Changes in muscle free carnitine and acetylcarnitine with increasing work intensity in the Thoroughbred horse

Treadmill exercise in Thoroughbred horses of 2 min duration and increasing intensity resulted in increased formation and accumulation of acetylcarnitine in the working middle gluteal muscle. At high work intensities a plateau in acetylcarnitine formation was reached corresponding to approximately 70% of the total carnitine pool (approx. 30 mmol.kg-1 dry muscle). Formation of acetylcarnitine was mirrored by an equal fall in the free carnitine content, which stabilised, at the highest work intensities, at around 8 mmol.kg-1 dry muscle. Acetylcarnitine and carnitine reached their point of maximum change at a work intensity just below that resulting in the rapid production and accumulation of lactate and glycerol 3-phosphate. It is possible that the formation of acetylcarnitine is important in the regulation of the intramitochondrial acetyl CoA/CoA ratio; equally these changes may represent a blocking mechanism aimed at preventing the transfer of unwanted free fatty acids (as acylcarnitines) into the mitochondria at work intensities where they could contribute little to energy production.     

Association between muscle acetyl-CoA and acetylcarnitine levels in the exercising horse

Treadmill exercise of 2-min duration and increasing intensity resulted in increased formation of acetyl-CoA and acetylcarnitine in working muscle of Thoroughbred horses. At high work intensities a plateau was reached for both acetyl-CoA (approximately 50 mumols/kg dry muscle) and acetylcarnitine (approximately 20 mmol/kg dry muscle). Postexercise concentrations were significantly (P less than 0.001) correlated; [acetylcarnitine] = 349.[acetyl-CoA] + 2.4. The results indicate that approximately 350 mumols acetylcarnitine were accumulated for every 1 mumol acetyl-CoA. Under the conditions of exercise used it is probable that most of the acetyl-CoA formed is generated through the intramitochondrial decarboxylation of pyruvate. The acetyl groups of acetyl-CoA are apparently redistributed throughout the whole cell through formation of acetylcarnitine, which readily transverses the mitochondrial membrane. Despite the redistribution, however, the close correlation between acetylcarnitine and acetyl-CoA would indicate that equilibrium was maintained and that neither acetylcarnitine transferase nor carnitine/acetylcarnitine translocase were rate limiting. There is some question as to whether the changes observed relate directly to exercise itself or to the state in muscle 10 s or more after exercise.     

The effects of diet on muscle pH and metabolism during high intensity exercise

Five healthy male subjects exercised for 3 min at a workload equivalent to 100% VO2max on two separate occasions. Each exercise test was performed on an electrically braked cycle ergometer after a four-day period of dietary manipulation. During each of these periods subjects consumed either a low carbohydrate (3 +/- 0%, mean +/- SD), high fat (73 +/- 2%), high protein (24 +/- 3%) diet (FP) or a high carbohydrate (82 +/- 1%), low fat (8 +/- 1%) low protein (10 +/- 1%) diet (CHO). The diets were isoenergetic and were assigned in a randomised manner. Muscle biopsy samples (Vastus lateralis) were taken at rest prior to dietary manipulation, immediately prior to exercise and immediately post-exercise for measurement of pH, glycogen, glucose 6-phosphate, fructose 1,6-diphosphate, triose phosphates, lactate and glutamine content. Blood acid-base status and selected metabolites were measured in arterialised venous samples at rest prior to dietary manipulation, immediately prior to exercise and at pre-determined intervals during the post-exercise period. There was no differences between the two treatments in blood acid-base status at rest prior to dietary manipulation; immediately prior to exercise plasma pH (p less than 0.01), blood PCO2 (p less than 0.01), plasma bicarbonate (p less than 0.001) and blood base-excess (p less than 0.001) values were all lower on the FP treatment. There were no major differences in blood acid-base variables between the two diets during the post-exercise period.(ABSTRACT TRUNCATED AT 250 WORDS)     

Endogenous opioids may modulate catecholamine secretion during high intensity exercise

