Physiological role and selectivity of the in situ potassium channel of the sarcoplasmic reticulum in skinned frog skeletal muscle fibers

Abstract

The role of K+ as a counterion during Ca2+ release from the sarcoplasmic reticulum (SR) has been investigated. An optical technique using the Ca2+-sensitive dye antipyrylazo III monitored Ca2+ release from skinned (sarcolemma removed) muscle fibers of the frog. Skinned fibers were used since the removal of the sarcolemma allows direct access to the SR membrane. Releases were stimulated by caffeine, which activates Ca2+ release directly by binding to a receptor on the SR. Two different methods were used to decrease the SR K+ conductance so that its effect on Ca2+ release could be assessed: (a) the SR K+ channel blocker, 1,10-bis-quanidino-n-decane (bisG10) was used to eliminate current pathways and (b) substitution of the impermeant ion choline for K+ was used to decrease charge carriers. Both bisG10 and choline substitution caused a concentration-dependent decrease in the Ca2+ release rate. Therefore we conclude that K+ is an important counterion for Ca2+ during its release from the SR. The selectivity of the in situ SR K+ channel to several monovalent cations was determined by substituting them for K+ and comparing their effect on Ca2+ release. The substituted ions were expected to affect Ca2+ release in proportion to their ability to support a counterion flux, which is, in turn, a function of their relative conductance through the SR K+ channel. The selectivity sequence determined by these experiments was K+ = Rb+ = Na+ greater than Cs+ greater than Li+ greater than choline.