Journal article
Live imaging molecular changes in junctional tension upon VE-cadherin in zebrafish
AK Lagendijk, GA Gomez, S Baek, D Hesselson, WE Hughes, S Paterson, DE Conway, HG Belting, M Affolter, KA Smith, MA Schwartz, AS Yap, BM Hogan
Nature Communications | NATURE PORTFOLIO | Published : 2017
Open access
Abstract
Forces play diverse roles in vascular development, homeostasis and disease. VE-cadherin at endothelial cell-cell junctions links the contractile acto-myosin cytoskeletons of adjacent cells, serving as a tension-transducer. To explore tensile changes across VE-cadherin in live zebrafish, we tailored an optical biosensor approach, originally established in vitro. We validate localization and function of a VE-cadherin tension sensor (TS) in vivo. Changes in tension across VE-cadherin observed using ratio-metric or lifetime FRET measurements reflect acto-myosin contractility within endothelial cells. Furthermore, we apply the TS to reveal biologically relevant changes in VE-cadherin tension that..
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Awarded by Australian Cancer Research Foundation
Funding Acknowledgements
A.K.L. was supported by a UQ Postdoctoral Fellowship, B.M.H. by an NHMRC/National Heart Foundation Career Development Fellowship (1083811) and A.S.Y. by an NHMRC Research Fellowship (1044041). This research was supported by an ARC Discovery Project grant (DP150104119) and NHMRC grants (1067405, 1037320). We thank Kylie Georgas for design of graphics in the manuscript. We thank Dr. Sean Warren for assistance with FLIM acquisition and kindly providing us with FLIMfit software<SUP>55</SUP> for lifetime analysis. We thank H. Gerhardt and L-K. Phng for kindly providing DNA constructs that we used to generate the Tg(flilep:lifeact-mCherry)<SUP>uq12bh</SUP> strain. We thank E. Scott for providing the Tg(10.5xUAS:GCaMP5G)<SUP>uq2Tg</SUP> stain<SUP>41</SUP>. Imaging was performed in the Australian Cancer Research Foundation's Dynamic Imaging Facility at IMB (established with the generous support of the ACRF), in the Queensland Brain Institute's Advanced Microscopy Facility and generously supported by ARC LIEF LE130100078 and in the Microscope Facility of the Garvan Institute of Medical Research.