A dysmorphic mouse model reveals developmental interactions of chondrocranium and dermatocranium

  1. Susan M Motch Perrine  Is a corresponding author
  2. M Kathleen Pitirri
  3. Emily L Durham
  4. Mizuho Kawasaki
  5. Hao Zheng
  6. Danny Z Chen
  7. Kazuhiko Kawasaki
  8. Joan T Richtsmeier  Is a corresponding author
  1. The Pennsylvania State University, United States
  2. University of Notre Dame, United States
  3. Pennsylvania State University, United States

Abstract

The cranial endo- and dermal skeletons, which comprise the vertebrate skull, evolved independently over 470 million years ago and form separately during embryogenesis. In mammals, much of the cartilaginous chondrocranium is transient, undergoing endochondral ossification or disappearing, so its role in skull morphogenesis is not well studied and it remains an enigmatic structure. We provide complete three-dimensional (3D) reconstructions of the laboratory mouse chondrocranium from embryonic day 13.5 through 17.5 using a novel methodology of uncertainty-guided segmentation of phosphotungstic enhanced 3D microcomputed tomography images with sparse annotation. We evaluate the embryonic mouse chondrocranium and dermatocranium in 3D and delineate the effects of a Fgfr2 variant on embryonic chondrocranial cartilages and on their association with forming dermal bones using the Fgfr2cC342Y/+ Crouzon syndrome mouse. We show that the dermatocranium develops outside of and in shapes that conform to the chondrocranium. Results reveal direct effects of the Fgfr2 variant on embryonic cartilage, on chondrocranium morphology, and on the association between chondrocranium and dermatocranium development. Histologically we observe a trend of relatively more chondrocytes, larger chondrocytes, and/or more matrix in the Fgfr2cC342Y/+ embryos at all timepoints before the chondrocranium begins to disintegrate at E16.5. The chondrocrania and forming dermatocrania of Fgfr2cC342Y/+ embryos are relatively large, but a contrasting trend begins at E16.5 and continues into early postnatal (P0 and P2) timepoints, with the skulls of older Fgfr2cC342Y/+ mice reduced in most dimensions compared to Fgfr2c+/+ littermates. Our findings have implications for the study and treatment of human craniofacial disease, for understanding the impact of chondrocranial morphology on skull growth, and potentially on the evolution of skull morphology.

Data availability

Due to the large size of the majority of these data, data have been made available through Penn State University Libraries ScholarSphere repository at DOI 10.26207/qgke-r185 and include: bone micro-CT images, PTA-e micro-CT images, 3D reconstruction examples of the chondrocrania of one unaffected (Fgfr2c+/+) and one affected (Fgfr2cC342Y/+) at E13.5, E14.5, E15.5, E16.5, and E17.5, bone volumes, histological images, histomorphometric data, 3D landmark coordinate data, correlation matrices estimated by MIBoot used in MI analyses, PCA output, and suture scores. Information on how to download the WinEDMA programs can be found at https://getahead.la.psu.edu/resources/edma and the EDMAinR programs are available on github (https://github.com/psolymos/EDMAinR). Code for automatic chondrocranium segmentation with very sparse annotation via uncertainty-guided self-training is available through https://github.com/ndcse-medical/CartSeg_UGST. PTA-e staining protocols for various embryonic ages of mice are available: https://doi.org/10.1002/dvdy.136

The following data sets were generated

Article and author information

Author details

  1. Susan M Motch Perrine

    Department of Anthropology, The Pennsylvania State University, University Park, United States
    For correspondence
    qzk2@psu.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3412-221X
  2. M Kathleen Pitirri

    Department of Anthropology, The Pennsylvania State University, University Park, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Emily L Durham

    Department of Anthropology, The Pennsylvania State University, University Park, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Mizuho Kawasaki

    Department of Anthropology, The Pennsylvania State University, University Park, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Hao Zheng

    Department of Computer Science and Engineering, University of Notre Dame, Notre Dame, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9790-7607
  6. Danny Z Chen

    Department of Computer Science and Engineering, University of Notre Dame, Notre Dame, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Kazuhiko Kawasaki

    Department of Anthropology, Pennsylvania State University, University Park, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Joan T Richtsmeier

