Effect of repeated oral administration of glucose and leucine immediately after exercise on plasma insulin concentration and glycogen synthesis in horses.

OBJECTIVE: To determine whether repeated oral administration of glucose and leucine during the period immediately after intense exercise would increase the release of insulin and thereby enhance glycogen synthesis in horses. ANIMALS: 12 Standardbred horses. PROCEDURES: In a crossover study design, after glycogen-depleting exercise, horses received oral boluses of glucose (1 g/kg at 0, 2, and 4 hours) and leucine (0.1 g/kg at 0 and 4 hours) or boluses of water (10 mL/kg at 0, 2, and 4 hours; control treatment). Blood samples for determination of glucose, insulin, and leucine concentrations were collected prior to and during a 6-hour period immediately after exercise. Biopsy specimens of a gluteus muscle were obtained before and immediately after exercise and at 3, 6, and 24 hours after exercise for measurement of glycogen concentration. RESULTS: When glucose and leucine were administered to the horses, plasma insulin concentration was significantly higher during the 6 hours immediately after exercise than it was when water was administered to the horses. Serum glucose concentration during the 4 hours immediately after exercise was significantly higher when glucose and leucine were administered than the serum glucose concentration when water was administered. Muscle glycogen concentrations did not differ between the 2 treatments during the 24 hours after exercise. CONCLUSIONS AND CLINICAL RELEVANCE: Synthesis of muscle glycogen after intense intermittent exercise was not enhanced by oral boluses of glucose and leucine after exercise despite pronounced increases in plasma insulin and serum glucose concentrations.

Repeated post-exercise administration with a mixture of leucine and glucose alters the plasma amino acid profile in Standardbred trotters.

BACKGROUND: The branched chain amino acid leucine is a potent stimulator of insulin secretion. Used in combination with glucose it can increase the insulin response and the post exercise re-synthesis of glycogen in man. Decreased plasma amino acid concentrations have been reported after intravenous or per oral administration of leucine in man as well as after a single per oral dose in horses. In man, a negative correlation between the insulin response and the concentrations of isoleucine, valine and methionine have been shown but results from horses are lacking. This study aims to determine the effect of repeated per oral administration with a mixture of glucose and leucine on the free amino acid profile and the insulin response in horses after glycogen-depleting exercise. METHODS: In a crossover design, after a glycogen depleting exercise, twelve Standardbred trotters received either repeated oral boluses of glucose, 1 g/kg body weight (BW) at 0, 2 and 4 h with addition of leucine 0.1 g/kg BW at 0 and 4 h (GLU+LEU), or repeated boluses of water at 0, 2 and 4 h (CON). Blood samples for analysis of glucose, insulin and amino acid concentrations were collected prior to exercise and over a 6 h post-exercise period. A mixed model approach was used for the statistical analyses. RESULTS: Plasma leucine, isoleucine, valine, tyrosine and phenylalanine concentrations increased after exercise. Post-exercise serum glucose and plasma insulin response were significantly higher in the GLU+LEU treatment compared to the CON treatment. Plasma leucine concentrations increased after supplementation. During the post-exercise period isoleucine, valine and methionine concentrations decreased in both treatments but were significantly lower in the GLU+LEU treatment. There was no correlation between the insulin response and the response in plasma leucine, isoleucine, valine and methionine. CONCLUSIONS: Repeated post-exercise administration with a mixture of leucine and glucose caused a marked insulin response and altered the plasma amino acid profile in horses in a similar manner as described in man. However, the decreases seen in plasma amino acids in horses seem to be related more to an effect of leucine and not to the insulin response as seen in man.

Postmortem changes in pork muscle protein phosphorylation in relation to the RN genotype.

Postmortem changes in pork muscle protein phosphorylation in relation to the RN(-) genotype were investigated using one-dimensional gel electrophoresis and a phosphor specific staining. The phosphorylation levels of several protein bands were found to be affected by the RN(-) genotype and to change during postmortem development. Glycogen phosphorylase, phosphofructokinase, and pyruvate kinase were found in protein bands affected by the RN(-) genotype, and the phosphorylation profile indicates that part of the increased rate and extended pH decline of the RN(-) genotype could be a consequence of phosphorylation of these key enzymes during the postmortem metabolism. The results illustrate that the protein phosphorylation level of the muscle proteins could be interpreted as a global metabolic fingerprint containing information about the activity status of the enzymes in the postmortem metabolism.

