Skeletal Muscle Mitochondrial Capacity Research Articles

Reduced oxidative capacity of skeletal muscle mitochondria is a fundamental consequence of adult ageing.

Reduced oxidative capacity of skeletal muscle mitochondria is a fundamental consequence of adult ageing.

Exercise Training Mitigates Inhibition of Skeletal Muscle Mitochondrial Respiration with Empagliflozin Treatment in Male and Female Mice

Sodium-glucose cotransporter-2 inhibitors (SGLT2i) are antihyperglycemic drugs designed to lower renal glucose reabsorption, yet recent evidence indicates that SGLT2i have off-target effects on skeletal muscle to inhibit mitochondrial respiration. Such inhibition raises important questions regarding the interaction of SGLT2i and exercise on regulation of skeletal muscle mitochondrial metabolism. The purpose of the current study was to test the hypothesis that SGLT2i treatment (empagliflozin, EMPA) would blunt improvements to skeletal muscle mitochondrial oxidation capacity following aerobic training. Male and female C57BL/6J mice consumed a western diet (40% kcal from fat) for 8 weeks, then performed an additional 8 weeks of either treadmill training (5 times per week progressing up to 23 m/min) or remained sedentary. During the intervention period, all mice consumed either western diet or western diet enriched with EMPA (10mg/kg diet) in a 2x2 design (n=10/group for each sex). We isolated mitochondria from quadriceps muscles collected 72 hours after the final training session (or rest) and measured ex vivo mitochondrial substrate oxidation capacity using high-resolution respirometry. There was an interaction between exercise and EMPA (p=0.037) on complex I-II respiration whereby sedentary mice had lower respiration following EMPA versus untreated (-17% p=0.01) while exercise-trained mice had similar respiration between EMPA and untreated. The interaction was less prominent in complex I-supported respiration (p=0.06) or complex-II alone (p=0.09). Our results show that, contrary to expectations, EMPA treatment did not impair mitochondrial adaptations to exercise training. Instead, exercise training prevented the suppression of mitochondrial respiration during EMPA treatment. These findings indicate that aerobic exercise has positive effects on the off-target inhibition of mitochondrial respiration by SGLT2i. Supported by funding from the Collins Medical Trust (Wilsonville, Oregon) awarded to Matthew Robinson. This is the full abstract presented at the American Physiology Summit 2024 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.

Comparison of skeletal muscle oxidative function in young, middle-aged, and older males and females with similar physical activity levels.

Optimal skeletal muscle oxidative function (microvascular reactivity and mitochondrial capacity) is an integral part of healthy aging and is related to physical function and quality of life. We aimed to extend upon the understanding of skeletal muscle oxidative function with healthy aging in males and females across the adult life span. Younger (N = 22; 11 males), middle-aged (N = 19; 10 males), and older (N = 21; 10 males) adults completed this study. Time spent in moderate and vigorous physical activity was self-reported and similar among groups. Near-infrared spectroscopy was used to investigate skeletal muscle microvascular reperfusion [oxyhemoglobin-myoglobin (O2Hb+Mb) half-time to peak hyperemia (T½)], mitochondrial capacity [muscle oxygen consumption (mV̇o2) recovery rate constant], and walking tissue oxygen saturation ([Formula: see text]) of the tibialis anterior (TA) muscle at seven incremental walking speeds. Mitochondrial capacity was not significantly different across groups (P = 0.07). Younger adults exhibited significantly slower T½ compared with older adults (P = 0.006) and middle-aged adults (P = 0.025). There were no observed sex differences for mitochondrial capacity (P = 0.442) or T½ (P = 0.402). Older adults exhibited significantly lower [Formula: see text] across all walking speeds compared with younger adults (P = 0.003). Mitochondrial capacity and microvascular reperfusion are maintained in middle and older age, with no sex differences in either outcome. However, in older adults whole body functional movement, such as walking, may place an additional demand on the TA as a compensatory response to lower functional reserve not evident in distinct measures of mitochondrial capacity and microvascular reperfusion.NEW & NOTEWORTHY Compared with younger adults, mitochondrial capacity and microvascular reperfusion of the tibialis anterior (TA) muscle are well maintained in similarly physically active middle-aged and older adults, with no sex differences observed in either outcome. However, greater tissue oxygen utilization in older adults during walking highlights how whole body functional movement may place an additional demand on the TA that reveals a potential compensatory response to lower functional reserve not evident in distinct measures of mitochondrial capacity and microvascular reperfusion.

