Amine Weak Bases Disrupt Vesicular Storage and Promote Exocytosis in Chromaffin Cells

Authors


  • Abbreviations used : AM, acetoxymethyl ester ; [Ca2+]i, intracellular Ca2+ concentration ; VMAT, vesicular monoamine transporter.

Address correspondence and reprint requests to Dr. R. M. Wightman at Department of Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-3290, U.S.A.

Abstract

Abstract : The vesicular contents in bovine chromaffin cells are maintained at high levels owing to the strong association of its contents, which is promoted by the low vesicular pH. The association is among the catecholamines, Ca2+, ATP, and vesicular proteins. It was found that transient application of a weak base, methylamine (30 mM), amphetamine (10 μM), or tyramine (10 μM), induced exocytotic release. Exposure to these agents was also found to increase both cytosolic catecholamine and intracellular Ca2+ concentration, as measured by amperometry and fura-2 fluorescence. Amphetamine, the most potent amine with respect to evoking exocytosis, was found to be effective even in buffer with out external Ca2+ ; however, the occurrence of spikes was suppressed when BAPTA-acetoxymethyl ester was used to complex intracellular Ca2+. Amphetamine-induced spikes in Ca2+-free medium were not suppressed by thapsigargin or ruthenium red, inhibitors of the sarco(endo)plasmic reticulum Ca2+-ATPase and mitochondrial Ca2+ stores. Atomic absorption measurements of amphetamine- and methylamine-treated vesicles reveal that intravesicular Ca2+ stores are decreased after a 15-min incubation. Taken together, these data indicate that amphetamine and methylamine can disrupt vesicular stores to a sufficient degree that Ca2+ can escape and trigger exocytosis.

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