Methods Mol Biol 2014 Jan 01;1183:355-69. doi: 10.1007/978-1-4939-1096-0_22.

Ion selectivity of pore-forming peptides and ion channels measured in Xenopus oocytes.

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The Xenopus laevis oocyte is a widely used system for heterologous expression of exogenous ion channel proteins. They are easy to obtain, mechanically and electrically stable, have a large size, enabling multiple types of electrophysiological recordings: two-electrode voltage clamp, single cell-attached or cell-free patch-clamp and macropatch recordings. The size of an oocyte (1 mm in diameter) also allows for the use of additional electrodes (from 1 to 3) for injection of diverse materials (Ca(2+) chelators, peptides, chemicals, antibodies, proteic-partners, etc.) before or during the course of the electrophysiological experiment.We have successfully used this system to analyze the biophysical properties of pore-forming peptides. Simple extracellular perfusion of these peptides induced the formation of channels in the oocyte plasma membrane; these channels can then be studied and characterized in diverse ionic conditions. The ease of the perfusion and the stability of the voltage-clamped oocyte make it a powerful tool for such analyses. Compared to artificial bilayers, oocytes offer a real animal plasma membrane where biophysical properties and toxicity can be studied in the stable environment.

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