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

FEBS Lett. 1998 Feb 6;422(3):339-42. doi: 10.1016/s0014-5793(98)00036-2.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • ATP-Binding Cassette Transporters / biosynthesis*
  • ATP-Binding Cassette Transporters / genetics
  • Amino Acid Transport System X-AG
  • Animals
  • Biological Transport
  • CHO Cells
  • Cell Size
  • Cricetinae
  • Doxycycline / pharmacology
  • Glutamic Acid / metabolism*
  • Sodium / metabolism
  • Transfection

Substances

  • ATP-Binding Cassette Transporters
  • Amino Acid Transport System X-AG
  • Glutamic Acid
  • Sodium
  • Doxycycline