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Louis-Jeantet Research Laboratories (V.L., K.R., P.A.H., J.-C.I.) and Department of Physiology (A.M.), University Medical Center, 1211 Geneva 4, Switzerland; and Department of Biochemistry and Molecular Biology and the Howard Hughes Medical Institute, University of Chicago (G.W., D.F.S.), Chicago, Illinois 60637
Address all correspondence and requests for reprints to: Dr. Valérie Lilla, Laboratoires de Recherche Louis-Jeantet, Centre Médical Universitaire, 1 rue Michel Servet, 1211 Geneva 4, Switzerland. E-mail: valerie.lilla{at}medecine.unige.ch.
To identify genes involved in regulated insulin secretion, we have established and characterized two sublines derived from the mouse pancreatic ß-cell line MIN6, designated B1 and C3. They have a similar insulin content, but differ in their secretory properties. B1 responded to glucose in a concentration- and cell confluence-dependent manner, whereas C3 did not. B1 cells were stimulated by phorbol 12-myristate 13-acetate, leucine, arginine, glibenclamide, isobutylmethylxanthine, and KCl, whereas C3 did not respond (leucine, arginine, and glibenclamide) or responded to a lesser extent (isobutylmethylxanthine, phorbol 12-myristate 13-acetate, and KCl). Although intracellular Ca2+ rose in response to glucose in B1 but not C3 cells, KCl increased intracellular Ca2+ in a similar manner in both sublines. GLUT-1, GLUT-2, Kir6.2, and SUR1 expression was not significantly different between B1 and C3 cells, whereas E-cadherin was more abundantly expressed in B1 cells. A more complete list of differentially expressed genes was established by suppression subtractive hybridization and high density (Affymetrix) oligonucleotide microarrays. Genes were clustered according to known or putative function. Those involved in metabolism, intracellular signaling, cytoarchitecture, and cell adhesion are of potential interest. These two sublines should be useful for identification of the genes and mechanisms involved in regulated insulin secretion of the pancreatic ß-cell.
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