Hamilton B J, Lennon D J, Im H K, Im W B, Seeburg P H, Carter D B
CNS Research, Upjohn Company, Kalamazoo, MI 49001.
Neurosci Lett. 1993 Apr 30;153(2):206-9. doi: 10.1016/0304-3940(93)90323-d.
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.
GABAA/苯二氮䓬受体 - 氯离子通道复合物的主要形式据信由三种不同的48 - 55 kDa亚基（α、β、γ）组成，其化学计量未知。将编码α1、β2和γ2的大鼠GABAA受体cDNA的质粒，与编码G418抗性的质粒一起共转染到先前用腺病毒5转化的人胚肾细胞（HEK - 293）中[《普通病毒学杂志》，36 (1977) 59 - 72]。发现4%的G418抗性菌落组成性表达所有三种GABAA亚基的mRNA。从其中一个α1β2γ2表达细胞系衍生出的单细胞克隆在超过25代中表现出稳定的电生理特性。该细胞系中的GABA激活的氯离子电流被印防己毒素和荷包牡丹碱阻断，并受到多种激动剂和反向激动剂配体的调节，包括地西泮、Ro 154513、唑吡坦和β - CCE。在12个月的时间里，该细胞系已成功用作筛选调节GABA介导的神经元细胞极化的新型药物的工具。
