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Wide and high resolution tension measurement using FRET in embryo

During embryonic development, physical force plays an important role in morphogenesis and differentiation. Stretch sensitive fluorescence resonance energy transfer (FRET) has the potential to provide non-invasive tension measurements inside living tissue. In this study, we introduced a FRET-based ac...

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Autores principales: Yamashita, Satoshi, Tsuboi, Takashi, Ishinabe, Nanako, Kitaguchi, Tetsuya, Michiue, Tatsuo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4917836/
https://www.ncbi.nlm.nih.gov/pubmed/27335157
http://dx.doi.org/10.1038/srep28535
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author Yamashita, Satoshi
Tsuboi, Takashi
Ishinabe, Nanako
Kitaguchi, Tetsuya
Michiue, Tatsuo
author_facet Yamashita, Satoshi
Tsuboi, Takashi
Ishinabe, Nanako
Kitaguchi, Tetsuya
Michiue, Tatsuo
author_sort Yamashita, Satoshi
collection PubMed
description During embryonic development, physical force plays an important role in morphogenesis and differentiation. Stretch sensitive fluorescence resonance energy transfer (FRET) has the potential to provide non-invasive tension measurements inside living tissue. In this study, we introduced a FRET-based actinin tension sensor into Xenopus laevis embryos and demonstrated that this sensor captures variation of tension across differentiating ectoderm. The actinin tension sensor, containing mCherry and EGFP connected by spider silk protein, was validated in human embryonic kidney (HEK) cells and embryos. It co-localized with actin filaments and changed FRET efficiencies in response to actin filament destruction, myosin deactivation, and osmotic perturbation. Time-lapse FRET analysis showed that the prospective neural ectoderm bears higher tension than the epidermal ectoderm during gastrulation and neurulation, and cells morphogenetic behavior correlated with the tension difference. These data confirmed that the sensor enables us to measure tension across tissues concurrently and with high resolution.
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spelling pubmed-49178362016-06-27 Wide and high resolution tension measurement using FRET in embryo Yamashita, Satoshi Tsuboi, Takashi Ishinabe, Nanako Kitaguchi, Tetsuya Michiue, Tatsuo Sci Rep Article During embryonic development, physical force plays an important role in morphogenesis and differentiation. Stretch sensitive fluorescence resonance energy transfer (FRET) has the potential to provide non-invasive tension measurements inside living tissue. In this study, we introduced a FRET-based actinin tension sensor into Xenopus laevis embryos and demonstrated that this sensor captures variation of tension across differentiating ectoderm. The actinin tension sensor, containing mCherry and EGFP connected by spider silk protein, was validated in human embryonic kidney (HEK) cells and embryos. It co-localized with actin filaments and changed FRET efficiencies in response to actin filament destruction, myosin deactivation, and osmotic perturbation. Time-lapse FRET analysis showed that the prospective neural ectoderm bears higher tension than the epidermal ectoderm during gastrulation and neurulation, and cells morphogenetic behavior correlated with the tension difference. These data confirmed that the sensor enables us to measure tension across tissues concurrently and with high resolution. Nature Publishing Group 2016-06-23 /pmc/articles/PMC4917836/ /pubmed/27335157 http://dx.doi.org/10.1038/srep28535 Text en Copyright © 2016, 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
Yamashita, Satoshi
Tsuboi, Takashi
Ishinabe, Nanako
Kitaguchi, Tetsuya
Michiue, Tatsuo
Wide and high resolution tension measurement using FRET in embryo
title Wide and high resolution tension measurement using FRET in embryo
title_full Wide and high resolution tension measurement using FRET in embryo
title_fullStr Wide and high resolution tension measurement using FRET in embryo
title_full_unstemmed Wide and high resolution tension measurement using FRET in embryo
title_short Wide and high resolution tension measurement using FRET in embryo
title_sort wide and high resolution tension measurement using fret in embryo
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4917836/
https://www.ncbi.nlm.nih.gov/pubmed/27335157
http://dx.doi.org/10.1038/srep28535
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