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Identification of a vesicular glutamate transporter that defines a glutamatergic phenotype in neurons

Nature volume 407, pages 189194 (14 September 2000) | Download Citation

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Abstract

Glutamate is the major excitatory neurotransmitter in the mammalian central nervous system. Synaptic vesicles are loaded with neurotransmitter by means of specific vesicular transporters. Here we show that expression of BNPI, a vesicle-bound transporter associated with sodium-dependent phosphate transport1,2,3, results in glutamate uptake by intracellular vesicles. Substrate specificity and energy dependence are very similar to glutamate uptake by synaptic vesicles. Stimulation of exocytosis—fusion of the vesicles with the cell membrane and release of their contents—resulted in quantal release of glutamate from BNPI-expressing cells. Furthermore, we expressed BNPI in neurons containing GABA (γ-aminobutyric acid) and maintained them as cultures of single, isolated neurons that form synapses to themselves. After stimulation of these neurons, a component of the postsynaptic current is mediated by glutamate as it is blocked by a combination of the glutamate receptor antagonists, but is insensitive to a GABAA receptor antagonist. We conclude that BNPI functions as vesicular glutamate transporter and that expression of BNPI suffices to define a glutamatergic phenotype in neurons.

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Acknowledgements

We thank J. Rettig for discussions and the help in the viral infection technique; D. Pommereit and Y. Stern-Bach for providing the non-desensitizing AMPA-receptor vectors GluR1L497Y and GluR2QL504Y–IRES–DsRed; M. Druminski, D. Diezmann, A. Bührmann, I. Herfort and N. Narajagan for their technical assistance; P. Holroyd for critical reading of this manuscript. We also thank The HHMI Biopolymer/W.M. Keck Foundation, Biotechnology Resource Laboratory at Yale University for amino-acid sequencing.

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  1. *Department of Neurobiology and

    • Shigeo Takamori
    •  & Reinhard Jahn
  2. †Department of Membrane Biophysics, Max-Planck-Institute for Biophysical Chemistry, Am Faβberg 11, D-37077 Göttingen, Germany

    • Jeong Seop Rhee
    •  & Christian Rosenmund

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Correspondence to Reinhard Jahn.

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https://doi.org/10.1038/35025070

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