Treadmill Running Capacity Research Articles

WASF3 disrupts mitochondrial respiration and may mediate exercise intolerance in myalgic encephalomyelitis/chronic fatigue syndrome

Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is characterized by various disabling symptoms including exercise intolerance and is diagnosed in the absence of a specific cause, making its clinical management challenging. A better understanding of the molecular mechanism underlying this apparent bioenergetic deficiency state may reveal insights for developing targeted treatment strategies. We report that overexpression of Wiskott-Aldrich Syndrome Protein Family Member 3 (WASF3), here identified in a 38-y-old woman suffering from long-standing fatigue and exercise intolerance, can disrupt mitochondrial respiratory supercomplex formation and is associated with endoplasmic reticulum (ER) stress. Increased expression of WASF3 in transgenic mice markedly decreased their treadmill running capacity with concomitantly impaired respiratory supercomplex assembly and reduced complex IV levels in skeletal muscle mitochondria. WASF3 induction by ER stress using endotoxin, well known to be associated with fatigue in humans, also decreased skeletal muscle complex IV levels in mice, while decreasing WASF3 levels by pharmacologic inhibition of ER stress improved mitochondrial function in the cells of the patient with chronic fatigue. Expanding on our findings, skeletal muscle biopsy samples obtained from a cohort of patients with ME/CFS showed increased WASF3 protein levels and aberrant ER stress activation. In addition to revealing a potential mechanism for the bioenergetic deficiency in ME/CFS, our study may also provide insights into other disorders associated with fatigue such as rheumatic diseases and long COVID.

Effects of high-resistance wheel running on hallmarks of endurance and resistance training adaptations in mice.

Exercise effectively promotes and preserves cardiorespiratory, neuromuscular, metabolic, and cognitive functions throughout life. The molecular mechanisms underlying the beneficial adaptations to exercise training are, however, still poorly understood. To improve the mechanistic study of specific exercise training adaptations, standardized, physiological, and well-characterized training interventions are required. Therefore, we performed a comprehensive interrogation of systemic changes and muscle-specific cellular and molecular adaptations to voluntary low-resistance wheel running (Run) and progressive high-resistance wheel running (RR) in young male mice. Following 10 weeks of training, both groups showed similar improvements in body composition and peak oxygen uptake (V̇O2peak ), as well as elevated mitochondrial proteins and capillarization markers in the M. plantaris. Run mice clearly outperformed RR mice in a forced treadmill running capacity test, while RR mice displayed increased grip strength as well as superior mass gains in the M. soleus, associated with distinct proteomic changes specifying the two paradigms. Thus, even though both training modalities induce overlapping adaptations, Run interventions preferably improve submaximal running performance, while progressive RR is a valid model to study training-induced gains in grip strength and plantar flexor hypertrophy.

The acute removal of metformin induces mitochondrial remodeling and increases mitochondrial respiration

