ± 0.10 fold (p = 0.006) while there was a trend for CIRP to decrease1.27 ± 0.14 fold (p = 0.059) from pre- to 3 h post-exercise. CIRP and RBM3 mRNA were not different between temperatures (p = 0.273 and p = 0.686, respectively). HSP70 mRNA was 2.27 ± 0.23 fold higher 3 h post-exercise when compared to pre-exercise (p = 0.002) but was not significantly different between temperatures (p = 0.103). HSP27, HSP90, and HSF1 mRNA did not change from pre- to post-exercise (p = 0.052, p = 0.324, p = 0.795) and were not different between temperatures (p = 0.247, p = 0.134, p = 0.808). These data indicate that exposure to mild heat and cold during aerobic exercise have limited effect on the skeletal muscle mRNA expression of heat shock and cold shock proteins. However, skeletal muscle mRNA of cold shock proteins decrease, while HSP70 mRNA increases in response to a low to moderate intensity aerobic exercise bout.

Skeletal Muscle mRNA Response to Hypobaric and Normobaric Hypoxia After Normoxic Endurance Exercise.

Background: The physiological effects of hypoxia may be influenced by how hypoxia is achieved. The purpose of this study was to determine the effects of recovery in hypobaric hypoxia (HH), normobaric hypoxia (NH), and normobaric normoxia (NN) after endurance exercise on gene expression related to mitochondrial biogenesis, myogenesis, and proteolysis. Methods: Fifteen recreationally trained subjects each cycled for 1 hour before recovering for 4 hours in NN (laboratory atmospheric conditions, 975 m), HH (depressurized to simulate 4420 m), and NH (fraction of O2 reduced to simulate 4420 m). Muscle biopsy samples were obtained before exercise and after 4 hours of recovery. Results: Blood oxygenation (SpO2) was lower in HH (76.02 ± 0.58%) than NH (79.45 ± 0.56, p < 0.001), which were both lower than in NN (96.3 ± 0.17, p < 0.001). Heart rate was higher in HH (82 ± 2 bpm) than NH (77 ± 1 bpm, p < 0.001), which were both higher than in NN (67 ± 1 bpm, p < 0.001). Mitochondrial transcription factor A (TFAM) mRNA was lower after NN than HH (p = 0.034) or NH (p = 0.005), but was not different between HH and NH (p = 0.460). Myostatin (MSTN) mRNA decreased from pre- to postexercise (p < 0.001) in all conditions and was lower in HH compared with NH (p = 0.035) and NN (p = 0.017). No other differences were noted in genes related to mitochondrial biogenesis, myogenesis, or proteolysis (p > 0.05). Conclusion:TFAM mRNA is lower with hypoxia exposure, but effected by the type of hypoxia. MSTN gene expression is lower after exposure to HH than NH or NN. These data support previous work and caution the translation of NH data obtained in a NH environment to a HH environment.

The effect of environmental temperature on exercise-dependent release of brain-derived neurotrophic factor.

Brain-derived neurotrophic factor (BDNF) is a biomarker of cognitive function that is released into the blood stream following exercise, and cognitive function is impaired by environmental temperatures that are hot and cold. Purpose: To evaluate the exercise-dependent release of BDNF in different environmental temperatures. Methods: Recreationally trained males each completed three trials consisting of cycling for 1 h at 60% Wmax at three different temperatures: 33°C (hot), 7°C (cold), and 20°C (moderate room temperature). Blood was taken from the antecubital vein pre-exercise, immediately post-exercise, and 3 h post-exercise. Respiratory gases were collected periodically throughout exercise and recovery. Results: BDNF was elevated immediately following an exercise bout (1711 ± 766 pg·ml-1) regardless of temperature from pre-exercise (1257 ± 653 pg·ml-1, p = 0.001) and returned to basal levels following 3 h of recovery (1289 ± 650 pg·ml-1, p = 0.786). There was no effect (p > 0.05) of temperature on BDNF following the exercise bout. Plasma glucose was elevated in hot (6.2 ± 0.9 mmol) over cold (5.3 ± 0.6 mmol, p = 0.035) and moderate room temperature (5.2 ± 0.5, p = 0.008). VO2 was elevated during exercise in hot (3.01 ± 0.45 L·min-1) over cold (2.67 ± 0.35 L·min-1, p = 0.005) and moderate room temperature (2.80 ± 0.38 L·min-1, p = 0.001). There was no relationship between BDNF and plasma glucose (p > 0.05) or VO2 across any time point or temperature (p > 0.05). Conclusion: With aerobic exercise, BDNF is elevated; however, the release of BDNF is not impacted by different environmental temperatures during exercise.

Impact of hot and cold exposure on human skeletal muscle gene expression.

Many human diseases lead to a loss of skeletal muscle metabolic function and mass. Local and environmental temperature can modulate the exercise-stimulated response of several genes involved in mitochondrial biogenesis and skeletal muscle function in a human model. However, the impact of environmental temperature, independent of exercise, has not been addressed in a human model. Thus, the purpose of this study was to compare the effects of exposure to hot, cold, and room temperature conditions on skeletal muscle gene expression related to mitochondrial biogenesis and muscle mass. Recreationally trained male subjects (n = 12) had muscle biopsies taken from the vastus lateralis before and after 3 h of exposure to hot (33 °C), cold (7 °C), or room temperature (20 °C) conditions. Temperature had no effect on most of the genes related to mitochondrial biogenesis, myogenesis, or proteolysis (p > 0.05). Core temperature was significantly higher in hot and cold environments compared with room temperature (37.2 ± 0.1 °C, p = 0.001; 37.1 ± 0.1 °C, p = 0.013; 36.9 ± 0.1 °C, respectively). Whole-body oxygen consumption was also significantly higher in hot and cold compared with room temperature (0.38 ± 0.01 L·min-1, p < 0.001; 0.52 ± 0.03 L·min-1, p < 0.001; 0.35 ± 0.01 L·min-1, respectively). In conclusion, these data show that acute temperature exposure alone does not elicit significant changes in skeletal muscle gene expression. When considered in conjunction with previous research, exercise appears to be a necessary component to observe gene expression alterations between different environmental temperatures in humans.

