Proton efflux in human skeletal muscle during recovery from exercise

In recovery from exercise, phosphocreatine resynthesis results in the net generation of protons, while the net efflux of protons restores pH?to resting values. Because proton efflux rate declines as pH?increases, it appears to have an approximately linear pH-dependence. We set out to examine this in detail using recovery data from human calf muscle. Proton efflux rates were calculated from changes in pH?and phosphocreatine concentration, measured by ³¹P magnetic resonance spectroscopy, after incremental dynamic exercise to exhaustion. Results were collected post hoc into five groups on the basis of end-exercise pH. Proton efflux rates declined approximately exponentially with time. These were rather similar in all groups, even when pH?changes were small, so that the apparent rate constant (the ratio of efflux rate to pH?change) varied widely. However, all groups showed a consistent pattern of decrease with time; the halftimes of both proton efflux rate and the apparent rate constant were longer at lower pH. At each time-point, proton efflux rates showed a?significant pH-dependence [slope 17 (3)?mmol?·?l^?1?· min^?1?·?pH?unit^?1 at the start of recovery, mean (SEM)], but also a significant intercept at resting pH?[16?(3)?mmol?·?l^?1?·?min^?1 at the start of recovery]. The intercept and the slope both decreased with time, with halftimes of 0.37?(0.06) and 1.4 (0.4)?min, respectively. We conclude that over a wide range of end-exercise pH, net proton efflux during recovery comprises pH-dependent and pH-independent components, both of which decline with time. Comparison with other data in the literature suggests that lactate/proton cotransport can be only a small component of this initial recovery proton efflux.     

Interstitial glycerol concentrations in human skeletal muscle and adipose tissue during graded exercise

AIM: It is not clear how lipolysis changes in skeletal muscle and adipose tissue during exercise of different intensities. We aimed at estimating this by microdialysis and muscle biopsy techniques. METHODS: Nine healthy, young men were kicking with both legs at 25% of maximal power (Wmax) for 45 min and then simultaneously with one leg at 65% and the other leg at 85% Wmax for 35 min. RESULTS: Glycerol concentrations in skeletal muscle and adipose tissue interstitial fluid and in arterial plasma increased (P<0.001) during low intensity exercise and increased (P<0.05) even more during moderate intensity exercise. The difference between interstitial muscle and arterial plasma water glycerol concentration, which indicates the direction of the glycerol flux, was positive (P<0.05) at rest (21 +/- 9 microM) and during exercise at 25% Wmax (18 +/- 6 microM). The difference decreased (P<0.05) with increasing exercise intensity and was not significantly different from zero during exercise at 65% (-11 +/- 17 microM) and 85% (-12 +/- 13 microM) Wmax. In adipose tissue, the difference between interstitial and arterial plasma water glycerol increased (P<0.001) with increasing intensity. The net triacylglycerol breakdown, measured chemically from the biopsy, did not differ significantly from zero at any exercise intensity although directional changes were similar to microdialysis changes. CONCLUSIONS: Skeletal muscle releases glycerol at rest and at low exercise intensity but not at higher intensities. This can be interpreted as skeletal muscle lipolysis peaking at low exercise intensities but could also indicate that glycerol is taken up in skeletal muscle at a rate which is increasing with exercise intensity.     

NADH in human skeletal muscle during short-term intense exercise

The influence of high-intensity bicycle exercise on the redox level and lactate accumulation in skeletal muscle (m. quadriceps femoris) of man has been investigated. Six subjects exercised to exhaustion at a load corresponding to 100% VO2max. Muscle content of NADH, determined by the bioluminescence technique, increased from (means +/- SEM) 0.089 +/- 0.007 mmol/kg dry wt. at rest to 0.190 +/- 0.031 after 2 min of exercise (P less than 0.05) and to 0.213 +/- 0.021 at exhaustion (P less than 0.05). Values after 2 min exercise and at exhaustion were not statistically different (P greater than 0.05). Muscle lactate was increased 13-fold after 2 min of exercise and 22-fold at exhaustion as compared to the resting value. After 10 min recovery NADH was restored back to the pre-exercise level whereas muscle lactate was still elevated. The increase of muscle NADH during exercise is in contrast to earlier studies on isolated animal muscles, where an oxidation of NADH was observed during contractions. The difference might be due to the experimental model (isolated muscle vs. in vivo) or to the analytical method (qualitative data by reflectance fluorimetri from the surface of intact muscle vs. quantitative data from muscle extracts). Calculations of the cytosolic NADH concentration from the lactate dehydrogenase equilibrium show that 95% or more of the NADH is confined to the mitochondrial compartment. The observed increase of muscle NADH therefore imply that the redox potential of the mitochondria is decreased during intense exercise.(ABSTRACT TRUNCATED AT 250 WORDS)     

