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

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.

Metrics

  • 5,603
    views
  • 734
    downloads
  • 46
    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. 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

Share this article

https://doi.org/10.7554/eLife.51221

Further reading

    1. Cell Biology
    Jittoku Ihara, Yibin Huang ... Koichi Yamamoto
    Research Article

    Chronic kidney disease (CKD) and atherosclerotic heart disease, frequently associated with dyslipidemia and hypertension, represent significant health concerns. We investigated the interplay among these conditions, focusing on the role of oxidized low-density lipoprotein (oxLDL) and angiotensin II (Ang II) in renal injury via G protein αq subunit (Gq) signaling. We hypothesized that oxLDL enhances Ang II-induced Gq signaling via the AT1 (Ang II type 1 receptor)-LOX1 (lectin-like oxLDL receptor) complex. Based on CHO and renal cell model experiments, oxLDL alone did not activate Gq signaling. However, when combined with Ang II, it significantly potentiated Gq-mediated inositol phosphate 1 production and calcium influx in cells expressing both LOX-1 and AT1 but not in AT1-expressing cells. This suggests a critical synergistic interaction between oxLDL and Ang II in the AT1-LOX1 complex. Conformational studies using AT1 biosensors have indicated a unique receptor conformational change due to the oxLDL-Ang II combination. In vivo, wild-type mice fed a high-fat diet with Ang II infusion presented exacerbated renal dysfunction, whereas LOX-1 knockout mice did not, underscoring the pathophysiological relevance of the AT1-LOX1 interaction in renal damage. These findings highlight a novel mechanism of renal dysfunction in CKD driven by dyslipidemia and hypertension and suggest the therapeutic potential of AT1-LOX1 receptor complex in patients with these comorbidities.

    1. Cell Biology
    Qi Zeng, Chen Yao ... Shuai Chen
    Research Article

    Mounting evidence has demonstrated the genetic association of ORMDL sphingolipid biosynthesis regulator 3 (ORMDL3) gene polymorphisms with bronchial asthma and a diverse set of inflammatory disorders. However, its role in type I interferon (type I IFN) signaling remains poorly defined. Herein, we report that ORMDL3 is a negative modulator of the type I IFN signaling by interacting with mitochondrial antiviral signaling protein (MAVS) and subsequently promoting the proteasome-mediated degradation of retinoic acid-inducible gene I (RIG-I). Immunoprecipitation coupled with mass spectrometry (IP-MS) assays uncovered that ORMDL3 binds to ubiquitin-specific protease 10 (USP10), which forms a complex with and stabilizes RIG-I through decreasing its K48-linked ubiquitination. ORMDL3 thus disrupts the interaction between USP10 and RIG-I, thereby promoting RIG-I degradation. Additionally, subcutaneous syngeneic tumor models in C57BL/6 mice revealed that inhibition of ORMDL3 enhances anti-tumor efficacy by augmenting the proportion of cytotoxic CD8 positive T cells and IFN production in the tumor microenvironment (TME). Collectively, our findings reveal the pivotal roles of ORMDL3 in maintaining antiviral innate immune responses and anti-tumor immunity.