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The Journal of Neuroscience, November 15, 1999, 19(22):9803-9812
Subcellular Localization of Tetanus Neurotoxin-Insensitive Vesicle-Associated Membrane Protein (VAMP)/VAMP7 in Neuronal Cells: Evidence for a Novel Membrane Compartment
Silvia Coco2, Graca Raposo1, Sonia Martinez1, Jean-Jacques Fontaine4, Shigeo Takamori3, Ahmed Zahraoui1, Reinhard Jahn3, Michela Matteoli2, Daniel Louvard1, and Thierry Galli1 1 Centre National de la Recherche Scientifique, Unité Mixte de Recherche 144, Compartimentation et Dynamique Cellulaires, Institut Curie, F-75248 Paris CEDEX 05, France, 2 Consiglio Nazionale delle Ricerche, Center of Cellular and Molecular Pharmacology and B. Ceccarelli Center, 20129 Milano, Italy, 3 Department of Neurobiology, Max-Planck-Institute for Biophysical Chemistry, Am Faßberg, D-37077 Göttingen, Germany, and 4 Laboratoire d'Anatomie Pathologique, Ecole Vétérinaire d'Alfort, F-94704 Maisons Alfort CEDEX, France
The clostridial neurotoxin-insensitive soluble N-ethylmaleimide-sensitive factor attachment protein (SNAP) receptors, tetanus neurotoxin-insensitive (TI)-vesicle-associated membrane protein (VAMP)/VAMP7, SNAP23, and syntaxin 3 have recently been implicated in transport of exocytotic vesicles to the apical plasma membrane of epithelial cells. This pathway had been shown previously to be insensitive to tetanus neurotoxin and botulinum neurotoxin F. TI-VAMP/VAMP7 is also a good candidate to be implicated in an exocytotic pathway involved in neurite outgrowth because tetanus neurotoxin does not inhibit this process in conditions in which it abolishes neurotransmitter release. We have now found that TI-VAMP/VAMP7 has a widespread distribution in the adult rat brain in which its localization strikingly differs from that of nerve terminal markers. TI-VAMP/VAMP7 does not enrich in synaptic vesicles nor in large dense-core granules but is associated with light membranes. In hippocampal neurons developing in vitro, TI-VAMP/VAMP7 localizes to vesicles in the axonal and dendritic outgrowths and concentrates into the leading edge of the growth cone, a region devoid of synaptobrevin 2, before synaptogenesis. After the onset of synaptogenesis, TI-VAMP/VAMP7 is found predominantly in the somatodendritic domain. In PC12 cells, TI-VAMP/VAMP7 does not colocalize with synaptobrevin 2, chromogranin B, or several markers of endocytic compartments. At the electron microscopic level, TI-VAMP/VAMP7 is mainly associated with tubules and vesicles. Altogether, these results suggest that TI-VAMP/VAMP7 defines a novel membrane compartment in neurite outgrowths and in the somatodendritic domain.
Key words: exocytosis; clostridial neurotoxin; neurite outgrowth; SNARE; TI-VAMP/VAMP7; synaptobrevin 2
Copyright © 1999 Society for Neuroscience 0270-6474/99/19229803-10$05.00/0
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