Metformin is a candidate to slow biological aging and extend healthspan, as it reduces the risk of morbidity and mortality in people with type 2 diabetes. However, it is not clear how metformin impacts individuals absent of chronic disease. This unknown is important to answer before prescribing metformin to healthy individuals to prolong the healthspan. It has been shown that metformin creates an inefficiency in energy transfer by accumulating within the mitochondria. Therefore, we hypothesized that the effects of metformin on healthspan are context specific; beneficial in the context of low energy demand/mitochondrial capacity but detrimental in the context of high energy demand/mitochondrial capacity. To test this hypothesis, we used 18-month, male rats selectively bred for low and high treadmill running capacity (LCR/HCR) as models of low and high intrinsic mitochondrial function. For both HCR and LCR, we used high-resolution respirometry (Oroboros O2K) on skeletal muscles from controls (CON), 4 weeks of metformin-treatment (MET), 4 weeks of metformin-treatment with a 48-hour metformin washout (MET-WO), and 48-hour washout with metformin added back to the respirometry chamber (MET-WO-SPIKE). A week prior to sacrifice, we labeled the rats with deuterium oxide to assess protein turnover as an indicator of mitochondrial remodeling. In gastrocnemius muscle, the HCR MET-WO had greater Complex I (CI), CI+Complex II (CII), and maximal respiration (MaxR) (115 ± 23, 164 ± 38, 157 ± 35 pmol/(s✕mg), respectively) compared to the CON (70 ± 24, p = 0.02, 114 ± 32, p = 0.01, 115 ± 40 pmol/(s✕mg), p = 0.04, respectively), and MET (63 ± 54, p = 0.007, 90 ± 50, p < 0.001, 91 ± 48 pmol/(s✕mg), p = 0.002, respectively). The HCR MET-SPIKE group had greater CII (83 ± 33 pmol/(s✕mg)) and MaxR (86 ± 35 pmol/(s✕mg)) compared to the MET (p = 0.007 and p = 0.002, respectively). In the soleus, MET-WO (4.15 ± 0.33%/day) had greater mitochondrial fractional synthesis rates (FSRs) compared to MET (3.12 ± 0.62%/day, p = 0.001) in the HCRs. In the LCRs, MET-WO (4.61 ± 0.42%/day) had greater FSRs compared to CON (p < 0.001, 3.07 ± 0.67%/day) and MET (3.12 ± 0.61%/day, p = 0.001). In the tibialis anterior, MET-WO (3.47 ± 0.80%/day) had greater FSRs compared to the MET (2.41 ± 0.47%/day, p = 0.04) within the HCRs with no differences in the LCR. These data show that metformin reduces mitochondrial respiration in HCR rats but not in LCR rats. The presence of metformin produced a greater subsequent response in HCR compared to LCR as shown by its acute removal revealing metformin-induced mitochondrial remodeling. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Biology: Motion is Function.

In 1966 Francis Crick declared that: “The ultimate aim of the modern movement in biology is to explain all biology in terms of physics and chemistry.” This motivated us to contemplate approaches that unify biology at a fundamental level. Exploration led us to consider the features of energy, entropy, and motion. Overall, it can be considered that motion of matter is the feature of life function. No motion. No function. In initial work we evaluated the hypothesis that the scope for biologic function is mediated mechanistically by a differential for energy transfer. Maximal treadmill running capacity served as a proxy for energy transfer. The span for capacity was estimated “biologically” by application of two-way artificial selection in rats for running capacity. Consistent with our “Energy Transfer Hypothesis” (ETH), low physical health and dysfunction segregated with low running capacity and high physical health and function segregated with high running capacity. The high energy yield of aerobic metabolism is also consonant with the ETH; that is, amongst the elements of the universe, oxygen is second only to fluorine in electronegativity. Although we deem these energy findings possibly correct, they are based on correlation and do not illuminate function via fundamental principles. For consideration of life, Entropy (2nd Law of thermodynamics) can be viewed as an open system that exchanges energy with the universe operating via nonequilibrium thermodynamics. The Principle of Maximal Entropy Production (MEP) states that: If a source of free energy is present, complex systems can intercept the free energy flow, and self-organize to enhance entropy production. The development of Benard convection cells in a water heat gradient demonstrate simplistic operation of MEP. A direct step forward would be to explain the mechanism of the obligatory motion of molecules for life function. Motion may be mediated by operation of “action at a distance” for molecules as considered by the Einstein-Podolsky-Rosen Paradox and confirmed by JS Bell. Magnetism, electricity, and gravity are also examples of action at a distance. We propose that some variant of “action at a distance” as directed by the property of Maximal Entropy Production (MEP) underwrites biologic motion.

Effects of different types of exercise training on angiogenic responses in the left ventricular muscle of aged rats.

We evaluated angiogenic responses in the left ventricular muscle and aerobic capacity according to exercise type (aerobic, resistance, combined) in aged rats. In total, 24 male Sprague-Dawley rats (100weeks old) were used. To investigate the effect of regular training, the rats were divided into non-exercise (NE), aerobic exercise (AE), resistance exercise (RE), and combined exercise (CE) groups (six rats per group). Regular training tailored to each exercise type was performed for 8weeks (five times a week, 1h per day). After 8weeks of training, aerobic capacity was evaluated by a treadmill running test. Left ventricular muscle tissue was collected and the protein levels of angiogenesis indicators (eNOS, HIF-1α, PGC-1α, VEGF, FLK-1, Ang-1, Ang-2) were analyzed by Western blotting. Capillaries were observed by immunohistochemical staining for CD31. Body weight, heart weight, and heart/body weight ratio showed no difference among the groups. The AE and CE groups showed higher treadmill running capacity than the NE and RE groups. The eNOS, VEGF, HIF-1α, PGC-1α, and Ang-2 protein levels were significantly higher in the AE than NE group. The PGC-1α and FLK-1 protein levels were significantly higher in the RE than NE group. In addition, in the CE group, the eNOS, FLK-1, and PGC-1α protein levels were significantly higher than in the NE group. Expression of CD31 in cardiac tissue was higher in the AE and CE groups than in the other groups. Taken together, the results suggest that regular exercise training, irrespective of exercise type, might improve cardiovascular function by inducing angiogenic responses in the aged myocardium; however, AE may be the most effective.

