Single-channel recording of ligand-gated ion channels.
Electrophysiological recording of single-channel currents is the most direct method available for obtaining detailed and precise information about the kinetic behavior of ion channels. A wide variety of cell types can be used for single-channel recording, but to obtain the highest resolution of the briefest channel opening and closing events, low-noise recordings, coupled with a minimal filtering frequency, are required. Here, we present a protocol designed to help those with some electrophysiological expertise who wish to explore the properties of native and recombinant single ligand-gated ion channels. We have focused on the practical aspects of recording single GABA channels from cell-attached and outside-out patches and also introduced some of the preliminary considerations that are necessary for the analysis of single-channel data, including an introduction to single-channel analysis software.
Mortensen
Martin
M
Department of Pharmacology, University College London, Gower Street, London WC1E 6BT, UK.
Smart
Trevor G
TG
D002460:Cell Line; D004553:Electric Conductivity; D006801:Humans; D015640:Ion Channel Gating; D007473:Ion Channels / Q000737:chemistry*; D008024:Ligands; D018408:Patch-Clamp Techniques / Q000379:methods*; D011963:Receptors, GABA-A / Q000737:chemistry; D005680:gamma-Aminobutyric Acid / Q000737:chemistry
D007473:Ion Channels; D008024:Ligands; D011963:Receptors, GABA-A; D005680:gamma-Aminobutyric Acid