No description available.

The purpose of this study was to evaluate muscle oxygenation, cardiorespiratory, and blood lactate responses to an acute bout of a high-intensity interval training (HIIT) bodyweight squat protocol (HIIT-squats) in comparison to (continuous) moderate intensity cycling exercise (MOD). On separate days, within a two week period, 15 recreationally active males (28 (4.6) years) performed: 1) incremental test to exhaustion on a cycle ergometer, 2) 30-minutes of moderate intensity cycling (MOD; 65% VO2max), and 3) HIIT-squats consisting of eight x 20 seconds of bodyweight squats performed at maximal cadence with 10-s rest intervals. During each exercise condition, oxygen consumption (VO2) and heart rate were monitored continuously, and muscle oxygenation (tissue saturation index, TSI) at the left vastus lateralis muscle was measured for 2 minutes pre-, throughout, and for 5 minutes post-exercise using Near-Infrared Spectroscopy (NIRS; Portalite, Artinis Medical Systems, Netherlands). Blood lactate was measured at pre- and one, three, and five minutes post-exercise. Mean and peak changes in TSI were similar in both HIIT-squats (mean = -14.6 (5.3)%, peak = -19.7 (5.2)%; p > 0.05) and MOD (mean = -13.2 (5.6)%, peak = -18.2 (7.6)%; p > 0.05), with peak changes in TSI occurring significantly faster in HIIT-squats (71.2 (95.2) seconds (s) after onset of exercise) than in MOD (1452.9 (647.8)s; p < 0.05). The half time of TSI recovery following HIIT-squats (T1/2TSI = 25 (7.9)s) was not significantly different post-MOD (25 (9.6)s). Mean VO2 during HIIT-squats (31.48 (4.58) ml.kg-1.min-1) was similar to MOD (33.76 (5.71) ml.kg-1.min-1), however minute ventilation (VE), respiratory exchange ratio (RER) and all post-exercise blood lactate concentrations were significantly higher in HIIT-squats compared to MOD (p < 0.05). Despite the different durations of HIIT-squats and MOD, mean and peak changes in aerobic metabolism during and after exercise were similar. Results provide evidence of both aerobic and anaerobic contributions to energy metabolism in response to HIIT-squats, and highlight possible mechanisms for the commonly reported improvements in aerobic power following chronic HIIT. / Graduate / akates@uvic.ca

The Thoroughbred racehorse is capable of maintaining speeds of approximately 17 m/s for distances of over a mile. This represents an average speed and the Thoroughbred can reach speeds in excess of 20 m/s over short distances. The present series of studies was undertaken to investigate further the metabolic response to high-intensity exercise in the Thoroughbred racehorse. Unlike man, high-intensity exercise in the horse results in an increase in packed cell volume. This in turn causes acute changes in the colligative properties of blood and plasma. The changes in these properties were investigated and the effect on calculation and distribution of metabolites in blood and muscle was determined. The horse has a high capacity for lactate production compared to man and existing methodology for the assessment of muscle buffering capacity in biopsy samples was further developed and investigated. The horse was found to have a significantly higher muscle buffering capacity compared with man and it was calculated that this could be explained wholly on the basis of the higher muscle carnosine content in the horse. The metabolic response to high-intensity exercise was investigated using several exercise models, including single and multiple field gallops and treadmill exercise. A consistent finding was that high-intensity exercise in the Thoroughbred racehorse was nearly always accompanied by a reduction in muscle ATP content. The nature of the ATP decrease was further investigated using a treadmill exercise model. Muscle ATP decrease was found to occur at a particular exercise intensity rather than show a gradual decrease with increasing intensity. The intensity at which muscle ATP content began to decline significantly varied between individual horses, but in each case appeared to coincide with muscle lactate contents of approximately 70 mmol/kg dry muscle. The significance of the decline in ATP is discussed.

