Synchronization of endothelial Dll4-Notch dynamics switch blood vessels from branching to expansion

  1. Benedetta Ubezio
  2. Raquel Agudo Blanco
  3. Ilse Geudens
  4. Fabio Stanchi
  5. Thomas Mathivet
  6. Martin L Jones
  7. Anan Ragab
  8. Katie Bentley
  9. Holger Gerhardt  Is a corresponding author
  1. London Research Institute, United Kingdom
  2. Vesalius Research Center, VIB, Belgium
  3. Harvard Medical School, United States

Abstract

Formation of a regularly branched blood vessel network is crucial in development and physiology. Here we show that the expression of the Notch ligand Dll4 fluctuates in individual endothelial cells within sprouting vessels in the mouse retina in vivo and in correlation with dynamic cell movement in mouse embryonic stem cell-derived sprouting assays. We also find that sprout elongation and branching associates with a highly differential phase pattern of Dll4 between endothelial cells. Stimulation with pathologically high levels of Vegf, or overexpression of Dll4, leads to Notch dependent synchronization of Dll4 fluctuations within clusters, both in vitro and in vivo. Our results demonstrate that the Vegf-Dll4/Notch feedback system normally operates to generate heterogeneity between endothelial cells driving branching, whilst synchronization drives vessel expansion. We propose that this sensitive phase transition in the behaviour of the Vegf-Dll4/Notch feedback loop underlies the morphogen function of Vegfa in vascular patterning.

Article and author information

Author details

  1. Benedetta Ubezio

    Vascular Biology Laboratory, London Research Institute, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  2. Raquel Agudo Blanco

    Vascular Biology Laboratory, London Research Institute, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  3. Ilse Geudens

    Vascular Patterning Laboratory, Vesalius Research Center, VIB, Leuven, Belgium
    Competing interests
    The authors declare that no competing interests exist.
  4. Fabio Stanchi

    Vascular Patterning Laboratory, Vesalius Research Center, VIB, Leuven, Belgium
    Competing interests
    The authors declare that no competing interests exist.
  5. Thomas Mathivet

    Vascular Patterning Laboratory, Vesalius Research Center, VIB, Leuven, Belgium
    Competing interests
    The authors declare that no competing interests exist.
  6. Martin L Jones

    Vascular Biology Laboratory, London Research Institute, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  7. Anan Ragab

    Vascular Biology Laboratory, London Research Institute, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  8. Katie Bentley

    Computational Biology Laboratory, Center for Vascular Biology Research, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Holger Gerhardt

    Vascular Biology Laboratory, London Research Institute, London, United Kingdom
    For correspondence
    holger.gerhardt@mdc-berlin.de
    Competing interests
    The authors declare that no competing interests exist.

Ethics

Animal experimentation: Mice were maintained at London Research Institute under standard husbandry conditions. All protocols were approved by the UK Home Office (P80/2391). Glioblastoma studies were performed at the Vesalius Research Center, VIB, KU Leuven where housing and all experimental animal procedures were performed in accordance with Belgian law on animal care and were approved by the Institutional Animal Care and Research Advisory Committee of the K. U. Leuven (P105/2012).

Reviewing Editor

  1. Tanya T Whitfield, University of Sheffield, United Kingdom

Publication history

  1. Received: October 7, 2015
  2. Accepted: April 11, 2016
  3. Accepted Manuscript published: April 13, 2016 (version 1)
  4. Version of Record published: June 6, 2016 (version 2)
  5. Version of Record updated: May 9, 2017 (version 3)

Copyright

© 2016, Ubezio 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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  1. Benedetta Ubezio
  2. Raquel Agudo Blanco
  3. Ilse Geudens
  4. Fabio Stanchi
  5. Thomas Mathivet
  6. Martin L Jones
  7. Anan Ragab
  8. Katie Bentley
  9. Holger Gerhardt
(2016)
Synchronization of endothelial Dll4-Notch dynamics switch blood vessels from branching to expansion
eLife 5:e12167.
https://doi.org/10.7554/eLife.12167
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