1. Cell Biology
  2. Developmental Biology

Limited column formation in the embryonic growth plate implies divergent growth mechanisms during pre- and postnatal bone development

  1. Sarah Rubin
  2. Ankit Agrawal  Is a corresponding author
  3. Anne Seewald
  4. Meng-Jia Lian
  5. Olivia Gottdenker
  6. Paul Villoutreix
  7. Adrian Baule
  8. Tomer Stern
  9. Elazar Zelzer  Is a corresponding author
  1. Weizmann Institute of Science, Israel
  2. University of Michigan-Ann Arbor, United States
  3. Aix Marseille Univ, INSERM, MMG, UMR1251, France
  4. Queen Mary University of London, United Kingdom
https://doi.org/10.7554/eLife.95289
Share this article
Cite this article
  1. Sarah Rubin
  2. Ankit Agrawal
  3. Anne Seewald
  4. Meng-Jia Lian
  5. Olivia Gottdenker
  6. Paul Villoutreix
  7. Adrian Baule
  8. Tomer Stern
  9. Elazar Zelzer
(2024)
Limited column formation in the embryonic growth plate implies divergent growth mechanisms during pre- and postnatal bone development
eLife 13:e95289.
https://doi.org/10.7554/eLife.95289
  2. Download BibTeX
  3. Download .RIS

Abstract

Chondrocyte columns, which are a hallmark of growth plate architecture, play a central role in bone elongation. Columns are formed by clonal expansion following rotation of the division plane, resulting in a stack of cells oriented parallel to the growth direction. In this work, we analyzed hundreds of Confetti multicolor clones in growth plates of mouse embryos using a pipeline comprising 3D imaging and algorithms for morphometric analysis. Surprisingly, analysis of the elevation angles between neighboring pairs of cells revealed that most cells did not display the typical stacking pattern associated with column formation, implying incomplete rotation of the division plane. Morphological analysis revealed that although embryonic clones were elongated, they formed clusters oriented perpendicular to the growth direction. Analysis of growth plates of postnatal mice revealed both complex columns, composed of ordered and disordered cell stacks, and small, disorganized clusters located in the outer edges. Finally, correlation between the temporal dynamics of the ratios between clusters and columns and between bone elongation and expansion suggests that clusters may promote expansion, whereas columns support elongation. Overall, our findings support the idea that modulations of division plane rotation of proliferating chondrocytes determines the formation of either clusters or columns, a multifunctional design that regulates morphogenesis throughout pre- and postnatal bone growth. Broadly, this work provides a new understanding of the cellular mechanisms underlying growth plate activity and bone elongation during development.

Data availability

The datasets generated and analyzed during the current study are available on Zenodo at the following links: https://doi.org/10.5281/zenodo.10440013, https://doi.org/10.5281/zenodo.10444731, https://doi.org/10.5281/zenodo.10446055, https://doi.org/10.5281/zenodo.10446092, https://doi.org/10.5281/zenodo.10446121, https://doi.org/10.5281/zenodo.10446131, https://doi.org/10.5281/zenodo.10446123, https://doi.org/10.5281/zenodo.10446145.

The following data sets were generated

Article and author information

Author details

  1. Sarah Rubin

    Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0601-8802

  2. Ankit Agrawal

    Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel
    For correspondence
    ankitbioinfo@gmail.com
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0009-0006-1700-2397

  3. Anne Seewald

    Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4904-2063

  4. Meng-Jia Lian

    Department of Biologic and Materials and Prosthodontics, University of Michigan-Ann Arbor, Ann Arbor, United States
    Competing interests
    The authors declare that no competing interests exist.

  5. Olivia Gottdenker

    Department of Biologic and Materials and Prosthodontics, University of Michigan-Ann Arbor, Ann Arbor, United States
    Competing interests
    The authors declare that no competing interests exist.

  6. Paul Villoutreix

    Turing Center for Living Systems, Aix Marseille Univ, INSERM, MMG, UMR1251, Marseille, France
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6333-5735

  7. Adrian Baule

    School of Mathematical Sciences, Queen Mary University of London, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.

  8. Tomer Stern

    Department of Biologic and Materials and Prosthodontics, University of Michigan-Ann Arbor, Ann Arbor, United States
    Competing interests
    The authors declare that no competing interests exist.