Analysis of acid-soluble glycogen in pork extracts of two PRKAG3 genotypes by 1H liquid-state NMR spectroscopy and biochemical methods.

Meat extracts with acid-soluble glycogen (macroglycogen) from M. longissmus dorsi of carriers and noncarriers of the PRKAG3 mutation (RN(-) and rn(+) genotype) were analyzed by both (1)H liquid-state NMR spectroscopy and a biochemical method. The (1)H NMR analysis revealed that shorter polymers (dimers, trimers, etc.) of α-1,4-linked glucose were generated 24-48 h post-mortem. This is not possible to elucidate with the biochemical method, by which only the total amount of hydrolyzed glucose residues is determined. The shorter polymers were primarily formed in carriers of the PRKAG3 mutation, suggesting different post-mortem glycogen degradation mechanisms in the two genotypes.

Muscle glycogen resynthesis, signalling and metabolic responses following acute exercise in exercise-trained pigs carrying the PRKAG3 mutation.

Hampshire pigs carrying the PRKAG3 mutation in the AMP-activated protein kinase (AMPK) γ3 subunit exhibit excessive skeletal muscle glycogen storage and an altered glycogen synthesis signalling response following exercise. AMPK plays an important role as a regulator of carbohydrate and fat metabolism in mammalian cells. Exercise-trained muscles are repeatedly exposed to glycogen degradation and resynthesis, to which the signalling pathways adapt. The aim of this study was to examine the effect of acute exercise on glycogen synthesis signalling pathways, and the levels of insulin and other substrates in blood in exercise-trained pigs with and without the PRKAG3 mutation. After 5 weeks of training, pigs performed two standardized treadmill exercise tests, and skeletal muscle biopsies were obtained immediately after exercise and 3 h postexercise in the first test, and 6 h postexercise in the second test. The PRKAG3 mutation carriers had higher glycogen storage, and resynthesis of glycogen was faster after 3 h but not after 6 h of recovery. Alterations in the concentrations of insulin, glucose, lactate and free fatty acids after exercise did not differ between the genotypes. The carriers showed a lower expression of AMPK and increased phosphorylation of Akt Ser(473) after exercise, compared with non-carriers. Acute exercise stimulated the phosphorylation of AS160 in both genotypes, and the phosphorylation of GSK3α Ser(21) and ACC Ser(79) in the non-carriers. In conclusion, exercise-trained pigs carrying the PRKAG3 mutation show an altered Akt and AMPK signalling response to acute exercise, indicating that glucose metabolism is associated with faster resynthesis of muscle glycogen in this group.

The influence of the PRKAG3 mutation on glycogen, enzyme activities and fibre types in different skeletal muscles of exercise trained pigs.