Role of GDF15 in active lifestyle induced metabolic adaptations and acute exercise response in mice

Physical activity is an important contributor to muscle adaptation and metabolic health. Growth differentiation factor 15 (GDF15) is established as cellular and nutritional stress-induced cytokine but its physiological role in response to active lifestyle or acute exercise is unknown. Here, we investigated the metabolic phenotype and circulating GDF15 levels in lean and obese male C57Bl/6J mice with long-term voluntary wheel running (VWR) intervention. Additionally, treadmill running capacity and exercise-induced muscle gene expression was examined in GDF15-ablated mice. Active lifestyle mimic via VWR improved treadmill running performance and, in obese mice, also metabolic phenotype. The post-exercise induction of skeletal muscle transcriptional stress markers was reduced by VWR. Skeletal muscle GDF15 gene expression was very low and only transiently increased post-exercise in sedentary but not in active mice. Plasma GDF15 levels were only marginally affected by chronic or acute exercise. In obese mice, VWR reduced GDF15 gene expression in different tissues but did not reverse elevated plasma GDF15. Genetic ablation of GDF15 had no effect on exercise performance but augmented the post exercise expression of transcriptional exercise stress markers (Atf3, Atf6, and Xbp1s) in skeletal muscle. We conclude that skeletal muscle does not contribute to circulating GDF15 in mice, but muscle GDF15 might play a protective role in the exercise stress response.

Early life undernutrition reduces maximum treadmill running capacity in adulthood in mice.

Undernutrition during early life causes chronic disease with specific impairments to the heart and skeletal muscle. The purpose of this study was to determine the effects of early life undernutrition on adult exercise capacity as a result of cardiac and skeletal muscle function. Pups were undernourished during gestation (GUN) or lactation (PUN) using a cross-fostering nutritive mouse model. At postnatal day 21, all mice were weaned and refed a control diet. At postnatal day 67, mice performed a maximal treadmill test. Echocardiography and Doppler blood flow analysis was performed at postnatal day 72, following which skeletal muscle cross-sectional area (CSA) and fiber type were determined. Maximal running capacity was reduced (diet: P = 0.0002) in GUN and PUN mice. Left ventricular mass (diet: P = 0.03) and posterior wall thickness during systole (diet × sex: P = 0.03) of GUN and PUN mice was reduced, causing PUN mice to have reduced (diet: P = 0.04) stroke volume. Heart rate of GUN mice showed a trend (diet: P = 0.07) towards greater resting values than other groups. PUN mice had greater CSA of soleus fibers. PUN had a reduced (diet: P = 0.03) proportion of type-IIX fibers in the extensor digitorum longus (EDL) and a greater (diet: P = 0.008) percentage of type-IIB fibers in the EDL. In conclusion, gestational and postnatal undernourishment impairs exercise capacity.

Effects of Early Life Undernutrition on Maximum Treadmill Running Capacity in Mice