  9. Elazar Zelzer

    Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel
    For correspondence
    eli.zelzer@weizmann.ac.il
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1584-6602

Funding

Israel Science Foundation (1387/23)

  • Elazar Zelzer

Weizmann - Sagol Institute for longetivity research

  • Elazar Zelzer

Julie and Eric Borman Family Research Funds

  • Elazar Zelzer

University of Michigan School of Dentistry startup funds

  • Tomer Stern

University of Michigan Oral Health Sciences PhD program

  • Meng-Jia Lian

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 pre-approved by and conducted according to the guidelines of the Institutional Animal Care and Use Committee (IACUC) of the Weizmann Institute (IACUC 01750221-1 and IACUC 05700723-2). All animals used in this study had access to food and water ad libitum and were maintained under controlled humidity and temperature (45-65%, 22 {plus minus} 2{degree sign}C, respectively).

Copyright

© 2024, Rubin 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

  • 1,049
    views
  • 163
    downloads
  • 0
    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. Sarah Rubin
  2. Ankit Agrawal
  3. Anne Seewald
  4. Meng-Jia Lian
  5. Olivia Gottdenker
  6. Paul Villoutreix
  7. Adrian Baule
  8. Tomer Stern
  9. Elazar Zelzer
(2024)
Limited column formation in the embryonic growth plate implies divergent growth mechanisms during pre- and postnatal bone development
eLife 13:e95289.
https://doi.org/10.7554/eLife.95289

Share this article

https://doi.org/10.7554/eLife.95289

Categories and tags

Research organism

Further reading

    1. Cell Biology

    Distinct trafficking routes of polarized and non-polarized membrane cargoes in Aspergillus nidulans

    Georgia Maria Sagia, Xenia Georgiou ... Sofia Dimou
    Research Article Updated

    Membrane proteins are sorted to the plasma membrane via Golgi-dependent trafficking. However, our recent studies challenged the essentiality of Golgi in the biogenesis of specific transporters. Here, we investigate the trafficking mechanisms of membrane proteins by following the localization of the polarized R-SNARE SynA versus the non-polarized transporter UapA, synchronously co-expressed in wild-type or isogenic genetic backgrounds repressible for conventional cargo secretion. In wild-type, the two cargoes dynamically label distinct secretory compartments, highlighted by the finding that, unlike SynA, UapA does not colocalize with the late-Golgi. In line with early partitioning into distinct secretory carriers, the two cargoes collapse in distinct ER-Exit Sites (ERES) in a sec31ts background. Trafficking via distinct cargo-specific carriers is further supported by showing that repression of proteins essential for conventional cargo secretion does not affect UapA trafficking, while blocking SynA secretion. Overall, this work establishes the existence of distinct, cargo-dependent, trafficking mechanisms, initiating at ERES and being differentially dependent on Golgi and SNARE interactions.

    1. Cell Biology
    2. Microbiology and Infectious Disease

    Zika virus remodels and hijacks IGF2BP2 ribonucleoprotein complex to promote viral replication organelle biogenesis

    Clément Mazeaud, Stefan Pfister ... Laurent Chatel-Chaix
    Research Article

    Zika virus (ZIKV) infection causes significant human disease that, with no approved treatment or vaccine, constitutes a major public health concern. Its life cycle entirely relies on the cytoplasmic fate of the viral RNA genome (vRNA) through a fine-tuned equilibrium between vRNA translation, replication, and packaging into new virions, all within virus-induced replication organelles (vROs). In this study, with an RNA interference (RNAi) mini-screening and subsequent functional characterization, we have identified insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2) as a new host dependency factor that regulates vRNA synthesis. In infected cells, IGF2BP2 associates with viral NS5 polymerase and redistributes to the perinuclear viral replication compartment. Combined fluorescence in situ hybridization-based confocal imaging, in vitro binding assays, and immunoprecipitation coupled to RT-qPCR showed that IGF2BP2 directly interacts with ZIKV vRNA 3’ nontranslated region. Using ZIKV sub-genomic replicons and a replication-independent vRO induction system, we demonstrated that IGF2BP2 knockdown impairs de novo vRO biogenesis and, consistently, vRNA synthesis. Finally, the analysis of immunopurified IGF2BP2 complex using quantitative mass spectrometry and RT-qPCR revealed that ZIKV infection alters the protein and RNA interactomes of IGF2BP2. Altogether, our data support that ZIKV hijacks and remodels the IGF2BP2 ribonucleoprotein complex to regulate vRO biogenesis and vRNA neosynthesis.

    1. Cell Biology

    Microbiome: Balancing microbial composition through diet

    L Catalina Acuff, Karen Guillemin
    Insight

    A reciprocal interaction between gut bacteria and gut cells affects protein absorption in the host.