Neuropeptide Y is released together with noradrenaline from the human heart during exercise and hypoxia

The myocardial release of neuropeptide Y-like immunoreactivity (NPY-LI) and noradrenaline (NA) during exercise with and without arterial hypoxia was measured in 18 healthy men by arterial (a) and coronary sinus (cs) catheterization. Exercise was performed in the supine position on a cycle ergometer at a load, selected to produce a heart rate during air breathing of 120 beats min-1. Coronary sinus blood flow (CSBF) was measured and a and cs samples for NPY-LI, NA, oxygen and lactate analyses were taken at rest and after 6 min exercise. The inspiratory gas was then switched to 15% (n = 8) or 12% (n = 10) oxygen in nitrogen, exercise continued at the same load and measurements repeated after 6 min. At rest no significant release and during normoxic exercise a very small myocardial release of NPY-LI and NA was detected. During hypoxia compared to normoxia the cardiac NPY-LI release increased four-fold and the NA net release doubled at the same time as the arterial NPY-LI remained unaltered or only slightly increased. Both the NPY-LI and the NA net release from the heart correlated with the heart rate and the arterial but not the cs oxygen tension. The NPY-LI release was correlated with the NA net release. The findings suggest that arterial hypoxia stimulates cardiac NPY together with NA release which derives from local sympathetic nerves. The release from the heart seems to be greater than from other tissues.     

Utilization of long-chain fatty acids in human skeletal muscle during exercise

Long-chain fatty acids (LCFA) are important sources of energy in contracting skeletal muscle: during the course of endurance exercise the contribution of LCFA in energy metabolism increases whereas when the intensity of exercise increases, the energy need is covered more and more by carbohydrates. Although this has been known for nearly 100 years, the mechanisms controlling fatty acid uptake and oxidation during various exercise modes are still not completely elucidated. Besides passive diffusion, data suggest that both membrane-associated and cytosolic fatty acid binding proteins are involved in the uptake of LCFA into skeletal muscle. However, data from human studies suggest that the regulation of fatty acid utilization in skeletal muscle during exercise lies mainly within the entrance into the mitochondria or metabolism within the mitochondria. Although possible compartmentalization within the cell makes definitive conclusions difficult, available evidence suggests that changes in malonyl CoA concentration in muscle do not play a major regulatory role in controlling LCFA oxidation during exercise in man. In contrast, it is suggested that the availability of free carnitine may play a major regulatory role in oxidation of LCFA during exercise.     

Regulation of skeletal muscle perfusion during exercise

For exercise to be sustained, it is essential that adequate blood flow be provided to skeletal muscle. The local vascular control mechanisms involved in regulating muscle perfusion during exercise include metabolic control, endothelium-mediated control, propagated responses, myogenic control, and the muscle pump. The primary determinant of muscle perfusion during sustained exercise is the metabolic rate of the muscle. Metabolites from contracting muscle diffuse to resistance arterioles and act directly to induce vasodilation, or indirectly to inhibit noradrenaline release from sympathetic nerve endings and oppose α-adrenoreceptor-mediated vasoconstriction. The vascular endothelium also releases vasodilator substances (e.g., prostacyclin and nitric oxide) that are prominent in establishing basal vascular tone, but these substances do not appear to contribute to the exercise hyperemia in muscle. Endothelial and smooth muscle cells may also be involved in propagating vasodilator signals along arterioles to parent and daughter vessels. Myogenic autoregulation does not appear to be involved in the exercise hyperemia in muscle, but the rhythmic propulsion of blood from skeletal muscle veins facilitates venous return to the heart and muscle perfusion. It appears that the primary determinants of sustained exercise hyperemia in skeletal muscle are metabolic vasodilation and increased vascular conductance via the muscle pump. Additionally, sympathetic neural control is important in regulating muscle blood flow during exercise.     

Absence of humoral mediated 5'AMP-activated protein kinase activation in human skeletal muscle and adipose tissue during exercise

5'AMP-activated protein kinase (AMPK) exists as a heterotrimer comprising a catalytic α subunit and regulatory β and γ subunits. The AMPK system is activated under conditions of cellular stress, indicated by an increase in the AMP/ATP ratio, as observed, e.g. in muscles during contractile activity. AMPK was originally thought to be activated only by local intracellular mechanisms. However, recently it has become apparent that AMPK in mammals is also regulated by humoral substances, e.g. catecholamines. We studied whether humoral factors released during exercise regulate AMPK activity in contracting and resting muscles as well as in abdominal subcutaneous adipose tissue in humans. In resting leg muscle and adipose tissue the AMPK activity was not up-regulated by humoral factors during one-legged knee extensor exercise even when arm cranking exercise, inducing a ∼20-fold increase in plasma catecholamine level, was added simultaneously. In exercising leg muscle the AMPK activity was increased by one-legged knee extensor exercise eliciting a whole body respiratory load of only 30%     but was not further increased by adding arm cranking exercise. In conclusion, during exercise with combined leg kicking and arm cranking, the AMPK activity in human skeletal muscle is restricted to contracting muscle without influence of marked increased catecholamine levels. Also, with this type of exercise the catecholamines or other humoral factors do not seem to be physiological regulators of AMPK in the subcutaneous adipose tissue.     

