1. Developmental Biology

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
https://doi.org/10.7554/eLife.76653
Share this article
Cite this article
  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
  2. Download BibTeX
  3. Download .RIS

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.

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.

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

  • 1,561
    views
  • 281
    downloads
  • 6
    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

Categories and tags

Research organism

Further reading

    1. Developmental Biology
    2. Neuroscience

    Aberrant FGF signaling promotes granule neuron precursor expansion in SHH subgroup infantile medulloblastoma

    Odessa R Yabut, Jessica Arela ... Samuel J Pleasure
    Research Article

    Mutations in Sonic Hedgehog (SHH) signaling pathway genes, for example, Suppressor of Fused (SUFU), drive granule neuron precursors (GNP) to form medulloblastomas (MBSHH). However, how different molecular lesions in the Shh pathway drive transformation is frequently unclear, and SUFU mutations in the cerebellum seem distinct. In this study, we show that fibroblast growth factor 5 (FGF5) signaling is integral for many infantile MBSHH cases and that FGF5 expression is uniquely upregulated in infantile MBSHH tumors. Similarly, mice lacking SUFU (Sufu-cKO) ectopically express Fgf5 specifically along the secondary fissure where GNPs harbor preneoplastic lesions and show that FGFR signaling is also ectopically activated in this region. Treatment with an FGFR antagonist rescues the severe GNP hyperplasia and restores cerebellar architecture. Thus, direct inhibition of FGF signaling may be a promising and novel therapeutic candidate for infantile MBSHH.

    1. Developmental Biology
    2. Genetics and Genomics

    Evaluating the transcriptional regulators of arterial gene expression via a catalogue of characterized arterial enhancers

    Svanhild Nornes, Susann Bruche ... Sarah De Val
    Research Article

    The establishment and growth of the arterial endothelium requires the coordinated expression of numerous genes. However, regulation of this process is not yet fully understood. Here, we combined in silico analysis with transgenic mice and zebrafish models to characterize arterial-specific enhancers associated with eight key arterial identity genes (Acvrl1/Alk1, Cxcr4, Cxcl12, Efnb2, Gja4/Cx37, Gja5/Cx40, Nrp1 and Unc5b). Next, to elucidate the regulatory pathways upstream of arterial gene transcription, we investigated the transcription factors binding each arterial enhancer compared to a similar assessment of non-arterial endothelial enhancers. These results found that binding of SOXF and ETS factors was a common occurrence at both arterial and pan-endothelial enhancers, suggesting neither are sufficient to direct arterial specificity. Conversely, FOX motifs independent of ETS motifs were over-represented at arterial enhancers. Further, MEF2 and RBPJ binding was enriched but not ubiquitous at arterial enhancers, potentially linked to specific patterns of behaviour within the arterial endothelium. Lastly, there was no shared or arterial-specific signature for WNT-associated TCF/LEF, TGFβ/BMP-associated SMAD1/5 and SMAD2/3, shear stress-associated KLF4 or venous-enriched NR2F2. This cohort of well characterized and in vivo-verified enhancers can now provide a platform for future studies into the interaction of different transcriptional and signalling pathways with arterial gene expression.

    1. Developmental Biology
    2. Genetics and Genomics

    Determining the effects of paternal obesity on sperm chromatin at histone H3 lysine 4 tri-methylation in relation to the placental transcriptome and cellular composition

    Anne-Sophie Pepin, Patrycja A Jazwiec ... Sarah Kimmins
    Research Article Updated

    Paternal obesity has been implicated in adult-onset metabolic disease in offspring. However, the molecular mechanisms driving these paternal effects and the developmental processes involved remain poorly understood. One underexplored possibility is the role of paternally induced effects on placenta development and function. To address this, we investigated paternal high-fat diet-induced obesity in relation to sperm histone H3 lysine 4 tri-methylation signatures, the placenta transcriptome, and cellular composition. C57BL6/J male mice were fed either a control or high-fat diet for 10 weeks beginning at 6 weeks of age. Males were timed-mated with control-fed C57BL6/J females to generate pregnancies, followed by collection of sperm, and placentas at embryonic day (E)14.5. Chromatin immunoprecipitation targeting histone H3 lysine 4 tri-methylation (H3K4me3) followed by sequencing (ChIP-seq) was performed on sperm to define obesity-associated changes in enrichment. Paternal obesity corresponded with altered sperm H3K4me3 at promoters of genes involved in metabolism and development. Notably, altered sperm H3K4me3 was also localized at placental enhancers. Bulk RNA-sequencing on placentas revealed paternal obesity-associated sex-specific changes in expression of genes involved in hypoxic processes such as angiogenesis, nutrient transport, and imprinted genes, with a subset of de-regulated genes showing changes in H3K4me3 in sperm at corresponding promoters. Paternal obesity was also linked to impaired placenta development; specifically, a deconvolution analysis revealed altered trophoblast cell lineage specification. These findings implicate paternal obesity effects on placenta development and function as one potential developmental route to offspring metabolic disease.