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Calcium regulates exocytosis at the level of single vesicles

Nature Neuroscience volume 6, pages 846–853 (2003)Cite this article

Abstract

Ca2+ microdomains that form during the opening of voltage-gated Ca2+ channels have been implicated in regulating the kinetics of hormone and transmitter release. Direct assessment of the interaction between a single Ca2+ microdomain and a single secretory vesicle has been impossible because of technical limitations. Using evanescent field imaging of near-membrane micromolar Ca2+ concentration ([Ca2+]) and fluorescently labeled vesicles, we have observed exocytosis of individual chromaffin dense-core vesicles that was triggered by Ca2+ microdomains. Ca2+ microdomains selectively triggered the release of vesicles that were docked within 300 nm. Not all vesicles exposed to a Ca2+ microdomain were released, indicating that some vesicles are docked but are not ready for release. In addition to its established role as a trigger for release, elevated near-membrane [Ca2+] reduced the distance between docked vesicles and Ca2+ entry sites. Our results suggest a new mechanism for stimulation-dependent facilitation of exocytosis, whereby vesicles are moved closer to Ca2+ entry sites, thereby increasing a Ca2+ microdomain's efficacy to trigger vesicle fusion.

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Figure 1: Dual-color evanescent field imaging of individual vesicles and Ca2+ microdomains.
Figure 2: Ca2+ microdomains require voltage-gated Ca2+ influx.
Figure 3: A nearby Ca2+ microdomain triggers release.
Figure 4: Factors determining the efficacy of a Ca2+ microdomain for triggering release.
Figure 5: Stimulation increases the degree of colocalization.
Figure 6: Vesicles are recruited during episodes of high near-membrane [Ca2+].

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Acknowledgements

We thank J. B. Sørensen for chromaffin cells and carbon-fiber electrodes, B. Barbour, K. Delaney, E. Neher and members of our labs for comments on the manuscript. Supported by the Ministère de la Recherche (ACI 'young investigator group' 5242 to M.O.), the Max-Planck Society and SFB 406 (to W.S. and T.M.) and the Alexander-von-Humboldt Foundation (to U.B. and M.O.).

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  1. Ute Becherer

    Present address: Physiologisches Institut, Geb 59, Universität des Saarlandes, D-66421, Homburg/Saar, Germany

Authors and Affiliations

  1. Max-Planck Institute for Experimental Medicine, Molecular Biology of Neuronal Signals, Hermann-Rein Str. 3, Göttingen, D-37075, Germany

    Ute Becherer & Walter Stühmer

  2. Department of Otolaryngology, University of Göttingen, Robert-Koch Str. 40, Göttingen, D-37075, Germany

    Tobias Moser

  3. Ecole Supérieure de Physique et Chimie Industrielles (ESPCI), CNRS fre 2500 – INSERM epi 0002, Neurophysiologie et Nouvelles Microscopies, 10 rue Vauquelin, Paris, F-75005, France

    Martin Oheim

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Correspondence to Martin Oheim.

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Becherer, U., Moser, T., Stühmer, W. et al. Calcium regulates exocytosis at the level of single vesicles. Nat Neurosci 6, 846–853 (2003). https://doi.org/10.1038/nn1087

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