Skeletal muscle mitochondrial capacity in patients with statin-associated muscle symptoms (SAMS)

ObjectiveThe objective of this article is to evaluate near-infrared spectroscopy (NIRS), a non-invasive technique to assess tissue oxygenation and mitochondrial function, as a diagnostic tool for statin-associated muscle symptoms (SAMS).MethodsWe...

Comprehensive interrogation of human skeletal muscle reveals a dissociation between insulin resistance and mitochondrial capacity.

Insulin resistance and blunted mitochondrial capacity in skeletal muscle are often synonymous, however, this association remains controversial. The aim of this study was to perform an in-depth multifactorial comparison of skeletal muscle mitochondrial capacity between individuals who were lean and active (Active, n = 9), individuals with obesity (Obese, n = 9), and individuals with obesity, insulin resistance, and type 2 diabetes (T2D, n = 22). Mitochondrial capacity was assessed by ex vivo mitochondrial respiration with fatty-acid and glycolytic-supported protocols adjusted for mitochondrial content (mtDNA and citrate synthase activity). Supercomplex assembly was measured by Blue Native (BN)-PAGE and immunoblot. Tricarboxylic (TCA) cycle intermediates were assessed with targeted metabolomics. Exploratory transcriptomics and DNA methylation analyses were performed to uncover molecular differences affecting mitochondrial function among the three groups. We reveal no discernable differences in skeletal muscle mitochondrial content, mitochondrial capacity, supercomplex assembly, TCA cycle intermediates, and mitochondrial molecular profiles between obese individuals with and without T2D that had comparable levels of confounding factors (body mass index, age, and aerobic capacity). We highlight that lean, active individuals have greater mitochondrial content, mitochondrial capacity, supercomplex assembly, and TCA cycle intermediates. These phenotypical changes are reflected at the level of DNA methylation and gene transcription. The collective observation of comparable muscle mitochondrial capacity in individuals with obesity and T2D (vs. individuals without T2D) underscores a dissociation from skeletal muscle insulin resistance. Clinical trial number: NCT01911104.NEW & NOTEWORTHY Whether impaired mitochondrial capacity contributes to skeletal muscle insulin resistance is debated. Our multifactorial analysis shows no differences in skeletal muscle mitochondrial content, mitochondrial capacity, and mitochondrial molecular profiles between obese individuals with and without T2D that had comparable levels of confounding factors (BMI, age, aerobic capacity). We highlight that lean, active individuals have enhanced skeletal muscle mitochondrial capacity that is also reflected at the level of DNA methylation and gene transcription.

Invasive and noninvasive markers of human skeletal muscle mitochondrial function.

Mitochondria are organelles that fuel cellular energy requirements by ATP formation via aerobic metabolism. Given the wide variety of methods to assess skeletal muscle mitochondrial capacity, we tested how well different invasive and noninvasive markers of skeletal muscle mitochondrial capacity reflect mitochondrial respiration in permeabilized muscle fibers. Nineteen young men (mean age: 24 ± 4 years) were recruited, and a muscle biopsy was collected to determine mitochondrial respiration from permeabilized muscle fibers and to quantify markers of mitochondrial capacity, content such as citrate synthase (CS) activity, mitochondrial DNA copy number, TOMM20, VDAC, and protein content for complex I-V of the oxidative phosphorylation (OXPHOS) system. Additionally, all participants underwent noninvasive assessments of mitochondrial capacity: PCr recovery postexercise (by 31 P-MRS), maximal aerobic capacity, and gross exercise efficiency by cycling exercise. From the invasive markers, Complex V protein content and CS activity showed the strongest concordance (Rc = 0.50 to 0.72) with ADP-stimulated coupled mitochondrial respiration, fueled by various substrates. Complex V protein content showed the strongest concordance (Rc = 0.72) with maximally uncoupled mitochondrial respiration. From the noninvasive markers, gross exercise efficiency, VO2max , and PCr recovery exhibited concordance values between 0.50 and 0.77 with ADP-stimulated coupled mitochondrial respiration. Gross exercise efficiency showed the strongest concordance with maximally uncoupled mitochondrial respiration (Rc = 0.67). From the invasive markers, Complex V protein content and CS activity are surrogates that best reflect skeletal muscle mitochondrial respiratory capacity. From the noninvasive markers, exercise efficiency and PCr recovery postexercise most closely reflect skeletal muscle mitochondrial respiratory capacity.