BACKGROUND: AMP-activated protein kinase (AMPK) plays an important role in the regulation of glucose and lipid metabolism in skeletal muscle. Many pigs of Hampshire origin have a naturally occurring dominant mutation in the AMPK γ3 subunit. Pigs carrying this PRKAG3 (R225Q) mutation have, compared to non-carriers, higher muscle glycogen levels and increased oxidative capacity in m. longissimus dorsi, containing mainly type II glycolytic fibres. These metabolic changes resemble those seen when muscles adapt to an increased physical activity level. The aim was to stimulate AMPK by exercise training and study the influence of the PRKAG3 mutation on metabolic and fibre characteristics not only in m. longissimus dorsi, but also in other muscles with different functions. METHODS: Eight pigs, with the PRKAG3 mutation, and eight pigs without the mutation were exercise trained on a treadmill. One week after the training period muscle samples were obtained after euthanisation from m. biceps femoris, m. longissimus dorsi, m. masseter and m. semitendinosus. Glycogen content was analysed in all these muscles. Enzyme activities were analysed on m. biceps femoris, m. longissimus dorsi, and m. semitendinosus to evaluate the capacity for phosphorylation of glucose and the oxidative and glycolytic capacity. Fibre types were identified with the myosin ATPase method and in m. biceps femoris and m. longissimus dorsi, immunohistochemical methods were also used. RESULTS: The carriers of the PRKAG3 mutation had compared to the non-carriers higher muscle glycogen content, increased capacity for phosphorylation of glucose, increased oxidative and decreased glycolytic capacity in m. longissimus dorsi and increased phosphorylase activity in m. biceps femoris and m. longissimus dorsi. No differences between genotypes were seen when fibre type composition was evaluated with the myosin ATPase method. Immunohistochemical methods showed that the carriers compared to the non-carriers had a higher percentage of type II fibres stained with the antibody identifying type IIA and IIX fibres in m. longissimus dorsi and a lower percentage of type IIB fibres in both m. biceps femoris and m. longissimus dorsi. In these muscles the relative area of type IIB fibres was lower in carriers than in non-carriers. CONCLUSIONS: In exercise-trained pigs, the PRKAG3 mutation influences muscle characteristics and promotes an oxidative phenotype to a varying degree among muscles with different functions.

Effects of exercise on muscle glycogen synthesis signalling and enzyme activities in pigs carrying the PRKAG3 mutation.

The dominant RN mutation in pigs results in excessive glycogen storage in skeletal muscle. The mutation is situated in the PRKAG3 gene, which encodes a muscle-specific isoform of the AMP-activated protein kinase (AMPK) gamma3 subunit. AMPK is an important regulator of carbohydrate and fat metabolism in mammalian cells. The aim of the present study was to examine the effect of exercise on glycogen synthesis signalling pathways in muscle and to study enzyme activities of importance in carbohydrate metabolism in pigs with or without the PRKAG3 mutation. Glycogen content, metabolic enzyme activities and expression or phosphorylation of signalling proteins were analysed in skeletal muscle specimens obtained at rest, after a single treadmill exercise bout and after 3 h recovery. The PRKAG3 mutation carriers had higher glycogen content, a tendency for lower expression of AMPK (P < 0.07) and higher hexokinase and phosphorylase activities, whereas citrate synthase, 3-hydroxyacyl-CoA dehydrogenase and glycogen synthase activities did not differ between genotypes. Carriers and non-carriers of the RN mutation showed a similar degradation of glycogen after exercise, whereas the rate of resynthesis was faster in the carriers. Acute exercise stimulated Akt phosphorylation on Ser(473) in both genotypes, and the effect was greater in the carriers than in the non-carriers. Acute exercise also stimulated phosphorylation of Akt substrate of 160 kDA and Glycogen synthase kinase 3 in the carriers and GSK3alpha in the non-carriers. In conclusion, the increased rate of glycogen synthesis following exercise in pigs carrying the PRKAG3 mutation correlates with an increased signalling response of Akt and its substrate, AS160, and a higher activity of hexokinase, indicating an increased glucose influx and phosphorylation of glucose, directed towards glycogen synthesis.

Effect of triheptanoin on muscle metabolism during submaximal exercise in horses.

OBJECTIVE: To compare effects of corn oil or a 7-carbon fat (triheptanoin) on acylcarnitine, lipid, and carbohydrate metabolism in plasma or muscle of exercising horses. ANIMALS: 8 Thoroughbred geldings. PROCEDURES: Horses received isocaloric diets containing 650 mL of oil (triheptanoin or corn oil)/d for 18 or 25 days in a crossover design with a 26-day washout period. On day 17 or 24 of each feeding period, the respective oil (217 mL) was nasogastrically administered; 120 minutes later, horses performed a 90-minute submaximal exercise test (SET). Blood and muscle samples were obtained before oil administration and immediately before (blood only), during (blood only), immediately after, and 24 hours after SETs. RESULTS: Compared with values before oil administration, triheptanoin administration increased plasma insulin and C7:0-, C5:0- and C3:0-acylcarnitine concentrations, whereas corn oil administration increased plasma NEFA concentrations. During SETs, plasma C7:0-, C5:0-, and C3:0-acylcarnitine concentrations were higher when triheptanoin, rather than corn oil, was administered to horses. Plasma glucose, NEFA, and C2:0-, C18:1-, and C18:2-acylcarnitine concentrations increased during SETs similarly for both oils. Respiratory quotient and muscle lactate, citrate, malate, glycogen, and ATP concentrations changed similarly from before to after SETs for both oils. Compared with muscle concentrations immediately after SETs, those for glucose-6-phosphate and citrate 24 hours after SETs were lower and for glycogen were similar to values before SETs. CONCLUSIONS AND CLINICAL RELEVANCE: Fatigue was not associated with depletion of citric acid cycle intermediates for either oil. Triheptanoin induced a significantly higher insulin secretion and did not appear to enhance muscle glycogen repletion.