ATP breakdown products in human skeletal muscle during prolonged exercise to exhaustion

To study changes in muscle energy state during prolonged exercise, especially in relation to fatigue, muscle biopsies were obtained from seven healthy males working until exhaustion on a cycle ergometer at 68% (63-74%) of their maximal oxygen uptake. Biopsies were taken at rest, after 15 and 45 min of exercise and at exhaustion, and analysed for ATP, ADP, AMP, inosine monophosphate (IMP) and hypoxanthine content by high performance liquid chromatography (HPLC), and for creatine phosphate (CP), lactate and glycogen by enzymatic fluorometric techniques. Glycogen content at exhaustion was approximately 30% of the pre-exercise level. The CP content decreased steeply during the first 15 min of exercise (P less than 0.01) and continued to decrease during the rest of the exercise period (P less than 0.05). Pronounced increases in contents of IMP (64% P less than 0.001) and hypoxanthine (69%, P less than 0.05) were found when exhaustion was approaching. Furthermore, energy charge [EC; (ATP + 0.5 ADP)/(ATP + ADP + AMP)] was decreased at exhaustion (P less than 0.05). The increases in IMP and hypoxanthine which occurred when exhaustion was approaching during prolonged submaximal exercise together with the decrease in EC during this phase of exercise suggest a failure of the exercising skeletal muscle to regenerate ATP at exhaustion.     

Effect of hypoxia on glucose metabolism in human skeletal muscle during exercise

The effect of respiratory hypoxia on muscle glucose metabolism during short-term dynamic exercise has been investigated. Eight men cycled for 5 min at 120 +/- 6 W (mean +/- SE), which corresponded to 50% of maximal O2 uptake during normoxia (N), breathing air (N) on one occasion and 11% O2 (hypoxia-H) on the other. Biopsies were taken from the quadriceps femoris muscle before and after exercise. Oxygen uptake during exercise was not affected by H. The arterial blood glucose concentration during N exercise remained constant, but increased from 4.62 +/- 0.11 mmol l(-1) at rest to 5.22 +/- 0.19 mmol l-1 at the end of H exercise (P less than 0.05 vs N exercise). The intracellular glucose content at rest was low and did not change during N exercise, but was four times higher after exercise during H vs N (P less than 0.01). Glucose 6-P increased under both conditions but significantly more during H (P less than 0.01), while glucose 1,6-P2 was not significantly different between treatments either at rest or after exercise. It is concluded that: (1) glucose uptake by skeletal muscle during short-term exercise. It is concluded that: (1) glucose uptake by skeletal muscle during short-term exercise during H is not associated with a stoichiometric glucose utilization; (2) the inhibition of hexokinase during H (evidenced by increase in muscle glucose) is due primarily to the increase in glucose 6-P; and (3) glucose 1,6-P2 is of minor importance for the regulation of contraction-mediated flux through hexokinase in human skeletal muscle.     

Mitochondrial function in human skeletal muscle is not impaired by high intensity exercise

 The hypothesis that high-intensity (HI) intermittent exercise impairs mitochondrial function was investigated with different microtechniques in human muscle samples. Ten male students performed three bouts of cycling at 130% of peak O₂ consumption (V ^·O_2,peak). Muscle biopsies were taken from the vastus lateralis muscle at rest, at fatigue and after 110 min recovery. Mitochondrial function was measured both in isolated mitochondria and in muscle fibre bundles made permeable with saponin (skinned fibres). In isolated mitochondria there was no change in maximal respiration, rate of adenosine 5’-triphosphate (ATP) production (measured with bioluminescence) and respiratory control index after exercise or after recovery. The ATP production per consumed oxygen (P/O ratio) also remained unchanged at fatigue but decreased by 4% (P<0.05) after recovery. In skinned fibres, maximal adenosine 5’-diphosphate (ADP)-stimulated respiration increased by 23% from rest to exhaustion (P<0.05) and remained elevated after recovery, whereas the respiratory rates in the absence of ADP and at 0.1 mM ADP (submaximal respiration) were unchanged. The ratio between respiration at 0.1 and 1 mM ADP (ADP sensitivity index) decreased at fatigue (P<0.05) but after the recovery period was not significantly different from that at rest. It is concluded that mitochondrial oxidative potential is maintained or improved during exhaustive HI exercise. The finding that the sensitivity of mitochondrial respiration to ADP is reversibly decreased after strenuous exercise may indicate that the control of mitochondrial respiration is altered. Received: 17 June 1998 / Received after revision: 11 November 1998 / Accepted: 26 November 1998     

Prostaglandin E release from dog skeletal muscle during restricted flow exercise.

