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Advancing mechanobiology by performing whole-cell patch clamp recording on mechanosensitive cells subjected simultaneously to dynamic stretch events.

iScience (2021-02-04)
Etienne de Coulon, Christian Dellenbach, Stephan Rohr
ABSTRACT

A comprehensive understanding of mechano-electrical coupling requires continuous intracellular electrical recordings being performed on cells undergoing simultaneously in vivo like strain events. Here, we introduce a linear strain single-cell electrophysiology (LSSE) system that meets these requirements by delivering highly reproducible unidirectional strain events with magnitudes up to 12% and strain rates exceeding 200%s-1 to adherent cells kept simultaneously in whole-cell patch-clamp recording configuration. Proof-of-concept measurements with NIH3T3 cells demonstrate that stable recording conditions are maintained over tens of strain cycles at maximal amplitudes and strain rates thereby permitting a full electrophysiological characterization of mechanically activated ion currents. Because mechano-electrical responses to predefined strain patterns can be investigated using any adherent wild-type or genetically modified cell type of interest, the LSSE system offers the perspective of providing advanced insights into mechanosensitive ion channel function that can finally be compared quantitatively among different types of channels and cells.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Vitamin B12, ≥98%
Sigma-Aldrich
Medium 199, With Hanks′ salts and sodium bicarbonate, without L-glutamine, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Collagen from rat tail, Bornstein and Traub Type I, powder, BioReagent, suitable for cell culture