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Chromaffin-cell stimulation triggers fast millimolar mitochondrial Ca2+ transients that modulate secretion

Nature Cell Biology volume 2, pages 57–61 (2000)Cite this article

Abstract

Activation of calcium-ion (Ca2+) channels on the plasma membrane and on intracellular Ca2+ stores, such as the endoplasmic reticulum, generates local transient increases in the cytosolic Ca2+ concentration that induce Ca2+ uptake by neighbouring mitochondria. Here, by using mitochondrially targeted aequorin proteins with different Ca2+ affinities, we show that half of the chromaffin-cell mitochondria exhibit surprisingly rapid millimolar Ca2+ transients upon stimulation of cells with acetylcholine, caffeine or high concentrations of potassium ions. Our results show a tight functional coupling of voltage-dependent Ca2+ channels on the plasma membrane, ryanodine receptors on the endoplasmic reticulum, and mitochondria. Cell stimulation generates localized Ca2+ transients, with Ca2+ concentrations above 20–40 µM, at these functional units. Protonophores abolish mitochondrial Ca2+ uptake and increase stimulated secretion of catecholamines by three- to fivefold. These results indicate that mitochondria modulate secretion by controlling the availability of Ca2+ for exocytosis.

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Figure 1: Effects of high K+ concentrations or caffeine on [Ca2+]M, measured with AEQ1 or AEQ2.
Figure 2: Effects of high K+ concentrations or caffeine on [Ca2+]M measured with AEQ3.
Figure 3: Effects of CGP37157, an inhibitor of the mitochondrial Na+/Ca2+ exchanger, and temperature on the rate of [Ca2+]M decrease after stimulation.
Figure 4: Dependence of the rate of mitochondrial Ca2+ uptake on [Ca2+]c.
Figure 5: CCCP enhances secretory responses evoked by acetylcholine, high K+ concentration or caffeine.
Figure 6: The complex functional unit responsible for the generation of local high [Ca2+]c transients and catecholamine secretion in a bovine chromaffin cell.

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Acknowledgements

We acknowledge financial support from the Dirección General de Enseñanza Superior (grant PM98/0142 to J.A. and grant PB97/0474 to J.G.-S.), from the Dirección General de Investigación Científica y Técnica (grant PB94/0150) and from Janssen-Cilag to A.G.G., and from Junta de Castilla y León (grant VA19/99 to J.A. and grant VA62/99 to M.T.A.). I.C.-I. and A.A. hold fellowships from the Ministerio de Educación y Ciencia. We thank C. González and T. Pozzan for helpful comments, and J. Fernández for technical assistance.

Correspondence and requests for materials should be addressed to J.A.

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Authors and Affiliations

  1. Departamento de Bioquímica y Biología Molecular y Fisiología, Instituto de Biología y Genética Molecular (IBGM), Universidad de Valladolid y Consejo Superior de Investigaciones Científicas (CSIC), Facultad de Medicina, Ramón y Cajal 7, Valladolid, E-47005 , Spain

    Mayte Montero, Maria Teresa Alonso, Estela Carnicero, Javier García-Sancho & Javier Alvarez

  2. Departamento de Farmacología y Terapéutica, Instituto de Farmacología Teófilo Hernando, Facultad de Medicina, Universidad Autónoma de Madrid, Arzobispo Morcillo 4, Madrid, E-28029 , Spain

    Inmaculada Cuchillo-Ibáñez, Almudena Albillos & Antonio G. García

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  1. Mayte Montero
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Correspondence to Javier Alvarez.

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Montero, M., Alonso, M., Carnicero, E. et al. Chromaffin-cell stimulation triggers fast millimolar mitochondrial Ca2+ transients that modulate secretion. Nat Cell Biol 2, 57–61 (2000). https://doi.org/10.1038/35000001

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