To determine the effect of endogenous opioids on catecholamine response during intense exercise [80% maximal oxygen uptake ( O_2max)], nine fit men [mean (SE) ( O_2max, 63.9 (1.7) ml · kg^–1 · min^–1; age 27.6 (1.6) years] were studied during two treadmill exercise trials. A double-blind experimental design was used with subjects undertaking the two exercise trials in counterbalanced order. Exercise trials were 20 min in duration and were conducted 7 days apart. One exercise trial was undertaken following administration of naloxone (N; 1.2 mmol · l^–1; 3 ml) and the other after receiving a placebo (P; 0.9% saline; 3 ml). Prior to each experimental trial a flexible catheter was placed into an antecubital vein and baseline blood samples were collected. Immediately afterwards, each subject received bolus injection of either N or P. Blood samples were also collected after 20 min of continuous exercise while running. Epinephrine and norepinephrine were higher (P < 0.05) in the N than P exercise trial with mean (SE) values of 1679 (196) versus 1196 (155) pmol · l^–1 and 24 (2.2) versus 20 (1.7) nmol · · l^–1 respectively. Glucose and lactate were higher (P < 0.05) in the N than P exercise trial with values of 7 (0.37) versus 5.9 (0.31) mmol · l^–1 and 6.9 (1.1) versus 5.3 (0.9) mmol · l^–1 respectively. These data suggest an opioid inhibition in the release of catecholamines during intense exercise.     

Training at high exercise intensity promotes qualitative adaptations of mitochondrial function in human skeletal muscle

This study explored mitochondrial capacities to oxidize carbohydrate and fatty acids and functional optimization of mitochondrial respiratory chain complexes in athletes who regularly train at high exercise intensity (ATH, n = 7) compared with sedentary (SED, n = 7). Peak O(2) uptake (Vo(2max)) was measured, and muscle biopsies of vastus lateralis were collected. Maximal O(2) uptake of saponin-skinned myofibers was evaluated with several metabolic substrates [glutamate-malate (V(GM)), pyruvate (V(Pyr)), palmitoyl carnitine (V(PC))], and the activity of the mitochondrial respiratory complexes II and IV were assessed using succinate (V(s)) and N,N,N',N'-tetramethyl-p-phenylenediamine dihydrochloride (V(TMPD)), respectively. Vo(2max) was higher in ATH than in SED (57.8 +/- 2.2 vs. 31.4 +/- 1.3 ml.min(-1).kg(-1), P < 0.001). V(GM) was higher in ATH than in SED (8.6 +/- 0.5 vs. 3.3 +/- 0.3 micromol O(2).min(-1).g dry wt(-1), P < 0.001). V(Pyr) was higher in ATH than in SED (8.7 +/- 1.0 vs. 5.5 +/- 0.2 micromol O(2).min(-1).g dry wt(-1), P < 0.05), whereas V(PC) was not significantly different (5.3 +/- 0.9 vs. 4.4 +/- 0.5 micromol O(2).min(-1).g dry wt(-1)). V(S) was higher in ATH than in SED (11.0 +/- 0.6 vs. 6.0 +/- 0.3 micromol O(2).min(-1).g dry wt(-1), P < 0.001), as well as V(TMPD) (20.1 +/- 1.0 vs. 16.2 +/- 3.4 micromol O(2).min(-1).g dry wt(-1), P < 0.05). The ratios V(S)/V(GM) (1.3 +/- 0.1 vs. 2.0 +/- 0.1, P < 0.001) and V(TMPD)/V(GM) (2.4 +/- 1.0 vs. 5.2 +/- 1.8, P < 0.01) were lower in ATH than in SED. In conclusion, comparison of ATH vs. SED subjects suggests that regular endurance training at high intensity promotes the enhancement of maximal mitochondrial capacities to oxidize carbohydrate rather than fatty acid and induce specific adaptations of the mitochondrial respiratory chain at the level of complex I.     

