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© Research
Publication : Molecular biology of the cell

Novel cryo-tomography workflow reveals nanometer-scale responses of epithelial cells to matrix stiffness and dimensionality.

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Molecular biology of the cell - 26 Oct 2022

Gaietta G, Kai F, Swift MF, Weaver VM, Volkmann N, Hanein D,

Link to Pubmed [PMID] – 36287913

Link to DOI – 10.1091/mbc.E22-03-0092

Mol Biol Cell 2022 Oct; (): mbcE22030092

Matrix stiffness and dimensionality were shown to be major determinants of cell behavior. However, a workflow for examining nanometer-scale responses of the associated molecular machinery is not available. Here, we describe a comprehensive, quantitative workflow that permits the analysis of cells responding to mechanical and dimensionality cues by cryogenic electron-tomography, in their native state at nanometer scale. Using this approach, we quantified distinct cytoskeletal nanoarchitectures and vesicle phenotypes induced in human mammary epithelial cells in response to stiffness and dimensionality of reconstituted basement membrane. Our workflow closely recapitulates the microenvironment associated with acinar morphogenesis and identified distinct differences in-situ at nanometer scale. Using drug treatment, we showed that molecular events and nanometer-scale rearrangements triggered by engagement of apical cell receptors with reconstituted basement membrane correspond to changes induced by reduction of cortical tension. Our approach is fully adaptable to any kind of stiffness regime, extracellular matrix composition, and drug treatment.

https://pubmed.ncbi.nlm.nih.gov/36287913