Hyperbaric oxygen rapidly improves tissue-specific insulin sensitivity and mitochondrial capacity in humans with type 2 diabetes: a randomised placebo-controlled crossover trial

Aims/hypothesisHyperbaric oxygen (HBO) therapy may improve hyperglycaemia in humans with type 2 diabetes, but underlying mechanisms are unclear. Our objective was to examine the glucometabolic effects of HBO on whole-body glucose disposal in humans with type 2 diabetes.MethodsIn a randomised placebo-controlled crossover trial located at the German Diabetes Center, 12 male individuals with type 2 diabetes (age 18–75 years, BMI <35 kg/m2, HbA1c 42–75 mmol/mol [6–9%]), randomly allocated by one person, underwent 2-h HBO, once with 100% (240 kPa; HBO) and once with 21% oxygen (240 kPa; control, CON). Insulin sensitivity was assessed by hyperinsulinaemic–euglycaemic clamps with d-[6,6-2H2]glucose, hepatic and skeletal muscle energy metabolism were assessed by 1H/31P-magnetic resonance spectroscopy, while high-resolution respirometry measured skeletal muscle and white adipose tissue (WAT) mitochondrial capacity. All participants and people assessing the outcomes were blinded.ResultsHBO decreased fasting blood glucose by 19% and increased whole-body, hepatic and WAT insulin sensitivity about one-third (p<0.05 vs CON). Upon HBO, hepatic γ-ATP concentrations doubled, mitochondrial respiratory control doubled in skeletal muscle and tripled in WAT (p<0.05 vs CON). HBO increased myocellular insulin-stimulated serine-473/threonine-308 phosphorylation of Akt but decreased basal inhibitory serine-1101 phosphorylation of IRS-1 and endoplasmic reticulum stress (p<0.05 vs CON).Conclusions/interpretationHBO-mediated improvement of insulin sensitivity likely results from decreased endoplasmic reticulum stress and increased mitochondrial capacity, possibly leading to low-dose reactive oxygen species-mediated mitohormesis in humans with type 2 diabetes.Trial registrationClinicalTrials.gov NCT04219215FundingGerman Federal Ministry of Health, German Federal Ministry of Education and Research, North-Rhine Westfalia Ministry of Culture and Science, European-Regional-Development-Fund, German-Research-Foundation (DFG), Schmutzler StiftungGraphical abstract

Aerobic training increases mitochondrial respiratory capacity in human skeletal muscle stem cells from sedentary individuals.

The impact of aerobic training on human skeletal muscle cell (HSkMC) mitochondrial metabolism is a significant research gap, critical to understanding the mechanisms by which exercise augments skeletal muscle metabolism. We therefore assessed mitochondrial content and capacity in fully differentiated CD56+ HSkMCs from lean active (LA) and sedentary individuals with obesity (OS) at baseline, as well as lean/overweight sedentary individuals (LOS) at baseline and following an 18-day aerobic training intervention. Participants had in vivo skeletal muscle PCr recovery rate by 31P-MRS (mitochondrial oxidative kinetics) and cardiorespiratory fitness (V̇o2max) assessed at baseline. Biopsies of the vastus lateralis were performed for the isolation of skeletal muscle stem cells. LOS individuals repeated all assessments posttraining. HSkMCs were evaluated for mitochondrial respiratory capacity by high-resolution respirometry. Data were normalized to two indices of mitochondrial content (CS activity and OXPHOS protein expression) and a marker of total cell count (quantity of DNA). LA individuals had significantly higher V̇o2max than OS and LOS-Pre training; however, no differences were observed in skeletal muscle mitochondrial capacity, nor in carbohydrate- or fatty acid-supported HSkMC respiratory capacity. Aerobic training robustly increased in vivo skeletal muscle mitochondrial capacity of LOS individuals, as well as carbohydrate-supported HSkMC respiratory capacity. Indices of mitochondrial content and total cell count were similar among the groups and did not change with aerobic training. Our findings demonstrate that bioenergetic changes induced with aerobic training in skeletal muscle in vivo are retained in HSkMCs in vitro without impacting mitochondrial content, suggesting that training improves intrinsic skeletal muscle mitochondrial capacity.

The Effect of Aerobic Exercise on the Oxidative Capacity of Skeletal Muscle Mitochondria in Mice with Impaired Glucose Tolerance.

