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An Advanced Automated Patch Clamp Protocol Design to Investigate Drug—Ion Channel Binding Dynamics

Standard high throughput screening projects using automated patch-clamp instruments often fail to grasp essential details of the mechanism of action, such as binding/unbinding dynamics and modulation of gating. In this study, we aim to demonstrate that depth of analysis can be combined with acceptab...

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Autores principales: Lukacs, Peter, Pesti, Krisztina, Földi, Mátyás C., Zboray, Katalin, Toth, Adam V., Papp, Gábor, Mike, Arpad
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8513526/
https://www.ncbi.nlm.nih.gov/pubmed/34658875
http://dx.doi.org/10.3389/fphar.2021.738260
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author Lukacs, Peter
Pesti, Krisztina
Földi, Mátyás C.
Zboray, Katalin
Toth, Adam V.
Papp, Gábor
Mike, Arpad
author_facet Lukacs, Peter
Pesti, Krisztina
Földi, Mátyás C.
Zboray, Katalin
Toth, Adam V.
Papp, Gábor
Mike, Arpad
author_sort Lukacs, Peter
collection PubMed
description Standard high throughput screening projects using automated patch-clamp instruments often fail to grasp essential details of the mechanism of action, such as binding/unbinding dynamics and modulation of gating. In this study, we aim to demonstrate that depth of analysis can be combined with acceptable throughput on such instruments. Using the microfluidics-based automated patch clamp, IonFlux Mercury, we developed a method for a rapid assessment of the mechanism of action of sodium channel inhibitors, including their state-dependent association and dissociation kinetics. The method is based on a complex voltage protocol, which is repeated at 1 Hz. Using this time resolution we could monitor the onset and offset of both channel block and modulation of gating upon drug perfusion and washout. Our results show that the onset and the offset of drug effects are complex processes, involving several steps, which may occur on different time scales. We could identify distinct sub-processes on the millisecond time scale, as well as on the second time scale. Automated analysis of the results allows collection of detailed information regarding the mechanism of action of individual compounds, which may help the assessment of therapeutic potential for hyperexcitability-related disorders, such as epilepsies, pain syndromes, neuromuscular disorders, or neurodegenerative diseases.
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spelling pubmed-85135262021-10-14 An Advanced Automated Patch Clamp Protocol Design to Investigate Drug—Ion Channel Binding Dynamics Lukacs, Peter Pesti, Krisztina Földi, Mátyás C. Zboray, Katalin Toth, Adam V. Papp, Gábor Mike, Arpad Front Pharmacol Pharmacology Standard high throughput screening projects using automated patch-clamp instruments often fail to grasp essential details of the mechanism of action, such as binding/unbinding dynamics and modulation of gating. In this study, we aim to demonstrate that depth of analysis can be combined with acceptable throughput on such instruments. Using the microfluidics-based automated patch clamp, IonFlux Mercury, we developed a method for a rapid assessment of the mechanism of action of sodium channel inhibitors, including their state-dependent association and dissociation kinetics. The method is based on a complex voltage protocol, which is repeated at 1 Hz. Using this time resolution we could monitor the onset and offset of both channel block and modulation of gating upon drug perfusion and washout. Our results show that the onset and the offset of drug effects are complex processes, involving several steps, which may occur on different time scales. We could identify distinct sub-processes on the millisecond time scale, as well as on the second time scale. Automated analysis of the results allows collection of detailed information regarding the mechanism of action of individual compounds, which may help the assessment of therapeutic potential for hyperexcitability-related disorders, such as epilepsies, pain syndromes, neuromuscular disorders, or neurodegenerative diseases. Frontiers Media S.A. 2021-09-28 /pmc/articles/PMC8513526/ /pubmed/34658875 http://dx.doi.org/10.3389/fphar.2021.738260 Text en Copyright © 2021 Lukacs, Pesti, Földi, Zboray, Toth, Papp and Mike. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Pharmacology
Lukacs, Peter
Pesti, Krisztina
Földi, Mátyás C.
Zboray, Katalin
Toth, Adam V.
Papp, Gábor
Mike, Arpad
An Advanced Automated Patch Clamp Protocol Design to Investigate Drug—Ion Channel Binding Dynamics
title An Advanced Automated Patch Clamp Protocol Design to Investigate Drug—Ion Channel Binding Dynamics
title_full An Advanced Automated Patch Clamp Protocol Design to Investigate Drug—Ion Channel Binding Dynamics
title_fullStr An Advanced Automated Patch Clamp Protocol Design to Investigate Drug—Ion Channel Binding Dynamics
title_full_unstemmed An Advanced Automated Patch Clamp Protocol Design to Investigate Drug—Ion Channel Binding Dynamics
title_short An Advanced Automated Patch Clamp Protocol Design to Investigate Drug—Ion Channel Binding Dynamics
title_sort advanced automated patch clamp protocol design to investigate drug—ion channel binding dynamics
topic Pharmacology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8513526/
https://www.ncbi.nlm.nih.gov/pubmed/34658875
http://dx.doi.org/10.3389/fphar.2021.738260
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