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The 'glial' glutamate transporter, EAAT2 (Glt-1) accounts for high affinity glutamate uptake into adult rodent nerve endings

Suchak, S., Baloyianni, N., Perkinton, M., Williams, R., Meldrum, B. and Rattray, M. (2003) The 'glial' glutamate transporter, EAAT2 (Glt-1) accounts for high affinity glutamate uptake into adult rodent nerve endings. Journal of Neurochemistry, 84 (3). pp. 522-532. ISSN 0022-3042

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To link to this article DOI: 10.1046/j.1471-4159.2003.01553.x

Abstract/Summary

The excitatory amino acid transporters (EAAT) removes neurotransmitters glutamate and aspartate from the synaptic cleft. Most CNS glutamate uptake is mediated by EAAT2 into glia, though nerve terminals show evidence for uptake, through an unknown transporter. Reverse-transcriptase PCR identified the expression of EAAT1, EAAT2, EAAT3 and EAAT4 mRNAs in primary cultures of mouse cortical or striatal neurones. We have used synaptosomes and glial plasmalemmal vesicles (GPV) from adult mouse and rat CNS to identify the nerve terminal transporter. Western blotting showed detectable levels of the transporters EAAT1 (GLAST) and EAAT2 (Glt-1) in both synaptosomes and GPVs. Uptake of [3H]D-aspartate or [3H]L-glutamate into these preparations revealed sodium-dependent uptake in GPV and synaptosomes which was inhibited by a range of EAAT blockers: dihydrokainate, serine-o-sulfate, l-trans-2,4-pyrrolidine dicarboxylate (PDC) (+/-)-threo-3-methylglutamate and (2S,4R )-4-methylglutamate. The IC50 values found for these compounds suggested functional expression of the 'glial, transporter, EAAT2 in nerve terminals. Additionally blockade of the majority EAAT2 uptake sites with 100 micro m dihydrokainate, failed to unmask any functional non-EAAT2 uptake sites. The data presented in this study indicate that EAAT2 is the predominant nerve terminal glutamate transporter in the adult rodent CNS.

Item Type:Article
Refereed:Yes
Divisions:Faculty of Life Sciences > School of Chemistry, Food and Pharmacy > School of Pharmacy > Division of Pharmacology
ID Code:15730
Uncontrolled Keywords:*Amino Acid Transport System X-AG Animals Biological Transport/physiology Blotting, Western Brain/cytology/metabolism Cell Membrane/chemistry/metabolism Cells, Cultured Excitatory Amino Acid Transporter 1/biosynthesis/genetics Excitatory Amino Acid Transporter 2/genetics/*metabolism Excitatory Amino Acid Transporter 3 Excitatory Amino Acid Transporter 4 Glutamate Plasma Membrane Transport Proteins Glutamic Acid/*metabolism/pharmacokinetics Mice Nerve Endings/chemistry/*metabolism Neuroglia/chemistry/metabolism Neurons/chemistry/cytology/metabolism RNA, Messenger/metabolism Receptors, Glutamate/biosynthesis/genetics Sodium/metabolism *Symporters Synaptosomes/chemistry/metabolism
Additional Information:Suchak, Sachin K Baloyianni, Nicoletta V Perkinton, Michael S Williams, Robert J Meldrum, Brian S Rattray, Marcus Research Support, Non-U.S. Gov't England Journal of neurochemistry J Neurochem. 2003 Feb;84(3):522-32.
Publisher:Wiley-Blackwell

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