Undernourishment in early life has been shown to impair cardiovascular function, which could potentially influence maximum exercise capacity in adulthood. PURPOSE: To determine the effects of early life undernutrition on maximal exercise capacity in adulthood. METHODS: Using a cross fostering model, pups were undernourished either during gestation (GUN) or lactation (PUN; PN1-21) by feeding the dam a low-protein diet (8% protein) to decrease milk production. Control pups were born and suckled to dams fed an isocaloric diet with 20% protein content. At postnatal day 21 (PN21), all mice were weaned and switched to a control diet. To assess exercise capacity, mice began a 5-day treadmill acclimation protocol at PN61. At PN67, mice underwent a maximum work test, which began at 10 meters/minute with a 10% grade. Speed was increased every two minutes until exhaustion. The amount of work completed by each mouse was calculated as: Maximum Work(J)= 9.8 x Maximum Speed (m/min) x grade(radians) x Time (min) x Weight (kg). A two-way ANOVA was used to determine differences in maximal work and cardiac parameters between groups with the effects of gender and diet. RESULTS: GUN (0.3312±0.037 J) mice performed higher than PUN (0.2527±0.050 J) and CON (0.2674±0.030 J) mice on a maximum work test (p<0.05). There was no gender effect. CONCLUSION: Undernourishment during lactation leads to lower work capacity, indicating that developmental programing during the first 21 days of life impairs work capacity during adulthood.

Loss of mitochondrial phosphate carrier in skeletal muscle: dissociation of muscle dysfunction from lower ADP phosphorylating potential

Many mutations in the mitochondrial genome and in nuclear genes that encode mitochondrial proteins hamper oxidative phosphorylation (oxphos) and lead to tissue dysfunction. Yet, whether an energy deficit is necessarily a direct cause of tissue dysfunction is not clear. We addressed this question in mice in which the mitochondrial phosphate carrier, PiC, was depleted from skeletal muscle (skm) by tamoxifen‐induced excision of nuclear gene SLC25A3 encoding PiC (Cre+). Three weeks after tamoxifen injections, PiC protein in skm was &lt;5% of levels found in control (Ctrl) wild‐type mice (tamoxifen‐treated). Oxphos, measured in mitochondria isolated from skm, was greatly suppressed to &lt;2%, 35% or 50% of Ctrl, depending on substrate, with essentially no oxphos when succinate (+rotenone) was supplied and the highest residual oxphos when fatty acid substrate and malate were supplied; the residual oxphos may be driven by alternate Pi uptake on the dicarboxylate carrier. Residual oxphos was similar at 3 and 9 wks of PiC depletion, suggesting that the oxphos defect was stable for this interval. Furthermore, subunits of the oxphos machinery were robustly upregulated (50–70% rise) at both time points. Nonetheless, treadmill running capacity worsened during this time. Time‐to‐exhaustion was ~50 mins in Cre+ mice with PiC absent for 3 wks, but ~25 mins when PiC was absent for 9 wks, whereas Ctrl mice ran for the scheduled 80 mins. Thus, deteriorating treadmill running in Cre+ mice seems unrelated to suppressed oxphos per se. Stress signaling via transcription factor ATF4 as well as elevated mTORC1 signaling have been associated with oxphos defects caused by mitochondrial DNA depletion in cell models and in mouse skm. These models have also suggested a causal relationship between persistent mTORC1 activation/stress signaling and cell dysfunction. Yet we found little evidence for stress signaling via ATF4 (or ATF2, ATF3, or ATF5) after 3 or 9 wks of PiC depletion, and evidence for only mild mTORC1 activation after 9 wks of PiC loss. Overall, these observations suggest that skm dysfunction is not related to an oxphos deficit in a simple way. Rather, the oxphos deficit may drive adaptive mechanisms that either become exhausted or turn maladaptive when ongoing over a longer term, or trigger additional processes that are deleterious for muscle function. Furthermore, the model presented here suggests that a mechanism other than stress signaling through ATFs is responsible for tissue dysfunction in the context of a severe oxphos defect.Support or Funding InformationFunding: R01 GM123771 to ELSThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Theoretical and Biological Evaluation of the Link between Low Exercise Capacity and Disease Risk.