The purpose of this thesis was to study the effects of a pre-exercise carbohydrate meal on metabolism, endurance capacity and performance during prolonged running when carbohydrate was, or was not consumed during exercise. The first study (Chapter 4) examined the effects on endurance running capacity of ingesting a carbohydrate-electrolyte solution during treadmill exercise to fatigue at 70% V02 max after subjects (10 males) had undergone an overnight fast (P+C), or when fed with a 2.5 g. kg-1 BW carbohydrate meal 3 hours before exercise (M+C). Exercise time to exhaustion was longer in the M+C (147.4 ± 9.6 min) and P+C (125.1 ± 7.0 min) trials compared with the control condition (P+P: 115.1 ± 17.6 min) (p< 0.01 and p< 0.05 respectively). Also, exercise time was longer in the M+C compared with the P+C trial (p< 0.01). The improvement in endurance capacity in the M+C trial occurred despite a higher carbohydrate oxidation rate during the first hour of exercise. The second study (Chapter 5) examined whether a pre-exercise carbohydrate meal (M+W) can improve endurance capacity, and further examined if the combination of a pre-exercise meal together with the ingestion of a carbohydrate-electrolyte solution during exercise (M+C) would be superior to the carbohydrate meal (M+W) alone. Ten males volunteered in this study. Although the consumption of the meal increased carbohydrate oxidation during the first hour of exercise, exercise time to fatigue at 70% V02 max was longer in the M+C (125.1 + 5.3 min) and M+W (111.9 + 5.6 min) trials compared with the control trial (P+W : 102.9 ± 7.9 min) (p< 0.01 and p< 0.05 respectively). Also, exercise time was longer (p< 0.05) in the M+C compared with the M+W trial. The third study (Chapter 6) investigated whether the high carbohydrate meal can influence muscle glycogen levels. Eight male subjects participated in the study. Three hours after the ingestion of the 2.5 g. kg-1 BW carbohydrate meal, muscle glycogen concentration was 10.6% higher (p< 0.05) in the vastus lateralis muscle (347.3 + 31.3 mmol. kg dw-1) compared with the muscle glycogen concentration before feeding (314 ± 33.9 mmol. kg dw-1). The fourth study (Chapter 7) examined the influence of ingesting a carbohydrate-electrolyte drink (M+C) on the muscle glycogen utilisation during 60 min running at 70% V02 max in subjects (8 males) who had consumed a carbohydrate meal 3 hours before exercise (M+W). Muscle glycogen concentrations were not different before (M+C : 321.9 ± 27.2 vs M+W : 338.8 ± 32.8 mmol. kg dw-1), as well as after exercise (M+C : 225.8 ± 26.7 vs M+W: 261 + 40.5 mmol. kg dw-1) between the two experimental trials. Neither was there any difference in the rate of muscle glycogen utilisation (M+C : 96.1 ± 22.1 vs M+W: 77.9 ± 11.7 mmol. kg dwl. h-1). The aim of the last study (Chapter 8) was to investigate whether, after an overnight fast, the ingestion of a carbohydrate-electrolyte solution during a 30 km self-paced treadmill run (C) would be as effective as the consumption of a carbohydrate meal (M) (2.0 g. kg-1 BW carbohydrate) 4 hours before exercise. Ten males volunteered for this study. The overall performance times in the M and C trials were identical (M: 121.8 ± 3.6 min vs C: 121.7 ± 4.1 min). No differences were found between the two trials in running speeds over each successive 5 km, or even when running speed was analysed every kilometre. Also, no reduction in the self-selected speeds of subjects was observed towards the end of the 30 km run in both conditions. The ingestion of a carbohydrate meal, providing 2.5 g. kg-1 BW carbohydrate, 3 hours before exercise increases muscle glycogen concentration and improves endurance running capacity, despite an elevated carbohydrate oxidation rate during the first hour of exercise. It seems that the amount of carbohydrate given before exercise compensates for the greater carbohydrate used. Furthermore, the combination of both a pre-exercise carbohydrate meal and a carbohydrate-electrolyte solution ingested during exercise further improves endurance capacity.

