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J Biol Chem, Vol. 274, Issue 52, 36883-36890, December 24, 1999

Glucose Regulation of Free Ca2+ in the Endoplasmic Reticulum of Mouse Pancreatic Beta Cells*

Anders Tengholm, Bo Hellman, and Erik GylfeDagger

From the Department of Medical Cell Biology, Uppsala University, SE-751 23 Uppsala, Sweden

Free Ca2+ was measured in organelles of individual mouse pancreatic beta cells loaded with the low affinity indicator furaptra. After removal of cytoplasmic indicator by controlled digitonin permeabilization the organelle Ca2+ was located essentially in the endoplasmic reticulum (ER), >90% being sensitive to inhibition of sarco(endo)plasmic reticulum Ca2+-ATPases. The Ca2+ accumulation in the ER of intact beta cells depended in a hyperbolic fashion on the glucose concentration with half-maximal and maximal filling at 5.5 and >20 mM, respectively. Also elevation of cytoplasmic Ca2+ by K+ depolarization significantly enhanced the Ca2+ accumulation. In permeabilized beta cells 1-3 mM ATP caused rapid Ca2+ filling of the ER reaching almost 500 µM. At 50 nM, Ca2+ ER became half-maximally filled at 45 µM ATP, whereas only 3.5 µM ATP was required at 200 nM Ca2+. Inositol 1,4,5-trisphosphate induced a rapid release of about 65% of the ER Ca2+, and its precursor phosphatidylinositol 4,5-bisphosphate was found to slowly mobilize 75% by another mechanism. It is concluded that glucose is an efficient stimulator of Ca2+ uptake in the ER of pancreatic beta cells both by increasing ATP and cytoplasmic Ca2+. Because physiological concentrations of cytoplasmic ATP are in the mM range, Ca2+ sequestration can be anticipated to be modulated by factors reducing its ATP sensitivity.

* This work was supported by Grants 12X-562 and 12X-6240 from the Swedish Medical Research Council, the Swedish Foundation for Strategic Research, the Swedish Diabetes Association, the Novo Nordisk Foundation, Novo Nordisk Pharma AB, Family Ernfors' Foundation, Åke Wiberg's Foundation, and the Swedish Society for Medical Research.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: Dept. of Medical Cell Biology, Uppsala University, Biomedicum, Box 571, SE-751 23 Uppsala, Sweden. Tel.: 46 18 471 44 28; Fax: 46 18 471 40 59; E-mail: erik.gylfe@medcellbiol.uu.se.

Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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