Planar patch clamp devices are finding their place in academic and industrial labs because of their ease of use and higher throughput as compared with conventional patch clamp. Planar borosilicate glass chips are used for performing whole cell, perforated patch, or cell attached electrophysiological recordings. High quality data is acquired with a high success rate for obtaining giga-seals (typically 60-80%). Given the design of the borosilicate glass chip, the internal recording solution is much more accessible to exchange. This means that experiments involving exchange of the internal solution can be performed with relative ease. Exemplar traces will be shown for block of KV1.3 channels by the internal perfusion of ions and small molecules. In addition, data will be presented showing planar lipid bilayers which were used to estimate the time required to fully exchange the internal solution. Since many ion channels are sensitive to temperature and some compounds have been shown to exhibit different pharmacology at different temperatures, temperature can become an important parameter to control in electrophysiological experiments. By heating the bath solution and the solution entering the bath of a planar patch clamp system, the temperature of the external solution can be constantly maintained at a given temperature. Data will be presented showing hERG recordings at room temperature and at physiological temperature, including a full concentration response curve to quinidine at physiological temperature. Not only can the amplitude and kinetics of currents be changed by temperature, indeed, some ion channels can actually be activated by elevated temperature. The TRPV1 channel, for example, can be activated by ligands such as capsaicin, but also has been reported to be activated by temperatures above ~42°C. Using a heated pipette, the temperature of the added solution can be increased and then rapidly applied to the cell. Very rapid changes in temperature can be achieved at the cell (within ms), from room temperature up to 60°C, whilst continuously recording. Exemplar traces will be presented showing heat activation of TRPV1 expressed in CHO cells using a range of temperatures. TRPV1 channels could prove to be important targets for the treatment of pain and, therefore, the temperature activation of TRPV1 could have important implications for drug design as well as investigations into physiological modes of action.
University College Dublin (2009) Proc Physiol Soc 15, PC163
Poster Communications: Expanding applications of automated planar patch clamp: A focus on internal exchange and heat activation
A. R. Haythornthwaite1, S. Stoelzle1, C. Haarmann1, C. Farre1, A. Brueggemann1, M. Kreir1, M. Beckler1, M. George1, N. Fertig1
1. Nanion Technologies, Munich, Germany.
View other abstracts by:
Where applicable, experiments conform with Society ethical requirements.