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Measuring mechanodynamics in an unsupported epithelial monolayer grown at an air–water interface

Actomyosin contraction and relaxation in a monolayer is a fundamental biophysical process in development and homeostasis. Current methods used to characterize the mechanodynamics of monolayers often involve cells grown on solid supports such as glass or gels. The results of these studies are fundame...

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Autores principales: Gullekson, Corinne, Walker, Matthew, Harden, James L., Pelling, Andrew E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5221614/
https://www.ncbi.nlm.nih.gov/pubmed/28035043
http://dx.doi.org/10.1091/mbc.E16-05-0300
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author Gullekson, Corinne
Walker, Matthew
Harden, James L.
Pelling, Andrew E.
author_facet Gullekson, Corinne
Walker, Matthew
Harden, James L.
Pelling, Andrew E.
author_sort Gullekson, Corinne
collection PubMed
description Actomyosin contraction and relaxation in a monolayer is a fundamental biophysical process in development and homeostasis. Current methods used to characterize the mechanodynamics of monolayers often involve cells grown on solid supports such as glass or gels. The results of these studies are fundamentally influenced by these supporting structures. Here we describe a new method for measuring the mechanodynamics of epithelial monolayers by culturing cells at an air–liquid interface. These model monolayers are grown in the absence of any supporting structures, removing cell–substrate effects. This method’s potential was evaluated by observing and quantifying the generation and release of internal stresses upon actomyosin contraction (800 ± 100 Pa) and relaxation (600 ± 100 Pa) in response to chemical treatments. Although unsupported monolayers exhibited clear major and minor strain axes, they were not correlated with nuclear alignment as observed when the monolayers were grown on soft deformable gels. It was also observed that both gels and glass substrates led to the promotion of long-range cell nuclei alignment not seen in the hanging-drop model. This new approach provides us with a picture of basal actomyosin mechanodynamics in a simplified system, allowing us to infer how the presence of a substrate affects contractility and long-range multicellular organization and dynamics.
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spelling pubmed-52216142017-03-16 Measuring mechanodynamics in an unsupported epithelial monolayer grown at an air–water interface Gullekson, Corinne Walker, Matthew Harden, James L. Pelling, Andrew E. Mol Biol Cell Articles Actomyosin contraction and relaxation in a monolayer is a fundamental biophysical process in development and homeostasis. Current methods used to characterize the mechanodynamics of monolayers often involve cells grown on solid supports such as glass or gels. The results of these studies are fundamentally influenced by these supporting structures. Here we describe a new method for measuring the mechanodynamics of epithelial monolayers by culturing cells at an air–liquid interface. These model monolayers are grown in the absence of any supporting structures, removing cell–substrate effects. This method’s potential was evaluated by observing and quantifying the generation and release of internal stresses upon actomyosin contraction (800 ± 100 Pa) and relaxation (600 ± 100 Pa) in response to chemical treatments. Although unsupported monolayers exhibited clear major and minor strain axes, they were not correlated with nuclear alignment as observed when the monolayers were grown on soft deformable gels. It was also observed that both gels and glass substrates led to the promotion of long-range cell nuclei alignment not seen in the hanging-drop model. This new approach provides us with a picture of basal actomyosin mechanodynamics in a simplified system, allowing us to infer how the presence of a substrate affects contractility and long-range multicellular organization and dynamics. The American Society for Cell Biology 2017-01-01 /pmc/articles/PMC5221614/ /pubmed/28035043 http://dx.doi.org/10.1091/mbc.E16-05-0300 Text en © 2017 Gullekson 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 for Cell Biology.
spellingShingle Articles
Gullekson, Corinne
Walker, Matthew
Harden, James L.
Pelling, Andrew E.
Measuring mechanodynamics in an unsupported epithelial monolayer grown at an air–water interface
title Measuring mechanodynamics in an unsupported epithelial monolayer grown at an air–water interface
title_full Measuring mechanodynamics in an unsupported epithelial monolayer grown at an air–water interface
title_fullStr Measuring mechanodynamics in an unsupported epithelial monolayer grown at an air–water interface
title_full_unstemmed Measuring mechanodynamics in an unsupported epithelial monolayer grown at an air–water interface
title_short Measuring mechanodynamics in an unsupported epithelial monolayer grown at an air–water interface
title_sort measuring mechanodynamics in an unsupported epithelial monolayer grown at an air–water interface
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5221614/
https://www.ncbi.nlm.nih.gov/pubmed/28035043
http://dx.doi.org/10.1091/mbc.E16-05-0300
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