Optogenetic targeting of cardiac myocytes and non-myocytes: tools, challenges and utility
| Autor: | Callum M. Johnston, Eike M. Wülfers, Peter Kohl, Thomas Knöpfel, Torsten Houwaart, Franziska Schneider-Warme, Eva A. Rog-Zielinska, Roland Nitschke, Angela Naumann, Urszula Siedlecka |
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| Přispěvatelé: | British Heart Foundation, Commission of the European Communities, National Institutes of Health |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Cell Biophysics Optogenetics Biology Bioinformatics 03 medical and health sciences Mice Channelrhodopsins medicine Myocyte Animals Myocytes Cardiac Molecular Biology Cell specific Ventricular wall 0601 Biochemistry And Cell Biology Coupling (electronics) Mice Inbred C57BL Electrophysiology 030104 developmental biology medicine.anatomical_structure Gene Expression Regulation Neuroscience Clearance |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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