Patch-clamp measurements on isolated stomatal protoplasts

Patch-clamp measurements on isolated guard cell protoplasts and vacuoles

by Raschke K., Hedrich R. (1989)

Klaus Raschke, Rainer Hedrich,

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In Methods in Enzymol. 174: 312–330 – https://doi.org/10.1016/0076-6879(89)74025-8 –

https://www.sciencedirect.com/science/article/pii/0076687989740258

Abstract

This chapter discusses the patch-clamp measurements on isolated guard cell protoplasts and vacuoles.

Guard cells can serve as models for other cells that export or import ions and break down carbohydrates to make organic acids for balancing cation fluxes. Such cells occur in roots, growing tissue, and in the pelvini of the leaves of plant species displaying nasty leaf movements.

It allows separate recordings of the activities of the transport systems in the plasmalemma and in the tonoplast.

Characterization becomes possible of electrogenic pumps, ion channels, and carders of very high turnover in any type of membrane. The basic requirement for the application of the technique is an absolutely clean membrane surface.

The chapter also describes the whole-vacuole configuration (analogous to whole cell) can be established by rupturing the membrane patch covering the tip of the patch pipette (starting from a vacuole-attached configuration).

Published by

Willem Van Cotthem

Honorary Professor of Botany, University of Ghent (Belgium). Scientific Consultant for Desertification and Sustainable Development.

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