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Originally published In Press as doi:10.1074/jbc.M202291200 on May 13, 2002

J. Biol. Chem., Vol. 277, Issue 29, 26233-26237, July 19, 2002
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Effects of Glucose, Exogenous Insulin, and Carbachol on C-peptide and Insulin Secretion from Isolated Perifused Rat Islets*

Walter S. ZawalichDagger and Kathleen C. Zawalich

From the Yale University School of Nursing, New Haven, Connecticut 06536-0740

Isolated perifused rat islets were stimulated with glucose, exogenous insulin, or carbachol. C-peptide and, where possible, insulin secretory rates were measured. Glucose (8-10 mM) induced dose-dependent and kinetically similar patterns of C-peptide and insulin secretion. The addition of 100 nM bovine insulin had no effect on C-peptide release in response to 8-10 mM glucose stimulation. The addition of 100 nM bovine insulin or 500 nM human insulin together with 3 mM glucose had no stimulatory effect on C-peptide secretion rates from perifused rat islets. Stimulation with carbachol plus 7 mM glucose enhanced both C-peptide and insulin secretion, and the further addition of 100 nM bovine insulin had no inhibitory effect on C-peptide secretory rates under this condition. Perifusion studies using pharmacologic inhibitors (genistein and wortmannin) of the kinases thought to be involved in insulin signaling potentiated 10 mM glucose-induced secretion. The results support the following conclusions. 1) C-peptide release rates accurately reflect insulin secretion rates from collagenase-isolated, perifused rat islets. 2) Exogenously added bovine insulin exerts no inhibitory effect on release to several agonists including glucose. 3) In the presence of 3 mM glucose, exogenously added bovine or human insulin do not stimulate endogenous insulin secretion.

* This work was supported by Grant 41230 from the NIDDK, National Institutes of Health and by the American Diabetes Association.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Yale University School of Nursing, P.O. Box 9740, 100 Church St. S., New Haven, CT 06536-0740. Tel.: 203-785-5522; Fax: 203-785-6455; E-mail: Walter.Zawalich@Yale.Edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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