Stable expression of cloned rat GABAA receptor subunits in a human kidney cell 
line

Hamilton, B. J.; Lennon, D. J.; Im, H. K.; Im, W. B.; Seeburg, Peter H.; Carter, D. B.

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Stable expression of cloned rat GABAA receptor subunits in a human kidney cell line

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Seeburg, Peter H.
Department of Molecular Neurobiology, Max Planck Institute for Medical Research, Max Planck Society;

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http://www.sciencedirect.com/science/article/pii/030439409390323D/pdf?md5=0a5abab7172571a29fdd2f9a15b86ee0&pid=1-s2.0-030439409390323D-main.pdf
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http://doi.org/10.1016/0304-3940(93)90323-D
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Hamilton, B. J., Lennon, D. J., Im, H. K., Im, W. B., Seeburg, P. H., & Carter, D. B. (1993). Stable expression of cloned rat GABAA receptor subunits in a human kidney cell line. Neuroscience Letters, 153(2), 206-209. Retrieved from https://www.ncbi.nlm.nih.gov/pubmed/7687050.

Cite as: https://hdl.handle.net/11858/00-001M-0000-0019-AA7A-E

Abstract

A predominant form of the GABAA/benzodiazepine receptor-Cl- channel complex is believed to consist of three different 48-55 kDa subunits (alpha, beta, gamma) with unknown stoichiometry. Plasmids containing the rat GABAA receptor cDNAs coding for alpha 1, beta 2, and gamma 2 were co-transfected, along with a plasmid encoding G418 resistance, into human embryonic kidney cells previously transformed with Adenovirus 5 (HEK-293) [J. Gen. Virol., 36 (1977) 59-72]. Four percent of the G418 resistant colonies were found to express mRNA for all three of the GABAA subunits constitutively. A single cell clone derived from one of the alpha 1 beta 2 gamma 2 expressors has demonstrated stable electrophysiological characteristics over 25 passages. The GABA-activated Cl- current in this cell line is blocked by picrotoxin and bicuculline, and is modulated by a variety of agonist and inverse agonist ligands including diazepam, Ro 154513, zolpidem, and beta-CCE. The cell line has been used successfully over a 12-month period as a screen for novel drugs modulating GABA-mediated polarization of neuronal cells.