Biochemical evaluation of mitochondrial respiratory chain enzymes in canine skeletal muscle.

OBJECTIVE: To perform respiratory chain enzymatic activity assays on canine skeletal muscle biopsy specimens and establish reference range values of skeletal muscle enzyme activities for dogs. SAMPLE POPULATION: Biopsy specimens from the vastus lateralis muscle were obtained from 24 dogs (8 sexually intact males and 14 sexually intact females) ranging from 15 months to 6 years of age. PROCEDURE: Mean values of citrate synthase, cytochrome-c oxidase, succinate dehydrogenase, succinate dehydrogenase-cytochrome-c reductase, nicotinamide adenine dinucleotide (NADH) dehydrogenase, and NADH dehydrogenase-cytochrome-c reductase activities were established by use of 6 standard spectrophotometric assays for respiratory chain enzyme analysis. RESULTS: Compared with published data for skeletal muscle enzyme activities in humans, skeletal muscle enzyme activities in dogs were 2- to 4-fold higher. Additionally, citrate synthase activity, a marker for mitochondrial volume, was positively correlated with age in dogs, suggesting that mitochondrial volume increases with age, although no apparent change in respiratory chain enzymatic activity with an increase in age was found. CONCLUSIONS AND CLINICAL RELEVANCE: Reference range values for skeletal muscle enzyme activities of dogs are needed to accurately interpret results of respiratory chain enzymatic activity assays. During investigation of metabolic myopathies, if skeletal muscle biopsy specimens are evaluated for respiratory chain enzyme kinetics, they should be performed and evaluated in concert with skeletal muscle biopsy specimens from clinically normal animals of the same species.

Effects of exercise on canine skeletal muscle proteolysis: an investigation of the ubiquitin-proteasome pathway and other metabolic markers.

The effects of long-term athletic training are associated with excessive skeletal muscle turnover attributable to increased rates of myofibrillar protein synthesis and proteolysis, which are mechanisms poorly understood in the athletic dog. A physiologic field study using 44 English pointers and Labrador retrievers that had been purposely bred for bird hunting and retrieving was conducted to examine changes in the ubiquitin-proteasome (UP) pathway, which has been implicated in exercise-induced proteolysis. Muscle biopsy samples were collected from all dogs in September (preseason, pretraining) and February (peak season, peak activity). Western blot analysis was used to assess changes in expression of various components of the UP pathway in the biopsy samples. Citrate synthase and glycogen levels were also measured in a subset of these samples. Results across the population indicated pronounced up-regulation of ubiquitinated conjugates and the p31 regulatory capping subunit during the peak hunting period compared with the preseason period. In contrast, the catalytic core of the proteasome (beta-subunits) showed no apparent up-regulation in response to increased physical activity. Increased physical activity during the hunting season was associated with increased muscle glycogen levels and citrate synthase activity in these dogs. Overall, up-regulation of specific components of the UP pathway was an indication that it plays a role in the proteolytic process associated with skeletal muscle turnover during long-term athletic training, as previously believed.

Effects of post-exercise supplements on glycogen repletion in skeletal muscle.

Post-exercise carbohydrate supplementation has been routinely used to enhance glycogen concentrations in skeletal muscle, particularly during multiple-day athletic events. Consumption of protein hydrolysates mixed with carbohydrate supplements during the post-exercise period may increase insulin response and cause glycogen repletion in skeletal muscle. A group of Alaskan sled dogs were selected to examine post-exercise supplementation in a paired crossover study design. The dogs were subjected to the same exercise regimen and provided one of three treatments-water, glucose polymers, or glucose polymers with protein hydrolysates-over a 2-month period. Parameters tested at various post-exercise time points included plasma insulin, glucagon and glucose concentrations, and skeletal muscle glycogen content to gain a better understanding of glucose metabolism and glycogen repletion. The results showed an enhanced insulin, glucose, and glucagon response immediately after supplementation and significantly increased glycogen concentrations in skeletal muscle within 24 hours when dogs received either of the glucose-containing supplements compared with water alone. There were no differences in the plasma parameters or skeletal muscle glycogen stores in dogs provided the glucose polymers alone or the glucose polymers plus protein hydrolysates. Thus, post-exercise carbohydrate supplementation increased muscle glycogen repletion, but inclusion of protein hydrolysates in the supplements provided no additional benefits.