Methods Male C57BL/6J mice were randomly divided into six different experimental groups (8 animals/group): (1) normal group (NOR), (2) normal control group (NC), (3) normal + exercise group (NE), (4) IGT group (IGT), (5) IGT control group (IC), and (6) IGT+ exercise group (IE).The exercise group received aerobic exercise for 8 weeks. After the intervention, a blood glucose meter was used to detect the level of glucose tolerance in the mouse's abdominal cavity; a biochemical kit was used to detect serum lipid metabolism indicators, malondialdehyde, and superoxide dismutase levels; the ELISA method was used to detect serum insulin and mouse gastrocnemius homogenate LDH, PDH, SDH, and CCO levels. Western blot method was used to detect the protein expression levels of NOX4, PGC-1α, and Mfn2 in the gastrocnemius muscle of mice. Results (1) Mice with high-fat diet for 30 weeks showed impaired glucose tolerance, insulin resistance, and lipid metabolism disorders. The level of LDH, PDH, SDH, and CCO in the gastrocnemius homogenate of mice was reduced. The expressions of NOX4 protein were significantly upregulated, while the expressions of PGC-1α and Mfn2 proteins were significantly downregulated. (2) 8-week aerobic exercise improved the disorders of glucose and lipid metabolism in IGT mice and increased homogenized LDH, PDH, SDH, and CCO levels, and the expressions of NOX4, PGC-1α, and Mfn2 proteins in the gastrocnemius muscle of mice were reversed. It is speculated that aerobic exercise can accelerate energy metabolism. Conclusion (1) C57BL/6 mice were fed high fat for 30 weeks and successfully constructed a mouse model of reduced diabetes; the mice with reduced diabetes have impaired glucose tolerance, insulin resistance, and lipid metabolism disorders; (2) 8 weeks of aerobic exercise improve glucose tolerance, reduce glucose tolerance in mice, reduce insulin resistance, improve lipid metabolism disorders, and reduce oxidative stress; (3) 8-week aerobic exercise reduces skeletal muscle NOX4 expression and increases glucose tolerance; reduces the expression of LDH, PDH, SDH, and CCO in mouse skeletal muscle; increases the expression level of mitochondrial fusion protein 2 and PGC-1α; improves glucose tolerance; reduces energy metabolism of mouse skeletal muscle; reduces oxidative stress; and reduces insulin resistance. It is speculated that aerobic exercise can accelerate energy metabolism. This process may involve two aspects: firstly, increase the expression level of oxidative metabolism enzymes and promote the tricarboxylic acid cycle; secondly, increase the expression of Mfn2 and accelerate mitochondria fission or fusion to regulate energy metabolism, thereby reducing oxidative stress and insulin resistance.

224-LB: Mitochondrial Capacity of Skeletal Muscle in Older Adults with Diabetes

Skeletal muscle mitochondrial capacity is reduced in both aging and diabetes. Whether diabetes further impairs mitochondrial capacity in older adults and how this relates to cardiorespiratory fitness (VO2peak) and glycemic control (HbA1c) remains uncertain. We examined muscle mitochondrial capacity of those with diabetes (self-reported, including Type 1 and 2) and without diabetes in the Study of Muscle Mobility and Aging (SOMMA) (273 men, 194 women; mean±SD age: 76.8±5.3 years) . Mitochondrial capacity was assessed by 31P magnetic resonance spectroscopy (ATPmax) and respirometry of vastus lateralis biopsy specimens. ATPmax (diabetes; 0.48 ± 0.1, nondiabetes; 0.54 ± 0.1 mM/sec, p=0.07) and fatty acid oxidation (FAO) supported OXPHOS (FAO: diabetes; 11.5 ± 4.5, nondiabetes; 12.5 ± 4.4 pmol/min/mg, p=0.01) were lower in those with diabetes compared to those without. VO2peak, assessed by a graded exercise test, was lower in those with diabetes (diabetes; 1478.8± 399.9, nondiabetes; 1495.5 ± 431.3 mL/min, p=0.01) . In all participants, ATPmax (r2=0.07, p=&amp;lt;0.01) and FAO (r2=0.04, p=0.14) explained a small proportion of variance in HbA1c. VO2peak significantly related to mitochondrial capacity (ATPmax: r2=0.66, p=&amp;lt;0.01, FAO: r2=0.64, p&amp;lt;0.01) , but the relationships do not vary by diabetes status. In summary, older adults with diabetes have marginally lower fitness and mitochondrial capacity. Mitochondrial capacity is related to both glycemic control and cardiorespiratory fitness regardless of diabetes status. Disclosure S. V. Ramos: None. P. M. Coen: None. G. Distefano: None. P. Cawthon: None. S. M. Marshall: None. P. Kramer: None. A. Molina: None. M. J. Jurczak: None. F. Toledo: None. B. H. Goodpaster: None. Funding Study of Muscle, Mobility and Aging (SOMMA) RO1 AG059416