The effects of high intensity exercise on muscle and plasma levels of alpha-ketoisocaproic acid

Alpha-ketoisocaproic acid (KIC) is the product of the transamination of the indispensable amino acid leucine, which is the first step in the complete degradation of leucine. To determine the effects of intense exercise on muscle and blood levels of KIC, 7 male volunteers performed cycle exercise to exhaustion. After pedaling at an intensity of 90 W for 3 min, the load was increased by 60 W every 3 min until volitional fatigue. Muscle biopsies were obtained prior to and immediately after exercise and rapidly frozen for later determination of KIC. During exercise, blood lactate levels increased as expected, while plasma KIC levels did not change. Following exercise, plasma KIC levels rose significantly with peak values occurring 15 min after exercise and did not return to pre-exercise values until 60 min after exercise. In contrast, muscle KIC levels increased during exercise from a pre-exercise mean of 49.4 +/- 4.1 mumol X kg-1 wet wt to 78.1 +/- 6.5 mumol X kg-1 after exercise, an average increase of 48% (P less than 0.05). These data indicate that during intense exercise, leucine transamination in muscle may continue at a faster rate than the decarboxylation of KIC. In addition, plasma levels of KIC did not reflect the intracellular accumulation of KIC during exercise, suggesting a delay in the diffusion of KIC from muscle.     

Fluid replacement drinks during high intensity exercise effects on minimizing exercise-induced disturbances in homeostasis

The purpose of these experiments was to examine the influence of various fluid replacement drinks on exercise-induced disturbances in homeostasis during heavy exercise. Nine trained cyclists performed constant load exercise on a cycle ergometer to fatigue on three occasions with 1-week separating experiments. The work rate was set initially at approximately 85% of VO2max (range 82-88%) with fatigue being defined as a 10% decline in power output below the initial value. During each experiment subjects consumed one of the following three beverages prior to and every 15 min during exercise: (1) non-electrolyte placebo (NEP; 31 mosmol.kg-1); (2) glucose polymer drink containing electrolytes (GP; 7% CHO, 231 mosmol.kg-1), and (3) electrolyte placebo drink without carbohydrate (EP; 48 mosmol.kg-1). Both the GP and EP beverage contained sodium citrate/citric acid (C) as a flavoring agent while C was not contained in the NEP drink. Although seven of nine subjects worked longer during the GP and EP treatment when compared with the NEP trial, the difference was not significant (P greater than 0.05). No differences (P greater than 0.05) existed between the GP and EP treatments in performance time. Exercise changes in rectal temperature, heart rate, delta % plasma volume and plasma concentrations of total protein, free fatty acids, glucose, lactate, potassium, chloride, calcium, and sodium did not differ (P greater than 0.05) between trials.(ABSTRACT TRUNCATED AT 250 WORDS)     

Equal volumes of high and low intensity of eccentric exercise in relation to muscle damage and performance

We examined differences in muscle damage and muscle performance perturbations in relation to the same volumes of high (HI) and low intensity (LI) of eccentric exercise. Untrained young healthy men (n = 12) underwent 2 isokinetic quadriceps eccentric exercise sessions, 1 on each randomly selected leg, separated by a 2-week interval. In the first session subjects performed HI exercise (i.e., 12 sets of 10 maximal voluntary efforts). In the second session, volunteers were subjected to continuous exercise of LI (50% of peak torque) until the total work done was approximately equal to that generated during HI. Muscle damage (serum creatine kinase concentration [CK], delayed onset of muscle soreness, and range of motion) and muscle performance (eccentric [EPT] and isometric peak torque [IPT]) indicators were assessed pre-exercise and 24, 48, 72, and 96 hours postexercise. Compared to baseline data, changes in muscle damage indicators were significantly different (p < 0.05) at almost all postexercise time points in both conditions. However, apart from the significant elevation of CK at 24 hours after HI (p < 0.05), no other significant differences were observed between the 2 exercise conditions (p > 0.05). The main finding in relation to muscle performance was that decrements following HI exercise were significantly greater (p < 0.05) compared to LI. Compared with baseline data, the EPT values following HI and LI exercise were as follows: 24 hours, 72.1% vs. 92%; 48 hours, 81.9% vs. 94.8%; 72 hours, 77.7% vs. 100.6%; 96 hours, 86.8% vs. 107.9%. The corresponding data for IPT were as follows: 24 hours, 86.4% vs. 102.8%; 48 hours, 84.2% vs. 107%; 72 hours, 84.8% vs. 109.2%; 96 hours, 86.8% vs. 114.4%. These results indicate that matching volumes of HI and LI eccentric exercise have similar effects on muscle damage, but HI has a more prominent effect on muscle performance.     

