Notochord vacuoles absorb compressive bone growth during zebrafish spine formation

  1. Jennifer Bagwell
  2. James Norman
  3. Kathryn L Ellis
  4. Brianna Peskin
  5. James Hwang
  6. Xiaoyan Ge
  7. Stacy Nguyen
  8. Sarah K McMenamin
  9. Didier YR Stainier
  10. Michel Bagnat  Is a corresponding author
  1. Duke University, United States
  2. University of California, San Francisco, United States
  3. Boston College, United States
  4. Max Planck Institute for Heart and Lung Research, Germany

Abstract

The vertebral column or spine assembles around the notochord rod which contains a core made of large vacuolated cells. Each vacuolated cell possesses a single fluid-filled vacuole, and loss or fragmentation of these vacuoles in zebrafish leads to spine kinking. Here, we identified a mutation in the kinase gene dstyk that causes fragmentation of notochord vacuoles and a severe congenital scoliosis-like phenotype in zebrafish. Live imaging revealed that Dstyk regulates fusion of membranes with the vacuole. We find that localized disruption of notochord vacuoles causes vertebral malformation and curving of the spine axis at those sites. Accordingly, in dstyk mutants the spine curves increasingly over time as vertebral bone formation compresses the notochord asymmetrically, causing vertebral malformations and kinking of the axis. Together, our data show that notochord vacuoles function as a hydrostatic scaffold that guides symmetrical growth of vertebrae and spine formation.

Data availability

All data generated or analyses during this study are included in the manuscript and supporting files. Source data files have been provided as indicated. Data has been deposited to Dryad, under the DOI: 10.5061/dryad.73n5tb2tb. Due to their large size, raw image files can be accessed upon request.

The following data sets were generated

Article and author information

Author details

  1. Jennifer Bagwell

    Department of Cell Biology, Duke University, Durham, United States
    Competing interests
    No competing interests declared.
  2. James Norman

    Department of Cell Biology, Duke University, Durham, United States
    Competing interests
    No competing interests declared.
  3. Kathryn L Ellis

    Department of Cell Biology, Duke University, Durham, United States
    Competing interests
    No competing interests declared.
  4. Brianna Peskin

    Department of Cell Biology, Duke University, Durham, United States
    Competing interests
    No competing interests declared.
  5. James Hwang

    Department of Cell Biology, Duke University, Durham, United States
    Competing interests
    No competing interests declared.
  6. Xiaoyan Ge

    Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United States
    Competing interests
    No competing interests declared.
  7. Stacy Nguyen

    Department of Biology, Boston College, Boston, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2641-3984
  8. Sarah K McMenamin

    Department of Biology, Boston College, Boston, United States
    Competing interests
    No competing interests declared.
  9. Didier YR Stainier

    Department of Developmental Genetics, Max Planck Institute for Heart and Lung Research, Bad Nauheim, Germany
    Competing interests
    Didier YR Stainier, Senior editor, eLife.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0382-0026
  10. Michel Bagnat

    Department of Cell Biology, Duke University, Durham, United States
    For correspondence
    michel.bagnat@duke.edu
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3829-0168

Funding

National Institutes of Health (R01AR065439)

  • Michel Bagnat

Howard Hughes Medical Institute (Faculty Scholars)

  • Michel Bagnat

National Institutes of Health (R01HL54737)

  • Didier YR Stainier

National Institutes of Health (R00GM105874 and R03HD091634)

  • Sarah K McMenamin

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

Ethics

Animal experimentation: Zebrafish (Danio rerio) were used in accordance with Duke University Institutional Animal Care and Use Committee (IACUC) guidelines and approved under our animal protocol A089-17-04

Reviewing Editor

  1. Lilianna Solnica-Krezel, Washington University School of Medicine, United States

Publication history

  1. Received: August 20, 2019
  2. Accepted: January 28, 2020
  3. Accepted Manuscript published: January 29, 2020 (version 1)
  4. Version of Record published: February 11, 2020 (version 2)

Copyright

© 2020, Bagwell 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.

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  1. Jennifer Bagwell
  2. James Norman
  3. Kathryn L Ellis
  4. Brianna Peskin
  5. James Hwang
  6. Xiaoyan Ge
  7. Stacy Nguyen
  8. Sarah K McMenamin
  9. Didier YR Stainier
  10. Michel Bagnat
(2020)
Notochord vacuoles absorb compressive bone growth during zebrafish spine formation
eLife 9:e51221.
https://doi.org/10.7554/eLife.51221

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