Assessing functional properties of ligand-gated ion channels with automated whole-cell patch clamp technology

著者: Hervør L Olsen, Rasmus B Jacobsen, Rikke Schrøder, Jonatan Kutchinsky, Morten Sunesen, Niels Willumsen

Most ligand-gated ion channels (LGICs) are characterized by fast transient currents in response to application of agonists. Typically the time constants for activation and subsequent desensitization amount to1-100 and 10-1000 ms, respectively. Consequently, recording of proper whole-cell LGIC currents with the patch-clamp technique requires a fast solution exchange system. Furthermore, characterization of LGIC blockers and modulators generally requires complex compound application protocols, because the effect of a test compound needs to be evaluated simultaneously with a tran- sient application of the agonist. These require- ments challenge a realistic electrophysiological characterization of LGICs. We employed the auto- mated QPatch 16 patch-clamp system, to charac- terize the effects of agonists, antagonists and acti- vators on two types of fast LGICs: (1) GABAA (γ- amino-butyric acid A) receptors and (2) ASIC (acid sensing ion channels) types 1a and 3.