Metabolism during anaesthesia and recovery in colic and healthy horses: a microdialysis study.

BACKGROUND: Muscle metabolism in horses has been studied mainly by analysis of substances in blood or plasma and muscle biopsy specimens. By using microdialysis, real-time monitoring of the metabolic events in local tissue with a minimum of trauma is possible. There is limited information about muscle metabolism in the early recovery period after anaesthesia in horses and especially in the colic horse. The aims were to evaluate the microdialysis technique as a complement to plasma analysis and to study the concentration changes in lactate, pyruvate, glucose, glycerol, and urea during anaesthesia and in the recovery period in colic horses undergoing abdominal surgery and in healthy horses not subjected to surgery. METHODS: Ten healthy university-owned horses given anaesthesia alone and ten client-owned colic horses subjected to emergency abdominal surgery were anaesthetised for a mean (range) of 230 min (193-273) and 208 min (145-300) respectively. Venous blood samples were taken before anaesthesia. Venous blood sampling and microdialysis in the gluteal muscle were performed during anaesthesia and until 24 h after anaesthesia. Temporal changes and differences between groups were analysed with an ANOVA for repeated measures followed by Tukey Post Hoc test or Planned Comparisons. RESULTS: Lactate, glucose and urea, in both dialysate and plasma, were higher in the colic horses than in the healthy horses for several hours after recovery to standing. In the colic horses, lactate, glucose, and urea in dialysate, and lactate in plasma increased during the attempts to stand. The lactate-to-pyruvate ratio was initially high in sampled colic horses but decreased over time. In the colic horses, dialysate glycerol concentrations varied considerably whereas in the healthy horses, dialysate glycerol was elevated during anaesthesia but decreased after standing. In both groups, lactate concentration was higher in dialysate than in plasma. The correspondence between dialysate and plasma concentrations of glucose, urea and glycerol varied. CONCLUSION: Microdialysis proved to be suitable in the clinical setting for monitoring of the metabolic events during anaesthesia and recovery. It was possible with this technique to show greater muscle metabolic alterations in the colic horses compared to the healthy horses in response to regaining the standing position.

Metabolic stress-like condition can be induced by prolonged strenuous exercise in athletes.

Few studies have examined energy metabolism during prolonged, strenuous exercise. We wanted therefore to investigate energy metabolic consequences of a prolonged period of continuous strenuous work with very high energy expenditure. Twelve endurance-trained athletes (6 males and 6 females) were recruited. They performed a 7-h bike race on high work-load intensity. Physiological, biochemical, endocrinological, and anthropometric muscular compartment variables were monitored before, during, and after the race. The energy expenditure was high, being 5557 kcal. Work-load intensity (% of VO(2) peak) was higher in females (77.7%) than in men (69.9%). Muscular glycogen utilization was pronounced, especially in type I fibres (>90%). Additionally, muscular triglyceride lipolysis was considerably accelerated. Plasma glucose levels were increased concomitantly with an unchanged serum insulin concentration which might reflect an insulin resistance state in addition to proteolytic glyconeogenesis. Increased reactive oxygen species (malondialdehyde (MDA)) were additional signs of metabolic stress. MDA levels correlated with glycogen utilization rate. A relative deficiency of energy substrate on a cellular level was indicated by increased intracellular water of the leg muscle concomitantly with increased extracellular levels of the osmoregulatory amino acid taurine. A kindred nature of a presumed insulin-resistant state with less intracellular availability of glucose for erythrocytes was also indicated by the findings of decreased MCV together with increased MCHC (haemoconcentration) after the race. This strenuous energy-demanding work created a metabolic stress-like condition including signs of insulin resistance and deteriorated intracellular glucose availability leading to compromised fuelling of ion pumps, culminating in a disturbed cellular osmoregulation indicated by taurine efflux and cellular swelling.

