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J. Biol. Chem., Vol. 276, Issue 37, 34530-34536, September 14, 2001
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From the Rolf Luft Center for Diabetes Research, Department of
Molecular Medicine, Karolinska Institutet, Karolinska Hospital,
S-171 76 Stockholm, Sweden
Using dual excitation and fixed emission
fluorescence microscopy, we were able to measure changes in cytoplasmic
free Ca2+ concentration
([Ca2+]i) and mitochondrial membrane potential
simultaneously in the pancreatic
Glucose-induced Oscillations in Cytoplasmic Free Ca2+
Concentration Precede Oscillations in Mitochondrial Membrane Potential
in the Pancreatic
-Cell*
§,,
-cell. The -cells were exposed
to a combination of the Ca2+ indicator fura-2/AM and
the indicator of mitochondrial membrane potential, rhodamine 123 (Rh123). Using simultaneous measurements of mitochondrial membrane
potential and [Ca2+]i during glucose stimulation,
it was possible to measure the time lag between the onset of
mitochondrial hyperpolarization and changes in
[Ca2+]i. Glucose-induced oscillations in
[Ca2+]i were followed by transient
depolarizations of mitochondrial membrane potential. These results are
compatible with a model in which nadirs in
[Ca2+]i oscillations are generated by a
transient, Ca2+-induced inhibition of mitochondrial
metabolism resulting in a temporary fall in the cytoplasmic ATP/ADP
ratio, opening of plasma membrane KATP channels,
repolarization of the plasma membrane, and thus transient closure of
voltage-gated L-type Ca2+ channels.
*
Financial support was obtained from the Swedish Medical
Research Council (Grants 03X-09890, 03XS-12708, 03X-09891, and
19X-00034), the Swedish Diabetes Association, the Juvenile Diabetes
Foundation International, the Nordic Insulin Foundation, the Novo
Nordisk Foundation, Funds of Karolinska Institutet, Clas Groschinskys Memorial Foundation, Magnus Bergvall's Foundation, funds of the Swedish Society of Medicine, Fredrik och Ingrid Thurings Stiftelse, United States Public Health Service Grant DK-35 914, Berth von Kantzows Foundation, and Tore Nilsson's Foundation.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.
These authors contributed equally to this study.
§
To whom correspondence should be addressed. Tel.:
46-8-5177-5731; Fax: 46-8-517-79450; E-mail:
henrik.kindmark@ks.se.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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