Work-induced potassium changes in skeletal muscle and effluent venous blood assessed by liquid ion-exchanger microelectrodes

Using liquid ion-exchanger semimicroelectrodes with a side pore, we measured changes of extracellular potassium concentration (K_e ⁺) in adult rabbit and cat gastrocnemius muscles and in venous effluent blood flowing from the cat gastrocnemius muscle during various bouts of activity induced by sciatic nerve stimulation.

Thermogenesis due to noradrenaline in muscles under different rates of perfusion

Vascularly isolated hind legs of cold acclimated rats were perfused with arterial blood either without noradrenaline (NA) or with a constant concentration of NA (10 ng·ml^–1) at different perfusion rates ranging from 2 to 14l·g^–1·min^–1. The oxygen consumption of the leg during perfusion both with or without NA was linearly related to the perfusion rate. The linear increase of leg oxygen consumption with respect to the perfusion was steeper after NA, which indicates that the same arterial concentration of NA may produce a greater thermogenic effect at higher blood flow rates (the difference between resting metabolic rate and the thermogenesis stimulated by NA, was 8.20 l O₂·g^–1·h^–1 at a blood flow of 3l·g^–1·min^–1, compared with 45.02 l O₂·g^–1·h^–1 at a blood flow of 14 l·g^–1·min^–1). These data confirm the important role of the extravascular influx rate of NA in the control of thermogenesis due to NA in muscles.     

Energy Metabolism of Rat Skeletal Muscle Modulated by the Rate of Perfusion Flow

In order to advance our understanding of the phenomenon of flow-induced increases in the metabolism of the relaxed muscle, the metabolic rate of the isolated rat gracilis muscle was investigated at 28 degrees C in vitro. The muscle was perfused with cell-free Krebs-Henseleit bicarbonate buffer containing 5% bovine serum albumin and 5 mM glucose, saturated with a gas mixture of 95% O2 and 5% CO2 and simultaneously superfused with a medium saturated with with a low O2 gas mixture (1% O2, 5% CO2 and 94% N2). Two different perfusion flow rates (0.054 and 0.100 ml min-1) have been used. Their influence on oxygen consumption and lactate production has been measured. After a 100 min perfusion period, the muscle was freeze-clamped and analysed for ATP, phosphocreatine, creatine, lactate, pyruvate, inorganic phosphate and glycogen content. The energy state of the cell and the proportions of glycolytic and mitochondrial fluxes of ATP synthesis were evaluated. During perfusion at the low flow rate of 0.054 ml min-1, the oxygen uptake was 45 +/- 9 nmol min-1 (g wet wt)-1, accompanied by a dominance of anaerobic glycolytic synthesis of ATP over mitochondrial ATP synthesis, even though the total delivery of oxygen to muscle was three times higher than oxygen consumption. Increasing the perfusion flow rate to 0.100 ml min-1 increased the oxygen uptake to 120 +/- 6 nmol min-1 (g wet wt)-1, thus leading to a prevalence of mitochondrial ATP synthesis over glycolytic ATP synthesis. The inner stores of glycogen served as the main substrate of energy metabolism and the role of exogenous substrates in the flow-stimulated increase of oxygen uptake was negligible. The increase in perfusion rate also enhanced the energy state of the muscle fibres, which was expressed either as the creatine charge or as the value of the change of Gibbs free energy of ATP hydrolysis. Data indicate that the change of perfusion flow rate per se, apart from oxygen and exogenous substrate supply, elicits changes in the regulation of energy metabolism within non-contracting skeletal muscle under open microcirculation.     

Work-Induced potassium changes in muscle venous effluent blood measured by ion-specific electrodes

Summary Modified Walker's liquid ion-exchanger microelectrodes were employed for measuring changes of K⁺ concentration in venous effluent blood from the cat gastrocnemius muscle during and after isometric tetani of various duration induced by indirect stimulation. The time course of these changes was obtained and the overall loss of K⁺ from a working muscle could thus be estimated. By comparing present results in the venous blood and previous findings of K⁺ concentration changes in the muscle extracellular space, a concentration gradient was found between the muscle and venous effluent blood.The authors wish to acknowledge the assistance of Dr. J. L. Walker in developing the ion-specific microelectrodes and for supplying them with silicone oil and Corning ion-exchanger.     

Substrate channelling in a creatine kinase system of rat skeletal muscle under various pH conditions

The aim of this study was to evaluate myofibrillar creatine kinase (CK) activity and to quantify the substrate channelling of ATP between CK and myosin ATPase under different pH conditions within the integrity of myofibrils. A pure myofibrillar fraction was prepared using differential centrifugation. The homogeneity of the preparation and the purity of the fraction were confirmed microscopically and by enzymatic assays for contaminant enzyme activities. The specific activity of myofibrillar CK reached 584 +/- 33 nmol PCr min(-1) mg(-1) at pH 6.75. Two methods were used to detect CK activity: (1) measurement of direct ATP production, and (2) measurement of PCr consumption. This method of evaluation has been tested in experiments with isolated creatine kinase. No discrepancy in CK activity between the methods was observed in the pH range tested (6.0-7.5). However, the same procedures resulted in a significant discrepancy between the amounts of reacted PCr and produced ATP within the pure myofibrillar fraction. This discrepancy represents the portion of ATP produced by the CK reaction, which is preferentially channelled to the myosin ATPase before diffusing into the bulk solution. The maximum evaluated difference reached 42.3 % at pH 6.95. The substrate channelling between myofibrillar-bound CK and myosin ATPase was evaluated under various pH levels within the physiological range and it reached a maximum value in a slightly acidic environment. These results suggest that ATP/ADP flux control by the CK system is more important at lower pH, corresponding to the physiological state of muscle fatigue.