Inhibition of arachidonate release by secretagogue-stimulated pancreatic islets suppresses both insulin secretion and the rise in beta-cell cytosolic calcium ion concentration.
Fuel secretagogues induce hydrolysis of esterified arachidonic acid from pancreatic islet cell phospholipids and accumulation of nonesterified arachidonate at concentrations up to 35 microM. Exogenous arachidonate (5-30 microM) amplifies depolarization-induced insulin secretion from islets. Fuel secretagogue-induced hydrolysis of arachidonate from islet phospholipids occurs in Ca(2+)-free medium, suggesting the possible involvement of a Ca(2+)-independent phospholipase. In the companion paper [Gross et al. (1993) Biochemistry (preceding paper in this issue)], we demonstrated that the major islet phospholipase A2 is Ca(2+)-independent, ATP-stimulated, and inhibited by the haloenol lactone suicide substrate (HELSS) (E)-6-(bromomethylene)-3-(1-naphthalenyl)-2H-tetrahydropyran-2-one. Here we demonstrate that HELSS suppressed both release of the arachidonate metabolite prostaglandin E2 and insulin secretion from islets stimulated with D-glucose and the muscarinic agonist carbachol. Both prostaglandin E2 release and insulin secretion were suppressed with similar concentration profiles and time courses. Islet oxidation of [14C]-glucose to [14C]CO2, activities of islet lactate dehydrogenase and alanine and aspartate aminotransferases, and carbachol-induced inositol phosphate accumulation in islets were all unaffected by HELSS. Depolarization of isolated beta-cells with 40 mM KCl induced a rise in cytosolic [Ca2+] that was also unaffected by HELSS. In contrast, the 17 mM D-glucose-induced rise in beta-cell [Ca2+] was inhibited by HELSS in a concentration-dependent manner, but that induced by exogenous arachidonate (15 microM) was not. These results suggest that fuel secretagogues activate the islet Ca(2+)-independent phospholipase A2, resulting in release of nonesterified arachidonate, which facilitates Ca2+ entry into beta-cells and promotes insulin secretion.
Ramanadham
S
S
Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
Gross
R W
RW
Han
X
X
Turk
J
J
D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D002217:Carbachol / Q000494:pharmacology; D003600:Cytosol / Q000378:metabolism; D015232:Dinoprostone / Q000378:metabolism; D005947:Glucose / Q000494:pharmacology; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D009281:Naphthalenes / Q000494:pharmacology*; D010741:Phospholipases A / Q000037:antagonists & inhibitors*; D054467:Phospholipases A2; D011189:Potassium Chloride / Q000494:pharmacology; D011753:Pyrones / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley
D007328:Insulin; D009281:Naphthalenes; D011753:Pyrones; D016718:Arachidonic Acid; D011189:Potassium Chloride; C068314:6-(bromomethylene)tetrahydro-3-(1-naphthaleneyl)-2H-pyran-2-one; D002217:Carbachol; D010741:Phospholipases A; D054467:Phospholipases A2; D005947:Glucose; D015232:Dinoprostone; D002118:Calcium