Nitric oxide and thermoregulation during exercise in the horse

Mills, Paul C., David J. Marlin, Caroline M. Scott, andNicola C. Smith. Nitric oxide and thermoregulation during exercise in the horse. J. Appl. Physiol. 82(4):1035-1039, 1997.The effect of inhibition of nitric oxideproduction on sweating rate (SR) and on core, rectal, and tail skintemperatures was measured in five Thoroughbred horses during exerciseof variable intensity on a high-speed treadmill. A standard exercisetest consisting of three canters [~55% maximumO₂ uptake(O_{2 max})], withwalking (~9%O_{2 max}) and trotting(~22% O_{2 max})between each canter, was performed twice (control or test), in randomorder, by each horse.N^G-nitro-L-arginine methyl ester(L-NAME; 20 mg/kg), acompetitive inhibitor of nitric oxide synthase, was infused into thecentral circulation and induced a significant reduction in the SRmeasured on the neck (31.6 ± 6.4 vs. 9.7 ± 4.2 g · min¹ · m²;69%) and rump (14.7 ± 5.2 vs. 4.8 ± 1.6 g · min¹ · m²;67%) of the horses during canter (P < 0.05). Significant increases in core, rectal, and tailskin temperatures were also measured (P < 0.05).L-Arginine (200 mg/kg iv)partially reversed the inhibitory effects ofL-NAME on SR, but core, rectal,and tail skin temperatures continued to increase(P < 0.05), suggesting a cumulationof body heat. The results support the contention that nitric oxidesynthase inhibition diminishes SR, resulting in elevated core andperipheral temperatures leading to deranged thermoregulation duringexercise. The inhibition of sweating byL-NAME may be related toperipheral vasoconstriction but may also involve the neurogenic controlof sweating.

     

Fat and carbohydrate metabolism during low intensity exercise: effects of the availability of muscle glycogen

Four subjects were studied during exercise at 50% of maximum oxygen uptake after a normal diet, after a low carbohydrate (CHO) diet following exercise-induced glycogen depletion, and after a high CHO diet. This regime has previously been shown to cause changes in the amount of glycogen stored in the exercising muscles. Metabolic and respiratory parameters were measured during the exercise. The respiratory exchange ratio, blood lactate, blood pyruvate, blood glucose and plasma triglycerides were lower than normal following the low CHO diet and higher than normal following the high CHO diet. Plasma free fatty acids and plasma glycerol were higher than normal after the low CHO diet and lower than normal after the high CHO diet. The contribution of CHO to metabolism was less than normal after the low CHO diet and greater than normal after the high CHO diet. The altered availability of FFA does not appear to be a result of the variations in the blood lactate content.     

Glucocorticoid receptor activation during exercise in muscle

This investigation was undertaken to evaluate whether endurance running of the type known to retard the muscle atrophy associated with glucocorticoid excess inhibits activation of glucocorticoid-receptor complexes to a DNA binding state. Female adrenalectomized rats received an injection (50 microCi/100 g body wt ip) of [3H]triamcinolone acetonide and remained sedentary or were immediately exercised by endurance running at 23 m/min for up to 90 min. Receptor activation, as quantified by binding to DNA-cellulose, steadily increased from 10-20% of the receptors capable of binding DNA in uninjected controls to 25-45% by 5 min and to 53-80% by 90 min after receiving the hormone in all muscles studied (fast-twitch red vastus lateralis, fast-twitch white vastus lateralis, slow-twitch soleus, mixed gastrocnemius, and heart). Exercise did not influence the time-course changes in percent activation. When activation was determined from changes in the conformational state of the receptor as measured by diethylaminoethyl-cellulose anion exchange chromatography, there was a similar time-dependent formation of activated receptor forms in all muscle types. However, exercise did not inhibit or delay the appearance of the activated receptor from the unactivated state. These results indicate that glucocorticoid receptor activation occurs at a rate that is independent of both fiber type and delivery of steroid to working muscles during exercise. If exercise alters receptor activation, a longer time period, beyond 90 min of running, or even additional training may be needed for inhibition to be expressed.     

