Glucose Stimulation of Insulin Release in the Absence of Extracellular Ca2+and in the Absence of any Increase in Intracellular Ca2+in Rat Pancreatic Islets

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 92; no. 23; pp. 10728 - 10732
• Main Authors: Komatsu, Mitsuhisa, Schermerhorn, Thomas, Aizawa, Toru, Geoffrey W. G. Sharp
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 07-11-1995; National Acad Sciences
• Subjects: Animals; Calcium - metabolism; Colforsin - pharmacology; Cyclic AMP-Dependent Protein Kinases - metabolism; Dose-Response Relationship, Drug; Drug Interactions; Exocytosis; Glucose - pharmacology; Insulin; Insulin - metabolism; Insulin Secretion; Islets of Langerhans; Islets of Langerhans - cytology; Islets of Langerhans - drug effects; Islets of Langerhans - enzymology; Islets of Langerhans - metabolism; Male; Metabolism; Norepinephrine; Physiological stimulation; Protein Kinase C - metabolism; Protein Kinases - metabolism; Rats; Rats, Sprague-Dawley; Secretion; Sulfoxides; Tetradecanoylphorbol Acetate - pharmacology; Washing
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.92.23.10728

Insulin secretion has been studied in isolated rat pancreatic islets under stringent Ca2+-depleted, Ca2+-free conditions. Under these conditions, the effect of 16.7 mM glucose to stimulate insulin release was abolished. Forskolin, which activates adenylyl cyclase, also failed to stimulate release in the presence of either low or high glucose concentrations. A phorbol ester (phorbol 12-myristate 13-acetate; PMA) increased the release rate slightly and this was further increased by 16.7 mM glucose. Remarkably, in the presence of both forskolin and PMA, 16.7 mM glucose strongly augmented insulin release. The augmentation was concentration dependent and monophasic and had a temporal profile similar to the "second phase" of glucose-stimulated insulin release, which is seen under normal conditions when Ca2+is present. Metabolism is required for the effect because mannoheptulose abolished the glucose response. Other nutrient secretagogues, α-ketoisocaproate, and the combination of leucine and glutamine augmented release under the same conditions. Norepinephrine, a physiological inhibitor of insulin secretion, totally blocked the stimulation of release by forskolin and PMA and the augmentation of release by glucose. Thus, under the stringent Ca2+-free conditions imposed, the stimulation of insulin release by forskolin and PMA, as well as the augmentation of release by glucose, is under normal physiological control. As no increase in intracellular [Ca2+] was observed, the results demonstrate that glucose can increase the rate of exocytosis and insulin release by pancreatic islets in a Ca2+-independent manner. This interesting pathway of stimulus-secretion coupling for glucose appears to exert its effect at a site beyond the usual elevation of intracellular [Ca2+] and is not due to an activation by glucose of protein kinase A or C.