Xenopus Oocytes as a Heterologous Expression System for Studying Ion Channels with the Patch-Clamp Technique
Oocytes from the Xenopus laevis represent one of the most widely used expression systems for functional characterization of ion channels. Their large size facilitates both injection of heterologous cRNA and subsequent electrophysiological recordings of ion channel currents. Furthermore, Xenopus oocytes translate cRNA very efficiently, resulting in the generation of a large number of ion channels in the plasma membrane. In this chapter, we outline methods for oocyte preparation and maintenance and describe procedures for patch-clamping of oocytes, with a special focus on the macropatch technique. We discuss some common problems associated with patch-clamping of oocytes and their use as an expression system for ion channels.
Key wordsXenopus laevis Oocytes Ion channels Patch-clamp Macropatch Inside-out Outside-out Patch-cramming Heterologous expression
We wish to thank Drs. Oscar Moran and Tim Craig for their critical reading of the manuscript and helpful comments. P.T. holds a Junior Research fellowship at Wolfson College, Oxford.
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