Prostaglandin E (PGE) release from the anterior calf muscles of anesthetized dogs was measured during and following exercise. Blood flow was held constant at 15.5 +/- 1.6 (SEM) ml.min-1. 100 g-1 and the muscles were stimulated for 20 min at a frequency of 4 Hz. PGE release dropped from a resting level of 11.4 +/- 3.8 ng.min-1.100 g-1 to 6.5 +/- 2.0 ng.min-1.100 g-1 during exercise (P less than 0.05). Following exercise, PGE release slowly returned to and eventually exceeded the resting level over a 60-min period. Return of vascular resistance to control was even more prolonged. Indomethacin (5 mg/kg) caused 1) an increase in resting resistance (40%), 2) a drop in PGE release (48% at rest), and 3) a more rapid return of vascular resistance to control following exercise. PGE release does not appear to contribute to the vasodilation during exercise, but can account for the portion of vascular resistance recovery not blocked by indomethacin. The remaining prolonged vasodilation could be explained by another as yet unidentified vasodilator(s). This preparation exhibits tonic prostaglandin release that causes a vasodilation at rest.     

Regulation of glycogen synthase in skeletal muscle during exercise

Glycogen synthase (GS) catalyses the incorporation of uridine diphosphate-glucose into glycogen in skeletal muscle. In concert with the glucose transport step, GS activity is thought to be rate-limiting in the disposal of glucose as muscle glycogen. Glycogen synthase is regulated by both allosteric factors (primarily glucose 6-phosphate) and covalent modification by reversible phosphorylation and dephosphorylation leading to inactivation and activation of GS, respectively. Exercise activates both stimulatory and inhibitory regulators of GS and it is thought that the resultant activity of GS during exercise depends on the relative strength of opposing signals. However, the mechanisms by which exercise regulates GS activity are not fully understood. Glycogen breakdown, the GM-protein phosphatase 1 complex and possibly cellular relocalization of GS may be considered important factors involved in the stimulation of GS activity during exercise, while adenosine monophosphate-activated protein kinase and plasma adrenaline (via protein kinase A) can be considered as essential for the exercise-induced inhibitory signals to GS.     

Signalling to glucose transport in skeletal muscle during exercise

Exercise-induced glucose uptake in skeletal muscle is mediated by an insulin-independent mechanism. Although the signalling events that increase glucose transport in response to muscle contraction are not fully elucidated, the aim of the present review is to briefly present the current understanding of the molecular signalling mechanisms involved. Glucose uptake may be regulated by Ca++-sensitive contraction-related mechanisms possibly involving protein kinase C, and by mechanisms that reflect the metabolic status of the muscle and may involve the AMP-activated protein kinase. Furthermore the p38 mitogen activated protein kinase may be involved. Still, the picture is incomplete and a substantial part of the exercise/contraction-induced signalling mechanism to glucose transport remains unknown.     

Albumin clearance from human skeletal muscle during prolonged steady-state running

This study was designed to find out if the lymph flow, indicated as albumin clearance, from active skeletal muscle is maintained constant during a prolonged steady-state exercise. 99mTc-labelled albumin was injected bilaterally into the vastus lateralis muscles of eight endurance-trained men. The radioactivity at the injection site was monitored by a gamma-camera before, frequently during, and after a 2 h run at a controlled steady intensity of 69 +/- 4% of the maximal heart rate. The fractional clearance rate of albumin was calculated for each monitoring interval, and was expressed as percentage clearance per minute (% min(-1)). During the first 15 min of exercise the clearance rate was five times higher than at rest before the exercise (0.29 +/- 0.12 vs. 0.06 +/- 0.05% min(-1), P = 0.001). During the next 25 min of running the clearance rate fell to 0.19 +/- 0.08% min(-1) (P = 0.02), from which level it was further attenuated, being 0.12 +/- 0.04% min(-1) at the end of the exercise. After the exercise the clearance rate fell rapidly to the level of 0.04 +/- 0.03% min(-1). The results showed that the albumin clearance from working muscles is not constant during steady-state exercise, suggesting that lymph flow from exercising skeletal muscle may not be constant despite lymphatic pumping being assumed to be unchanged during the course of exercise (i.e. constant exercise intensity and running speed).