Amplitude histogram-based method of analysis of patch clamp recordings that involve extreme changes in channel activity levels

摘要

Many ion channels show low basal activity which is increased hundreds fold by the relevant gating factor. A classical example is the activation G protein activated K⁺ channels (GIRK) by Gβγ subunit dimer. The extent of activation (relative to basal current), Ra, is an important physiological parameter, usually readily estimated from whole cell recordings. However, calculation of Ra often becomes non-trivial in multichannel patches, because of extreme changes in activity upon activation, from a seemingly single-channel pattern to a macroscopic one. In such cases calculation of the net current flowing through the channels in the patch, Ī, before and after activation may require different methods of analysis. To address this problem, we utilized neuronal GIRK channels activated by purified Gβγ in excised patches of Xenopus oocytes. Channels were expressed at varying densities, from a few to several hundreds per patch. We present a simple and fast method of calculating Ī using amplitude histogram analysis, and establish its accuracy by comparing with Ī calculated from event lists.This method allows the analysis of extreme changes in Ī in multichannel patches, that would be impossible using the standard methods of idealization and event list generation.