Sheard, T., Hurley, M., Smith, A. et al. (3 more authors) (2022) Three-dimensional visualization of the cardiac ryanodine receptor clusters and the molecular-scale fraying of dyads. Philosophical Transactions of the Royal Society B: Biological Sciences, 377 (1864). ISSN 0962-8436
Abstract
Clusters of ryanodine receptor calcium channels (RyRs) form the primary molecular machinery of intracellular calcium signalling in cardiomyocytes. While a range of optical super-resolution microscopy techniques have revealed the nanoscale structure of these clusters, the three-dimensional (3D) nanoscale topologies of the clusters have remained mostly unresolved. In this paper, we demonstrate the exploitation of molecular-scale resolution in enhanced expansion microscopy (EExM) along with various 2D and 3D visualization strategies to observe the topological complexities, geometries and molecular sub-domains within the RyR clusters. Notably, we observed sub-domains containing RyR-binding protein junctophilin-2 (JPH2) occupying the central regions of RyR clusters in the deeper interior of the myocytes (including dyads), while the poles were typically devoid of JPH2, lending to a looser RyR arrangement. By contrast, peripheral RyR clusters exhibited variable co-clustering patterns and ratios between RyR and JPH2. EExM images of dyadic RyR clusters in right ventricular (RV) myocytes isolated from rats with monocrotaline-induced RV failure revealed hallmarks of RyR cluster fragmentation accompanied by breaches in the JPH2 sub-domains. Frayed RyR patterns observed adjacent to these constitute new evidence that the destabilization of the RyR arrays inside the JPH2 sub-domains may seed the primordial foci of dyad remodelling observed in heart failure.
This article is part of the theme issue ‘The cardiomyocyte: new revelations on the interplay between architecture and function in growth, health, and disease’.
Metadata
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Copyright, Publisher and Additional Information: | © 2022 The Authors. Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, provided the original author and source are credited. | ||||
Keywords: | dyad; 3D visualisation; expansion microscopy; ryanodine receptor; junctophilin; right ventricular failure | ||||
Dates: |
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Institution: | The University of Sheffield | ||||
Academic Units: | The University of Sheffield > Faculty of Science (Sheffield) > School of Biosciences (Sheffield) | ||||
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Depositing User: | Symplectic Sheffield | ||||
Date Deposited: | 06 Jun 2022 11:38 | ||||
Last Modified: | 10 Feb 2023 14:13 | ||||
Status: | Published | ||||
Publisher: | Royal Society, The | ||||
Refereed: | Yes | ||||
Identification Number: | https://doi.org/10.1098/rstb.2021.0316 |