title: Antagonism of rat beta-cell voltage-dependent K+ currents by exendin 4 requires dual activation of the cAMP/protein kinase A and phosphatidylinositol 3-kinase signaling pathways.
abstract: Antagonism of voltage-dependent K+ (Kv) currents in pancreatic beta-cells may contribute to the ability of glucagon-like peptide-1 (GLP-1) to stimulate insulin secretion. The mechanism and signaling pathway regulating these currents in rat beta-cells were investigated using the GLP-1 receptor agonist exendin 4. Inhibition of Kv currents resulted from a 20-mV leftward shift in the voltage dependence of steady-state inactivation. Blocking cAMP or protein kinase A (PKA) signaling (Rp-cAMP and H-89, respectively) prevented the inhibition of currents by exendin 4. However, direct activation of this pathway alone by intracellular dialysis of cAMP or the PKA catalytic subunit (cPKA) could not inhibit currents, implicating a role for alternative signaling pathways. A number of phosphorylation sites associated with phosphatidylinositol 3 (PI3)-kinase activation were up-regulated in GLP-1-treated MIN6 insulinoma cells, and the PI3 kinase inhibitor wortmannin could prevent antagonism of beta-cell currents by exendin 4. Antagonists of Src family kinases (PP1) and the epidermal growth factor (EGF) receptor (AG1478) also prevented current inhibition by exendin 4, demonstrating a role for Src kinase-mediated trans-activation of the EGF tyrosine kinase receptor. Accordingly, the EGF receptor agonist betacellulin could replicate the effects of exendin 4 in the presence of elevated intracellular cAMP. Downstream, the PKCzeta pseudosubstrate inhibitor could prevent current inhibition by exendin 4. Therefore, antagonism of beta-cell Kv currents by GLP-1 receptor activation requires both cAMP/PKA and PI3 kinase/PKCzeta signaling via trans-activation of the EGF receptor. This represents a novel dual pathway for the control of Kv currents by G protein-coupled receptors.
authors.affiliation: Department of Physiology, University of Toronto, Toronto, Ontario M1H 1E6, Canada.
authors.forename: Patrick E
authors.initials: PE
authors.lastname: MacDonald
authors.forename: Xiaolin
authors.initials: X
authors.lastname: Wang
authors.forename: Fuzhen
authors.initials: F
authors.lastname: Xia
authors.forename: Wasim
authors.initials: W
authors.lastname: El-kholy
authors.forename: Elisha D
authors.initials: ED
authors.lastname: Targonsky
authors.forename: Robert G
authors.initials: RG
authors.lastname: Tsushima
authors.forename: Michael B
authors.initials: MB
authors.lastname: Wheeler
mesh_terms: D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D015153:Blotting, Western; D002384:Catalysis; D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D004594:Electrophysiology; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D066246:ErbB Receptors / Q000378:metabolism; D000077270:Exenatide; D005934:Glucagon / Q000737:chemistry; D052216:Glucagon-Like Peptide 1; D005947:Glucose / Q000378:metabolism; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007700:Kinetics; D008297:Male; D051379:Mice; D010446:Peptide Fragments / Q000737:chemistry; D010455:Peptides / Q000737:chemistry / Q000502:physiology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D010766:Phosphorylation; D011188:Potassium / Q000737:chemistry* / Q000378:metabolism; D011233:Precipitin Tests; D011498:Protein Precursors / Q000737:chemistry; D011799:Quinazolines; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction; D013997:Time Factors; D015533:Transcriptional Activation; D020032:Tyrphostins / Q000494:pharmacology; D015854:Up-Regulation; D014688:Venoms / Q000737:chemistry; D000077191:Wortmannin
chemical_list: D000730:Androstadienes; D004791:Enzyme Inhibitors; D007328:Insulin; D010446:Peptide Fragments; D010455:Peptides; D011498:Protein Precursors; D011799:Quinazolines; D020032:Tyrphostins; D014688:Venoms; C101044:RTKI cpd; D052216:Glucagon-Like Peptide 1; D005934:Glucagon; D000077270:Exenatide; D000242:Cyclic AMP; D019869:Phosphatidylinositol 3-Kinases; D066246:ErbB Receptors; D017868:Cyclic AMP-Dependent Protein Kinases; D005947:Glucose; D011188:Potassium; D000077191:Wortmannin