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A Protocol for Dual Calcium-Voltage Optical Mapping in Murine Sinoatrial Preparation With Optogenetic Pacing
Among the animal models for studying the molecular basis of atrial and sinoatrial node (SAN) biology and disease, the mouse is a widely used species due to its feasibility for genetic modifications in genes encoding ion channels or calcium handling and signaling proteins in the heart. It is therefor...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6698704/ https://www.ncbi.nlm.nih.gov/pubmed/31456689 http://dx.doi.org/10.3389/fphys.2019.00954 |
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author | Dong, Ruirui Mu-u-min, Razik Reith, Alastair J. M. O’Shea, Christopher He, Shicheng Duan, Kaizhong Kou, Kun Grassam-Rowe, Alexander Tan, Xiaoqiu Pavlovic, Davor Ou, Xianhong Lei, Ming |
author_facet | Dong, Ruirui Mu-u-min, Razik Reith, Alastair J. M. O’Shea, Christopher He, Shicheng Duan, Kaizhong Kou, Kun Grassam-Rowe, Alexander Tan, Xiaoqiu Pavlovic, Davor Ou, Xianhong Lei, Ming |
author_sort | Dong, Ruirui |
collection | PubMed |
description | Among the animal models for studying the molecular basis of atrial and sinoatrial node (SAN) biology and disease, the mouse is a widely used species due to its feasibility for genetic modifications in genes encoding ion channels or calcium handling and signaling proteins in the heart. It is therefore highly valuable to develop robust methodologies for studying SAN and atrial electrophysiological function in this species. Here, we describe a protocol for performing dual calcium-voltage optical mapping on mouse sinoatrial preparation (SAP), in combination with an optogenetic approach, for studying SAP membrane potential, intracellular Ca(2+) transients, and pacemaker activity. The protocol includes the details for preparing the intact SAP, robust tissue dual-dye loading, light-programmed pacing, and high-resolution optical mapping. Our protocol provides an example of use of the combination of optogenetic and optical mapping techniques for investigating SAP membrane potential and intracellular Ca(2+) transients and pacemaker activity with high temporal and spatial resolution in specific cardiac tissues. Thus, our protocol provides a useful tool for studying SAP physiology and pathophysiology in mice. |
format | Online Article Text |
id | pubmed-6698704 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-66987042019-08-27 A Protocol for Dual Calcium-Voltage Optical Mapping in Murine Sinoatrial Preparation With Optogenetic Pacing Dong, Ruirui Mu-u-min, Razik Reith, Alastair J. M. O’Shea, Christopher He, Shicheng Duan, Kaizhong Kou, Kun Grassam-Rowe, Alexander Tan, Xiaoqiu Pavlovic, Davor Ou, Xianhong Lei, Ming Front Physiol Physiology Among the animal models for studying the molecular basis of atrial and sinoatrial node (SAN) biology and disease, the mouse is a widely used species due to its feasibility for genetic modifications in genes encoding ion channels or calcium handling and signaling proteins in the heart. It is therefore highly valuable to develop robust methodologies for studying SAN and atrial electrophysiological function in this species. Here, we describe a protocol for performing dual calcium-voltage optical mapping on mouse sinoatrial preparation (SAP), in combination with an optogenetic approach, for studying SAP membrane potential, intracellular Ca(2+) transients, and pacemaker activity. The protocol includes the details for preparing the intact SAP, robust tissue dual-dye loading, light-programmed pacing, and high-resolution optical mapping. Our protocol provides an example of use of the combination of optogenetic and optical mapping techniques for investigating SAP membrane potential and intracellular Ca(2+) transients and pacemaker activity with high temporal and spatial resolution in specific cardiac tissues. Thus, our protocol provides a useful tool for studying SAP physiology and pathophysiology in mice. Frontiers Media S.A. 2019-08-06 /pmc/articles/PMC6698704/ /pubmed/31456689 http://dx.doi.org/10.3389/fphys.2019.00954 Text en Copyright © 2019 Dong, Mu-u-min, Reith, O’Shea, He, Duan, Kou, Grassam-Rowe, Tan, Pavlovic, Ou and Lei. http://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 | Physiology Dong, Ruirui Mu-u-min, Razik Reith, Alastair J. M. O’Shea, Christopher He, Shicheng Duan, Kaizhong Kou, Kun Grassam-Rowe, Alexander Tan, Xiaoqiu Pavlovic, Davor Ou, Xianhong Lei, Ming A Protocol for Dual Calcium-Voltage Optical Mapping in Murine Sinoatrial Preparation With Optogenetic Pacing |
title | A Protocol for Dual Calcium-Voltage Optical Mapping in Murine Sinoatrial Preparation With Optogenetic Pacing |
title_full | A Protocol for Dual Calcium-Voltage Optical Mapping in Murine Sinoatrial Preparation With Optogenetic Pacing |
title_fullStr | A Protocol for Dual Calcium-Voltage Optical Mapping in Murine Sinoatrial Preparation With Optogenetic Pacing |
title_full_unstemmed | A Protocol for Dual Calcium-Voltage Optical Mapping in Murine Sinoatrial Preparation With Optogenetic Pacing |
title_short | A Protocol for Dual Calcium-Voltage Optical Mapping in Murine Sinoatrial Preparation With Optogenetic Pacing |
title_sort | protocol for dual calcium-voltage optical mapping in murine sinoatrial preparation with optogenetic pacing |
topic | Physiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6698704/ https://www.ncbi.nlm.nih.gov/pubmed/31456689 http://dx.doi.org/10.3389/fphys.2019.00954 |
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