Measurements of calcium transients in the soma, neurite, and growth cone of single cultured neurons

Abstract

Voltage-gated changes in cytosolic free calcium ion concentration were measured in single, differentiated cells of mouse neuroblastoma clone N1E-115 using the calcium-sensitive dye arsenazo III (AIII). In cells bathed in normal medium containing 10 mM calcium, the changes in AIII absorbance during a single action potential indicated an increase of 1.4 nM in cytosolic calcium. When 10 mM tetraethylammonium (TEA) was added to the bath, the action potential became prolonged and the change in cytosolic calcium increased to 3.9 nM. Under these conditions, repetitive stimulation at 0.5 Hz or faster caused a gradual decline in the amplitude and duration of the action potential and a gradual decline of the change in cytosolic calcium associated with each action potential. The amplitude of the prolonged after-hyperpolarization (AHP) that follows the action potential was found to reflect the magnitude of the change in cytosolic calcium. An action potential elicited in the cell soma caused an increase in cytosolic calcium in the soma, neurite, and growth cone regions of a single cell, indicating that the membrane of all three regions possesses voltage-gated calcium channels. Estimation of calcium flux per unit area of membrane suggests a distinct topographical organization of calcium channels. Calcium channel densities in the growth cone and cell soma regions are similar and significantly higher than that in the neurite.