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Notch signalling limits angiogenic cell behaviour in developing zebrafish arteries

Nature volume 445, pages 781–784 (2007)Cite this article

Abstract

Recent evidence indicates that growing blood-vessel sprouts consist of endothelial cells with distinct cell fates and behaviours1,2; however, it is not clear what signals determine these sprout cell characteristics. Here we show that Notch signalling is necessary to restrict angiogenic cell behaviour to tip cells in developing segmental arteries in the zebrafish embryo. In the absence of the Notch signalling component Rbpsuh (recombining binding protein suppressor of hairless) we observed excessive sprouting of segmental arteries, whereas Notch activation suppresses angiogenesis. Through mosaic analysis we find that cells lacking Rbpsuh preferentially localize to the terminal position in developing sprouts. In contrast, cells in which Notch signalling has been activated are excluded from the tip-cell position. In vivo time-lapse analysis reveals that endothelial tip cells undergo a stereotypical pattern of proliferation and migration during sprouting. In the absence of Notch, nearly all sprouting endothelial cells exhibit tip-cell behaviour, leading to excessive numbers of cells within segmental arteries. Furthermore, we find that flt4 (fms-related tyrosine kinase 4, also called vegfr3) is expressed in segmental artery tip cells and becomes ectopically expressed throughout the sprout in the absence of Notch. Loss of flt4 can partially restore normal endothelial cell number in Rbpsuh-deficient segmental arteries. Finally, loss of the Notch ligand dll4 (delta-like 4) also leads to an increased number of endothelial cells within segmental arteries. Together, these studies indicate that proper specification of cell identity, position and behaviour in a developing blood-vessel sprout is required for normal angiogenesis, and implicate the Notch signalling pathway in this process.

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Figure 1: Notch signalling influences positional cell fates in segmental artery sprouts.
Figure 2: Confocal time lapses of wild-type and Rbpsuh-deficient embryos.
Figure 3: Endothelial cell numbers in segmental artery sprouts and expression of flt4 depend on Notch signalling.
Figure 4: Loss of dll4 phenocopies Rbpsuh loss of function.

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Acknowledgements

We would like to thank C. Sagerström and B. Weinstein for critical reading of the manuscript. We thank J. Polli and M. Kacergis for fish care and maintenance. We would also like to thank members of the Lawson laboratory for helpful discussions, and C. Lange for the Rbpsuh in situ analysis. We are grateful to J. Leslie and J. Lewis for sharing data before publication. This work was supported in part by a grant awarded to N.D.L. from the National Heart Lung and Blood Institute, NIH.

Author Contributions A.F.S. and N.D.L. designed and carried out the experiments, analysed the data and wrote the paper.

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  1. Program in Gene Function and Expression, University of Massachusetts Medical School, Lazare Research Building, 364 Plantation Street, Worcester, Massachusetts 01605, USA,

    Arndt F. Siekmann & Nathan D. Lawson

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  1. Arndt F. Siekmann
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  2. Nathan D. Lawson
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Correspondence to Nathan D. Lawson.

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Reprints and permissions information is available at www.nature.com/reprints. The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Methods, Supplementary Figures 1-7 with legends and Supplementary Tables 1-5 with legends, additionl references and legends to Supplementary Movies 1-3. (PDF 1930 kb)

Supplementary Movie 1

This file contains Time lapse Supplementary Movie 1 of transplanted endothelial cells injected with Rbpsuh morpholino enter segmental artery sprouts (MOV 701 kb)

Supplementary Movie 2

This file contains Time-lapse Supplementary Movie 2 of Tg(fli1:negfp)y7 embryo (MOV 2207 kb)

Supplementary Movie 3

This file contains Time-lapse Supplementary Movie 3 of Tg(fli1:negfp)y7 embryo injected with Rbpsuh morpholino (MOV 2323 kb)

Corrigendum

This file describes an error in the Supplementary Information that was posted for this paper on 15 February 2007. (DOC 26 kb)

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Siekmann, A., Lawson, N. Notch signalling limits angiogenic cell behaviour in developing zebrafish arteries. Nature 445, 781–784 (2007). https://doi.org/10.1038/nature05577

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