Excitation-induced exchange of Na+, K+, and Cl− in rat EDL muscle in vitro and in vivo: Physiology and pathophysiology

Abstract

In skeletal muscle, excitation leads to increased [Na+]i, loss of K+, increased [K+]o, depolarization, and Cl− influx. This study quantifies these changes in rat extensor digitorum longus (EDL) muscles in vitro and in vivo using flame photometric determination of Na+ and K+ and 36Cl as a tracer for Cl−. In vitro, 5-Hz stimulation for 300 s increased intracellular Na+ content by 4.6 ± 1.2 µmol/g wet wt (P < 0.002) and decreased intracellular K+ content by 5.5 ± 2.3 µmol/g wet wt (P < 0.03). This would increase [K+]o by 28 ± 12 mM, sufficient to cause severe loss of excitability as the result of inactivation of Na+ channels. In rat EDL, in vivo stimulation at 5 Hz for 300 s or 60 Hz for 60 s induced significant loss of K+ (P < 0.01), sufficient to increase [K+]o by 71 ± 22 mM and 73 ± 15 mM, respectively. In spite of this, excitability may be maintained by the rapid and marked stimulation of the electrogenic Na+,K+ pumps already documented. This may require full utilization of the transport capacity of Na+,K+ pumps, which then becomes a limiting factor for physical performance. In buffer containing 36Cl, depolarization induced by increasing [K+]o to 40–80 mM augmented intracellular 36Cl by 120–399% (P < 0.001). Stimulation for 120–300 s at 5–20 Hz increased intracellular 36Cl by 100–188% (P < 0.001). In rats, Cl− transport in vivo was examined by injecting 36Cl, where electrical stimulation at 5 Hz for 300 s or 60 Hz for 60 s increased 36Cl uptake by 81% (P < 0.001) and 84% (P < 0.001), respectively, indicating excitation-induced depolarization. Cl− influx favors repolarization, improving K+ clearance and maintenance of excitability. In conclusion, excitation-induced fluxes of Na+, K+, and Cl− can be quantified in vivo, providing new evidence that in working muscles, extracellular accumulation of K+ is considerably higher than previously observed and the resulting depression of membrane excitability may be a major cause of muscle fatigue.