Computer modeling of whole-cell voltage-clamp analyses to delineate guidelines for good practice of manual and automated patch-clamp

Abstract The patch-clamp technique and more recently the high throughput patch-clamp technique have contributed to major advances in the characterization of ion channels. However, the whole-cell voltage-clamp technique presents certain limits that need to be considered for robust data generation. On...

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Autores principales:	Jérôme Montnach, Maxime Lorenzini, Adrien Lesage, Isabelle Simon, Sébastien Nicolas, Eléonore Moreau, Céline Marionneau, Isabelle Baró, Michel De Waard, Gildas Loussouarn
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/fc995879801249e6af526213e9a3435d
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spelling	oai:doaj.org-article:fc995879801249e6af526213e9a3435d2021-12-02T14:26:54ZComputer modeling of whole-cell voltage-clamp analyses to delineate guidelines for good practice of manual and automated patch-clamp10.1038/s41598-021-82077-82045-2322https://doaj.org/article/fc995879801249e6af526213e9a3435d2021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-82077-8https://doaj.org/toc/2045-2322Abstract The patch-clamp technique and more recently the high throughput patch-clamp technique have contributed to major advances in the characterization of ion channels. However, the whole-cell voltage-clamp technique presents certain limits that need to be considered for robust data generation. One major caveat is that increasing current amplitude profoundly impacts the accuracy of the biophysical analyses of macroscopic ion currents under study. Using mathematical kinetic models of a cardiac voltage-gated sodium channel and a cardiac voltage-gated potassium channel, we demonstrated how large current amplitude and series resistance artefacts induce an undetected alteration in the actual membrane potential and affect the characterization of voltage-dependent activation and inactivation processes. We also computed how dose–response curves are hindered by high current amplitudes. This is of high interest since stable cell lines frequently demonstrating high current amplitudes are used for safety pharmacology using the high throughput patch-clamp technique. It is therefore critical to set experimental limits for current amplitude recordings to prevent inaccuracy in the characterization of channel properties or drug activity, such limits being different from one channel type to another. Based on the predictions generated by the kinetic models, we draw simple guidelines for good practice of whole-cell voltage-clamp recordings.Jérôme MontnachMaxime LorenziniAdrien LesageIsabelle SimonSébastien NicolasEléonore MoreauCéline MarionneauIsabelle BaróMichel De WaardGildas LoussouarnNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-16 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jérôme Montnach Maxime Lorenzini Adrien Lesage Isabelle Simon Sébastien Nicolas Eléonore Moreau Céline Marionneau Isabelle Baró Michel De Waard Gildas Loussouarn Computer modeling of whole-cell voltage-clamp analyses to delineate guidelines for good practice of manual and automated patch-clamp
description	Abstract The patch-clamp technique and more recently the high throughput patch-clamp technique have contributed to major advances in the characterization of ion channels. However, the whole-cell voltage-clamp technique presents certain limits that need to be considered for robust data generation. One major caveat is that increasing current amplitude profoundly impacts the accuracy of the biophysical analyses of macroscopic ion currents under study. Using mathematical kinetic models of a cardiac voltage-gated sodium channel and a cardiac voltage-gated potassium channel, we demonstrated how large current amplitude and series resistance artefacts induce an undetected alteration in the actual membrane potential and affect the characterization of voltage-dependent activation and inactivation processes. We also computed how dose–response curves are hindered by high current amplitudes. This is of high interest since stable cell lines frequently demonstrating high current amplitudes are used for safety pharmacology using the high throughput patch-clamp technique. It is therefore critical to set experimental limits for current amplitude recordings to prevent inaccuracy in the characterization of channel properties or drug activity, such limits being different from one channel type to another. Based on the predictions generated by the kinetic models, we draw simple guidelines for good practice of whole-cell voltage-clamp recordings.
format	article
author	Jérôme Montnach Maxime Lorenzini Adrien Lesage Isabelle Simon Sébastien Nicolas Eléonore Moreau Céline Marionneau Isabelle Baró Michel De Waard Gildas Loussouarn
author_facet	Jérôme Montnach Maxime Lorenzini Adrien Lesage Isabelle Simon Sébastien Nicolas Eléonore Moreau Céline Marionneau Isabelle Baró Michel De Waard Gildas Loussouarn
author_sort	Jérôme Montnach
title	Computer modeling of whole-cell voltage-clamp analyses to delineate guidelines for good practice of manual and automated patch-clamp
title_short	Computer modeling of whole-cell voltage-clamp analyses to delineate guidelines for good practice of manual and automated patch-clamp
title_full	Computer modeling of whole-cell voltage-clamp analyses to delineate guidelines for good practice of manual and automated patch-clamp
title_fullStr	Computer modeling of whole-cell voltage-clamp analyses to delineate guidelines for good practice of manual and automated patch-clamp
title_full_unstemmed	Computer modeling of whole-cell voltage-clamp analyses to delineate guidelines for good practice of manual and automated patch-clamp
title_sort	computer modeling of whole-cell voltage-clamp analyses to delineate guidelines for good practice of manual and automated patch-clamp
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/fc995879801249e6af526213e9a3435d
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Computer modeling of whole-cell voltage-clamp analyses to delineate guidelines for good practice of manual and automated patch-clamp

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