Abstract
We have developed a novel method to examine [ 3H]-noradrenaline and [ 14C]-glutamate release from the same sample of streptolysin-O (SLO) perforated rat cortical synaptosomes. Ca 2+-dependent [ 3H]-noradrenaline and [ 14C]-glutamate release was examined at different temperatures and was found to be greater at 30°C than at 25°C. Ca 2+-dependent release of [ 3H]-noradrenaline is more ATP dependent than Ca 2+-dependent release of [ 14C]-glutamate. No significant reuptake of either neurotransmitter by the perforated synaptosomes was detected, indicating all the synaptosomes were indeed perforated. Incubations with 1 mM ouabain, a specific Na +,K +-ATPase inhibitor, slightly increased Ca 2+-dependent release of both neurotransmitters. [ 3H]-noradrenaline is released from large dense-core vesicles and [ 14C]-glutamate is released from small clear synaptic vesicles, so one can directly compare and contrast neurotransmitter release mechanisms between large dense-core vesicles and small clear synaptic vesicles using this preparation.
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