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Originally published In Press as doi:10.1074/jbc.C100465200 on September 10, 2001

J. Biol. Chem., Vol. 276, Issue 43, 40293-40297, October 26, 2001
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Mitochondrial [Ca2+] Oscillations Driven by Local High [Ca2+] Domains Generated by Spontaneous Electric Activity*,

Carlos Villalobos, Lucía Núñez, Pablo Chamero, María Teresa Alonso, and Javier García-SanchoDagger

From the Instituto de Biología y Genética Molecular (IBGM), Universidad de Valladolid and Consejo Superior de Investigaciones Científicas, Departamento de Fisiología y Bioquímica, Facultad de Medicina, E-47005 Valladolid, Spain

Mitochondria take up calcium during cell activation thus shaping Ca2+ signaling and exocytosis. In turn, Ca2+ uptake by mitochondria increases respiration and ATP synthesis. Targeted aequorins are excellent Ca2+ probes for subcellular analysis, but single-cell imaging has proven difficult. Here we combine virus-based expression of targeted aequorins with photon-counting imaging to resolve dynamics of the cytosolic, mitochondrial, and nuclear Ca2+ signals at the single-cell level in anterior pituitary cells. These cells exhibit spontaneous electric activity and cytosolic Ca2+ oscillations that are responsible for basal secretion of pituitary hormones and are modulated by hypophysiotrophic factors. Aequorin reported spontaneous [Ca2+] oscillations in all the three compartments, bulk cytosol, nucleus, and mitochondria. Interestingly, a fraction of mitochondria underwent much larger [Ca2+] oscillations, which were driven by local high [Ca2+] domains generated by the spontaneous electric activity. These oscillations were large enough to stimulate respiration, providing the basis for local tune-up of mitochondrial function by the Ca2+ signal.

* This work was supported by the Spanish Dirección General de Enseñanza Superior (DGES; Grants PB97-0474, APC1999-011, and 1FD97-1725-C02-02). C. Villalobos and L. Núñez hold postdoctoral fellowships from the Spanish DGES, and P. Chamero holds a predoctoral fellowship from the Basque Government.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.

The on-line version of this article (available at http://www.jbc.org) contains Movie 1 and Movie 2.

Dagger To whom correspondence should be addressed: IBGM, Dept. Fisiología, Facultad de Medicina, E-47005 Valladolid, Spain. Tel.: 34-983-423085; Fax: 34-983-423588; E-mail: jgsancho@ibgm.uva.es.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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