Diabetes, Vol 34, Issue 2 115-120, Copyright © 1985 by American Diabetes Association

ARTICLES

Perifusion of a clonal cell line of Simian virus 40-transformed beta cells. Insulin secretory dynamics in response to glucose, 3-isobutyl-1-methylxanthine, and potassium

RS Hill and AE Boyd

A perifusion system for the study of insulin secretory dynamics of a clonal, Simian virus 40-transformed hamster pancreatic beta cell line (HIT cells) is described. After a change from glucose-free to higher glucose levels in the perifusate, insulin secretion increased rapidly in a dose-dependent manner. The pattern of glucose-stimulated insulin release was monophasic and was not sustained during a continued glucose stimulus. Perifusing the cells with low glucose (0.3 mg/ml) before a glucose stimulus of 3.5 mg/ml resulted in more rapid insulin release with lower peak secretory rates than those seen after a glucose-free period. The combined stimulus of high glucose and 100 microM 3-isobutyl-1-methylxanthine (IBMX), a phosphodiesterase inhibitor, significantly enhanced the acute insulin secretory response and also resulted in a biphasic secretory pattern that was sustained throughout the 60-min stimulation period. Insulin secretion stimulated by IBMX required a nonstimulatory level of glucose in the perifusing media, and, if this requirement was met, the immediate release of insulin was similar to that evoked by high glucose alone. High potassium (40 mM) also triggered a monophasic release of insulin. These studies demonstrate that glucose or high K+, which depolarizes the plasma membrane, and IBMX, an agent presumed to increase intracellular cyclic AMP levels, can signal the acute release of insulin from these beta cells. This cell line is a unique model system for studying the mechanism of insulin secretion.
CiteULike    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				G. C. Yaney, J. M. Fairbanks, J. T. Deeney, H. M. Korchak, K. Tornheim, and B. E. Corkey Potentiation of insulin secretion by phorbol esters is mediated by PKC-alpha and nPKC isoforms Am J Physiol Endocrinol Metab, November 1, 2002; 283(5): E880 - E888. [Abstract] [Full Text] [PDF]

				G. C. Yaney, H. M. Korchak, and B. E. Corkey Long-Chain Acyl CoA Regulation of Protein Kinase C and Fatty Acid Potentiation of Glucose-Stimulated Insulin Secretion in Clonal {beta}-Cells Endocrinology, June 1, 2000; 141(6): 1989 - 1998. [Abstract] [Full Text] [PDF]

				C. Schofl, J. Borger, T. Mader, M. Waring, A. von zur Muhlen, and G. Brabant Tolbutamide and diazoxide modulate phospholipase C-linked Ca2+ signaling and insulin secretion in beta -cells Am J Physiol Endocrinol Metab, April 1, 2000; 278(4): E639 - E647. [Abstract] [Full Text] [PDF]

				W. Zhang, A. Efanov, S.-N. Yang, G. Fried, S. Kolare, H. Brown, S. Zaitsev, P.-O. Berggren, and B. Meister Munc-18 Associates with Syntaxin and Serves as a Negative Regulator of Exocytosis in the Pancreatic beta -Cell J. Biol. Chem., December 22, 2000; 275(52): 41521 - 41527. [Abstract] [Full Text] [PDF]