Single-cell measurements of quantal secretion induced by alpha-latrotoxin from rat adrenal chromaffin cells: dependence on extracellular Ca2+
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Methods for Cell-attached Capacitance Measurements in Mouse Adrenal Chromaffin Cell
Published on: October 22, 2014
1Department of Medicine, Box 8217, Washington University School of Medicine, St. Louis, MO-63110, USA.
Abstract:
alpha-Latrotoxin (alpha-LT), from black widow spider venom, is a potent enhancer of the spontaneous quantal release of neurotransmitter from a variety of nerve terminals and clonal neurosecretory cells. Using electrochemical amperometry and estimation of membrane impedance by phase detection, we present evidence that alpha-LT induces exocytosis of catecholamines from rat adrenal chromaffin cells beginning as rapidly as 30 s after close application of the toxin. This release is largely dependent on adequate levels of extracellular Ca2+ ([Ca2+]o). Lowering [Ca2+]o from 2 mM to = 10-20 microM reduces the alpha-LT-induced rise in membrane capacitance by at least sixfold, on average, and nearly abolishes alpha-LT-induced quantal amperometric events, while still permitting insertion of non-selective cation channels. Based on these experiments, we argue that the rapid onset of alpha-LT action in promoting massive quantal release from chromaffin cells is primarily due to an increase in the Ca2+ permeability of the plasma membrane through non-selective cation channels.