Irisin and Fibronectin Type III Domain-Containing 5 Responses to Exercise in Different Environmental Conditions.

Fibronectin type III domain-containing 5 (FNDC5) is a skeletal muscle membrane-bound precursor to the myokine irisin. Irisin is involved in stimulating adipose tissue to become more metabolically active in order to produce heat. The purpose of this study was to determine the effects of exercise in a hot (33 °C), cold (7 °C), and room temperature (RT, 20 °C) environment on the skeletal muscle gene expression of FNDC5 and the plasma concentrations of irisin. Twelve recreationally trained males completed three separate, 1 h cycling bouts at 60% of Wmax in a hot, cold, and RT environment followed by three hours of recovery at room temperature. Blood samples were taken from the antecubital vein and muscle biopsies were taken from the vastus lateralis pre-, post-, and 3 h post-exercise. Plasma concentrations of irisin did not change from pre- (9.23 ± 2.68 pg·mL-1) to post-exercise (9.6 ± 0.2 pg·mL-1, p = 0.068), but did decrease from post-exercise to 3 h post-exercise (8.9 ± 0.5 pg·mL-1, p = 0.047) regardless of temperature. However, when plasma volume shifts were considered, no differences were found in irisin (p = 0.086). There were no significant differences between trials for irisin plasma concentrations (p > 0.05). No significant differences in FNDC5 were observed between the hot, cold, or RT or pre-, post-, or 3 h post-exercise time points (p > 0.05). These data indicate that the temperature in which exercise takes place does not influence FNDC5 transcription or circulating irisin in a human model.

Leptin, adiponectin, and ghrelin responses to endurance exercise in different ambient conditions.

Excessive positive energy balance is a major factor leading to obesity. The ability to alter the appetite-regulating hormones leptin, adiponectin, and ghrelin may help decrease excessive energy intake. Exercise and exposure to extreme temperatures can independently affect these appetite-regulating hormones. PURPOSE: To determine the effect of exercising in different environmental conditions on the circulating concentrations of leptin, adiponectin, and ghrelin. METHODS: Eleven recreationally-trained male participants completed 3 separate 1 h cycling bouts at 60% Wmax in hot, cold, and room temperature conditions (33°C, 7°C, 20°C), followed by a 3 h recovery at room temperature. Blood was drawn pre-exercise, post-exercise, and 3 h post-exercise. Hematocrit and hemoglobin were measured to account for change in plasma volume. RESULTS: Leptin concentrations were lower at post and 3 h post-exercise compared with pre-exercise, with and without correction for plasma volume shifts, regardless of temperature (p < 0.05). Adiponectin was higher post-exercise compared with pre-exercise (p = 0.021) but not 3 h post-exercise (p = 0.084) without correction for plasma volume shifts. However, adiponectin concentrations were not different at any time point when plasma volume shifts were accounted for (p > 0.05). Total ghrelin and acylated ghrelin concentrations were not affected at post and 3 h post-exercise compared with pre-exercise, with and without correcting for plasma volume shifts, regardless of ambient temperature (p > 0.05). No differences in leptin, adiponectin, or ghrelin were found between trials (p > 0.05). CONCLUSION: Temperature does not affect the circulating concentrations of appetite-regulating hormones during an acute bout of endurance exercise.

Transcriptional control, but not subcellular location, of PGC-1α is altered following exercise in a hot environment.

The purpose of this study was to determine mitochondrial biogenesis-related mRNA expression, binding of transcription factors to the peroxisome proliferator-activated receptor-γ coactivator 1-α (PGC-1α) promoter, and subcellular location of PGC-1α protein in human skeletal muscle following exercise in a hot environment compared with a room temperature environment. Recreationally trained males (n = 11) completed two trials in a temperature- and humidity-controlled environmental chamber. Each trial consisted of cycling in either a hot (H) or room temperature (C) environment (33 and 20°C, respectively) for 1 h at 60% of maximum wattage (Wmax) followed by 3 h of supine recovery at room temperature. Muscle biopsies were taken from the vastus lateralis pre-, post-, and 3 h postexercise. PGC-1α mRNA increased post (P = 0.039)- and 3 h postexercise in C (P = 0.002). PGC-1α, estrogen-related receptor-α (ERRα), and nuclear respiratory factor 1 (NRF-1) mRNA was all lower in H than C post (P = 0.038, P < 0.001, and P = 0.030, respectively)- and 3 h postexercise (P = 0.035, P = 0.007, and P < 0.001, respectively). Binding of cAMP response element-binding protein (CREB) (P = 0.005), myocyte enhancer factor 2 (MEF2) (P = 0.047), and FoxO forkhead box class-O1 (FoxO1) (P = 0.010) to the promoter region of the PGC-1α gene was lower in H than C. Nuclear PGC-1α protein increased postexercise in both H and C (P = 0.029) but was not different between trials (P = 0.602). These data indicate that acute exercise in a hot environment blunts expression of mitochondrial biogenesis-related mRNA, due to decreased binding of CREB, MEF2, and FoxO1 to the PGC-1α promoter.