Abstract
The aim of the present study was to investigate possible changes of inositol 1,4,5-trisphosphate (IP3) mass in Torpedo cholinergic synaptosomes in conditions promoting stimulated acetylcholine (ACh) release. For this purpose, we used a radioreceptor IP3 mass assay and a chemiluminescent method for ACh detection. Torpedo cholinergic synaptosomes have consistent IP3 mass levels under resting conditions. The IP3 mass was neither modified by changes in external Ca2+ nor by a Ca2+-free medium containing EGTA. IP3 mass and ACh release, measured in the same conditions and in parallel, were increased by depolarization with high K+ and by the ionophores A-23187 and gramicidin-D in a manner dependent on external Ca2+ emphasizing that Ca2+ entry, independently of the influx mechanism involved, leads to an IP3 increase. The phospholipase Cβ inhibitors U-73122 and U-73343 reduced K+-stimulated IP3 levels while K+-evoked ACh release was almost completely blocked suggesting an additional effect of these drugs on depolarization–neurotransmitter secretion coupling. The effect reported showing an increase of IP3 by agents that stimulate ACh release may suggest a possible link between IP3 metabolism and the neurotransmitter release mechanism. However, such a link is probably not a direct one as implied by the results obtained with the inhibitors of phospholipase C.
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