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 the N terminus facing the cytoplasm and the C terminus residing in the synaptic vesicle lumen. Antibodies recognizing the C terminus of VGAT (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.
| Original language | English |
|---|---|
| Pages (from-to) | 13125-13131 |
| Number of pages | 7 |
| Journal | Journal of Neuroscience |
| Volume | 28 |
| Issue number | 49 |
| DOIs | |
| Publication status | Published - Dec 3 2008 |
Fingerprint
Keywords
- Endocytosis
- Fluorescence detection
- Live cell imaging
- Synaptic plasticity
- Synaptic vesicle
- Vesicular GABA transporter
ASJC Scopus subject areas
- Neuroscience(all)
- Medicine(all)
Cite this
Unique luminal localization of VGAT-C terminus allows for selective labeling of active cortical GABAergic synapses. / 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.
In: Journal of Neuroscience, Vol. 28, No. 49, 03.12.2008, p. 13125-13131.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Unique luminal localization of VGAT-C terminus allows for selective labeling of active cortical GABAergic synapses
AU - Martens, Henrik
AU - Weston, Matthew C.
AU - Boulland, Jean Luc
AU - Grønborg, Mads
AU - Grosche, Jens
AU - Kacza, Johannes
AU - Hoffmann, Anke
AU - Matteoli, Michela
AU - Takamori, Shigeo
AU - Harkany, Tibor
AU - Chaudhry, Farrukh A.
AU - Rosenmund, Christian
AU - Erck, Christian
AU - Jahn, Reinhard
AU - Härtig, Wolfgang
PY - 2008/12/3
Y1 - 2008/12/3
N2 - 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 the N terminus facing the cytoplasm and the C terminus residing in the synaptic vesicle lumen. Antibodies recognizing the C terminus of VGAT (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.
AB - 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 the N terminus facing the cytoplasm and the C terminus residing in the synaptic vesicle lumen. Antibodies recognizing the C terminus of VGAT (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.
KW - Endocytosis
KW - Fluorescence detection
KW - Live cell imaging
KW - Synaptic plasticity
KW - Synaptic vesicle
KW - Vesicular GABA transporter
UR - http://www.scopus.com/inward/record.url?scp=58149399397&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=58149399397&partnerID=8YFLogxK
U2 - 10.1523/JNEUROSCI.3887-08.2008
DO - 10.1523/JNEUROSCI.3887-08.2008
M3 - Article
C2 - 19052203
AN - SCOPUS:58149399397
VL - 28
SP - 13125
EP - 13131
JO - Journal of Neuroscience
JF - Journal of Neuroscience
SN - 0270-6474
IS - 49
ER -