Journal
SCIENTIFIC REPORTS
Volume 11, Issue 1, Pages -Publisher
NATURE RESEARCH
DOI: 10.1038/s41598-021-82077-8
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Funding
- Agence Nationale de la Recherche [ANR-15-CE14-0006-01, ANR-16-CE92-0013-01, ANR-11-LABX-0015]
- Federation Francaise de Cardiologie
- Fondation Leducq
- New Team of the Region Pays de la Loire
- European FEDER grant
- National Research Agency Grant [ANR-18-CE19-0024-01]
- Groupe de Reflexion sur la Recherche Cardiovasculaire-Societe Francaise de Cardiologie predoctoral fellowship [SFC/GRRC2018]
- Agence Nationale de la Recherche (ANR) [ANR-16-CE92-0013, ANR-15-CE14-0006] Funding Source: Agence Nationale de la Recherche (ANR)
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The whole-cell voltage-clamp technique requires consideration of the impact of current amplitude on data accuracy, especially in high throughput patch-clamp applications to prevent inaccuracies. Mathematical kinetic models can predict how large current amplitudes and series resistance artifacts affect voltage-dependent activation and inactivation processes, providing simple guidelines for practice.
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.
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