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Endoglin controls blood vessel diameter through endothelial cell shape changes in response to haemodynamic cues

Nature Cell Biology volume 19, pages 653665 (2017) | Download Citation

Abstract

The hierarchical organization of properly sized blood vessels ensures the correct distribution of blood to all organs of the body, and is controlled via haemodynamic cues. In current concepts, an endothelium-dependent shear stress set point causes blood vessel enlargement in response to higher flow rates, while lower flow would lead to blood vessel narrowing, thereby establishing homeostasis. We show that during zebrafish embryonic development increases in flow, after an initial expansion of blood vessel diameters, eventually lead to vessel contraction. This is mediated via endothelial cell shape changes. We identify the transforming growth factor beta co-receptor endoglin as an important player in this process. Endoglin mutant cells and blood vessels continue to enlarge in response to flow increases, thus exacerbating pre-existing embryonic arterial–venous shunts. Together, our data suggest that cell shape changes in response to biophysical cues act as an underlying principle allowing for the ordered patterning of tubular organs.

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Acknowledgements

This work was funded by the Max Planck Society (A.F.S.), the Deutsche Forschungsgemeinschaft (DFG SI-1374/3-2; DFG SI-1374/4-1; DFG SI-1374/5-1; A.F.S.), and a European Research Council (ERC) starting grant (260794-ZebrafishAngio; A.F.S.). This work was supported by the Deutsche Forschungsgemeinschaft (DFG) Cells-in-Motion Cluster of Excellence (EXC 1003-CIM), University of Münster, Germany. J.B. was supported by a long-term EMBO post-doctoral fellowship. L.J. was supported by The Swedish Research Council, The Swedish Cancer Society and Karolinska Institutet. We thank H. Arthur for providing the Engflox/flox mice and R. Adams for the Cdh5(PAC)-CreERT2 mice.

Author information

Author notes

    • Jeroen Bussmann

    Present address: Department of Supramolecular & Biomaterials Chemistry, Leiden Institute of Chemistry (LIC), Gorlaeus Laboratories, Leiden University, PO Box 9502, 2300 RA Leiden, The Netherlands.

Affiliations

  1. Max Planck Institute for Molecular Biomedicine, Roentgenstrasse 20, D-48149 Muenster, Germany

    • Wade W. Sugden
    • , Roman Tsaryk
    • , Elvin V. Leonard
    • , Jeroen Bussmann
    • , Mailin J. Hamm
    • , Wiebke Herzog
    •  & Arndt F. Siekmann
  2. Cells-in-Motion Cluster of Excellence (EXC 1003 – CiM), University of Muenster, D-48149 Muenster, Germany

    • Wade W. Sugden
    • , Robert Meissner
    • , Roman Tsaryk
    • , Elvin V. Leonard
    • , Mailin J. Hamm
    • , Wiebke Herzog
    • , Cornelia Denz
    •  & Arndt F. Siekmann
  3. Institute of Applied Physics and Center for Nonlinear Science (CeNoS), University of Muenster, D-48149 Muenster, Germany

    • Robert Meissner
    •  & Cornelia Denz
  4. Institute of Molecular Life Sciences, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland

    • Tinri Aegerter-Wilmsen
  5. University of Muenster, D-48149 Muenster, Germany

    • Mailin J. Hamm
    •  & Wiebke Herzog
  6. Karolinska Institute, Department of Medical Biochemistry and Biophysics, SE 171 77 Stockholm, Sweden

    • Yi Jin
    •  & Lars Jakobsson

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Contributions

W.W.S. and A.F.S. designed and interpreted the experiments and wrote the manuscript. J.B. identified the zebrafish eng homologue and generated the Tg(klf2a:YFP)mu107 zebrafish. R.T. performed cell culture experiments. E.V.L. performed FACS sorting of zebrafish embryos and performed qPCR experiments. R.M. and C.D. performed particle velocimetry, the optical rail experiments and determined flow parameters. T.A.-W. performed zebrafish cell shape analysis. M.J.H. and W.H. generated Tg(fli1a:lifeactEGFP)mu240 zebrafish. Y.J. and L.J. determined cell shape changes in eng mutant mice.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Arndt F. Siekmann.

Integrated supplementary information

Supplementary information

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  1. 1.

    Supplementary Information

    Supplementary Information

Videos

  1. 1.

    Distal blood flow in WT fin regenerate at 5 dpa.

    Brightfield movie of blood flow in the distal regenerating fin of WT fish at 5 dpa. Numbers label individual bony rays. Arrows indicate flow direction, while dashed lines denote arteries (red) and veins (blue). Flow from a centrally located artery distributes through a plexus to two veins at the end of each ray. Note weak flow reversal in lateral vein of the first ray, arcing over an inactive artery.

  2. 2.

    AVMs and disturbed flow in engmu130 fin regenerate at 5 dpa.

    Brightfield movie of blood flow in the distal regenerating fin of engmu130 mutant at 5 dpa. Numbers label individual bony rays. Arrows indicate flow direction, while dashed lines denote arteries (red) and veins (blue). Note prominent AVMs in second ray from the left. In ray #3, more proximally located AVMs either stop or reverse blood flow in the veins, and the central artery is not active. Ray #4 has strong flow reversal in the left vein, arcing over an inactive central artery. Note bleedings at distal end of all rays.

  3. 3.

    ISV flow in 72 hpf sibling embryo.

    Brightfield movie of RBC flow in the trunk of sibling embryo at 72 hpf. Nearly all ISVs on both sides of the embryo carry RBC flow.

  4. 4.

    ISV flow in 72 hpf engmu130 embryo.

    Brightfield movie of RBC flow in the trunk of engmu130 embryo at 72 hpf. Note that only 1 or 2 ISVs have persistent RBC flow, while most have no or only very intermittent RBC flow.

  5. 5.

    Optical rail restores RBC flow in capillaries of engmu130 mutants.

    Brightfield movie of RBC flow in the trunk of engmu130 embryo at 72 hpf. Holographic optical tweezers (HOT) are focused at entrance to an aISV, but laser is turned off. About halfway through movie the laser is turned on (red circle indicates HOT focal point), slows down RBCs that pass through the laser, and diverts them from the main arterial blood flow into the capillary.

  6. 6.

    Time-lapse of cell shape changes in DA between 48 and 72 hpf in sibling embryo.

    Maximum intensity projection of confocal z-stacks of Tg(fli1a:lifeactEGFP)mu240; Tg(fli1a:nEGFP)y7, and Tg(-0.8flt1:RFP)hu5333 in sibling embryo between 48 and 72 hpf. Acquisitions were made at 40 min intervals, shown at 5 frames per second. Note decrease in DA diameter between 48 and 72 hpf, and elongation of ECs.

  7. 7.

    Time-lapse of cell shape changes in DA between 48 and 72 hpf in engmu130 embryo.

    Maximum intensity projection of confocal z-stacks of Tg(fli1a:lifeactEGFP)mu240; Tg(fli1a:nEGFP)y7z, and Tg(-0.8flt1:RFP)hu5333 in engmu130 embryo between 48 and 72 hpf. Acquisitions were made at 40 min intervals, shown at 5 frames per second. Note a similar DA diameter at 48 hpf as sibling in Supplementary Video 4 that fails to decrease appreciably by 72 hpf. Note, additionally, the dramatic increase in EC sizes compared to siblings during the movie.

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DOI

https://doi.org/10.1038/ncb3528

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