Skeletal Muscle Mitochondrial Capacity in Patients with Statin‐Associated Muscle Symptoms (SAMS)

BackgroundMitochondrial dysfunction may be a key mechanism underlying SAMS. Near infrared spectroscopy (NIRS) is a non‐invasive technique of assessing tissue oxygenation and mitochondrial function and has never been evaluated as a diagnostic tool for SAMS.Methods and ResultsSAMS was confirmed using an 8‐week randomized, double‐blind crossover trial of simvastatin 20 mg / day and placebo separated by a 4‐week washout phase in 39 adults with self‐reported SAMS (Age: 65 ± 6 y; 25 female, 14 male). Tissue oxygenation was measured using NIRS during a handgrip exercise protocol at increasing intensities of maximal voluntary contraction (MVC) before and after statin or placebo treatment. Patients were considered to have SAMS only if they had muscle symptoms on simvastatin and not placebo. Similar to our previous trials, 17/39 (44%) patients were confirmed as having SAMS. There was a significant interaction (p &lt; 0.01) between placebo vs. statin, pre‐ vs. post‐treatment, and SAMS/not SAMS. Tissue oxygenation for average (20‐40%) MVC was significantly lower with placebo when symptoms were present compared with when participants did not have symptoms on a statin (15.66% ± 28.21% vs. 2.35% ± 19.13%, respectively) (p = 0.02). Neither group changed tissue oxygenation with statin therapy (both p &gt; 0.22; SAMS: 28.0 ± 17.6% vs. 22.6 ± 11.9%; No SAMS: 30.5 ± 18.8% vs. 31.0 ± 16.4%, respectively), but the decrease was numerically greater in SAMS patients. There were no significant differences in MVC change score based on symptoms however, patients with SAMS experienced more variable MVC change score responses compared with patients who did not have symptoms.ConclusionsThese results indicate that NIRS may be effective for differentiating patients with SAMS from those with non‐specific muscle pain, however this must be verified with a larger sample.

Impact of aging and exercise on skeletal muscle mitochondrial capacity, energy metabolism, and physical function

The relationship between the age-associated decline in mitochondrial function and its effect on skeletal muscle physiology and function remain unclear. In the current study, we examined to what extent physical activity contributes to the decline in mitochondrial function and muscle health during aging and compared mitochondrial function in young and older adults, with similar habitual physical activity levels. We also studied exercise-trained older adults and physically impaired older adults. Aging was associated with a decline in mitochondrial capacity, exercise capacity and efficiency, gait stability, muscle function, and insulin sensitivity, even when maintaining an adequate daily physical activity level. Our data also suggest that a further increase in physical activity level, achieved through regular exercise training, can largely negate the effects of aging. Finally, mitochondrial capacity correlated with exercise efficiency and insulin sensitivity. Together, our data support a link between mitochondrial function and age-associated deterioration of skeletal muscle.

Preserved skeletal muscle oxidative capacity in older adults despite decreased cardiorespiratory fitness with ageing.

Healthy older adults exhibit lower cardiorespiratory fitness ( ) than young in the absence of any age-related difference in skeletal muscle mitochondrial capacity, suggesting central haemodynamics plays a larger role in age-related declines in . Total physical activity did not differ by age, but moderate-to-vigorous physical activity was lower in older compared to young adults. Moderate-to-vigorous physical activity is associated with and muscle oxidative capacity, but physical inactivity cannot entirely explain the age-related reduction in . Declining fitness ( ) is a hallmark of ageing and believed to arise from decreased oxygen delivery and reduced muscle oxidative capacity. Physical activity is a modifiable lifestyle factor that is critical when evaluating the effects of age on parameters of fitness and energy metabolism. The objective was to evaluate the effects of age and sex on , muscle mitochondrial physiology, and physical activity in young and older adults. An additional objective was to assess the contribution of skeletal muscle oxidative capacity to age-related reductions in and determine if age-related variation in and muscle oxidative capacity could be explained on the basis of physical activity levels. In 23 young and 52 older men and women measurements were made of , mitochondrial physiology in permeabilized muscle fibres, and free-living physical activity by accelerometry. Regression analyses were used to evaluate associations between age and , mitochondrial function, and physical activity. Significant age-related reductions were observed for (P<0.001), but not muscle mitochondrial capacity. Total daily step counts did not decrease with age, but older adults showed lower moderate-to-vigorous physical activity, which was associated with (R2 =0.323, P<0.001) and muscle oxidative capacity (R2 =0.086, P=0.011). After adjusting for sex and physical activity, age was negatively associated with but not muscle oxidative capacity. Healthy older adults exhibit lower but preserved mitochondrial capacity compared to young. Physical activity, particularly moderate-to-vigorous, is a key factor in observed age-related changes in fitness and muscle oxidative capacity, but cannot entirely explain the age-related reduction in .

