Inducible expression of the GLT-1 glutamate transporter in a CHO cell line selected for low endogenous glutamate uptake

Inducible expression of the mammalian glial cell glutamate transporter GLT-1 has been established in a CHO cell line selected for low endogenous Na +-dependent glutamate uptake by [ 3H]aspartate suicide selection. Culturing the cells in doxycycline-containing medium, to activate GLT-1 expression via...

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Bibliographic Details
Published in:FEBS letters 1998-02, Vol.422 (3), p.339-342
Main Authors: Levy, Line M, Attwell, David, Hoover, Frank, Ash, John F, Bjørås, Magnar, Danbolt, Niels Christian
Format: Article
Language:English
Subjects:
Amino Acid Transport System X-AG
Animals
ATP-Binding Cassette Transporters - biosynthesis
ATP-Binding Cassette Transporters - genetics
Biological Transport
Cell Size
CHO cell
CHO Cells
Cricetinae
Doxycycline - pharmacology
Glutamate
Glutamate transporter
Glutamic Acid - metabolism
Inducible expression
Sodium - metabolism
Transfection
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Summary:Inducible expression of the mammalian glial cell glutamate transporter GLT-1 has been established in a CHO cell line selected for low endogenous Na +-dependent glutamate uptake by [ 3H]aspartate suicide selection. Culturing the cells in doxycycline-containing medium, to activate GLT-1 expression via the Tet-On system, increased uptake of the GLT-1 substrate d-aspartate 280-fold, and increased cell size. Applying glutamate to whole-cell clamped, doxycycline-treated cells evoked a transporter-mediated current with characteristics appropriate for GLT-1. This cell line provides a useful tool for further examination of the electrical, biochemical and pharmacological properties of GLT-1, the most abundant glutamate transporter in the brain.
ISSN:0014-5793
1873-3468
DOI:10.1016/S0014-5793(98)00036-2