Characterization of convergent thickening, a major convergence force producing morphogenic movement in amphibians

  1. David R Shook  Is a corresponding author
  2. Jason WH Wen
  3. Ana Rolo
  4. Michael O'Hanlon
  5. Brian Francica
  6. Destiny Dobbins
  7. Paul Skoglund
  8. Douglas W DeSimone
  9. Rudolf Winklbauer
  10. Raymond E Keller
  1. University of Virginia, United States
  2. University of Toronto, Canada
  3. King's College London, United Kingdom
  4. Aduro Biotech, United States
  5. Independent Researcher, United States

Abstract

The morphogenic process of convergent thickening (CT) was originally described as the mediolateral convergence and radial thickening of the explanted ventral involuting marginal zone (IMZ) of Xenopus gastrulae (Keller and Danilchik 1988). Here we show that CT is expressed in all sectors of the pre-involution IMZ, which transitions to expressing convergent extension (CE) after involution. CT occurs without CE and drives symmetric blastopore closure in ventralized embryos. Assays of tissue affinity and tissue surface tension measurements suggest CT is driven by increased interfacial tension between the deep IMZ and the overlying epithelium. The resulting minimization of deep IMZ surface area drives a tendency to shorten the mediolateral (circumblastoporal) aspect of the IMZ, thereby generating tensile force contributing to blastopore closure (Shook et al. 2018). These results establish CT as an independent force-generating process of evolutionary significance and provide the first clear example of an oriented, tensile force generated by an isotropic, Holtfreterian/Steinbergian tissue affinity change.

Data availability

No large-scale data set were generated. Data upon which figures are based is included as source data for those figures; specifically, there are files for each of Figure 2C-E; Figure 3C,D; Figure 3-figure supplement 1C,D; Figure 3-figure supplement 2B; Figure 4C; Figure 5C,F; Figure 5-figure supplement 2B-D; Figure 5-figure supplement 2E; Figure 5-figure supplement 3F-J; Figure 6B,C; Figure 7B; Figure 7C; Figure 7D. Additionally, there is also a source data file with the data supporting a statement within the results section.

Article and author information

Author details

  1. David R Shook

    Department of Biology, University of Virginia, Charlottesville, United States
    For correspondence
    drs6j@virginia.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0131-1834
  2. Jason WH Wen

    Department of Cell, University of Toronto, Toronto, Canada
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7402-5073
  3. Ana Rolo

    Centre for Craniofacial and Regenerative Biology, King's College London, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  4. Michael O'Hanlon

    Department of Cell Biology, University of Virginia, Charlottesville, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Brian Francica

    Aduro Biotech, Berkeley, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Destiny Dobbins

    Independent Researcher, Philadelphia, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Paul Skoglund

    Department of Biology, University of Virginia, Charlottesville, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Douglas W DeSimone

    Department of Cell Bioloy, University of Virginia, Charlottesville, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Rudolf Winklbauer

    Department of Cell and Systems Biology, University of Toronto, Toronto, Canada
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0628-0897
  10. Raymond E Keller

    Department of Biology, University of Virginia, Charlottesville, United States
    Competing interests
    The authors declare that no competing interests exist.

Funding

Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD R37 HD025594 MERIT award)

  • Raymond E Keller

National Institute of General Medical Sciences (NIH RO1 GM099108)

  • Paul Skoglund

National Institute of General Medical Sciences (NIH RO1 GM094793)

  • Douglas W DeSimone

National Institute of General Medical Sciences (R35 GM131865)

  • Douglas W DeSimone

Canadian Institutes of Health Research (CIHR MOP-53075)

  • Rudolf Winklbauer

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

Reviewing Editor

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

Ethics

Animal experimentation: This study was performed in strict accordance with the recommendations in the 8th Edition of the Guide for the Care and Use of Laboratory Animals, of the National Institutes of Health. All of the animals were manipulated according to an approved institutional animal care and use committee (IACUC) protocols of the University of Virginia. The protocols were approved by the Animal Care and Use Committee of the University of Virginia (protocols #2581 and #1830). All surgery was performed under Tricaine anesthesia, and every effort was made to minimize suffering. The animal care and use program is accredited by the Association for Assessment and Accreditation of Laboratory Animal Care, International. The University of Virginia has a PHS Assurance on file with the Office of Laboratory Animal Welfare (OLAW) (PHS Assurance #A3245-01). The University of Virginia is a USDA registered research facility(USDA Registration # 52-R-0011).

