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Endothelial cells dynamically compete for the tip cell position during angiogenic sprouting

Nature Cell Biology volume 12, pages 943–953 (2010)Cite this article

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Abstract

Sprouting angiogenesis requires the coordinated behaviour of endothelial cells, regulated by Notch and vascular endothelial growth factor receptor (VEGFR) signalling. Here, we use computational modelling and genetic mosaic sprouting assays in vitro and in vivo to investigate the regulation and dynamics of endothelial cells during tip cell selection. We find that endothelial cells compete for the tip cell position through relative levels of Vegfr1 and Vegfr2, demonstrating a biological role for differential Vegfr regulation in individual endothelial cells. Differential Vegfr levels affect tip selection only in the presence of a functional Notch system by modulating the expression of the ligand Dll4. Time-lapse microscopy imaging of mosaic sprouts identifies dynamic position shuffling of tip and stalk cells in vitro and in vivo, indicating that the VEGFR–Dll4–Notch signalling circuit is constantly re-evaluated as cells meet new neighbours. The regular exchange of the leading tip cell raises novel implications for the concept of guided angiogenic sprouting.

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Figure 1: VEGFR levels in mosaic-tip-cell selection in silico, in vitro and in vivo.
Figure 2: Dll4 and Notch signalling in competition.
Figure 3: Notch regulates Dll4, VEGFR2 and VEGFR1 in sprouting embryoid bodies.
Figure 4: VEGFR-mediated Dll4 expression dictates tip/stalk phenotype in a cell–cell dependent manner.
Figure 5: Dynamic observations of tip cell shuffling in sprouting angiogenesis.
Figure 6: Computational modelling suggests regulation of cell shuffling by Notch.
Figure 7: Dynamic position shuffling in vivo.

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Acknowledgements

We would like to thank the Cancer Research UK (CRUK) Light Microscopy facility, P. Jordan, the Protein Purification lab, S. Kjaer , J. Yang and J. Ure. We thank D. Barr and I. Moal for simulation and analysis work performed in the Biomolecular Modelling Laboratory CRUK, D. Sauvaget and J. Babbage for technical assistance and B. Thompson for comments on the manuscript. The authors are supported by CRUK, the Lister Institute of Preventive Medicine, the European Molecular Biology Organization (EMBO) Young Investigator Programme, the Fondation Leducq Transatlantic Network of Excellence ARTEMIS (H.G.), the Leukaemia Research Fund (to A.M.) and an EMBO long-term post-doctoral fellowship (to L.J.). We thank J. van Rheenen for discussions and assistance on the SP5 microscope (Leica Microsystems; equipment grant from the Dutch Organization of Scientific Research; NOW, 175.010.2007.007). We also thank A. de Graaff (M.Sc.) and the Hubrecht Imaging Centre (HIC) for imaging support. B.P. and S.S.-M. were supported by the Nederlandse Akademie van Wetenschappen (KNAW).

Author information

Author notes

  1. Holger Gerhardt

    Present address: Independent consultant for: Vascular Patterning Laboratory, Vesalius Research Center, Campus Gasthuisberg, O&N1, Herestraat 49-B912, B-3000 Leuven, Belgium.,

  2. Lars Jakobsson

    Present address: Vascular Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Scheeles Väg 2, SE171 77 Stockholm, Sweden.,

Authors and Affiliations

  1. Vascular Biology Laboratory, London Research Institute, Cancer Research UK, 44 Lincoln's Inn Fields, London, WC2A 3PX, UK

    Lars Jakobsson, Claudio A. Franco, Katie Bentley, Russell T. Collins, Irene M. Aspalter, Marta Busse & Holger Gerhardt

  2. Hubrecht Institute, Uppsalalaan 8, Utrecht, 3584 CT, Netherlands

    Bas Ponsioen & Stefan Schulte-Merker

  3. Transgenic Services, London Research Institute, Cancer Research UK, Clare Hall Laboratories, Blanche Lane, South Mimms, Potters Bar, Hertfordshire, EN6 3LD, UK

    Ian Rosewell

  4. Regeneron Pharmaceuticals, 777 Old Saw Mill River Road, Tarrytown, 10591, NY, USA

    Gavin Thurston

  5. MRC, Centre for Regenerative Medicine, Chancellor's Building, 49 Little France Crescent, Edinburgh, EH16 4SB, UK

    Alexander Medvinsky

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Jakobsson, L., Franco, C., Bentley, K. et al. Endothelial cells dynamically compete for the tip cell position during angiogenic sprouting. Nat Cell Biol 12, 943–953 (2010). https://doi.org/10.1038/ncb2103

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