Large-scale epidemiological studies show that low exercise capacity is the highest risk factor for all-cause morbidity and mortality relative to other conditions including diabetes, hypertension, and obesity. This led us to formulate the energy transfer hypothesis (ETH): Variation in capacity for energy transfer is the central mechanistic determinant of the divide between disease and health. As a test of this hypothesis, we predicted that two-way selective breeding of genetically heterogeneous rats for low and high intrinsic treadmill running capacity (a surrogate for energy transfer) would also produce rats that differ for disease risks. The lines are termed low-capacity runners (LCRs) and high-capacity runners (HCRs) and, after 36 generations of selection, they differ by more than eightfold in running capacity. Consistent with the ETH, the LCRs score high for developing disease risks, including metabolic syndrome, neurodegeneration, cognitive impairment, fatty liver disease, susceptibility to cancer, and reduced longevity. The HCRs are resistant to the development of these disease risks. Here we synthesize ideas on nonequilibrium thermodynamics and evolution from Ilya Prigogine, Hans Krebs, and Peter Mitchell to formulate theoretic explanations for the ETH. First, at every moment in time, the atoms and molecules of organisms are reorganizing to pursue avenues for energy transfer. Second, this continuous organization is navigating in a constantly changing environment such that "strategies" are perpetually in flux and do not leave a simple footprint (evolution). Third, as a consequence, human populations demonstrate a wide variation in capacity for energy transfer that mirrors mechanistically the divide between disease and health.

The AMPK activator R419 improves exercise capacity and skeletal muscle insulin sensitivity in obese mice

ObjectiveSkeletal muscle AMP-activated protein kinase (AMPK) is important for regulating glucose homeostasis, mitochondrial content and exercise capacity. R419 is a mitochondrial complex-I inhibitor that has recently been shown to acutely activate AMPK in myotubes. Our main objective was to examine whether R419 treatment improves insulin sensitivity and exercise capacity in obese insulin resistant mice and whether skeletal muscle AMPK was important for mediating potential effects.MethodsGlucose homeostasis, insulin sensitivity, exercise capacity, and electron transport chain content/activity were examined in wildtype (WT) and AMPK β1β2 muscle-specific null (AMPK-MKO) mice fed a high-fat diet (HFD) with or without R419 supplementation.ResultsThere was no change in weight gain, adiposity, glucose tolerance or insulin sensitivity between HFD-fed WT and AMPK-MKO mice. In both HFD-fed WT and AMPK-MKO mice, R419 enhanced insulin tolerance, insulin-stimulated glucose disposal, skeletal muscle 2-deoxyglucose uptake, Akt phosphorylation and glucose transporter 4 (GLUT4) content independently of alterations in body mass. In WT, but not AMPK-MKO mice, R419 improved treadmill running capacity. Treatment with R419 increased muscle electron transport chain content and activity in WT mice; effects which were blunted in AMPK-MKO mice.ConclusionsTreatment of obese mice with R419 improved skeletal muscle insulin sensitivity through a mechanism that is independent of skeletal muscle AMPK. R419 also increases exercise capacity and improves mitochondrial function in obese WT mice; effects that are diminished in the absence of skeletal muscle AMPK. These findings suggest that R419 may be a promising therapy for improving whole-body glucose homeostasis and exercise capacity.

Mitochondrial and performance adaptations to exercise training in mice lacking skeletal muscle LKB1

LKB1 and its downstream targets of the AMP-activated protein kinase family are important regulators of many aspects of skeletal muscle cell function, including control of mitochondrial content and capillarity. LKB1 deficiency in skeletal and cardiac muscle (mLKB1-KO) greatly impairs exercise capacity. However, cardiac dysfunction in that genetic model prevents a clear assessment of the role of skeletal muscle LKB1 in the observed effects. Our purposes here were to determine whether skeletal muscle-specific knockout of LKB1 (skmLKB1-KO) decreases exercise capacity and mitochondrial protein content, impairs accretion of mitochondrial proteins after exercise training, and attenuates improvement in running performance after exercise training. We found that treadmill and voluntary wheel running capacity was reduced in skmLKB1-KO vs. control (CON) mice. Citrate synthase activity, succinate dehydrogenase activity, and pyruvate dehydrogenase kinase content were lower in KO vs. CON muscles. Three weeks of treadmill training resulted in significantly increased treadmill running performance in both CON and skmLKB1-KO mice. Citrate synthase activity increased significantly with training in both genotypes, but protein content and activity for components of the mitochondrial electron transport chain increased only in CON mice. Capillarity and VEGF protein was lower in skmLKB1-KO vs. CON muscles, but VEGF increased with training only in skmLKB1-KO. Three hours after an acute bout of muscle contractions, PGC-1α, cytochrome c, and VEGF gene expression all increased in CON but not skmLKB1-KO muscles. Our findings indicate that skeletal muscle LKB1 is required for accretion of some mitochondrial proteins but not for early exercise capacity improvements with exercise training.

