Sulfonylurea Blockade of KATP Channels Unmasks a Distinct Type of Glucose-Induced Ca2+ Decrease in Pancreatic β-Cells.

Abstract

This study aimed to explore how sulfonylurea blockade of KATP channels affects the early Ca signals for glucose generation of insulin release. Cytoplasmic Ca was measured with ratiometric microfluorometry in isolated mouse islets loaded with Fura-PE3. After sulfonylurea blockade of the KATP channels (50 μM-1 mM tolbutamide or 1 μM-1 mM gliclazide), increase of glucose from 3 to 20 mM resulted in suppression of elevated Ca during a 3- to 5-minute period. The Ca decrease was shorter after inhibition of the Na/K pump with ouabain (10 and 100 μM) but prolonged when the α2A adrenoceptors were activated with clonidine (1 and 10 nM) or epinephrine (10 nM). Inhibition of the sarco/endoplasmic reticulum Ca-ATPase pump with 10 μM cyclopiazonic acid counteracted the action of 10 nM clonidine, making the Ca decrease shorter than in controls. Extended superfusion of islets with a medium containing 20 mM glucose and 1 mM tolbutamide sometimes resulted in delayed appearance of Ca oscillations mediated by periodic interruption of elevated Ca. Increase of glucose generates prompt suppression of cytoplasmic Ca in β-cells lacking functional KATP channels. Activation of α2A adrenoceptors markedly prolongs the period of glucose-induced Ca decrease, an effect counteracted by cyclopiazonic acid.