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.
Tammaro
Paolo
P
Department of Physiology, Anatomy and Genetics, University of Oxford, UK.
Shimomura
Kenju
K
Proks
Peter
P
D000818:Animals; D018118:Chloride Channels / Q000302:isolation & purification / Q000502:physiology*; D004594:Electrophysiology / Q000379:methods; D005260:Female; D007473:Ion Channels / Q000235:genetics / Q000502:physiology*; D054086:KATP Channels / Q000302:isolation & purification / Q000502:physiology*; D009865:Oocytes / Q000502:physiology*; D018408:Patch-Clamp Techniques; D018075:RNA, Complementary / Q000235:genetics; D014982:Xenopus laevis
D018118:Chloride Channels; D007473:Ion Channels; D054086:KATP Channels; D018075:RNA, Complementary