Calcium dependent gating of thel-glutamate activated, excitatory synaptic channel on crayfish muscle

Abstract

Excitatory, glutamate-activated single channel currents were measured in outside-out patches of crayfish muscle. The open times of single channel openings, and the durations and rates of bursts were evaluated. These kinetic parameters were not appreciably affected by replacement of extracellular Na+ by Li+ or choline. Changes in extracellular Ca2+ concentration Cao also did not influence the duration of single openings. However the mean burst duration decreased for Cao less than 13.5 mM and the rate of bursts declined with a power of almost 2 in low Cao. At Cao less than 1 mM practically no channel openings were observed in presence of glutamate. In order to exclude more rapid desensitization of the glutamate receptors in low Cao as the cause of disappearance of channel openings, glutamate was applied in short pulses with a liquid-filament switch. In 0 Cao also a glutamate pulse did not trigger channel openings. In presence of 13.5mM Cao, the inorganic Ca-channel blockers La3+ and Cd2+ diminished the duration and rate of bursts of channel openings in a similar manner as low Cao. The effects of low Cao and of Cd2+ were tested also on quantal postsynaptic currents, EPSCs, which were recorded through a perfused macro-patch-clamp electrode. At 1.4 mM Cao in the perfused electrode tip, spontaneous EPSCs were reduced at least by a factor of 4, and elicited EPSCs by a factor of 16. Application of Cd2+ had similarly strong effects on the EPSCs. Also the decay of EPSCs was shortened substantially in 1.4 mM Cao or 5 mM Cd2+. The inhibitory Cl(-)-channel of crayfish muscle, activated by glutamate or GABA, also was studied in outside-out patches. The openings of this channel persisted in 0 Cao solutions; the block of channel openings in low Cao thus is a specific property of the excitatory channel. The action of Cao on the excitatory channel may be described as that of a cofactor to glutamate. A possible reaction scheme is proposed.