The process of fatigue during 30 s of high-intensity exercise results in rapid and substantial mechanical, electrical and biochemical changes in muscle fibres. This thesis describes a series of experiments, performed upon a friction-loaded cycle ergometer, which investigated whether the magnitude of the mechanical and biochemical changes is affected by changes in average pedalling rate. The ability to generate peak power in a subsequent sprint of 6 s duration was used to assess the magnitude of the mechanical changes. Changes in the concentrations of blood and muscle metabolites pre- and post-exercise permitted some of the biochemical changes to be measured.

Continual intake of high-fat foods, coupled with limited physical activity, can lead to metabolic inflexibility. Eventually, this may lead to significant health issues such as obesity, insulin resistance, cardiovascular disease, and other chronic diseases. Metabolic flexibility of human skeletal muscles is influenced by changes to mitochondrial, nuclear, and cytosolic proteins, in part as a result of posttranslational modifications (PTMs). O-linked B-D-N-acetylglucosamine, known as O-GlcNAc, has recently been identified as an important posttranslational modification that responds to nutrient sensing and cellular stress. Unlike other PTMs, O-GlcNAc has only two cycling enzymes. Because of its novelty, little research has been performed on the role of O-GlcNAc in human skeletal muscle and metabolic flexibility. The purpose of the current study was to establish the effects of a 5-day high-fat diet on skeletal muscle O-GlcNAcylation. In the proposed study, 13 non-obese, sedentary, college-aged males consumed a controlled diet for two weeks followed by a high-fat diet composed of 55% fat, 30% carbohydrate, and 15% protein. Muscle biopsies were taken from the vastus lateralis both fasted and four hours after a high-fat meal, following both the control diet and the high-fat diet. Western blot analysis was used to assess global O-GlcNAc and protein concentrations of O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA) in whole-homogenates and isolated mitochondria from skeletal muscle. Results were analyzed using independent, two-tailed t-tests and 2-way ANOVA analysis with repeated measures and Bonferroni corrections; a p-value was set to α less than or equal to 0.05. It was found that O-GlcNAc and OGT levels remained stable, although fasting levels of OGA significantly decreased after the 5-day high-fat diet. It is possible that healthy individuals are capable of maintaining normal levels of O-GlcNAc and its cycling enzymes, but there is still more to learn about O-GlcNAc and its role in metabolic flexibility. / Master of Science

Alzheimer’s disease (AD), the most common form of dementia in the elderly, is a global issue affecting about 24 million individuals. Because AD is a systemic pathology, dementia is not the only leading factor contributing to loss of independence in AD patients. AD may also impair skeletal muscle metabolism and function. Creatine (CR) supplementation may enhance skeletal muscle hypertrophy/mass and function in sarcopenia and muscular dystrophies, but has yet to be studied in AD. This study examined the effect of oral CR on muscle metabolism in a triple-transgenic (3xTg) AD mouse model. Twenty-four, 3×Tg AD mice (~8 month-old) were randomly assigned to control (CON) or CR (3% w/w) diet. Bodyweights and feed intakes were measured throughout the 8-week study. Lower limb (quadriceps muscle; QM and gastrocnemius; GM) and upper limb muscles (triceps; TM) were collected to analyze levels of CR, total protein, DNA, RNA, amino acids (AA), adenosine triphosphate (ATP), adenosine diphosphate (ADP), total and phosphorylated p70 ribosomal S6 kinase (p70S6K). Data (mean ± SEM) were assessed by analysis of variance (ANOVA) and Fisher’s least significant difference (LSD) post hoc test. In comparison to the CON group, CR supplementation increased CR content in both GM (p=0.002) and QM (p=0.037), with higher (p=0.032) ATP/ADP ratio in CR in comparison with CON in QM. A higher protein concentration (p<0.0001) was notable in GM of CR supplemented group vs. CON. Total branched-chain AA levels in QM increased 2-fold (p< 0.0001) in CR groups. Additionally, CR resulted in a higher (p<0.05) protein/DNA ratio; an index of muscle cell size, in both QM and GM for CR groups. The index of cell capacity for protein synthesis (RNA/DNA ratio) in GM was also higher (p=0.001) in CR groups. However, phosphorylation (activation) level of p70S6K, an integral component in protein synthesis signalling pathway, did not show any significant differences in female (p=0.161) and male (p=0.292) CR supplemented groups compared with CON. To conclude, CR supplementation is capable of inducing muscle hypertrophy/growth parameters in the 3×Tg AD mouse model, thereby enhancing protein synthesis capacity in skeletal muscles, thus possibly promoting muscle function in AD. / May 2016