Version history

  1. Preprint posted: February 23, 2018 (view preprint)
  2. Received: April 7, 2020
  3. Accepted: April 10, 2022
  4. Accepted Manuscript published: April 11, 2022 (version 1)
  5. Version of Record published: May 3, 2022 (version 2)

Copyright

© 2022, Shook 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

  • 1,003
    Page views
  • 177
    Downloads
  • 6
    Citations

Article citation count generated by polling the highest count across the following sources: PubMed Central, Crossref, Scopus.

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. David R Shook
  2. Jason WH Wen
  3. Ana Rolo
  4. Michael O'Hanlon
  5. Brian Francica
  6. Destiny Dobbins
  7. Paul Skoglund
  8. Douglas W DeSimone
  9. Rudolf Winklbauer
  10. Raymond E Keller
(2022)
Characterization of convergent thickening, a major convergence force producing morphogenic movement in amphibians
eLife 11:e57642.
https://doi.org/10.7554/eLife.57642

Share this article

https://doi.org/10.7554/eLife.57642

Further reading

    1. Developmental Biology
    2. Immunology and Inflammation
    Amir Hossein Kayvanjoo, Iva Splichalova ... Elvira Mass
    Research Article Updated

    During embryogenesis, the fetal liver becomes the main hematopoietic organ, where stem and progenitor cells as well as immature and mature immune cells form an intricate cellular network. Hematopoietic stem cells (HSCs) reside in a specialized niche, which is essential for their proliferation and differentiation. However, the cellular and molecular determinants contributing to this fetal HSC niche remain largely unknown. Macrophages are the first differentiated hematopoietic cells found in the developing liver, where they are important for fetal erythropoiesis by promoting erythrocyte maturation and phagocytosing expelled nuclei. Yet, whether macrophages play a role in fetal hematopoiesis beyond serving as a niche for maturing erythroblasts remains elusive. Here, we investigate the heterogeneity of macrophage populations in the murine fetal liver to define their specific roles during hematopoiesis. Using a single-cell omics approach combined with spatial proteomics and genetic fate-mapping models, we found that fetal liver macrophages cluster into distinct yolk sac-derived subpopulations and that long-term HSCs are interacting preferentially with one of the macrophage subpopulations. Fetal livers lacking macrophages show a delay in erythropoiesis and have an increased number of granulocytes, which can be attributed to transcriptional reprogramming and altered differentiation potential of long-term HSCs. Together, our data provide a detailed map of fetal liver macrophage subpopulations and implicate macrophages as part of the fetal HSC niche.

    1. Developmental Biology
    2. Neuroscience
    Smrithi Prem, Bharati Dev ... Emanuel DiCicco-Bloom
    Research Article Updated

    Autism spectrum disorder (ASD) is defined by common behavioral characteristics, raising the possibility of shared pathogenic mechanisms. Yet, vast clinical and etiological heterogeneity suggests personalized phenotypes. Surprisingly, our iPSC studies find that six individuals from two distinct ASD subtypes, idiopathic and 16p11.2 deletion, have common reductions in neural precursor cell (NPC) neurite outgrowth and migration even though whole genome sequencing demonstrates no genetic overlap between the datasets. To identify signaling differences that may contribute to these developmental defects, an unbiased phospho-(p)-proteome screen was performed. Surprisingly despite the genetic heterogeneity, hundreds of shared p-peptides were identified between autism subtypes including the mTOR pathway. mTOR signaling alterations were confirmed in all NPCs across both ASD subtypes, and mTOR modulation rescued ASD phenotypes and reproduced autism NPC-associated phenotypes in control NPCs. Thus, our studies demonstrate that genetically distinct ASD subtypes have common defects in neurite outgrowth and migration which are driven by the shared pathogenic mechanism of mTOR signaling dysregulation.