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Three-dimensional morphogenesis of MDCK cells induced by cellular contractile forces on a viscous substrate

Substrate physical properties are essential for many physiological events such as embryonic development and 3D tissue formation. Physical properties of the extracellular matrix such as viscoelasticity and geometrical constraints are understood as factors that affect cell behaviour. In this study, we...

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Autores principales: Imai, Misako, Furusawa, Kazuya, Mizutani, Takeomi, Kawabata, Kazushige, Haga, Hisashi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4571640/
https://www.ncbi.nlm.nih.gov/pubmed/26374384
http://dx.doi.org/10.1038/srep14208
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author Imai, Misako
Furusawa, Kazuya
Mizutani, Takeomi
Kawabata, Kazushige
Haga, Hisashi
author_facet Imai, Misako
Furusawa, Kazuya
Mizutani, Takeomi
Kawabata, Kazushige
Haga, Hisashi
author_sort Imai, Misako
collection PubMed
description Substrate physical properties are essential for many physiological events such as embryonic development and 3D tissue formation. Physical properties of the extracellular matrix such as viscoelasticity and geometrical constraints are understood as factors that affect cell behaviour. In this study, we focused on the relationship between epithelial cell 3D morphogenesis and the substrate viscosity. We observed that Madin-Darby Canine Kidney (MDCK) cells formed 3D structures on a viscous substrate (Matrigel). The structures appear as a tulip hat. We then changed the substrate viscosity by genipin (GP) treatment. GP is a cross-linker of amino groups. Cells cultured on GP-treated-matrigel changed their 3D morphology in a substrate viscosity-dependent manner. Furthermore, to elucidate the spatial distribution of the cellular contractile force, localization of mono-phosphorylated and di-phosphorylated myosin regulatory light chain (P-MRLCs) was visualized by immunofluorescence. P-MRLCs localized along the periphery of epithelial sheets. Treatment with Y-27632, a Rho-kinase inhibitor, blocked the P-MRLCs localization at the edge of epithelial sheets and halted 3D morphogenesis. Our results indicate that the substrate viscosity, the substrate deformation, and the cellular contractile forces induced by P-MRLCs play crucial roles in 3D morphogenesis.
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spelling pubmed-45716402015-09-28 Three-dimensional morphogenesis of MDCK cells induced by cellular contractile forces on a viscous substrate Imai, Misako Furusawa, Kazuya Mizutani, Takeomi Kawabata, Kazushige Haga, Hisashi Sci Rep Article Substrate physical properties are essential for many physiological events such as embryonic development and 3D tissue formation. Physical properties of the extracellular matrix such as viscoelasticity and geometrical constraints are understood as factors that affect cell behaviour. In this study, we focused on the relationship between epithelial cell 3D morphogenesis and the substrate viscosity. We observed that Madin-Darby Canine Kidney (MDCK) cells formed 3D structures on a viscous substrate (Matrigel). The structures appear as a tulip hat. We then changed the substrate viscosity by genipin (GP) treatment. GP is a cross-linker of amino groups. Cells cultured on GP-treated-matrigel changed their 3D morphology in a substrate viscosity-dependent manner. Furthermore, to elucidate the spatial distribution of the cellular contractile force, localization of mono-phosphorylated and di-phosphorylated myosin regulatory light chain (P-MRLCs) was visualized by immunofluorescence. P-MRLCs localized along the periphery of epithelial sheets. Treatment with Y-27632, a Rho-kinase inhibitor, blocked the P-MRLCs localization at the edge of epithelial sheets and halted 3D morphogenesis. Our results indicate that the substrate viscosity, the substrate deformation, and the cellular contractile forces induced by P-MRLCs play crucial roles in 3D morphogenesis. Nature Publishing Group 2015-09-16 /pmc/articles/PMC4571640/ /pubmed/26374384 http://dx.doi.org/10.1038/srep14208 Text en Copyright © 2015, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Imai, Misako
Furusawa, Kazuya
Mizutani, Takeomi
Kawabata, Kazushige
Haga, Hisashi
Three-dimensional morphogenesis of MDCK cells induced by cellular contractile forces on a viscous substrate
title Three-dimensional morphogenesis of MDCK cells induced by cellular contractile forces on a viscous substrate
title_full Three-dimensional morphogenesis of MDCK cells induced by cellular contractile forces on a viscous substrate
title_fullStr Three-dimensional morphogenesis of MDCK cells induced by cellular contractile forces on a viscous substrate
title_full_unstemmed Three-dimensional morphogenesis of MDCK cells induced by cellular contractile forces on a viscous substrate
title_short Three-dimensional morphogenesis of MDCK cells induced by cellular contractile forces on a viscous substrate
title_sort three-dimensional morphogenesis of mdck cells induced by cellular contractile forces on a viscous substrate
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4571640/
https://www.ncbi.nlm.nih.gov/pubmed/26374384
http://dx.doi.org/10.1038/srep14208
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