No description available.

Hypoxia is a major stressor at high altitudes that limits tissue oxygen availability. High altitude environments are also extremely cold which increases thermogenic demand. Small mammals living at high altitude face the competing energetic challenge of maintaining thermogenesis in a hypoxic environment that can impair aerobic ATP supply. It has been suggested that hypoxia-induced impairments in ATP synthesis capacity and cold-induced increases in thermogenic demand could be counteracted by an increase in tissue oxidative capacity and/or fuel selection. As the organelle that consumes oxygen to produce ATP, changes in mitochondrial physiology can help offset physiological impairments at high altitudes. We explored this hypothesis in North American deer mice (Peromyscus maniculatus), from populations native to high and low altitude. We compared mitochondrial volume densities, intracellular distribution, respiratory capacities, enzyme activities of the mitochondrial complexes, capillarity, and fibre-type distribution in skeletal and cardiac muscles. To examine potential changes to mitochondrial physiology at high altitudes deer mice (P. maniculatus) were acclimated to: warm (25°C) normoxia; warm hypoxia (simulated altitude of 4300m); cold (5°C) normoxia; and cold + hypoxia. In skeletal muscle, highlanders had higher mitochondrial volume densities than lowlanders, entirely due to an increased abundance of mitochondria in a subsarcolemmal location next to capillaries. Mitochondria from highland mice also had higher mitochondrial respiratory capacities and cytochrome c oxidase activity in control conditions, but these values converged after hypoxia acclimation. Cold acclimation restored pyruvate and fatty acid respiratory capacity to control levels in highland mice, which also showed an increase in mitochondrial uncoupling. Cold increased respiratory capacities in lowland mice. Acclimation to cold+hypoxia did not change mitochondrial physiology beyond cold alone and appeared to counteract the effects of hypoxia on highland mice. In cardiac muscle highland mice had higher respiratory capacities, but after hypoxia acclimation lowland mice significantly increased respiratory capacities. In response to hypoxia, highland mice increased the relative capacity to oxidize carbohydrates compared to fatty acids. Our results suggest that both highland ancestry and plasticity affect mitochondrial physiology, and likely contributes to performance at high altitudes. / Thesis / Doctor of Philosophy (PhD)

