Tammaro Paolo, Shimomura Kenju, Proks Peter
Department of Physiology, Anatomy and Genetics, University of Oxford, UK.
Methods Mol Biol. 2008;491:127-39. doi: 10.1007/978-1-59745-526-8_10.
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.
非洲爪蟾的卵母细胞是用于离子通道功能特性表征的最广泛使用的表达系统之一。它们的大尺寸便于注射异源cRNA以及随后对离子通道电流进行电生理记录。此外，非洲爪蟾卵母细胞能非常高效地翻译cRNA，从而在质膜中产生大量离子通道。在本章中，我们概述了卵母细胞制备和维护的方法，并描述了卵母细胞膜片钳的操作步骤，特别着重于巨膜片技术。我们讨论了与卵母细胞膜片钳及其作为离子通道表达系统使用相关的一些常见问题。
