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Optogenetic targeting of cardiac myocytes and non-myocytes: tools, challenges and utility

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PBMB_manuscript_Johnston_et al._revised_highlighted_changes.pdfAccepted version2.6 MBAdobe PDFView/Open
1-s2.0-S007961071730189X-main.pdfPublished version4.79 MBAdobe PDFView/Open
Title: Optogenetic targeting of cardiac myocytes and non-myocytes: tools, challenges and utility
Authors: Johnston, CM
Rog-Zielinska, EA
Wulfers, EM
Houwaart, T
Siedlecka, U
Knopfel, T
Kohl, P
Schneider-Warme, F
Item Type: Journal Article
Abstract: In optogenetics, light-activated proteins are used to monitor and modulate cellular behaviour with light. Combining genetic targeting of distinct cellular populations with defined patterns of optical stimulation enables one to study specific cell classes in complex biological tissues. In the current study we attempted to investigate the functional relevance of heterocellular electrotonic coupling in cardiac tissue in situ. In order to do that, we used a Cre-Lox approach to express the light-gated cation channel Channelrhodopsin-2 (ChR2) specifically in either cardiac myocytes or non-myocytes. Despite high specificity when using the same Cre driver lines in a previous study in combination with a different optogenetic probe, we found patchy off-target ChR2 expression in cryo-sections and extended z-stack imaging through the ventricular wall of hearts cleared using CLARITY. Based on immunohistochemical analysis, single-cell electrophysiological recordings and whole-genome sequencing, we reason that non-specificity is caused on the Cre recombination level. Our study highlights the importance of careful design and validation of the Cre recombination targets for reliable cell class specific expression of optogenetic tools.
Issue Date: 15-Sep-2017
Date of Acceptance: 9-Sep-2017
URI: http://hdl.handle.net/10044/1/54812
DOI: https://dx.doi.org/10.1016/j.pbiomolbio.2017.09.014
ISSN: 0079-6107
Publisher: Elsevier
Start Page: 140
End Page: 149
Journal / Book Title: Progress in Biophysics and Molecular Biology
Volume: 130
Issue: Part B
Copyright Statement: © 2017 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
Sponsor/Funder: British Heart Foundation
Commission of the European Communities
British Heart Foundation
British Heart Foundation
National Institutes of Health
Funder's Grant Number: FS/05/089/19373
323099
FS/12/17/29532
FS/15/3/31047
1U01NS099573-01
Keywords: 0601 Biochemistry And Cell Biology
Biophysics
Publication Status: Published
Appears in Collections:National Heart and Lung Institute
Department of Medicine



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