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 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.
Levy
L M
LM
Department of Anatomy, University of Oslo, Norway.
Attwell
D
D
Hoover
F
F
Ash
J F
JF
Bjørås
M
M
Danbolt
N C
NC
D018528:ATP-Binding Cassette Transporters / Q000096:biosynthesis* / Q000235:genetics; D027322:Amino Acid Transport System X-AG; D000818:Animals; D001692:Biological Transport; D016466:CHO Cells; D048429:Cell Size; D006224:Cricetinae; D004318:Doxycycline / Q000494:pharmacology; D018698:Glutamic Acid / Q000378:metabolism*; D012964:Sodium / Q000378:metabolism; D014162:Transfection
D018528:ATP-Binding Cassette Transporters; D027322:Amino Acid Transport System X-AG; D018698:Glutamic Acid; D012964:Sodium; D004318:Doxycycline