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Journal of Pharmacology And Experimental Therapeutics Fast Forward
First published on January 21, 2003; DOI: 10.1124/jpet.102.046334

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Vol. 305, Issue 1, 271-278, April 2003

Cav1.3 Is Preferentially Coupled to Glucose-Stimulated Insulin Secretion in the Pancreatic beta -Cell Line INS-1

Guohong Liu , Nejmi Dilmac, Nathan Hilliard and Gregory H. Hockerman

Department of Medicinal Chemistry and Molecular Pharmacology (G.L., N.D., N.H., G.H.H.) and Neuroscience Graduate Program (G.L.), Purdue University, West Lafayette, Indiana

L-Type Ca2+ channel blockers inhibit glucose and KCl-stimulated insulin secretion by pancreatic beta  cells. However, the role of the two distinct L-type channels expressed by beta  cells, Cav1.2 and Cav1.3, in this process is not clear. Therefore, we stably transfected INS-1 cells with two mutant channel constructs, Cav1.2DHPi or Cav1.3 DHPi. Whole-cell patch-clamp recordings demonstrated that both mutant channels are insensitive to dihydropyridines (DHPs), but are blocked by diltiazem. INS-1 cells expressing Cav1.3/DHPi maintained glucose- and KCl-stimulated insulin secretion in the presence of DHPs, whereas cells expressing Cav1.2/DHPi demonstrated DHP resistance to only KCl-induced secretion. INS-1 cells were also stably transfected with cDNAs encoding the intracellular loop between domains II and III of either Cav1.2 or Cav1.3 (Cav1.2/II-III or Cav1.3/II-III). Glucose- and KCl-stimulated insulin secretion in Cav1.2/II-III cells were not different from untransfected INS-1 cells. However, glucose-stimulated insulin secretion was completely inhibited and KCl-stimulated secretion was substantially resistant to inhibition by DHPs, but sensitive to omega -agatoxin IVA in Cav1.3/II-III cells. Moreover, the L-type channel agonist FPL 64176 markedly enhanced KCl-stimulated secretion by Cav1.3/II-III cells. Together, our results suggest that Ca2+ influx via Cav1.3 is preferentially coupled to glucose-stimulated insulin secretion in INS-1 cells.

0022-3565/03/3051-0271$07.00/0
THE JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS
Copyright © 2003 by The American Society for Pharmacology and Experimental Therapeutics


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