详细信息:Recombinant HEK293 cell line expressing the human GABAA α5/β3/γ2 chloride ion channel. HEK293 cells expressing hGABAA α5/β3/γ2 channels were characterized in terms of their pharmacological properties using whole-cell PatchXpress and IonWorks automated patch clamp platforms.
The cell-line was shown to be selectively expressing hGABAA α5/β3/γ2 currents since large inward currents were seen upon exposure to GABA in the extracellular solution in cells held at -40 mV. These currents were blocked by bicuculline, a specific GABAA channel antagonist. In addition, an EC20 application of GABA elicited currents that were reduced by the negative allosteric modulator α5IA, and this effect was blocked by flumazenil.
Functional channel expression over time was monitored using IonWorks HT. Channel expression is robust over at least 26 passages: 100 % of cells expressed hGABAA α5/β3/γ2 currents at passage 26 giving a mean current amplitude at –40 mV of 6.4 ± 1.4 nA (n=94).
