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- W2046669165 abstract "Background: The cytoskeletal protein vimentin is involved in the determination of cell mechanical properties, shape and motile behavior. We are investigating how vimentin participates in the response to external mechanical cues. Methods: Normal and mouse embryo fibroblasts (mEF) harvested from the vimentin-knockout mouse are grown on fibronectin-coated polyacrylamide gels (Fn-PAA) of 0.5-40 kPa. Protein expression levels are assayed by Western blotting, and vimentin network distribution by immunofluorescence. Rates of vimentin subunit turnover are assayed by fluorescence recovery after photobleaching (FRAP) experiments; and atomic force microscopy (AFM) is used to measure cells’ elastic and viscoelastic-plastic properties. Results: Vimentin protein expression levels do not change in response to substrate stiffness. Whereas FRAP results in other cell types - which are less phenotypically responsive to substrate stiffness - show no difference in the rate of vimentin subunit turnover across various substrates, early results suggest that subunit turnover increases when fibroblasts are grown on physiological-range stiffnesses (∼6 kPa). Also, many short vimentin filaments and squiggles (<15 µm in length) are evident that appear not to be connected to the extended vimentin network, and vimentin is more detergent-soluble under these conditions. The responses (e.g. shape, motility, etc.) of vimentin-null fibroblasts confirm vimentin's participation in these processes across the range of substrate stiffnesses. Finally, vimentin-null mEF are less stiff than mEF on ∼6 kPa Fn-PAA, and initial experiments show that whether vimentin makes cells softer or stiffer depends upon the substrate elastic modulus. Conclusion: Vimentin is expressed at consistent levels across conditions that cause changes in cell stiffness, shape and motility, yet the response of vimentin-knockout cells to the same conditions demonstrates vimentin's role in response to mechanical stimuli. We hypothesize that modulation of vimentin's assembly state underlies its contribution to cell mechanics. NIH-NHLBI NRSA, NIH PO1GM096971" @default.
- W2046669165 created "2016-06-24" @default.
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- W2046669165 date "2013-01-01" @default.
- W2046669165 modified "2023-09-27" @default.
- W2046669165 title "Vimentin Filament Assembly is Altered by Substrate Elastic Modulus" @default.
- W2046669165 doi "https://doi.org/10.1016/j.bpj.2012.11.802" @default.
- W2046669165 hasPublicationYear "2013" @default.
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