Effects of monensin and veratridine on acetylcholine release and cytosolic free Ca2+ levels in cerebrocortical synaptosomes of rats

Abstract

Monensin (10(-8)-10(-4) M) caused a dose-dependent increase in the release of [3H]acetylcholine ([3H]ACh) from purified rat cerebrocortical synaptosomes, with an EC50 of approximately 1.6 x 10(-6) M. Extracellular Na+, but not Ca2+, was required for a monensin-induced increase in the release of [3H]ACh. Monensin also increased the cytosolic free Ca2+ concentration ([Ca2+]i) and uptake of 22Na+ in a dose-dependent manner. Monensin continued to cause a dose-dependent increase in [Ca2+]i in the absence of extracellular Ca2+, although an approximately 50% reduction was noted at concentrations of greater than 10(-5) M. The EC50 for the monensin-induced increase in [Ca2+]i was similar to that noted in the release of [3H]ACh. Veratridine exhibited effects similar to those of monensin, but a large portion of the increase in [Ca2+]i and [3H]ACh release was dependent on extracellular Ca2+. Measurements of rhodamine 6G fluorescence indicated that monensin and veratridine caused synaptosomal hyperpolarization and depolarization, respectively. Tetrodotoxin (10(-6) M) completely blocked all the effects of veratridine but had no effect on the activity of monensin. These results suggest that monensin increases the release of ACh at least in part by increasing [Ca2+]i, resulting from the increase in the Na+ influx through tetrodotoxin-insensitive mechanisms in rat cerebrocortical synaptosomes.