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J Biol Chem, Vol. 274, Issue 52, 36883-36890, December 24, 1999
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.
Glucose Regulation of Free Ca2+ in the Endoplasmic
Reticulum of Mouse Pancreatic Beta Cells*
*
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.
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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