Glycogenesis in muscle and liver during exercise

We hypothesized that glycogenesis increases in muscle during exercise before significant glycogen depletion occurs. Therefore, rats ran for 15 or 90 min at speeds of 8-22 m/min. D-[5-3H]glucose (10 microCi/100 g body wt) was administered 10 min before the end of exercise. Hindlimb muscles [soleus (SOL), plantaris (PL), extensor digitorum longus (EDL), and red (RG) and white gastrocnemius (WG)] and a portion of liver were analyzed for glycogen concentrations and rates of glycogen synthesis (i.e., D-[3H]glucose incorporated into glycogen). At rest, marked differences were observed among muscles in their rates of glucose incorporation into glycogen: i.e., SOL = 24.3 +/- 3.1, RG = 5.4 +/- 1.9, PL = 2.8 +/- 1.1, EDL = 0.54 +/- 0.10, WG = 0.12 +/- 0.02 (SE) dpm.micrograms glycogen-1.10 min-1 (P less than 0.05 between respective muscles). Compared with the glucose incorporation into glycogen at rest, increments in the PL (272%), RG (189%), WG (400%), EDL (274%), and liver (175%) were observed after 90 min of exercise (P less than 0.05, all data). In contrast, a decrease in glucose incorporation into glycogen (-62%) occurred in the SOL at min 15 (P less than 0.05), but this returned to the rates observed at rest after 90 min of exercise. This measure for rates of net glycogen synthesis (dpm.microgram glycogen-1.10 min-1) was weakly related to the ambient glycogen levels in most muscles; the exception was the SOL (r = -0.79; P less than 0.05). There was up to a 50-fold difference in glycogen synthesis among muscles.(ABSTRACT TRUNCATED AT 250 WORDS)     

Changes in muscle oxygenation during weight-lifting exercise

The quantitative analysis of haemoglobin oxygenation of contracting human muscle during weight-lifting exercise was studied noninvasively and directly using near-infrared spectroscopy. This method was developed as a three-wavelength method which confirmed the volume changes in oxygenated haemoglobin (oxy-Hb), deoxygenated haemoglobin (deoxy-Hb) and blood volume (total-Hb; Oxy-Hb + deoxy-Hb). Nine healthy adult men with various levels of training experience took part in the study. Ten repetition maximum (10 RM) one-arm curl exercise was performed by all the subjects. Results showed that at the beginning of the 10-RM exercise, rapid increases of deoxy-Hb and decreases of oxy-Hb were observed. In addition, total-Hb gradually increased during exercise. These results corresponded to the condition of arm blood flow experimentally restricted using a tourniquet in contact with the shoulder joint, and they showed the restriction of venous blood flow and an anoxic state occurring in the dynamically contracted muscle. In three sets of lifting exercise with short rest periods, these tendencies were accelerated in each set, while total-Hb volume did not return to the resting state after the third set for more than 90 s. These results would suggest that a training regimen emphasizing a moderately high load and a high number of repetitions, and a serial set with short rest periods such as usually performed by bodybuilders, caused a relatively long-term anoxic state in the muscle.     

Acute exercise and GLUT4 expression in human skeletal muscle: influence of exercise intensity.

