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- Title
Unique Luminal Localization of VGAT-C Terminus Allows for Selective Labeling of Active Cortical GABAergic Synapses.
- Authors
Martens, Henrik; Weston, Matthew C.; Boulland, Jean-Luc; Grønborg, Mads; Grosche, Jens; Kacza, Johannes; Hoffmann, Anke; Matteoli, Michela; Takamori, Shigeo; Harkany, Tibor; Chaudhry, Farrukh A.; Rosenmund, Christian; Erck, Christian; Jahn, Reinhard; Härtig, Wolfgang
- Abstract
Neurotransmitter uptake into synaptic vesicles is mediated by vesicular neurotransmitter transporters. Although these transporters belong to different families, they all are thought to share a common overall topology with an even number of transmembrane domains. Using epitope-specific antibodies and mass spectrometry we show that the vesicular GABA transporter (VGAT) possesses an uneven number of transmembrane domains, with theNterminus facing the cytoplasm and the C terminus residing in the synaptic vesicle lumen. Antibodies recognizing the C terminus ofVGAT(anti-VGAT-C) selectively label GABAergic nerve terminals of live cultured hippocampal and striatal neurons as confirmed by immunocytochemistry and patch-clamp electrophysiology. Injection of fluorochromated anti- VGAT-C into the hippocampus of mice results in specific labeling of GABAergic synapses in vivo. Overall, our data open the possibility of studying novel GABA release sites, characterizing inhibitory vesicle trafficking, and establishing their contribution to inhibitory neurotransmission at identified GABAergic synapses.
- Subjects
NEUROTRANSMITTERS; NEURAL transmission; MASS spectrometry; IMMUNOGLOBULINS; ELECTROPHYSIOLOGY; IMMUNOCYTOCHEMISTRY
- Publication
Journal of Neuroscience, 2008, Vol 28, Issue 49, p13125
- ISSN
0270-6474
- Publication type
Article
- DOI
10.1523/JNEUROSCI.3887-08.2008