详细信息:Recombinant HEK293 cell line expressing the human GABAA α3/β3/γ2 chloride ion channel. The pharmacological properties of HEK293 cells expressing human GABAA α3, β3 and γ2 subunits were principally characterized on the automated planar array electrophysiology platforms PatchXpress™ and IonWorks™. A number of manual patch clamp experiments were also performed. Bath application of GABA evoked large inward currents at -40 mV that were completely suppressed by 100 ™M of the GABAA receptor antagonist bicuculline. The response to GABA was dose-dependent and gave EC50 values of around 9-11 ™M using automated electrophysiology. These EC50 values are approximately 2-3 fold higher than values obtained using the hGABAA α1/β3/γ2-HEK293 cell line (CYL3053), consistent with published data describing a reduced affinity of GABA for α3 in comparison to α1. The non-sedating anxiolytic L-838,417 potentiated the α3 EC20 GABA response but had no effect on α1 responses, providing convincing evidence for selective functional expression of α3 subunits in this cell line. Functional channel expression over time was monitored using IonWorks™ HT. Channel expression was consistently >80% over at least 41 passages with mean outward GABA evoked currents consistently >4 nA.