A High Activity Level Is Required for Augmented Muscle Capillarization in Older Women.

This study aimed to evaluate the influence of lifelong regular physical activity on skeletal muscle capillarization in women. Postmenopausal women, 61±4 yr old, were divided according to self-reported physical activity level over the past 20 yrs: sedentary (SED; n = 14), moderately active (MOD; n = 12), and very active (VERY; n = 15). Leg blood flow (LBF) was determined by ultrasound Doppler, and blood samples were drawn from the femoral artery and vein for calculation of leg oxygen uptake (LVO2) at rest and during one-legged knee extensor exercise. A skeletal muscle biopsy was obtained from the vastus lateralis and analyzed for capillarization and vascular endothelial growth factor (VEGF) and mitochondrial OXPHOS proteins. Platelets were isolated from venous blood and analyzed for VEGF content and effect on endothelial cell proliferation. The exercise-induced rise in LBF and LVO2 was faster (P = 0.008) in VERY compared with SED and MOD. Steady-state LBF and LVO2 were lower (P < 0.04) in MOD and VERY compared with SED. Capillary-fiber ratio and capillary density were greater (P < 0.03) in VERY (1.65 ± 0.48 and 409.3 ± 57.5) compared with MOD (1.30 ± 0.19 and 365.0 ± 40.2) and SED (1.30 ± 0.30 and 356.2 ± 66.3). Skeletal muscle VEGF and OXPHOS complexes I, II, and V were ~1.6-fold and ~1.25-fold (P < 0.01) higher, respectively, in VERY compared with SED. Platelets from all groups induced an approximately nine-fold (P < 0.001) increase in endothelial cell proliferation. A very active lifestyle is associated with superior skeletal muscle exercise hemodynamics and greater potential for oxygen extraction concurrent with a higher skeletal muscle capillarization and mitochondrial capacity.

25 Race training improves skeletal muscle mitochondrial volume density, function, and capacity in Thoroughbreds

Racehorses are among the most elite athletes in the equine industry. However, little is known about mitochondrial adaptations to race training. To test the hypothesis that race training increases mitochondrial biogenesis and capacity, gluteus medius samples were collected from Thoroughbred horses (11 geldings, 2 mares) before and after 14 wk of early race training. Horses entered wk 0 at varying levels of fitness, and therefore, were grouped accordingly: 1) galloping on track (n = 5; mean ± SD V200 = 10.6 ± 0.9 m/s); 2) jogging and cantering on track (n = 5; V200 = 9.8 ± 0.6 m/s); and 3) breezed one furlong at sale (n = 3; unknown V200). All horses increased in workload from a 6-furlong canter (7–9 m/s) to a 12-furlong slow gallop (9–11 m/s). Once horses reached proper fitness, fast work was added weekly in the form of a one-furlong fast gallop (11–13 m/s) or 2–3 furlongs breeze (13–15 m/s). There was no difference in fitness level between groups at wk 14 (V200 = 10.9 ± 0.7 m/s; P = 0.6). Muscle mitochondrial capacities were determined by high resolution respirometry, and mitochondrial enzyme activities were determined by colorimetry. Data were analyzed using linear models in SAS v9.4 with repeated measures. Main effects included time, wk 0 fitness level (trt) and the time × trt interaction; horse(trt) was as a random effect. Mitochondrial volume density (citrate synthase activity; CS) and integrative (per mg protein) mitochondrial function (cytochrome c oxidase activity; CCO) increased ( P ≤ 0.05), while intrinsic CCO (relative to CS activity) decreased ( P = 0.005) from wk 0 to 14. Integrative (per mg tissue) oxidative phosphorylation (P) capacities with complex I (P CI ) and complex II (P CI+II ), as well as maximum electron transport capacity (E CI+II ) increased from wk 0 to 14 ( P ≤ 0.05) but none were affected by wk 0 fitness level. Incoming fitness level only affected intrinsic mitochondrial respiration. Horses with intermediate fitness at wk 0 (group 2) had greater intrinsic P CI , P CI+II , E CI+II , and electron transport capacity with CII only (E CII ) at wk 0 than group 1 or 3 ( P ≤ 0.007) but these measures were not different between groups at wk 14. Intrinsic E CII decreased in all horses from wk 0 to 14 ( P = 0.01) but no other intrinsic mitochondrial capacity measure was affected overall by time. The contribution of P CI to total E CI+II tended to decrease ( P = 0.07) and of E CII to total E CI+II decreased ( P < 0.0001) from wk 0 to 14. This is the first report of mitochondrial adaptations to early race training, which may assist in developing precision management programs to enhance performance and well-being in elite racehorses.

