Heterogeneity of cell membrane structure studied by single molecule tracking

Heterogeneity in cell membrane structure, typified by microdomains with different biophysical and biochemical properties, is thought to impact on a variety of cell functions. Integral membrane proteins act as nanometre-sized probes of the lipid environment and their thermally-driven movements can be...

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Bibliographic Details
Published in:Faraday discussions 2021-12, Vol.232, p.358-374
Main Authors: Mashanov, Gregory I, Nenasheva, Tatiana A, Mashanova, Alla, Lape, Remigijus, Birdsall, Nigel J. M, Sivilotti, Lucia, Molloy, Justin E
Format: Article
Language:English
Subjects:
Cell Membrane
Cell Membrane Structures
Cell membranes
Chemistry
Fluorescence
Heterogeneity
Lipids
Membrane Proteins
Membrane structures
Microscopy, Fluorescence
Molecular structure
Proteins
Single Molecule Imaging
Statistical tests
Tracking
Viscosity
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Summary:Heterogeneity in cell membrane structure, typified by microdomains with different biophysical and biochemical properties, is thought to impact on a variety of cell functions. Integral membrane proteins act as nanometre-sized probes of the lipid environment and their thermally-driven movements can be used to report local variations in membrane properties. In the current study, we have used total internal reflection fluorescence microscopy (TIRFM) combined with super-resolution tracking of multiple individual molecules, in order to create high-resolution maps of local membrane viscosity. We used a quadrat sampling method and show how statistical tests for membrane heterogeneity can be conducted by analysing the paths of many molecules that pass through the same unit area of membrane. We describe experiments performed on cultured primary cells, stable cell lines and ex vivo tissue slices using a variety of membrane proteins, under different imaging conditions. In some cell types, we find no evidence for heterogeneity in mobility across the plasma membrane, but in others we find statistically significant differences with some regions of membrane showing significantly higher viscosity than others. We use total internal reflection fluorescence microscopy combined with super-resolution tracking of multiple individual molecules, in order to create high-resolution maps of local membrane viscosity.
ISSN:1359-6640
1364-5498
DOI:10.1039/d1fd00035g