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OptoGap is an optogenetics-enabled assay for quantification of cell–cell coupling in multicellular cardiac tissue
Intercellular electrical coupling is an essential means of communication between cells. It is important to obtain quantitative knowledge of such coupling between cardiomyocytes and non-excitable cells when, for example, pathological electrical coupling between myofibroblasts and cardiomyocytes yield...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8085001/ https://www.ncbi.nlm.nih.gov/pubmed/33927252 http://dx.doi.org/10.1038/s41598-021-88573-1 |
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author | Boyle, Patrick M. Yu, Jinzhu Klimas, Aleksandra Williams, John C. Trayanova, Natalia A. Entcheva, Emilia |
author_facet | Boyle, Patrick M. Yu, Jinzhu Klimas, Aleksandra Williams, John C. Trayanova, Natalia A. Entcheva, Emilia |
author_sort | Boyle, Patrick M. |
collection | PubMed |
description | Intercellular electrical coupling is an essential means of communication between cells. It is important to obtain quantitative knowledge of such coupling between cardiomyocytes and non-excitable cells when, for example, pathological electrical coupling between myofibroblasts and cardiomyocytes yields increased arrhythmia risk or during the integration of donor (e.g., cardiac progenitor) cells with native cardiomyocytes in cell-therapy approaches. Currently, there is no direct method for assessing heterocellular coupling within multicellular tissue. Here we demonstrate experimentally and computationally a new contactless assay for electrical coupling, OptoGap, based on selective illumination of inexcitable cells that express optogenetic actuators and optical sensing of the response of coupled excitable cells (e.g., cardiomyocytes) that are light-insensitive. Cell–cell coupling is quantified by the energy required to elicit an action potential via junctional current from the light-stimulated cell(s). The proposed technique is experimentally validated against the standard indirect approach, GapFRAP, using light-sensitive cardiac fibroblasts and non-transformed cardiomyocytes in a two-dimensional setting. Its potential applicability to the complex three-dimensional setting of the native heart is corroborated by computational modelling and proper calibration. Lastly, the sensitivity of OptoGap to intrinsic cell-scale excitability is robustly characterized via computational analysis. |
format | Online Article Text |
id | pubmed-8085001 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-80850012021-04-30 OptoGap is an optogenetics-enabled assay for quantification of cell–cell coupling in multicellular cardiac tissue Boyle, Patrick M. Yu, Jinzhu Klimas, Aleksandra Williams, John C. Trayanova, Natalia A. Entcheva, Emilia Sci Rep Article Intercellular electrical coupling is an essential means of communication between cells. It is important to obtain quantitative knowledge of such coupling between cardiomyocytes and non-excitable cells when, for example, pathological electrical coupling between myofibroblasts and cardiomyocytes yields increased arrhythmia risk or during the integration of donor (e.g., cardiac progenitor) cells with native cardiomyocytes in cell-therapy approaches. Currently, there is no direct method for assessing heterocellular coupling within multicellular tissue. Here we demonstrate experimentally and computationally a new contactless assay for electrical coupling, OptoGap, based on selective illumination of inexcitable cells that express optogenetic actuators and optical sensing of the response of coupled excitable cells (e.g., cardiomyocytes) that are light-insensitive. Cell–cell coupling is quantified by the energy required to elicit an action potential via junctional current from the light-stimulated cell(s). The proposed technique is experimentally validated against the standard indirect approach, GapFRAP, using light-sensitive cardiac fibroblasts and non-transformed cardiomyocytes in a two-dimensional setting. Its potential applicability to the complex three-dimensional setting of the native heart is corroborated by computational modelling and proper calibration. Lastly, the sensitivity of OptoGap to intrinsic cell-scale excitability is robustly characterized via computational analysis. Nature Publishing Group UK 2021-04-29 /pmc/articles/PMC8085001/ /pubmed/33927252 http://dx.doi.org/10.1038/s41598-021-88573-1 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Boyle, Patrick M. Yu, Jinzhu Klimas, Aleksandra Williams, John C. Trayanova, Natalia A. Entcheva, Emilia OptoGap is an optogenetics-enabled assay for quantification of cell–cell coupling in multicellular cardiac tissue |
title | OptoGap is an optogenetics-enabled assay for quantification of cell–cell coupling in multicellular cardiac tissue |
title_full | OptoGap is an optogenetics-enabled assay for quantification of cell–cell coupling in multicellular cardiac tissue |
title_fullStr | OptoGap is an optogenetics-enabled assay for quantification of cell–cell coupling in multicellular cardiac tissue |
title_full_unstemmed | OptoGap is an optogenetics-enabled assay for quantification of cell–cell coupling in multicellular cardiac tissue |
title_short | OptoGap is an optogenetics-enabled assay for quantification of cell–cell coupling in multicellular cardiac tissue |
title_sort | optogap is an optogenetics-enabled assay for quantification of cell–cell coupling in multicellular cardiac tissue |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8085001/ https://www.ncbi.nlm.nih.gov/pubmed/33927252 http://dx.doi.org/10.1038/s41598-021-88573-1 |
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