Mitochondrial Capacity and Muscle Endurance in Individuals with Parkinson’s Disease

Introduction: Parkinson's disease (PD) is associated with loss of motor control and difficulty exercising. This study measured skeletal muscle mitochondrial capacity and endurance in individuals with and without PD using novel non-invasive methods. We hypothesized that individuals with PD will have decreased mitochondrial capacity, reduced oxygen recovery, and decreased endurance compared to controls. Methods: Eight participants with PD and nine healthy controls were tested. Mitochondrial capacity was measured as the rate of recovery of muscle metabolism after electrical stimulation using near-infrared spectroscopy (NIRS) and repeated short arterial occlusions. Oxygen recovery was measured as the half time of recovery of oxygen levels after 5 minutes of ischemia. Muscle endurance was determined from changes in twitch contraction acceleration during electrical stimulation at 2, 4, and 6 Hz. Results: Mitochondrial capacity was lower in individuals with PD compared to controls (1.5±0.1min-1 vs. 1.7±0.1min-1, p=0.02). Individuals with PD had slower oxygen recovery after ischemia compared to controls (8.9±2.3s vs. 5.4±0.8s, p=0.01). Endurance was not different between groups at 6 Hz (PD vs controls: 58±23% vs. 69±16%, p=0.34). The effect sizes for mitochondrial capacity and oxygen recovery were large (Cohen's d &gt;0.8). The Cohen's d for endurance was 1.11. Conclusion: Individuals with PD had slight impairments in mitochondrial capacity and blood flow but did not have reduced muscle endurance. While our study suggests that muscle metabolic dysfunction may play a minor role in exercise intolerance in people with PD, it demonstrates the use of noninvasive technologies to evaluate muscle function in people with neurological disorders.

108 President Oral Presentation Pick: Select skeletal muscle mitochondrial measures in Thoroughbred weanlings are related to race earnings and sire

Abstract Quantifiable, cellular differences of individuals are not widely used in breeding and training decisions in the equine industry. Our objective was to determine if mitochondrial parameters in weanling Thoroughbreds were related to sire or lifetime race earnings. We hypothesized that weanling skeletal muscle mitochondrial capacity would be positively correlated with race earnings. Gluteus medius muscle samples were collected from racing-bred Thoroughbred weanlings (n = 139; mean ± SD; 6.0 ± 0.4 mo) from 40 different sires over 3 years at 5 different farms and evaluated for mitochondrial volume density (citrate synthase activity; CS) and function (cytochrome c oxidase activity) by colorimetry, and oxidative (P) and electron transport system (E) capacities by high resolution respirometry; two- and three-year-old race earnings were available for a subset of 13 horses. Data were analyzed using mixed linear models with sire, sex, year of collection, and farm as fixed effects. Correlations between lifetime race earnings and mitochondrial measures were determined using Pearson Correlation Statistics. A main effect of sire was observed for weanling intrinsic (relative to CS activity) P with complex I substrates (P = 0.04) and the contribution of mitochondrial leak to O2 consumption (FCRL; P = 0.04). A trend was observed for the effect of sire on intrinsic mitochondrial leak (P = 0.09). Race earnings were positively correlated with integrated (relative to mg protein) leak (r = 0.7684, P = 0.009) and FCRL (r = 0.7035; P = 0.02). A trend for a negative correlation between E with complex II substrates and race earnings was also observed (r = -0.4775, P = 0.09), No other measures were influenced by sire, nor were correlated with race earnings. Our previous work has indicated a preferential use of complex I in breeds bred for short-duration racing (Thoroughbreds) compared to distance racing breeds (Standardbreds) that relied more heavily on complex II. Mitochondrial measures in weanlings may be utilized to inform future breeding and training decisions in horses.

An improvement in skeletal muscle mitochondrial capacity with short-term aerobic training is associated with changes in Tribbles 1 expression.