Skeletal muscle can perceive cellular energy status and substrate availability and demonstrates remarkable plasticity in response to environmental changes. Nonetheless, how skeletal muscle and its resident stem cells (satellite cells; SCs) sense and respond to nutrient flux remains largely undefined. The dynamic post-translational modification O-GlcNAcylation has been shown to serve as a cellular nutrient sensor in a wide range of cells and tissues, yet its role in skeletal muscle and SCs remains unexplored. Here, we ablated skeletal muscle O-GlcNAc transferase (OGT), and thus O-GlcNAcylation, and found the knockout mice exhibited enhanced glucose uptake, insulin sensitivity, and resistance to high-fat diet induced obesity. Additionally, mKO mice had a 3-fold increase in circulating levels of interleukin-15 (IL-15), a potent anti-obesity cytokine, potentially through epigenetic regulation of Il15 by OGT. To further investigate if there was a causal relationship between OGT ablation and the lean phenotype, we generated muscle specific OGT and interleukin-15 receptor alpha (IL-15ra) double knockout mice (mDKO). As a result, mDKO mice had blunted IL-15 secretion and minimal protection against HFD-induced obesity. Together, these data indicate the skeletal muscle OGT-IL15 axis plays an essential role in the maintenance of skeletal muscle and whole-body metabolic homeostasis. As satellite cells (SCs) play an indispensable role in postnatal muscle growth and adult regenerative myogenesis, we investigated the role of O-GlcNAcylation in SC function. To this end, we conditionally ablated OGT in SCs (cKO) and found cKO mice had impaired SC proliferation, in vivo cycling properties, population stability, metabolic regulation, and adult regenerative myogenesis. Together these findings show that SCs require O-GlcNAcylation, presumably to gauge nutritional signals, for proper function and metabolic homeostasis. Another critical yet often neglected player in myogenesis are mitochondria. Traditionally depicted as a power plant in cells, mitochondria are critical for numerous nonconventional, energy-independent cellular process. To investigate the role of both mitochondrial energy production and alternative mitochondrial functions in myogenic regulation, we ablated ATP synthase subunit beta (ATP5b) and ubiquinol-cytochrome c reductase (UQCRFS1) in C2C12 myoblasts to disrupt mitochondrial ATP production and mitochondrial membrane potential, respectively. Ablation of UQCRFS1, but not ATP5b, impaired myoblast proliferation, although lack of either gene compromised myoblast fusion. Interestingly, addition of the potent myogenic stimulator IGF-1 rescued ATP5b fusion but could not override UQCRFS1 knockout effects on proliferation or differentiation. These data demonstrate mitochondrial ATP production is not the "metabolic switch" that governs myogenic progression but rather an alternative mitochondrial function. In summary, skeletal muscle and their resident stem cell population (SCs) both use O-GlcNAcylation, feasibly to sense and respond to nutritional cues, for the maintenance of metabolic homeostasis and normal physiology. A deeper understand of both muscle and SC metabolic regulation may provide therapeutic targets to improve global metabolism and muscle growth. / Doctor of Philosophy / Skeletal muscle is responsible for approximately 20% of basal energy expenditure and 70-90% of insulin-mediated glucose disposal, and as such changes in skeletal muscle metabolism and insulin sensitivity have profound impacts on whole body metabolism. Skeletal muscle is a plastic tissue that can perceive nutrient availability, which permits metabolic adaptations to environmental changes. Deletion of the nutrient sensing pathway O-GlcNAcylation in skeletal muscle (mKO) protected mice from high-fat diet induced obesity and ameliorates whole-body insulin sensitivity. Skeletal muscle can secrete myokines to elicit endocrine effects on other tissues in the body, and as such, we proposed perturbation of this nutrient sensing pathway in skeletal muscle alters myokine secretion to elicit responses in other metabolically active tissues to support its energy requirements. Indeed, circulating levels of interleukin-15, a potent anti-obesity myokine, increased 3-fold in mKO mice. To determine the contribution of IL-15 to the mKO phenotype, we used a genetic approach to blunt IL-15 secretion from skeletal muscle (mDKO), which partially negated the lean mKO phenotype. Our findings show the ability of skeletal muscle to "sense" changes in nutrients through O-GlcNAcylation is necessary for proper muscle and whole-body metabolism. Moreover, this nutrient sensing mechanism is also important for proper muscle stem cell function, also known as satellite cells (SCs). Loss of O-GlcNAcylation in SCs impairs their ability to regenerate muscle after injury, which can be attributed to a reduced capacity to proliferate and an inability to maintain a healthy SC population. Interestingly, SCs lacking O-GlcNAcylation have a greater mitochondrial content. Using a myoblast cell line, we investigated the contribution of mitochondria to myogenesis, the formation of muscle, and found mitochondrial energy production is dispensable in the myogenic process. Our studies show skeletal muscle and SCs rely on highly integrated signaling cascades that sense and respond to intrinsic metabolic changes and extrinsic nutritional cues to function properly.