Ghrelin Attenuates cAMP-PKA Signaling to Evoke Insulinostatic Cascade in Islet β-Cells

• DOI: 10.2337/db11-0368
• Publication Date: 2011-07-26T04:36:54Z
• Authors: Katsuya Dezaki; Boldbaatar Damdin...; Hideyuki Sone; Oleg Dyachok; Anders Tengholm; Erik Gylfe; Tomoyuki Kurashina; Masashi Yoshida; Masafumi Kakei; Toshihiko Yada
• Keywords: Male; 0303 health sciences; Cyclic AMP-Dependent Protein Kinases; Ghrelin; Rats; Mice; 03 medical and health sciences; Glucose; Islet Studies; Insulin-Secreting Cells; Insulin Secretion; Cyclic AMP; Animals; Insulin; Rats, Wistar; Signal Transduction

OBJECTIVEGhrelin reportedly restricts insulin release in islet β-cells via the Gαi2 subtype of G-proteins and thereby regulates glucose homeostasis. This study explored whether ghrelin regulates cAMP signaling and whether this regulation induces insulinostatic cascade in islet β-cells.RESEARCH DESIGN AND METHODSInsulin release was measured in rat perfused pancreas and isolated islets and cAMP production in isolated islets. Cytosolic cAMP concentrations ([cAMP]i) were monitored in mouse MIN6 cells using evanescent-wave fluorescence imaging. In rat single β-cells, cytosolic protein kinase-A activity ([PKA]i) and Ca2+ concentration ([Ca2+]i) were measured by DR-II and fura-2 microfluorometry, respectively, and whole cell currents by patch-clamp technique.RESULTSGhrelin suppressed glucose (8.3 mmol/L)-induced insulin release in rat perfused pancreas and isolated islets, and these effects of ghrelin were blunted in the presence of cAMP analogs or adenylate cyclase inhibitor. Glucose-induced cAMP production in isolated islets was attenuated by ghrelin and enhanced by ghrelin receptor antagonist and anti-ghrelin antiserum, which counteract endogenous islet-derived ghrelin. Ghrelin inhibited the glucose-induced [cAMP]i elevation and [PKA]i activation in MIN6 and rat β-cells, respectively. Furthermore, ghrelin potentiated voltage-dependent K+ (Kv) channel currents without altering Ca2+ channel currents and attenuated glucose-induced [Ca2+]i increases in rat β-cells in a PKA-dependent manner.CONCLUSIONSGhrelin directly interacts with islet β-cells to attenuate glucose-induced cAMP production and PKA activation, which lead to activation of Kv channels and suppression of glucose-induced [Ca2+]i increase and insulin release.