| Literature DB >> 32865751 |
Marbely C Fernández1,2,3, Ramona A Kopton1,2,3, Ana Simon-Chica1,2, Josef Madl1,2, Ingo Hilgendorf2,4, Callum M Zgierski-Johnston1,2, Franziska Schneider-Warme5,6.
Abstract
Optogenetic approaches have evolved as potent means to investigate cardiac electrophysiology, with research ranging from the study of arrhythmia mechanisms to effects of cardiac innervation and heterocellular structural and functional interactions, both in healthy and diseased myocardium. Most commonly, these studies use channelrhodopsin-2 (ChR2)-expressing murine models that enable light-activated depolarization of the target cell population. However, each newly generated mouse line requires thorough characterization, as cell-type specific ChR2 expression cannot be taken for granted, and the electrophysiological response of its activation in the target cell should be evaluated. In this chapter, we describe detailed protocols for assessing ChR2 specificity using immunohistochemistry, isolation of specific cell populations to analyze electrophysiological effects of ChR2 activation with the patch-clamp technique, and whole-heart experiments to assess in situ effects of optical stimulation.Entities:
Keywords: Cardiac optogenetics; Cardiomyocytes; Cell-type specific gene targeting; Electrocardiogram; Electrophysiology; Fibroblasts; Macrophages; Optical pacing; Transgenic mice
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Year: 2021 PMID: 32865751 DOI: 10.1007/978-1-0716-0830-2_17
Source DB: PubMed Journal: Methods Mol Biol ISSN: 1064-3745