    Department of Anthropology, Pennsylvania State University, University Park, United States
    For correspondence
    jta10@psu.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0239-5822

Funding

National Institute of Dental and Craniofacial Research (R01DE027677)

  • Joan T Richtsmeier

Eunice Kennedy Shriver National Institute of Child Health and Human Development (P01HD078233)

  • Joan T Richtsmeier

National Institute of Dental and Craniofacial Research (R01 DE031439)

  • Joan T Richtsmeier

National Science Foundation (CCF-1617735)

  • Danny Z Chen

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

Reviewing Editor

  1. Min Zhu, Institute of Vertebrate Paleontology and Paleoanthropology, Chinese Academy of Sciences, China

Ethics

Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocols (#446558) of the Pennsylvania State University.

Version history

  1. Preprint posted: November 25, 2021 (view preprint)
  2. Received: December 23, 2021
  3. Accepted: June 14, 2022
  4. Accepted Manuscript published: June 15, 2022 (version 1)
  5. Version of Record published: July 6, 2022 (version 2)

Copyright

© 2022, Motch Perrine 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

  • 1,373
    views
  • 264
    downloads
  • 5
    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. Susan M Motch Perrine
  2. M Kathleen Pitirri
  3. Emily L Durham
  4. Mizuho Kawasaki
  5. Hao Zheng
  6. Danny Z Chen
  7. Kazuhiko Kawasaki
  8. Joan T Richtsmeier
(2022)
A dysmorphic mouse model reveals developmental interactions of chondrocranium and dermatocranium
eLife 11:e76653.
https://doi.org/10.7554/eLife.76653

Share this article

https://doi.org/10.7554/eLife.76653

Further reading

    1. Developmental Biology
    Zhimin Xu, Zhao Wang ... Yingchuan B Qi
    Research Article

    Precise developmental timing control is essential for organism formation and function, but its mechanisms are unclear. In C. elegans, the microRNA lin-4 critically regulates developmental timing by post-transcriptionally downregulating the larval-stage-fate controller LIN-14. However, the mechanisms triggering the activation of lin-4 expression toward the end of the first larval stage remain unknown. We demonstrate that the transmembrane transcription factor MYRF-1 is necessary for lin-4 activation. MYRF-1 is initially localized on the cell membrane, and its increased cleavage and nuclear accumulation coincide with lin-4 expression timing. MYRF-1 regulates lin-4 expression cell-autonomously and hyperactive MYRF-1 can prematurely drive lin-4 expression in embryos and young first-stage larvae. The tandem lin-4 promoter DNA recruits MYRF-1GFP to form visible loci in the nucleus, suggesting that MYRF-1 directly binds to the lin-4 promoter. Our findings identify a crucial link in understanding developmental timing regulation and establish MYRF-1 as a key regulator of lin-4 expression.

    1. Developmental Biology
    2. Structural Biology and Molecular Biophysics
    Samuel C Griffiths, Jia Tan ... Hsin-Yi Henry Ho
    Research Article Updated

    The receptor tyrosine kinase ROR2 mediates noncanonical WNT5A signaling to orchestrate tissue morphogenetic processes, and dysfunction of the pathway causes Robinow syndrome, brachydactyly B, and metastatic diseases. The domain(s) and mechanisms required for ROR2 function, however, remain unclear. We solved the crystal structure of the extracellular cysteine-rich (CRD) and Kringle (Kr) domains of ROR2 and found that, unlike other CRDs, the ROR2 CRD lacks the signature hydrophobic pocket that binds lipids/lipid-modified proteins, such as WNTs, suggesting a novel mechanism of ligand reception. Functionally, we showed that the ROR2 CRD, but not other domains, is required and minimally sufficient to promote WNT5A signaling, and Robinow mutations in the CRD and the adjacent Kr impair ROR2 secretion and function. Moreover, using function-activating and -perturbing antibodies against the Frizzled (FZ) family of WNT receptors, we demonstrate the involvement of FZ in WNT5A-ROR signaling. Thus, ROR2 acts via its CRD to potentiate the function of a receptor super-complex that includes FZ to transduce WNT5A signals.