title: Wide and high resolution tension measurement using FRET in embryo.
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.
authors.affiliation: Department of Life Sciences (Biology), Graduate School of Arts and Sciences, The University of Tokyo, Tokyo, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Yamashita
authors.affiliation: Department of Life Sciences (Biology), Graduate School of Arts and Sciences, The University of Tokyo, Tokyo, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Tsuboi
authors.affiliation: Department of Life Sciences (Biology), Graduate School of Arts and Sciences, The University of Tokyo, Tokyo, Japan.
authors.forename: Nanako
authors.initials: N
authors.lastname: Ishinabe
authors.affiliation: Cell Signaling Group, WASEDA Bioscience Research Institute in Singapore (WABIOS), 11 Biopolis Way #05-02 Helios, Singapore 138667, Singapore.|Organization for University Research Initiatives, Waseda University, #304, Block 120-4, 513 Wasedatsurumaki-cho, Shinjuku-ku, Tokyo, 162-0041, Japan.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Kitaguchi
authors.affiliation: Department of Life Sciences (Biology), Graduate School of Arts and Sciences, The University of Tokyo, Tokyo, Japan.
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Michiue
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000185:Actinin / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D004475:Ectoderm / Q000378:metabolism; D005349:Fibroins / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D054262:Gastrulation / Q000502:physiology; D057809:HEK293 Cells; D006801:Humans; D009024:Morphogenesis / Q000502:physiology*; D009218:Myosins / Q000378:metabolism; D014982:Xenopus laevis
chemical_list: D000185:Actinin; D005349:Fibroins; D009218:Myosins