J. Biol. Chem., Vol. 263, Issue 17, 8157-8161, 06, 1988

Muscarinic stimulation of calcium influx and norepinephrine release in PC12 cells

K Inoue and JG Kenimer
Laboratory of Cellular Physiology, Center for Biologics Evaluation and Research, Bethesda, Maryland 20892.

Muscarinic cholinergic receptor stimulation evokes catecholamine secretion from some cell types, but the mechanism has not been well characterized. Using pheochromocytoma (PC12) cells, we show that the muscarinic agonist methacholine stimulates 45Ca2+ influx and [3H]norepinephrine release in a dose-dependent manner. Experiments performed in Na+-free medium or with inhibitors of voltage-dependent Ca2+ channels suggest the involvement of a receptor-activated Ca2+ channel which differs significantly from the voltage-dependent Ca2+ channel involved in nicotinic receptor-stimulated release. Furthermore, both influx and release were inhibited by pertussis toxin (0.5-2.0 ng/ml, 21 h) with a dose dependency which paralleled the dose dependency of pertussis toxin-dependent in vivo ADP-ribosylation of a 41-kDa protein. These experiments provide the first evidence that muscarinic stimulation evokes neurotransmitter secretion by opening a receptor-activated Ca2+ channel which is controlled by a pertussis toxin-sensitive protein.
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