Control of endothelial cell polarity and sprouting angiogenesis by non-centrosomal microtubules

  1. Maud Martin
  2. Alexandra Veloso
  3. Jingchao Wu
  4. Eugene A Katrukha
  5. Anna Akhmanova  Is a corresponding author
  1. Utrecht University, Netherlands
  2. University of Liège, Belgium

Abstract

Microtubules control different aspects of cell polarization. In cells with a radial microtubule system, a pivotal role in setting up asymmetry is attributed to the relative positioning of the centrosome and the nucleus. Here, we show that centrosome loss had no effect on the ability of endothelial cells to polarize and move in 2D and 3D environments. In contrast, non-centrosomal microtubules stabilized by the microtubule minus-end-binding protein CAMSAP2 were required for directional migration on 2D substrates and for the establishment of polarized cell morphology in soft 3D matrices. CAMSAP2 was also important for persistent endothelial cell sprouting during in vivo zebrafish vessel development. In the absence of CAMSAP2, cell polarization in 3D could be partly rescued by centrosome depletion, indicating that in these conditions the centrosome inhibited cell polarity. We propose that CAMSAP2-protected non-centrosomal microtubules are needed for establishing cell asymmetry by enabling microtubule enrichment in a single cell protrusion.

Article and author information

Author details

  1. Maud Martin

    Department of Biology, Utrecht University, Utrecht, Netherlands
    Competing interests
    No competing interests declared.
  2. Alexandra Veloso

    Interdisciplinary Cluster for Applied Genoproteomics (GIGA-R), University of Liège, Liège, Belgium
    Competing interests
    No competing interests declared.
  3. Jingchao Wu

    Department of Biology, Utrecht University, Utrecht, Netherlands
    Competing interests
    No competing interests declared.
  4. Eugene A Katrukha

    Department of Biology, Utrecht University, Utrecht, Netherlands
    Competing interests
    No competing interests declared.
  5. Anna Akhmanova

    Department of Biology, Utrecht University, Utrecht, Netherlands
    For correspondence
    a.akhmanova@uu.nl
    Competing interests
    Anna Akhmanova, Senior editor, eLife.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9048-8614

Funding

European Research Council (Synergy 609822)

  • Anna Akhmanova

Nederlandse Organisatie voor Wetenschappelijk Onderzoek (ALW Open Program grant 824.15.017)

  • Anna Akhmanova

Marie Sklodowska-Curie Actions (IEF fellowship)

  • Maud Martin

China Scholarship Council (PhD fellowship)

  • Jingchao Wu

Fonds De La Recherche Scientifique - FNRS (FRIA fellowship)

  • Alexandra Veloso

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All animal experiments were approved by the animal welfare committee of the University of Liege (protocol number 14-1556, laboratory agreement number LA 1610002).

Copyright

© 2018, Martin 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.

Metrics

  • 6,828
    views
  • 1,104
    downloads
  • 75
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Maud Martin
  2. Alexandra Veloso
  3. Jingchao Wu
  4. Eugene A Katrukha
  5. Anna Akhmanova
(2018)
Control of endothelial cell polarity and sprouting angiogenesis by non-centrosomal microtubules
eLife 7:e33864.
https://doi.org/10.7554/eLife.33864

Share this article

https://doi.org/10.7554/eLife.33864

Further reading

    1. Cell Biology
    Kaili Du, Hongyu Chen ... Dan Li
    Research Article

    Niemann–Pick disease type C (NPC) is a devastating lysosomal storage disease characterized by abnormal cholesterol accumulation in lysosomes. Currently, there is no treatment for NPC. Transcription factor EB (TFEB), a member of the microphthalmia transcription factors (MiTF), has emerged as a master regulator of lysosomal function and promoted the clearance of substrates stored in cells. However, it is not known whether TFEB plays a role in cholesterol clearance in NPC disease. Here, we show that transgenic overexpression of TFEB, but not TFE3 (another member of MiTF family) facilitates cholesterol clearance in various NPC1 cell models. Pharmacological activation of TFEB by sulforaphane (SFN), a previously identified natural small-molecule TFEB agonist by us, can dramatically ameliorate cholesterol accumulation in human and mouse NPC1 cell models. In NPC1 cells, SFN induces TFEB nuclear translocation via a ROS-Ca2+-calcineurin-dependent but MTOR-independent pathway and upregulates the expression of TFEB-downstream genes, promoting lysosomal exocytosis and biogenesis. While genetic inhibition of TFEB abolishes the cholesterol clearance and exocytosis effect by SFN. In the NPC1 mouse model, SFN dephosphorylates/activates TFEB in the brain and exhibits potent efficacy of rescuing the loss of Purkinje cells and body weight. Hence, pharmacological upregulating lysosome machinery via targeting TFEB represents a promising approach to treat NPC and related lysosomal storage diseases, and provides the possibility of TFEB agonists, that is, SFN as potential NPC therapeutic candidates.

    1. Cell Biology
    2. Developmental Biology
    Sarah Y Coomson, Salil A Lachke
    Insight

    A study in mice reveals key interactions between proteins involved in fibroblast growth factor signaling and how they contribute to distinct stages of eye lens development.