Changes in amino acid concentration in plasma and type I and type II fibres during resistance exercise and recovery in human subjects.

Eight male subjects performed leg press exercise, 4 x 10 repetitions at 80% of their maximum. Venous blood samples were taken before, during exercise and repeatedly during 2 h of recovery. From four subjects, biopsies were taken from the vastus lateralis muscle prior to, immediately after and following one and 2 h of recovery. Samples were freeze-dried, individual muscle fibres were dissected out and identified as type I or type II. Resistance exercise led to pronounced reductions in the glutamate concentration in both type I (32%) and type II fibres (70%). Alanine concentration was elevated 60-75% in both fibre types and 29% in plasma. Glutamine concentration remained unchanged after exercise; although 2 h later the concentrations in both types of fibres were reduced 30-35%. Two hours after exercise, the plasma levels of glutamate and six of the essential amino acids, including the branched-chain amino acids were reduced 5-30%. The data suggest that glutamate acts as an important intermediate in muscle energy metabolism during resistance exercise, especially in type II fibres.

Blood concentrations of the cytokines IL-1beta, IL-6, IL-10, TNF-alpha and IFN-gamma during experimentally induced swine dysentery.

BACKGROUND: Knowledge of the cytokine response at infection with Brachyspira hyodysenteriae can help understanding disease mechanism involved during swine dysentery. Since this knowledge is still limited the aim of the present study was to induce dysentery experimentally in pigs and to monitor the development of important immunoregulatory cytokines in blood collected at various stages of the disease. METHODS: Ten conventional pigs (~23 kg) were orally inoculated with Brachyspira hyodysenteriae B204T. Eight animals developed muco-haemorrhagic diarrhoea with impaired general body condition. Blood was sampled before inoculation and repeatedly during acute dysentery and recovery periods and cytokine levels of IL-1beta, IL-6, Il-10, TNF-alpha and IFN-gamma were measured by ELISA. RESULTS: IL-1beta was increased at the beginning of the dysentery period and coincided with the appearance of Serum amyloid A and clinical signs of disease. TNF-alpha increased in all animals after inoculation, with a peak during dysentery, and IL-6 was found in 3 animals during dysentery and in the 2 animals that did not develop clinical signs of disease. IL-10 was found in all sick animals during the recovery period. IFN-gamma was not detected on any occasion. CONCLUSION: B. hyodysenteriae inoculation induced production of systemic levels of IL-1beta during the dysentery period and increased levels of IL-10 coincided with recovery from dysentery.

Metabolism before, during and after anaesthesia in colic and healthy horses.

BACKGROUND: Many colic horses are compromised due to the disease state and from hours of starvation and sometimes long trailer rides. This could influence their muscle energy reserves and affect the horses' ability to recover. The principal aim was to follow metabolic parameter before, during, and up to 7 days after anaesthesia in healthy horses and in horses undergoing abdominal surgery due to colic. METHODS: 20 healthy horses given anaesthesia alone and 20 colic horses subjected to emergency abdominal surgery were anaesthetised for a mean of 228 minutes and 183 minutes respectively. Blood for analysis of haematology, electrolytes, cortisol, creatine kinase (CK), free fatty acids (FFA), glycerol, glucose and lactate was sampled before, during, and up to 7 days after anaesthesia. Arterial and venous blood gases were obtained before, during and up to 8 hours after recovery. Gluteal muscle biopsy specimens for biochemical analysis of muscle metabolites were obtained at start and end of anaesthesia and 1 h and 1 day after recovery. RESULTS: Plasma cortisol, FFA, glycerol, glucose, lactate and CK were elevated and serum phosphate and potassium were lower in colic horses before anaesthesia. Muscle adenosine triphosphate (ATP) content was low in several colic horses. Anaesthesia and surgery resulted in a decrease in plasma FFA and glycerol in colic horses whereas levels increased in healthy horses. During anaesthesia muscle and plasma lactate and plasma phosphate increased in both groups. In the colic horses plasma lactate increased further after recovery. Plasma FFA and glycerol increased 8 h after standing in the colic horses. In both groups, plasma concentrations of CK increased and serum phosphate decreased post-anaesthesia. On Day 7 most parameters were not different between groups. Colic horses lost on average 8% of their initial weight. Eleven colic horses completed the study. CONCLUSION: Colic horses entered anaesthesia with altered metabolism and in a negative oxygen balance. Muscle oxygenation was insufficient during anaesthesia in both groups, although to a lesser extent in the healthy horses. The post-anaesthetic period was associated with increased lipolysis and weight loss in the colic horses, indicating a negative energy balance during the first week post-operatively.

