Wide and high resolution tension measurement using FRET in embryo

Satoshi Yamashita, Takashi Tsuboi, Nanako Ishinabe, Tetsuya Kitaguchi, Tatsuo Michiue

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Abstract

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.

Original languageEnglish
Article number28535
JournalScientific Reports
Volume6
DOIs
Publication statusPublished - 2016 Jun 23

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Yamashita, S., Tsuboi, T., Ishinabe, N., Kitaguchi, T., & Michiue, T. (2016). Wide and high resolution tension measurement using FRET in embryo. Scientific Reports, 6, [28535]. https://doi.org/10.1038/srep28535