Comparison of cell expression formats for the characterization of GABA(A) channels using a microfluidic patch clamp system.

Assay and Drug Development Technologies
Qin ChenCharles W Emala

Abstract

Ensemble recording and microfluidic perfusion are recently introduced techniques aimed at removing the laborious nature and low recording success rates of manual patch clamp. Here, we present assay characteristics for these features integrated into one automated electrophysiology platform as applied to the study of GABA(A) channels. A variety of cell types and methods of GABA(A) channel expression were successfully studied (defined as I(GABA)>500 pA), including stably transfected human embryonic kidney (HEK) cells expressing α(1)β(3)γ(2) GABA(A) channels, frozen ready-to-assay (RTA) HEK cells expressing α(1)β(3)γ(2) or α(3)β(3)γ(2) GABA(A) channels, transiently transfected HEK293T cells expressing α(1)β(3)γ(2) GABA(A) channels, and immortalized cultures of human airway smooth muscle cells endogenously expressing GABA(A) channels. Current measurements were successfully studied in multiple cell types with multiple modes of channel expression in response to several classic GABA(A) channel agonists, antagonists, and allosteric modulators. We obtained success rates above 95% for transiently or stably transfected HEK cells and frozen RTA HEK cells expressing GABA(A) channels. Tissue-derived immortalized cultures of airway smooth musc...Continue Reading

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Citations

Oct 28, 2014·ACS Chemical Neuroscience·Lik-Wei WongBrett A Cromer

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