Abstract
Synaptic transmission involves the regulated exocytosis of vesicles filled with neurotransmitter. Classical transmitters are synthesized in the cytoplasm, and so must be transported into synaptic vesicles. Although the vesicular transporters for monoamines and acetylcholine have been identified, the proteins responsible for packaging the primary inhibitory and excitatory transmitters, γ-aminobutyric acid (GABA) and glutamate remain unknown1,2. Studies in the nematode Caenorhabditis elegans have implicated the gene unc-47 in the release of GABA3. Here we show that the sequence of unc-47 predicts a protein with ten transmembrane domains, that the gene is expressed by GABA neurons, and that the protein colocalizes with synaptic vesicles. Further, a rat homologue of unc-47 is expressed by central GABA neurons and confers vesicular GABA transport in transfected cells with kinetics and substrate specificity similar to those previously reported for synaptic vesicles from the brain. Comparison of this vesicular GABA transporter (VGAT) with a vesicular transporter for monoamines shows that there are differences in the bioenergetic dependence of transport, and these presumably account for the differences in structure. Thus VGAT is the first of a new family of neurotransmitter transporters.
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Acknowledgements
We thank B. Westlund for unpublished unc-47 map data; E. King for help with the confocal microscope; H. Rausch and K. Knobel for integrating the unc-47::GFP transcriptional fusion construct; Y. Jin for the n2476 allele; D. P. Morse and B. Bamber for the gift of RNA and cDNA; R. Barstead and P. Okkema for the C. elegans cDNA libraries; J. Boulter and D. Julius for the mammalian cDNA libraries; D. Rice and D. Eisenberg for the analysis of hydrophobic moment; A. Tobin and S. Baekkeskov for the GAD-67 cDNA; Y. Liu and P. Tan for assistance in the isolation of PC12 cell clones and the preparation of membranes; J. Hell and P. Finn for suggestions about the measurement of transport activity; S. Craven and D. Bredt for assistance with the primary hippocampal cultures; and M. Horner, E.Kofoid, C. Bargmann and members of the Edwards and Jorgensen laboratories for discussions. This work was supported by the Gallo Center (S.L.M.), the Giannini Foundation (R.J.R.), an NIH Developmental Biology training grant (K.S.), the Klingenstein Foundation (E.M.J.), NINDS (S.L.M., R.H.E., E.M.J.) and NIMH (R.H.E.).
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McIntire, S., Reimer, R., Schuske, K. et al. Identification and characterization of the vesicular GABA transporter. Nature 389, 870–876 (1997). https://doi.org/10.1038/39908
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DOI: https://doi.org/10.1038/39908
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