MINFLUX microscopy resolves subunits of the cardiac ryanodine receptor and its 3D orientation in cells.
Publisher DOI
PubMed ID
41423693
Abstract
The cardiac ryanodine receptor (RyR2) constitutes the molecular basis of the process of calcium-induced calcium release where activation of RyR2s can be locally regenerative. Here, we present purely optical data of RyR2 distribution with sub-molecular resolution by applying 3D MINFLUX microscopy. Using single-domain antibodies and DNA-PAINT we determine the location of individual RyR2 subunits with high precision (~3 nm) and resolve the 3D orientations of RyR2s in-situ. We measured labeling efficiencies of ~50%, implying RyR2 tetramer detection probability approaching 95%. In HEK293 cells, RyR2 expression was dense, with some clusters containing several hundred RyR2s. Ventricular myocytes from mice contained large clusters containing many tens of close-packed RyR2s, resolving apparent discrepancies between electron microscopy and previous super-resolution microscopy data. The methodology developed here reveals the full 3D morphological complexity of RyR2 channels and is applicable to other multi-subunit complexes in a variety of cell types.
Date Issued
2025-12-21
Publication Type
Article
Subject(s)
Language(s)
en
Author(s)
Jansen, Isabelle | |
Jones, Peter P | |
Louch, William E |
Journal
Nature Communications
Publisher
Nature Research
ISSN
2041-1723
Access(Rights)
open.access