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Substrate stiffness regulates solubility of cellular vimentin

The intermediate filament protein vimentin is involved in the regulation of cell behavior, morphology, and mechanical properties. Previous studies using cells cultured on glass or plastic substrates showed that vimentin is largely insoluble. Although substrate stiffness was shown to alter many aspec...

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Detalles Bibliográficos
Autores principales: Murray, Maria E., Mendez, Melissa G., Janmey, Paul A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3873896/
https://www.ncbi.nlm.nih.gov/pubmed/24173714
http://dx.doi.org/10.1091/mbc.E13-06-0326
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author Murray, Maria E.
Mendez, Melissa G.
Janmey, Paul A.
author_facet Murray, Maria E.
Mendez, Melissa G.
Janmey, Paul A.
author_sort Murray, Maria E.
collection PubMed
description The intermediate filament protein vimentin is involved in the regulation of cell behavior, morphology, and mechanical properties. Previous studies using cells cultured on glass or plastic substrates showed that vimentin is largely insoluble. Although substrate stiffness was shown to alter many aspects of cell behavior, changes in vimentin organization were not reported. Our results show for the first time that mesenchymal stem cells (hMSCs), endothelial cells, and fibroblasts cultured on different-stiffness substrates exhibit biphasic changes in vimentin detergent solubility, which increases from nearly 0 to 67% in hMSCs coincident with increases in cell spreading and membrane ruffling. When imaged, the detergent-soluble vimentin appears to consist of small fragments the length of one or several unit-length filaments. Vimentin detergent solubility decreases when these cells are subjected to serum starvation, allowed to form cell–cell contacts, after microtubule disruption, or inhibition of Rac1, Rho-activated kinase, or p21-activated kinase. Inhibiting myosin or actin assembly increases vimentin solubility on rigid substrates. These data suggest that in the mechanical environment in vivo, vimentin is more dynamic than previously reported and its assembly state is sensitive to stimuli that alter cellular tension and morphology.
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spelling pubmed-38738962014-03-16 Substrate stiffness regulates solubility of cellular vimentin Murray, Maria E. Mendez, Melissa G. Janmey, Paul A. Mol Biol Cell Articles The intermediate filament protein vimentin is involved in the regulation of cell behavior, morphology, and mechanical properties. Previous studies using cells cultured on glass or plastic substrates showed that vimentin is largely insoluble. Although substrate stiffness was shown to alter many aspects of cell behavior, changes in vimentin organization were not reported. Our results show for the first time that mesenchymal stem cells (hMSCs), endothelial cells, and fibroblasts cultured on different-stiffness substrates exhibit biphasic changes in vimentin detergent solubility, which increases from nearly 0 to 67% in hMSCs coincident with increases in cell spreading and membrane ruffling. When imaged, the detergent-soluble vimentin appears to consist of small fragments the length of one or several unit-length filaments. Vimentin detergent solubility decreases when these cells are subjected to serum starvation, allowed to form cell–cell contacts, after microtubule disruption, or inhibition of Rac1, Rho-activated kinase, or p21-activated kinase. Inhibiting myosin or actin assembly increases vimentin solubility on rigid substrates. These data suggest that in the mechanical environment in vivo, vimentin is more dynamic than previously reported and its assembly state is sensitive to stimuli that alter cellular tension and morphology. The American Society for Cell Biology 2014-01-01 /pmc/articles/PMC3873896/ /pubmed/24173714 http://dx.doi.org/10.1091/mbc.E13-06-0326 Text en © 2014 Murray et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society of Cell Biology.
spellingShingle Articles
Murray, Maria E.
Mendez, Melissa G.
Janmey, Paul A.
Substrate stiffness regulates solubility of cellular vimentin
title Substrate stiffness regulates solubility of cellular vimentin
title_full Substrate stiffness regulates solubility of cellular vimentin
title_fullStr Substrate stiffness regulates solubility of cellular vimentin
title_full_unstemmed Substrate stiffness regulates solubility of cellular vimentin
title_short Substrate stiffness regulates solubility of cellular vimentin
title_sort substrate stiffness regulates solubility of cellular vimentin
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3873896/
https://www.ncbi.nlm.nih.gov/pubmed/24173714
http://dx.doi.org/10.1091/mbc.E13-06-0326
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