1. Developmental Biology
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Live-imaging of endothelial Erk activity reveals dynamic and sequential signalling events during regenerative angiogenesis

  1. Kazuhide S Okuda
  2. Mikaela S Keyser
  3. David B Gurevich
  4. Caterina Sturtzel
  5. Elizabeth A Mason
  6. Scott Paterson
  7. Huijun Chen
  8. Mark Scott
  9. Nicholas D Condon
  10. Paul Martin
  11. Martin Distel
  12. Benjamin M Hogan  Is a corresponding author
  1. Peter MacCallum Cancer Centre, Australia
  2. University of Queensland, Australia
  3. University of Bristol, United Kingdom
  4. St. Anna Kinderkrebsforschung, Children's Cancer Research Institute, Austria
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Cite this article as: eLife 2021;10:e62196 doi: 10.7554/eLife.62196

Abstract

The formation of new blood vessel networks occurs via angiogenesis during development, tissue repair and disease. Angiogenesis is regulated by intracellular endothelial signalling pathways, induced downstream of Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs). A major challenge in understanding angiogenesis is interpreting how signalling events occur dynamically within endothelial cell populations during sprouting, proliferation and migration. Erk is a central downstream effector of Vegf-signalling and reports the signalling that drives angiogenesis. We generated a vascular Erk biosensor transgenic line in zebrafish using a kinase translocation reporter that allows live-imaging of Erk-signalling dynamics. We demonstrate the utility of this line to live-image Erk activity during physiologically relevant angiogenic events. Further, we reveal dynamic and sequential endothelial cell Erk-signalling events following blood vessel wounding. Initial signalling is dependent upon Ca2+ in the earliest responding endothelial cells, but is independent of Vegfr-signalling and local inflammation. The sustained regenerative response however, involves a Vegfr-dependent mechanism that initiates concomitant with the wound inflammatory response. This work reveals a highly dynamic sequence of signalling events in regenerative angiogenesis and validates a new resource for the study of vascular Erk-signalling in real-time.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files.

Article and author information

Author details

  1. Kazuhide S Okuda

    Program in Organogenesis and Cancer, Peter MacCallum Cancer Centre, Melbourne, Australia
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8060-0377
  2. Mikaela S Keyser

    Institute for Molecular Bioscience, University of Queensland, Brisbane, Australia
    Competing interests
    The authors declare that no competing interests exist.
  3. David B Gurevich

    Biochemistry, University of Bristol, Bristol, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  4. Caterina Sturtzel

    Innovative Cancer Models, St. Anna Kinderkrebsforschung, Children's Cancer Research Institute, Vienna, Austria
    Competing interests
    The authors declare that no competing interests exist.
  5. Elizabeth A Mason

    Program in Organogenesis and Cancer, Peter MacCallum Cancer Centre, Melbourne, Australia
    Competing interests
    The authors declare that no competing interests exist.
  6. Scott Paterson

    Program in Organogenesis and Cancer, Peter MacCallum Cancer Centre, Melbourne, Australia
    Competing interests
    The authors declare that no competing interests exist.
  7. Huijun Chen

    Institute for Molecular Bioscience, University of Queensland, Brisbane, Australia
    Competing interests
    The authors declare that no competing interests exist.
  8. Mark Scott

    Institute for Molecular Bioscience, University of Queensland, Brisbane, Australia
    Competing interests
    The authors declare that no competing interests exist.
  9. Nicholas D Condon

    Institute for Molecular Bioscience, University of Queensland, Brisbane, Australia
    Competing interests
    The authors declare that no competing interests exist.
  10. Paul Martin

    Biochemistry, University of Bristol, Bristol, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  11. Martin Distel

    Innovative Cancer Models, St. Anna Kinderkrebsforschung, Children's Cancer Research Institute, Vienna, Austria
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5942-0817
  12. Benjamin M Hogan

    Program in Organogenesis and Cancer, Peter MacCallum Cancer Centre, Melbourne, Australia
    For correspondence
    ben.hogan@petermac.org
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0651-7065

Funding

National Health and Medical Research Council (1164734)

  • Benjamin M Hogan

National Health and Medical Research Council (1165117)

  • Benjamin M Hogan

Austrian Research Promotion Agency (7640628)

  • Martin Distel

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All zebrafish work was conducted in accordance with the guidelines of the animal ethics committees at the University of Queensland (AE54297), University of Melbourne, Peter MacCallum Cancer Centre (E634 and E643), University of Bristol (3003318), and the Children's Cancer Research Institute (GZ:565304/2014/6 and GZ:534619/2014/4).

Reviewing Editor

  1. Saulius Sumanas

Publication history

  1. Received: August 17, 2020
  2. Accepted: May 17, 2021
  3. Accepted Manuscript published: May 18, 2021 (version 1)
  4. Version of Record published: June 3, 2021 (version 2)

Copyright

© 2021, Okuda et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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