Identification and characterization of a cDNA encoding a neuronal glutamate transporter from Drosophila melanogaster

Rebecca P. Seal, Gwynn M. Daniels, William J. Wolfgang, Michael Forte, Susan G. Amara

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

Sodium-dependent glutamate transporters influence neurotransmission in the central nervous system by removing synaptically released glutamate from the extracellular space and by maintaining extracellular glutamate concentrations below neurotoxic levels. In insects, glutamate also serves as the neurotransmitter at the neuromuscular junction, but the mechanism for neurotransmitter clearance at this synapse has not been well-established. Here we report the cloning and characterization of a sodium-dependent glutamate transporter, dEAAT, from Drosophila melanogaster. The 479 amino acid dEAAT gene product is 40-50% homologous to mammalian members of this carrier family. A 3.3 kilobase (kb) transcript for dEAAT was detected in adult fly heads and to a lesser extent in bodies by Northern-blot analysis and was also localized to neurons in the central nervous system by in situ hybridization. The transport activity observed following expression of dEAAT in Xenopus oocytes or COS-7 cells shows a high affinity for L-glutamate, L-aspartate and D-aspartate, an absolute dependence on external sodium ions, and considerable stereoselectivity for the transport of L-glutamate over D-glutamate. As has been observed for the human carriers, EAAT 4 and EAAT 5, a significant component of the current activated by L-glutamate application to dEAAT-expressing oocytes appears to arise from the activation of a chloride channel associated with the carrier.

Original languageEnglish (US)
Pages (from-to)51-64
Number of pages14
JournalReceptors and Channels
Volume6
Issue number1
StatePublished - 1998

Fingerprint

Amino Acid Transport System X-AG
Drosophila melanogaster
Glutamic Acid
Complementary DNA
Glutamate Plasma Membrane Transport Proteins
Sodium
Neurology
Oocytes
Neurotransmitter Agents
Central Nervous System
D-Aspartic Acid
Stereoselectivity
Chloride Channels
Neuromuscular Junction
Cloning
COS Cells
Extracellular Space
Xenopus
Aspartic Acid
Synaptic Transmission

Keywords

  • Chloride conductance
  • Drosophila
  • Glutamate
  • Transport

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Endocrinology
  • Cell Biology

Cite this

Identification and characterization of a cDNA encoding a neuronal glutamate transporter from Drosophila melanogaster. / Seal, Rebecca P.; Daniels, Gwynn M.; Wolfgang, William J.; Forte, Michael; Amara, Susan G.

In: Receptors and Channels, Vol. 6, No. 1, 1998, p. 51-64.

Research output: Contribution to journalArticle

Seal, Rebecca P. ; Daniels, Gwynn M. ; Wolfgang, William J. ; Forte, Michael ; Amara, Susan G. / Identification and characterization of a cDNA encoding a neuronal glutamate transporter from Drosophila melanogaster. In: Receptors and Channels. 1998 ; Vol. 6, No. 1. pp. 51-64.
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