Exercise training and physical activity are known to be associated with high mitochondrial content and oxidative capacity in skeletal muscle. Metabolic diseases including obesity and insulin resistance are associated with low mitochondrial capacity in skeletal muscle. Certain transcriptional factors such as PGC‐1α are known to mediate the exercise response; however, the precise molecular mechanisms involved in the adaptation to exercise are not completely understood. We performed multiple measurements of mitochondrial capacity both in vivo and ex vivo in lean or overweight individuals before and after an 18‐day aerobic exercise training regimen. These results were compared to lean, active individuals. Aerobic training in these individuals resulted in a marked increase in mitochondrial oxidative respiratory capacity without an appreciable increase in mitochondrial content. These adaptations were associated with robust transcriptome changes. This work also identifies the Tribbles pseudokinase 1, TRIB1, as a potential mediator of the exercise response in human skeletal muscle.

Nicotinamide riboside does not alter mitochondrial respiration, content or morphology in skeletal muscle from obese and insulin-resistant men.

This is the first long-term human clinical trial to report on effects of nicotinamide riboside (NR) on skeletal muscle mitochondrial function, content and morphology. NR supplementation decreases nicotinamide phosphoribosyltransferase (NAMPT) protein abundance in skeletal muscle. NR supplementation does not affect NAD metabolite concentrations in skeletal muscle. Respiration, distribution and quantity of muscle mitochondria are unaffected by NR. NAMPT in skeletal muscle correlates positively with oxidative phosphorylation Complex I, sirtuin 3 and succinate dehydrogenase. Preclinical evidence suggests that the nicotinamide adenine dinucleotide (NAD+ ) precursor nicotinamide riboside (NR) boosts NAD+ levels and improves diseases associated with mitochondrial dysfunction. We aimed to determine if dietary NR supplementation in middle-aged, obese, insulin-resistant men affects mitochondrial respiration, content and morphology in skeletal muscle. In a randomized, placebo-controlled clinical trial, 40 participants received 1000mg NR or placebo twice daily for 12weeks. Skeletal muscle biopsies were collected before and after the intervention. Mitochondrial respiratory capacity was determined by high-resolution respirometry on single muscle fibres. Protein abundance and mRNA expression were measured by Western blot and quantitative PCR analyses, respectively, and in a subset of the participants (placebo n=8; NR n=8) we quantified mitochondrial fractional area and mitochondrial morphology by laser scanning confocal microscopy. Protein levels of nicotinamide phosphoribosyltransferase (NAMPT), an essential NAD+ biosynthetic enzyme in skeletal muscle, decreased by 14% with NR. However, steady-state NAD+ levels as well as gene expression and protein abundance of other NAD+ biosynthetic enzymes remained unchanged. Neither respiratory capacity of skeletal muscle mitochondria nor abundance of mitochondrial associated proteins were affected by NR. Moreover, no changes in mitochondrial fractional area or network morphology were observed. Our data do not support the hypothesis that dietary NR supplementation has significant impact on skeletal muscle mitochondria in obese and insulin-resistant men. Future studies on the effects of NR on human skeletal muscle may include both sexes and potentially provide comparisons between young and older people.

Lipid accumulation and mitochondrial abnormalities are associated with fiber atrophy in the skeletal muscle of rats with collagen-induced arthritis

Rheumatoid arthritis (RA) has a negative impact on muscle mass, and reduces patient's mobility and autonomy. Furthermore, RA is associated with metabolic comorbidities, notably in lipid homeostasis by unknown mechanisms. To understand the links between the loss in muscle mass and the metabolic abnormalities, arthritis was induced in male Sprague Dawley rats (n = 11) using the collagen-induced arthritis model. Rats immunized with bovine type II collagen were compared to a control group of animals (n = 11) injected with acetic acid and complete Freund's adjuvant. The clinical severity of the ensuing arthritis was evaluated weekly by a semi-quantitative score. Skeletal muscles from the hind limb were used for the histological analysis and exploration of mitochondrial activity, lipid accumulation, metabolism and regenerative capacities. A significant atrophy in tibialis anterior muscle fibers was observed in the arthritic rats despite a non-significant decrease in the weight of the muscles. Despite moderate inflammation, accumulation of triglycerides (P < 0.05), reduced mitochondrial DNA copy number (P < 0.05) and non-significant dysfunction in mitochondrial cytochrome c oxidase activity were found in the gastrocnemius muscle. Concomitantly, our results suggested an activation of the muscle specific E3 ubiquitin ligases MuRF-1 and MAFbx. Finally, the adipose tissue from the arthritic rats exhibited decreased PPARγ mRNA suggesting reduced adipogenic capacities. In conclusion, the reduced adipose tissue adipogenic capacity and skeletal muscle mitochondrial capacity are probably involved in the activation of protein catabolism, inhibition of myogenesis, accumulation of lipids and fiber atrophy in the skeletal muscle during RA.