MINFLUX microscopy resolves subunits of the cardiac ryanodine receptor and its 3D orientation in cells
Year: 2025
Type: article
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.
Source: Nature Communications
Language: English
Subfield: Biophysics
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $6,990
Awards: Molecular control of ryanodine receptor dispersion in cardiac myocytes – a new mechanism that affects calcium release in health and disease, A MINFLUX optical super-resolution microscope to investigate molecular scale structure-function relationships, G311218137, G4060900387, G7289894738, G8690818240
OpenAlex ID: W7116632277