Dependence of intracellular free calcium and tension on membrane potential and intracellular pH in single crayfish muscle fibres

Abstract

The dependence of intracellular free calcium ([Ca2+]i) and tension on membrane potential and intracellular pH (pHi) was studied in single isolated fibres of the crayfish claw-opener muscle using ion-selective microelectrodes. Tension (T) was quantified as a percentage of the maximum force, or as force per cross-sectional area (N/cm2). In resting fibres, pHi had a mean value of 7.06. Contractions evoked by an increase extracellular potassium [( K+]0) produced a fall in pHi of 0.01-0.05 units. The lowest measured levels of resting [Ca2+]i corresponded to a pCai (= -log [Ca2+]i) of 6.8. Intracellular Ca2+ transients recorded during K(+)-induced contractions did not reveal any distinct threshold for force development. Both the resting [Ca2+]i and resting tension were decreased by an intracellular alkalosis and increased by an acidosis. The sensitivity of resting tension to a change in pHi (quantified as -dT/dpHi) showed a progressive increase during a fall in pHi within the range examined (pHi 6.2-7.5). The pHi/[Ca2+]i and pHi/tension relationships were monotonic throughout the multiphasic pHi change caused by NH4Cl. A fall of 0.5-0.6 units in pHi did not produce a detectable shift in the pCai/tension relationship at low levels of force development. The results indicate that resting [Ca2+]i is slightly higher than the level required for contractile activation. They also show that the dependence of tension on pHi in crayfish muscle fibres is attributable to a direct H+ and Ca2+ interaction at the level of Ca2+ sequestration and/or transport. Finally, the results suggest that in situ, the effect of pH on the Ca2+ sensitivity of the myofibrillar system is not as large as could be expected on the basis of previous work on skinned crustacean muscle fibres.