Pancreatic β-cells communicate via intermittent release of ATP

Abstract

The role of external ATP for intercellular communication was studied in glucose-stimulated pancreatic β-cells isolated from ob/ob mice. Digital image analyses with fura-2 revealed spontaneous transients of cytoplasmic Ca²⁺appearing in synchrony in the absence of cell contacts. After removal of slow oscillations with methoxyverapamil, addition of ATP (0.1–100 μM) resulted in prompt firing of a transient, followed by suppression of the generation and synchronization of spontaneously occurring transients. It was possible to trigger transients during the suppressive phase by raising the concentration of ATP. The dual action of ATP was mimicked by ADP or 2-methylthio-ATP but not by AMP or UTP. The number of spontaneous transients and their synchronization were reduced in the presence of the dephosphorylating agent apyrase. Additional evidence that intermittent release of ATP participates in the generation of spontaneous Ca²⁺transients was obtained from the suppression observed from use of antagonists of the purinoceptors [suramin (0.3–30 μM), pyridoxalphosphate-6-azophenyl-2,4-disulfonic acid (PPADS; 10–30 μM) and 2-deoxy- N-methyladenosine (MRS 2179; 0.3–30 μM)] or from counteracting β-cell release of ATP by inhibiting exocytosis with 100 nM epinephrine, 100 nM somatostatin, or lowering the temperature below 30°C. The data indicate that ATP has time-dependent actions (prompt stimulation followed by inhibition) on the generation of Ca²⁺transients mediated by P2Y receptors. It is proposed that β-cells both receive a neural ATP signal with coordinating effects on their Ca²⁺oscillations and propagate this message to adjacent cells via intermittent release of ATP combined with gap junction coupling.