Vascularity of myocardium and gastrocnemius muscle in rats selectively bred for endurance running capacity

We tested the hypothesis that changes in the arteriolar branching architecture contributed to increased running capacity of rats subjected to two-way artificial selection for intrinsic aerobic endurance treadmill running capacity resulting in strains of low-capacity and high-capacity endurance rats. Hearts and gastrocnemius muscles were harvested from each strain, and the microvasculature's branching geometry measured from micro-CT images. The vascular branching geometry of the hearts and skeletal muscle from the high capacity was indistinguishable from low-capacity rats. Our hypothesis was not supported. Neither remodeling nor an increase in arteriolar microvasculature branching appears to play a role in the enhanced performance of the high capacity rats. We are led to speculate that endothelial tolerance for shear stress and/or increased coupling of myocardial muscle fiber metabolic-to-contractile function is increased in the high-capacity runner strain to the effect of allowing either higher flow rate per unit volume of muscle or more efficient use of oxygen and nutrients in the high-capacity endurance rats.

Construction of two novel reciprocal conplastic rat strains and characterization of cardiac mitochondria

Because of the lack of appropriate animal models, the potentially causal contributions of inherited mitochondrial genomic factors to complex traits are less well studied compared with inherited nuclear genomic factors. We previously detected variations between the mitochondrial DNA (mtDNA) of the Dahl salt-sensitive (S) rat and the spontaneously hypertensive rat (SHR). Specifically, multiple variations were detected in mitochondrial genes coding for subunits of proteins essential for electron transport, in mitochondrial reactive oxygen species production, and within the D-loop region. To evaluate the effects of these mtDNA variations in the absence of the corresponding nuclear genomic factors as confounding variables, novel reciprocal strains of S and SHR were constructed and characterized. When compared with that of the S rat, the heart tissue from the S.SHR(mt) conplastic strain wherein the mtDNA of the S rat was substituted with that of the SHR had a significant increase in mtDNA copy number and decrease in mitochondrial reactive oxygen species production. A corresponding increase in aerobic treadmill running capacity and a significant increase in survival that was not related to changes in blood pressure were observed in the S.SHR(mt) rats compared with the S rat. The reciprocal SHR.S(mt) rats did not differ from the SHR in any phenotype tested, suggesting lower penetrance of the S mtDNA on the nuclear genomic background of the SHR. These novel conplastic strains serve as invaluable tools to further dissect the relationship between heart function, aerobic fitness, cardiovascular disease progression, and mortality.

Novel conplastic strains reveal direct and independent effects of mitochondrial genomic variants on intrinsic aerobic fitness

Due to lack of appropriate animal models, the potential contributions of inherited mitochondrial genomic factors to complex traits are less well studied compared to inherited nuclear genomic factors. To unambiguously quantify the effect of the inherited mitochondrial factors on aerobic fitness and cardiovascular disease, we developed and characterized novel reciprocal conplastic strains by substitution of different mitochondrial genomes (mtDNA) on the same nuclear genetic background permissive to the development of cardiovascular disease. Conplastic strains of the Dahl S rat nuclear genomes with mtDNA from the Spontaneously Hypertensive Rat (SHR), with nonsynonymous variants in proteins of oxidative phosphorylation demonstrated increased aerobic treadmill running capacity (p&lt;0.05) and corresponding decreased levels of pro‐oxidant reactive oxygen species (p&lt;0.05) compared to the inbred Dahl S rat with S rat mitochondrial genome. Thus, increased pro‐oxidant stress caused by the mtDNA of S rat, which demonstrated lower aerobic running capacity, can be restored by substitution with the mtDNA of SHR. These results provide the first direct evidence linking naturally occurring variation in the mtDNA, independent of variation in the nuclear genome, to inherited variation in aerobic fitness. These novel conplastic strains serve as invaluable tools to further dissect the relationship between aerobic fitness and cardiovascular disease progression

