Abstract
Astrocytes establish rapid cell-to-cell communication through the release of chemical transmitters. The underlying mechanisms and functional significance of this release are, however, not well understood. Here we identify an astrocytic vesicular compartment that is competent for glutamate exocytosis. Using postembedding immunogold labeling of the rat hippocampus, we show that vesicular glutamate transporters (VGLUT1/2) and the vesicular SNARE protein, cellubrevin, are both expressed in small vesicular organelles that resemble synaptic vesicles of glutamatergic terminals. Astrocytic vesicles, which are not as densely packed as their neuronal counterparts, can be observed in small groups at sites adjacent to neuronal structures bearing glutamate receptors. Fluorescently tagged VGLUT-containing vesicles were studied dynamically in living astrocytes by total internal reflection fluorescence (TIRF) microscopy. After activation of metabotropic glutamate receptors, astrocytic vesicles underwent rapid (milliseconds) Ca2+- and SNARE-dependent exocytic fusion that was accompanied by glutamate release. These data document the existence of a Ca2+-dependent quantal glutamate release activity in glia that was previously considered to be specific to synapses.
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Acknowledgements
We thank H. Stubbe, Y. Gomez, K. Hüttmann and Centre de Microscopie Electronique, University of Lausanne for experimental support; J.-Y. Chatton, T. Coppola, P. Jourdain, G. Knott, T. Lang and R. Stoop for scientific discussions; R. Jahn, J. Storm-Mathisen for insights at various stages of this work and P. Clarke, J. Meldolesi, R. Regazzi and J. Storm-Mathisen for comments on the manuscript. This work was supported by grants OFES 00.0553 and FNRS 3100A0-100850/1 to A.V. and by Deutsche Forschungsgemeinschaft (SFB-TR3) and Fonds der Chemischen Industrie to C.S. V.G. is a visiting fellow within the European Community grant QLG3-CT-2001–2004 and recipient of a fellowship from the Norwegian Research Council.
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Supplementary Fig. 1
Cumulative number of AO flashes plotted against time during a 3 min application of LTx (12 nM; n = 20; Supplementary Fig. 1a) or IONO (10 μM; n = 38; Supplementary Fig. 1b). (JPG 27 kb)
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Bezzi, P., Gundersen, V., Galbete, J. et al. Astrocytes contain a vesicular compartment that is competent for regulated exocytosis of glutamate. Nat Neurosci 7, 613–620 (2004). https://doi.org/10.1038/nn1246
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DOI: https://doi.org/10.1038/nn1246
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