Prolonged Time Course of Glutamate Release from Nerve Terminals: Relationship Between Stimulus Duration and the Secretory Event

Abstract

The kinetics of synaptosomal [3H]glutamate release were measured on a subsecond time scale to study the relationship between the length of depolarization and the duration of the secretory event. The time course of release evoked by elevated K+ was complex, proceeding for several seconds after a 200-ms depolarization. We developed a protocol for depolarizing excitable membranes on a millisecond time scale to deliver brief depolarizations, termed the synthetic action potential, by using batrachotoxin to activate Na+ channels. Depolarization is achieved by superfusing with solutions containing elevated concentrations of Na+, and the duration of the depolarization is limited by including tetrodotoxin (TTX) in the superfusion solution to block Na+ current and membrane depolarizations were made in batrachotoxin-treated sensory neurons using patch clamp recording methods. Rapid increases in Na+ and TTX concentrations produced transient increases in inward Na+ current that decayed with a time course proportional to TTX concentration. Current clamp measurements indicated that, with 10 microM TTX, depolarizations last approximately 30 ms. Nonetheless, synaptosomal release of [3H]glutamate triggered by the synthetic action potential remained prolonged. Brief neuronal action potentials at some synapses may trigger transmitter release that persists for several seconds.