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Spatial regulation of VEGF receptor endocytosis in angiogenesis

Nature Cell Biology volume 15, pages 249–260 (2013)Cite this article

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Abstract

Activities as diverse as migration, proliferation and patterning occur simultaneously and in a coordinated fashion during tissue morphogenesis. In the growing vasculature, the formation of motile, invasive and filopodia-carrying endothelial sprouts is balanced with the stabilization of blood-transporting vessels. Here, we show that sprouting endothelial cells in the retina have high rates of VEGF uptake, VEGF receptor endocytosis and turnover. These internalization processes are opposed by atypical protein kinase C activity in more stable and mature vessels. aPKC phosphorylates Dab2, a clathrin-associated sorting protein that, together with the transmembrane protein ephrin-B2 and the cell polarity regulator PAR-3, enables VEGF receptor endocytosis and downstream signal transduction. Accordingly, VEGF receptor internalization and the angiogenic growth of vascular beds are defective in loss-of-function mice lacking key components of this regulatory pathway. Our work uncovers how vessel growth is dynamically controlled by local VEGF receptor endocytosis and the activity of cell polarity proteins.

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Figure 1: High VEGF receptor turnover in the angiogenic front.
Figure 2: Spatial differences in VEGF uptake in the retina.
Figure 3: Identification of Dab2 and PAR-3 as interactors of ephrin-B2 and VEGF receptors.
Figure 4: Dab2 and PAR-3 control VEGF receptor internalization.
Figure 5: Endothelial Dab2 and PAR-3 regulate angiogenic vessel growth.
Figure 6: Negative regulation of VEGF receptor internalization by aPKC.
Figure 7: Increased VEGF uptake and sprouting in the PrkciiΔEC central retina.
Figure 8: Spatial regulation of VEGFR endocytosis.

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Acknowledgements

We thank K. Ebnet and T. Nishioka for reagents and discussions. The Max Planck Society, the German Research Foundation program SFB 629 (R.H.A.), the EMBO LTF programme, the Japan Society for the Promotion of Science (M.N.), and the US National Institutes of Health (J.A.C) provided funding.

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Authors and Affiliations

  1. Department of Tissue Morphogenesis, Max Planck Institute for Molecular Biomedicine, and University of Muenster, Faculty of Medicine, D-48149 Muenster, Germany

    Masanori Nakayama, Akiko Nakayama, Max van Lessen, Hiroyuki Yamamoto, Sarah Hoffmann & Ralf H. Adams

  2. Max Planck Institute for Molecular Biomedicine, Bioanalytical Mass Spectrometry Facility, D-48149 Muenster, Germany

    Hannes C. A. Drexler

  3. Department of Cell Pharmacology, Nagoya University Graduate School of Medicine, Nagoya, Aichi 466-8550, Japan

    Norimichi Itoh & Kozo Kaibuchi

  4. Department of Molecular Biology, Yokohama City University School of Medicine, 236-0004 Yokohama, Japan

    Tomonori Hirose & Shigeo Ohno

  5. Institute of Pathology, Medical Faculty Carl Gustav Carus, Dresden University of Technology, D-01307 Dresden, Germany

    Georg Breier

  6. Department of Vascular Cell Biology, Max Planck Institute for Molecular Biomedicine, D-48149 Muenster, Germany

    Dietmar Vestweber

  7. Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA

    Jonathan A. Cooper

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Contributions

M.N., K.K. and R.H.A. designed the study. M.N., H.Y., H.C.A.D. and N.I. carried out the identification of ephrin-B2 interacting proteins. M.v.L. purified PAR-3 antibodies and GST-fusion proteins. T.H., S.O., G.B., D.V. and J.A.C. generated mouse mutants or lines. All other experiments were carried out by M.N., A.N. and S.H.; M.N. and R.H.A. wrote the manuscript.

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Correspondence to Masanori Nakayama or Ralf H. Adams.

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Nakayama, M., Nakayama, A., van Lessen, M. et al. Spatial regulation of VEGF receptor endocytosis in angiogenesis. Nat Cell Biol 15, 249–260 (2013). https://doi.org/10.1038/ncb2679

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