Exercise pattern influences skeletal muscle hybrid fibers of runners and nonrunners.

PURPOSE: To determine whether relationships between skeletal muscle hybrid fiber composition and whole-body exercise patterns help to elucidate their transitional capacity or a fine-tuning response to functional demands. METHODS: This study investigated hybrid fibers from vastus lateralis biopsies of runners (N= 13) and nonrunners (N = 9) and related hybrid fiber occurrence and distribution of myosin heavy-chain isoforms (MHC) within hybrid fibers to exercise patterns. MHC composition of single fibers was identified by SDS-PAGE. RESULTS: Runners had more fibers expressing only MHC I, fewer expressing MHC IIx, and fewer IIa/IIx hybrid fibers (P < 0.05). Hybrid IIa/IIx and I/IIa fibers were, respectively, negatively and positively related to training volume or average preferred racing distance (PRDA) in runners (P < 0.05). The relationship between IIa/IIx hybrid fibers and PRDA was more exponential (R(2) = 0.88) than linear (R(2) = 0.69). Only IIa/IIx hybrid fibers correlated negatively with exercise hours in nonrunners (P < 0.05). Their IIa/IIx hybrid fibers had MHC IIa content ranging from 1 to 99%, with most between 41 and 60%. Runners favoring longer distances (PRDA > 8 km or training > 70 km x wk(-1)) had no IIa/IIx hybrid fibers with MHC IIa proportion > 60%. In these runners, MHC I within hybrid I/IIa fibers was skewed toward higher proportions (> 60%), whereas MHC I proportions were skewed oppositely in runners favoring shorter training or racing distances. CONCLUSIONS: Training volume influences both IIa/IIx and I/IIa hybrid fiber proportions in runners, but only the former in nonrunners. Hybrid IIa/IIx fiber proportions were modulated by racing distance. Distinctly different distributions of MHC isoforms within the hybrid fibers were seen in runners favoring longer distances versus those favoring shorter distances.

Do skeletal muscle phenotypic characteristics of Xhosa and Caucasian endurance runners differ when matched for training and racing distances?

Although East African black athletes dominate endurance running events, it is unknown whether black and white endurance runners with similar racing ability, matched for training, may differ in their skeletal muscle biochemical phenotype. Thirteen Xhosa (XR) and 13 Caucasian (CR) endurance runners were recruited and matched for 10-km performance, average preferred racing distance (PRD(A)), and training volume. Submaximal and maximal exercise tests were done, and vastus lateralis muscle biopsies were taken. XR were significantly lighter and shorter than CR athletes but had similar maximum oxygen consumption corrected for body weight and peak treadmill speed (PTS). XR had lower plasma lactate concentrations at 80% PTS (P < 0.05) compared with CR. Also, XR had more type IIA (42.4 +/- 9.2 vs. 31.3 +/- 11.5%, P < 0.05) and less type I fibers (47.8 +/- 10.9 vs. 63.1 +/- 13.2%, P < 0.05), although oxidative enzyme activities did not differ. Furthermore, XR compared with CR had higher lactate dehydrogenase (LDH) activity in homogenate muscle samples (383 +/- 99 vs. 229 +/- 85 mumol.min(-1).g dry weight(-1), P < 0.05) and in both type IIa (P < 0.05) and type I (P = 0.05) single-fiber pools. A marked difference (P < 0.05) in the composition of LDH isoform content was found between the two groups with XR having higher levels of LDH(5-4) isoforms (skeletal muscle isozymes; LDH-M) than CR, which was not accounted for by fiber-type differences alone. These results confirm differences in muscle phenotype and physiological characteristics, particularly associated with high-intensity running.

