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

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

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium Channels / metabolism*
  • Cell Separation / methods
  • Electrophysiology / methods
  • Female
  • Oocytes / drug effects
  • Oocytes / metabolism*
  • Patch-Clamp Techniques / methods*
  • Peptides / pharmacology*
  • Xenopus laevis / metabolism*

Substances

  • Calcium Channels
  • Peptides