Kinetic characteristics of calcium-dependent, cholinergic receptor controlled ATP secretion from adrenal medullary chromaffin cells

Abstract

Adrenal chromaffin cells secrete catecholamines (CA) and ATP in response ot acetylcholine (ACh) and high [K +] o. The relase proces is relatively fast making it difficult to measure the early phase of the secretory response. Recently we were able to resolve the time course of the secretory response by measuring the release of ATP using luciferin-luciferase included in the extracellular medium. For the three secretagogues studied, ACh, nicotine and high [K + o, the early phase of release followed a complex kinetics. Allowing for an initial delay of the secretory response, the kinetics could be described as the sum of two power exponential processes. Increasing the temperature from 23 to 37°C induced a marked decrease in the two time constants needed to fit the early time course of the ATP secretion. The activation energies, estimated from Arrhenius plots, were approx. 20 and 16 kcal/mol for both phases of ATP release induced by either cholinergic agonists or high [K + o. These results suggest that cholinergic receptor activation and membrane depolarization induce ATP (and CA) secretion through a common pathway. The initial delay in the onset of the secretory response decreased with increasing doses of secretagogue and with temperature. We propose that the delay preceding the actual onset of ATP release represents the time required for generation of intracellular second messengers. The effective concentration attained by these messengers depend apparently on both receptor occupancy by the agonist and the ensuing Ca 2+ channel activation.