Proglycogen, macroglycogen, glucose, and glucose-6-phosphate concentrations in skeletal muscles of horses with polysaccharide storage myopathy performing light exercise.

OBJECTIVE: To determine concentrations of proglycogen (PG), macroglycogen (MG), glucose, and glucose-6-phosphate (G-6-P) in skeletal muscle of horses with polysaccharide storage myopathy (PSSM) before and after performing light submaximal exercise. ANIMALS: 6 horses with PSSM and 4 control horses. PROCEDURES: Horses with PSSM completed repeated intervals of 2 minutes of walking followed by 2 minutes of trotting on a treadmill until muscle cramping developed. Four untrained control horses performed a similar exercise test for up to 20 minutes. Serum creatine kinase (CK) activity was measured before and 4 hours after exercise. Concentrations of total glycogen (G(t)), PG, MG, G-6-P, free glucose, and lactate were measured in biopsy specimens of gluteal muscle obtained before and after exercise. RESULTS: Mean serum CK activity was 26 times higher in PSSM horses than in control horses after exercise. Before exercise, muscle glycogen concentrations were 1.5, 2.2, and 1.7 times higher for PG, MG, and G(t), respectively, in PSSM horses, compared with concentrations in control horses. No significant changes in G(t), PG, MG, G-6-P, and lactate concentrations were detected after exercise. However, free glucose concentrations in skeletal muscle increased significantly in PSSM horses after exercise. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis of the results suggests that glucose uptake in skeletal muscle is augmented in horses with PSSM after light exercise. There is excessive storage of PG and MG in horses with PSSM, and high concentrations of the 2 glycogen fractions may affect functional interactions between glycogenolytic and glycogen synthetic enzymes and glycosomes.

Effect of exercise on proglycogen and macroglycogen content in skeletal muscles of pigs with the Rendement Napole mutation.

OBJECTIVE: To investigate influence of the Rendement Napole (RN-) mutation on proglycogen (PG) and macroglycogen (MG) content in skeletal muscles before and after exercise and evaluate glycogen concentrations within various muscle fiber types. ANIMALS: 5 pigs with the RN- mutation and 3 noncarrier pigs. PROCEDURE: Pigs performed 2 exercise tests on a treadmill. In the first, pigs (mean body weight, 27 kg) ran a distance of approximately 800 m. In the second, pigs (mean body weight, 63 kg) ran until fatigued. Biopsy specimens (biceps femoris muscle) for determination of PG and MG contents were obtained before and after exercise, 24 hours after the first test, and 3 hours after the second test. Histochemical analysis was performed on specimens obtained before and after the second test. RESULTS: Before exercise, PG stores did not differ markedly between groups, but MG stores were twice as high in pigs with the RN- mutation, compared with noncarrier pigs. The MG content decreased to a similar extent in both groups after exercise. Resynthesis of MG was greater in pigs with the RN- mutation than in noncarrier pigs by 3 hours after exercise. A low glycogen content after exercise was observed in many type I and type IIA fibers and in some type lIB fibers. CONCLUSIONS AND CLINICAL RELEVANCE: The RN- mutation was associated with high MG stores in skeletal muscle that did not influence exercise performance. The RN- mutation did not impair glycogenolysis during exercise but may induce faster resynthesis of MG after exercise.

Effects of submaximal exercise on adenine nucleotide concentrations in skeletal muscle fibers of horses with polysaccharide storage myopathy.

