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Original Articles

In Situ Characterization of Nonmitochondrial Ca2+ Stores in Individual Pancreatic β-Cells

  1. Anders Tengholm,
  2. Charlotte Hagman,
  3. Erik Gylfe and
  4. Bo Hellman
  1. Department of Medical Cell Biology, Uppsala University Uppsala, Sweden
  1. Address correspondence and reprint requests to Prof. Bo Hellman, Department of Medical Cell Biology, Biomedicum, Box 571, S-751 23 Uppsala, Sweden. E-mail: bo.hellman{at}medcellbiol.uu.se
Diabetes 1998 Aug; 47(8): 1224-1230. https://doi.org/10.2337/diab.47.8.1224
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Abstract

Free Ca2+ was measured in intracellular stores of individual mouse pancreatic β-cells using dual-wavelength microfluorometry and the low-affinity Ca2+ indicator furaptra. Controlled permeabilization of the plasma membrane with 4 umol/1 digitonin revealed that 22% of the furaptra was trapped in intracellular nonnuclear compartments. When 3 mmol/l ATP and 200 nmol/1 Ca2+ were simultaneously present, this cation rapidly accumulated in the organelle pool, reaching an average concentration of 200–500 umol/1. Whereas agents affecting the mitochondrial function (5 mmol/l succinate, 2 umol/1 ruthenium red, or 10 umol/1 antimycin A + 2 ug/ml oligomycin) had little effects, the Ca2+-ATPase inhibitor thapsigargin released 92% of the Ca2+ mobilizable with the ionophore Br-A23187. Digital imaging revealed regional differences in the organelle Ca2+. The regions with the highest Ca2+ concentration were particularly responsive to inositol 1,4,5-trisphosphate (IP3). IP3 mobilized Ca2+ in a dose-dependent way with half-maximal and maximal effects at about 1 and 5 umol/1, respectively. High concentrations of IP3 released about half of the thapsigargin-sensitive Ca2+, but there were no responses to agents known to activate ryanodine receptors, such as 10 mmol/l caffeine, 0.1–1 umol/1 ryanodine, or 1–5 umol/1 cyclic ADP ribose. The results reinforce the concept that mobilization of intracellular Ca2+ in the pancreatic β-cell is mediated by IP3 receptors rather than ryanodine receptors.

  • Received January 20, 1998.
  • Revision received May 6, 1998.
  • Accepted May 6, 1998.
  • Copyright © 1998 by the American Diabetes Association

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August 1998, 47(8)
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In Situ Characterization of Nonmitochondrial Ca2+ Stores in Individual Pancreatic β-Cells
Anders Tengholm, Charlotte Hagman, Erik Gylfe, Bo Hellman
Diabetes Aug 1998, 47 (8) 1224-1230; DOI: 10.2337/diab.47.8.1224

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In Situ Characterization of Nonmitochondrial Ca2+ Stores in Individual Pancreatic β-Cells
Anders Tengholm, Charlotte Hagman, Erik Gylfe, Bo Hellman
Diabetes Aug 1998, 47 (8) 1224-1230; DOI: 10.2337/diab.47.8.1224
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