Intrinsic Aerobic Capacity Sets a Divide for Aging and Longevity

Low aerobic exercise capacity is a powerful predictor of premature morbidity and mortality for healthy adults as well as those with cardiovascular disease. For aged populations, poor performance on treadmill or extended walking tests indicates closer proximity to future health declines. Together, these findings suggest a fundamental connection between aerobic capacity and longevity. Through artificial selective breeding, we developed an animal model system to prospectively test the association between aerobic exercise capacity and survivability (aerobic hypothesis). Laboratory rats of widely diverse genetic backgrounds (N:NIH stock) were selectively bred for low or high intrinsic (inborn) treadmill running capacity. Cohorts of male and female rats from generations 14, 15, and 17 of selection were followed for survivability and assessed for age-related declines in cardiovascular fitness including maximal oxygen uptake (VO(2max)), myocardial function, endurance performance, and change in body mass. Median lifespan for low exercise capacity rats was 28% to 45% shorter than high capacity rats (hazard ratio, 0.06; P<0.001). VO(2max), measured across adulthood was a reliable predictor of lifespan (P<0.001). During progression from adult to old age, left ventricular myocardial and cardiomyocyte morphology, contractility, and intracellular Ca(2+) handling in both systole and diastole, as well as mean blood pressure, were more compromised in rats bred for low aerobic capacity. Physical activity levels, energy expenditure (Vo(2)), and lean body mass were all better sustained with age in rats bred for high aerobic capacity. These data obtained from a contrasting heterogeneous model system provide strong evidence that genetic segregation for aerobic exercise capacity can be linked with longevity and are useful for deeper mechanistic exploration of aging.

Rats Bred For Low Aerobic Capacity Become Rapidly Fatigued And Have Slow Metabolic Recovery After Stimulated Muscle Contractions

Good aerobic capacity has a hypothesized role in increased health and longevity. Since impaired oxidative metabolism underlies several complex diseases, such as type 2 diabetes and cardiovascular disease, it is generally thought that the ability to increase oxygen transport and utilization capacity by exercise training stimulus helps prevent one from developing the aforementioned diseases. However, the magnitude of exercise capacity differs substantially between individuals. Primary assumption is that this heterogeneity is the consequence of variation distributed across a large number of genes for both the intrinsic (untrained) aerobic capacity and that accrued from exercise training. To address the intrinsic component, Koch and Britton (Physiol Genomics, 2001) developed two rat strains by artificial selection for low and high endurance treadmill running capacity in the untrained condition. These contrasting rat models, termed low capacity runner (LCR) and high capacity runner (HCR), are excellent biological models for evaluating the interplay of genetic and environmental factors as determinants of high health and aerobic capacity. PURPOSE: Our long-term aim is to evaluate the influence of life-long aerobic exercise training on aging and longevity using the LCR/HCR model system. Such studies require the application of non-invasive functional measures across time. Here we report on our first measurements of skeletal muscle energy metabolism in non-trained LCR and HCR rats. METHODS: Muscle function was studied in electrically stimulated contracting gastrocnemius muscle using 1H-magnetic resonance imaging (MRI) and 31P-MR spectroscopy. The simultaneous force output was measured with an ergometer connected to a foot pedal. RESULTS: Our results from mechanical measurement (force output) show a significant difference between LCR and HCR groups for the decrease in the isometric force during transcutaneous stimulation, the decrease being greater in LCR rats (P<0,001). 31P-MR spectroscopy results demonstrated that phosphocreatine resynthesis after stimulation was significantly slower in LCR group (P<0,02). CONCLUSIONS: Consistent with diminished treadmill running capacity, this study shows that skeletal muscle in LCR rats becomes fast fatigued during maximal muscle stimulation. 31P-MR spectroscopy seems to be a valid non-invasive method to investigate consecutive differentials in skeletal muscle energy metabolism between the LCR and HCR rats, making longitudinal studies feasible. Supported by the Ministry of Education and Culture (Finland) and the National Center for Research Resources (a component of the NIH).