OBJECTIVE: To determine whether disruption of adenine triphosphate (ATP) regeneration and subsequent adenine nucleotide degradation are potential mechanisms for rhabdomyolysis in horses with polysaccharide storage myopathy (PSSM) performing submaximal exercise. ANIMALS: 7 horses with PSSM and 4 control horses. PROCEDURES: Horses with PSSM performed 2-minute intervals of a walk and trot exercise on a treadmill until muscle cramping developed. Control horses exercised similarly for 20 minutes. Serum creatine kinase (CK) activity was measured 4 hours after exercise. Citrate synthase (CS), 3-OH-acylCoA dehydrogenase, and lactate dehydrogenase activities prior to exercise and glucose-6-phosphate (G-6-P) and lactate concentrations before and after exercise were measured in gluteal muscle specimens. Adenine triphosphate, diphosphate (ADP), monophosphate (AMP), and inosine monophosphate (IMP) concentrations were measured before and after exercise in whole muscle, single muscle fibers, and pooled single muscle fibers. RESULTS: Serum CK activity ranged from 255 to 22,265 U/L in horses with PSSM and 133 to 278 U/L in control horses. Muscle CS activity was lower in horses with PSSM, compared with control horses. Muscle G-6-P lactate, ATP, ADP, and AMP concentrations in whole muscle did not change with exercise in any horses. Concentration of IMP increased with exercise in whole muscle, pooled muscle fibers, and single muscle fibers in horses with PSSM. Large variations in ATP and IMP concentrations were observed within single muscle fibers. CONCLUSIONS AND CLINICAL RELEVANCE: Increased IMP concentration without depletion of ATP in individual muscle fibers of horses with PSSM during submaximal exercise indicates an energy imbalance that may contribute to the development of exercise intolerance and rhabdomyolysis.

The effects of spontaneous and mechanical ventilation on central cardiovascular function and peripheral perfusion during isoflurane anaesthesia in horses.

OBJECTIVE: To compare the effects of spontaneous breathing and mechanical ventilation on haemodynamic variables, including muscle and skin perfusion measured with laser Doppler flowmetery, in horses anaesthetized with isoflurane. STUDY DESIGN: Prospective controlled study. ANIMALS: Ten warm-blood trotter horses (five males, five females). Mean mass was 492 kg (range 420-584 kg) and mean age was 5 years (range 4-8 years). MATERIALS AND METHODS: After pre-anaesthetic medication with detomidine (10 microg kg(-1)) anaesthesia was induced with intravenous (IV) guaifenesin and thiopental (4-5 mg kg(-1) IV) and maintained using isoflurane in oxygen. The horses were positioned in dorsal recumbency. In five animals breathing was initially spontaneous (SB) while the lungs of the other five were ventilated mechanically using intermittent positive pressure ventilation (IPPV). Total anaesthesia time was 4 hours with the ventilatory mode changed after 2 hours. During anaesthesia, heart rate (HR) cardiac output (Qt) stroke volume (SV) systemic arterial blood pressures (sAP), and pulmonary arterial pressure (pAP) were recorded. Peripheral perfusion was measured in the semimembranosus and gluteal muscles and on the tail skin using laser Doppler flowmetry. Arterial (a) and mixed venous (v) blood gases, pH, haemoglobin concentration [Hb], haematocrit (Hct), plasma lactate concentration and muscle temperature were measured. Oxygen content, venous admixture (s/Qt) oxygen delivery (DO(2)) and oxygen consumption (VO(2)) were calculated. RESULTS: During mechanical ventilation, HR, sAP, pAP, Qt, SV, Qs/Qt and PaCO(2) were lower and PaO(2) was higher compared with spontaneous breathing. There were no differences between the modes of ventilation in the level of perfusion, DO(2), VO(2), [Hb], (Hct), or plasma lactate concentration. After the change from IPPV to SB, left semimembranosus muscle and skin perfusion improved, while muscle perfusion tended to decrease when SB was changed to IPPV. Low-frequency flow motion was seen twice as frequently during IPPV compared with SB. CONCLUSIONS: Mechanical ventilation impaired cardiovascular function compared with SB in horses during isoflurane anaesthesia. Muscle and skin perfusion changes occurred with ventilation, although further studies are needed to elucidate the underlying mechanisms.