To examine the influence of exercise intensity on the increases in vastus lateralis GLUT4 mRNA and protein after exercise, six untrained men exercised for 60 min at 39 +/- 3% peak oxygen consumption (V(O2 peak)) (Lo) or 27 +/- 2 min at 83 +/- 2% V(O2 peak) (Hi) in counterbalanced order. Preexercise muscle glycogen levels were not different between trials (Lo: 408 +/- 35 mmol/kg dry mass; Hi: 420 +/- 43 mmol/kg dry mass); however, postexercise levels were lower (P < 0.05) in Hi (169 +/- 18 mmol/kg dry mass) compared with Lo (262 +/- 35 mmol/kg dry mass). Thus calculated muscle glycogen utilization was greater (P < 0.05) in Hi (251 +/- 24 mmol/kg) than in Lo (146 +/- 34). Exercise resulted in similar increases in GLUT4 gene expression in both trials. GLUT4 mRNA was increased immediately at the end of exercise (approximately 2-fold; P < 0.05) and remained elevated after 3 h of postexercise recovery. When measured 3 h after exercise, total crude membrane GLUT4 protein levels were 106% higher in Lo (3.3 +/- 0.7 vs. 1.6 +/- 0.3 arbitrary units) and 61% higher in Hi (2.9 +/- 0.5 vs. 1.8 +/- 0.5 arbitrary units) relative to preexercise levels. A main effect for exercise was observed, with no significant differences between trials. In conclusion, exercise at approximately 40 and approximately 80% V(O2 peak), with total work equal, increased GLUT4 mRNA and GLUT4 protein in human skeletal muscle to a similar extent, despite differences in exercise intensity and duration.     

VO2 kinetics in the horse during moderate and heavy exercise

Langsetmo, I., G. E. Weigle, M. R. Fedde, H. H. Erickson, T. J. Barstow, and D. C. Poole.O₂ kinetics in thehorse during moderate and heavy exercise. J. Appl.Physiol. 83(4): 1235-1241, 1997.The horse is asuperb athlete, achieving a maximalO₂ uptake (~160ml · min¹ · kg¹)approaching twice that of the fittest humans. Although equine O₂ uptake(O₂) kinetics arereportedly fast, they have not been precisely characterized, nor hastheir exercise intensity dependence been elucidated. To addressthese issues, adult male horses underwent incremental treadmill testingto determine their lactate threshold (T_lac) and peakO₂(O_{2 peak}),and kinetic features of their O₂ response to"square-wave" work forcings were resolved using exercisetransitions from 3 m/s to abelow-T_lac speed of 7 m/s or anabove-T_lac speed of 12.3 ± 0.7 m/s (i.e., between T_lac and O_{2 peak}) sustainedfor 6 min. O₂ andCO₂ output were measured using anopen-flow system: pulmonary artery temperature was monitored, and mixedvenous blood was sampled for plasma lactate.O₂ kinetics at work levelsbelow T_lac were well fit by atwo-phase exponential model, with a phase2 time constant(₁ = 10.0 ± 0.9 s) thatfollowed a time delay (TD₁ = 18.9 ± 1.9 s). TD₁ was similar tothat found in humans performing leg cycling exercise, but the timeconstant was substantially faster. For speeds aboveT_lac,TD₁ was unchanged (20.3 ± 1.2 s); however, the phase 2 time constantwas significantly slower (₁ = 20.7 ± 3.4 s, P < 0.05) than for exercise belowT_lac. Furthermore, in four of fivehorses, a secondary, delayed increase inO₂ became evident135.7 ± 28.5 s after the exercise transition. This "slowcomponent" accounted for ~12% (5.8 ± 2.7 l/min) of the netincrease in exercise O₂. Weconclude that, at exercise intensities below and aboveT_lac, qualitative features ofO₂ kinetics in the horseare similar to those in humans. However, at speeds belowT_lac the fast component of theresponse is more rapid than that reported for humans, likely reflectingdifferent energetics of O₂utilization within equine muscle fibers.

     

Skeletal muscle oxygenation during incremental exercise

The purpose of this study was to investigate the relationship between muscle oxygenation level at exhaustion and maximal oxygen uptake (VO2max) in an incremental cycling exercise. Nine male subjects took part in an incremental exhaustive cycling exercise, and then cuff occlusion was performed. Changes in oxy-(deltaHbO2) and deoxy-(deltaHb) hemoglobin concentrations in the vastus lateralis muscle were measured with a near infrared spectroscopy (NIRS). Muscle oxygenation during incremental exercise was expressed as a percentage (%Moxy) of the maximal range observed during an arterial occlusion as the lower reference point. A systematic decrease was observed in %Moxy with increasing intensity. A significant relationship was observed between %Moxy at exhaustion and VO2max (p < 0.01). We concluded that the one of the limiting factor of VO2max is the muscle oxygen diffusion capacity, and %Moxy during exercise could be one of the indexes of muscle oxygen diffusion capacity.