Peripheral oxygen transport and utilization in rats following continued selective breeding for endurance running capacity

Untrained rats selectively bred for either high (HCR) or low (LCR) treadmill running capacity previously demonstrated divergent physiological traits as early as the seventh generation (G7). We asked whether continued selective breeding to generation 15 (G15) would further increase the divergence in skeletal muscle capillarity, morphometry, and oxidative capacity seen previously at G7. At G15, mean body weight was significantly lower (P < 0.001) in the HCR rats (n = 11; 194 +/- 3 g) than in LCR (n = 12; 259 +/- 9 g) while relative medial gastrocnemius muscle mass was not different (0.23 +/- 0.01 vs. 0.22 +/- 0.01% total body weight). Normoxic (Fi(O(2)) = 0.21) Vo(2max) was 50% greater (P < 0.001) in HCR despite the lower absolute muscle mass, and skeletal muscle O(2) conductance (measured in hypoxia; Fi(O(2)) = 0.10) was 49% higher in HCR (P < 0.001). Muscle oxidative enzyme activities were significantly higher in HCR (citrate synthase: 16.4 +/- 0.4 vs. 14.0 +/- 0.6; beta-hydroxyacyl-CoA dehydrogenase: 5.2 +/- 0.2 vs. 4.2 +/- 0.2 mmol.kg(-1).min(-1)). HCR rats had approximately 36% more total muscle fibers and also 36% more capillaries in the medial gastrocnemius. Because average muscle fiber area was 35% smaller, capillary density was 36% higher in HCR, but capillary-to-fiber ratio was the same. Compared with G7, G15 HCR animals showed 38% greater total fiber number with an additional 25% decrease in mean fiber area. These data suggest that many of the skeletal muscle structural and functional adaptations enabling greater O(2) utilization in HCR at G7 continue to progress following additional selective breeding for endurance capacity. However, the largest changes at G15 relate to O(2) delivery to skeletal muscle and not to the capacity of skeletal muscle to use O(2).

It's all in the genes, so pick your parents wisely

Aerobic exercise capacity is a complex phenotype influenced by a myriad of environmental and genetic factors. At one end of the continuum, elite endurance athletes represent a rare union of genetic potential that, given appropriate environmental stimulus (e.g., training, nutrition) can culminate in

The Effect of Carbohydrate-Protein Mixtures on Muscle Glycogen Resynthesis following Prolonged Treadmill Running

The ingestion of both carbohydrate and protein during recovery from prolonged exercise has previously been demonstrated to result in a more rapid restoration of treadmill running capacity than ingestion of carbohydrate alone. Evidence based on prolonged cycling suggests that this ergogenic effect might be related to an increased rate of muscle glycogen resynthesis during recovery. PURPOSE: To determine whether the addition of protein to a carbohydrate solution accelerates the rate of muscle glycogen resynthesis during recovery from prolonged treadmill running. METHODS: Six recreationally active male volunteers (age 22 ± 1.3 y, body mass 73.8 ± 6.7 kg, VO2max 60.5 ± 6.3 ml·kg−1mm−1; mean ± SD) participated in 2 trials separated by 14 days. Each main trial involved a glycogen lowering run (R1; 90 min at 70% VO max) followed by a 4 hour recovery. At 30 min intervals during recovery, participants ingested either a solution containing 0.8 g CHO· kg−1· h−1 (CHO trial) or the same solution plus an additional 0.3 g· kg−1· h−1 of protein (CHO-PRO trial). Muscle biopsies were obtained from the vastus lateralis at the beginning and end of recovery to asses the rate of muscle glycogen resynthesis over the intervening period. Furthermore, a third muscle biopsy was taken following a repeated treadmill run subsequent to recovery (R2; 60 min at 70% VO2max). RESULTS: Despite a higher insulinemic response to the CHO-PRO solution than to the CHO solution (P <0.05), rates of muscle glycogen resynthesis during recovery were not different between treatments (CHO: 12.3 ± 2.2 and CHO-PRO: 12.0 ± 2.7 mmol glucosyl units-kg dm·h−1; mean ± SEM). Similarly, the additional protein did not influence the rate of muscle glycogen degradation during subsequent exercise (CHO: 2.2 ±0.3 and CHO: 2.0 ±0.1 mmol glucosyl units·kg dm−1h−1; mean± SEM). CONCLUSIONS: Combined ingestion of carbohydrate and protein did not increase muscle glycogen storage during recovery from prolonged treadmill running nor spare this carbohydrate store during a repeated exercise session. It is possible that, in contrast to prolonged cycling, the proposed benefit of ingesting carbohydrate-protein following running might involve mechanisms other than improved muscle glycogen status.