PrecisION hGABA_A α3/β3/γ2-HEK Recombinant Cell Line

PrecisION hGABA_A α3/β3/γ2-HEK Recombinant Cell Line

PrecisION

2 mL

Recombinant HEK293 cell line expressing the human GABA_A α3/β3/γ2 chloride ion channel

GABA is the major inhibitory neurotransmitter in the CNS, binding to fast-acting ionotropic GABA_A receptors to cause inward flux of Cl- resulting in membrane hyperpolarization and thus reducing membrane excitability (Nutt et al., 2006). Excessive stimulation of these receptors can lead to sedation and ataxia whereas attenuation leads to arousal, insomnia and anxiety. Since modulation of these receptors has such profound physiological effects they have become important drug targets for the treatment of many conditions e.g. anxiety, epilepsy, sleep disorders and for anesthesia (Rudolph and Mohler, 2006). GABA_A receptors are pentameric structures typically consisting of α, β and γ2 subunits in a stoichiometry of 2:2:1. The specific subunit composition is especially important since various combinations mediate different effects (Rudolph and Mohler, 2006). For example, α1-containing receptors, accounting for 60% of all GABA_A receptors, mediate the sedative/hypnotic effects of benzodiazepines (BZPs) whereas the anxiolytic effects of these drugs are mediated by receptors containing α2 and α3 subunits (Korpi and Sinkkonen, 2006). Hence, developing selective allosteric modulators for these latter subunits should ultimately lead to anxiolytic drugs devoid of unwanted sedative effects.

The pharmacological properties of HEK293 cells expressing human GABA_A 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 µM of the GABA_A receptor antagonist bicuculline. The response to GABA was dose-dependent and gave EC50 values of around 9-11 µM using automated electrophysiology. These EC50 values are approximately 2-3 fold higher than values obtained using the hGABA_A 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.

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

GABA, the major inhibitory neurotransmitter in the vertebrate brain, mediates neuronal inhibition by binding to the GABA/benzodiazepine receptor and opening an integral chloride channel.

Human

Mycoplasma Testing:
The cell line has been screened to confirm the absence of Mycoplasma species

HEK

Ion Channel Cell Lines

Pack contains 2 vials of mycoplasma-free cells, 1 ml per vial

Upstate

293 cell line

2 x 1 ml aliquots

hGABA_A α3/β3/γ2

Chloride

GABRA3

Ion Channels

PrecisION hGABA_A α3/β3/γ2-HEK Recombinant Cell Line