TGFβ signaling is critical for maintenance of the tendon cell fate

  1. Guak-Kim Tan
  2. Brian A Pryce
  3. Anna Stabio
  4. John V Brigande
  5. ChaoJie Wang
  6. Zheng Xia
  7. Sara F Tufa
  8. Douglas R Keene
  9. Ronen Schweitzer  Is a corresponding author
  1. Shriners Hospitals for Children, United States
  2. Oregon Health and Science University, United States

Abstract

Studies of cell fate focus on specification, but little is known about maintenance of the differentiated state. In this study we find that the mouse tendon cell fate requires continuous maintenance in vivo and identify an essential role for TGFβ signaling in maintenance of the tendon cell fate. To examine the role of TGFβ signaling in tenocyte function the TGFb type II receptor (Tgfbr2) was targeted in the Scleraxis-expressing cell lineage using the ScxCre deletor. Tendon development was not disrupted in mutant embryos, but shortly after birth tenocytes lost differentiation markers and reverted to a more stem/progenitor state. Viral reintroduction of Tgfbr2 to mutants prevented and even rescued tenocyte dedifferentiation suggesting a continuous and cell autonomous role for TGFβ signaling in cell fate maintenance. These results uncover the critical importance of molecular pathways that maintain the differentiated cell fate and a key role for TGFβ signaling in these processes.

Data availability

All data generated or analyzed during this study are included in the manuscript and Supplementary Files.Single cell RNA-Seq data has been deposited onto GEO under accession code GSE139558.

The following data sets were generated

Article and author information

Author details

  1. Guak-Kim Tan

    Research Division, Shriners Hospitals for Children, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Brian A Pryce

    Research Division, Shriners Hospitals for Children, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Anna Stabio

    Research Division, Shriners Hospitals for Children, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. John V Brigande

    Oregon Hearing Research Center, Oregon Health and Science University, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. ChaoJie Wang

    Computational Biology Program, Oregon Health and Science University, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Zheng Xia

    Computational Biology Program, Oregon Health and Science University, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Sara F Tufa

    Research Division, Shriners Hospitals for Children, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Douglas R Keene

    Research Division, Shriners Hospitals for Children, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Ronen Schweitzer

    Research Division, Shriners Hospitals for Children, Portland, United States
    For correspondence
    schweitz@ohsu.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7425-5028

Funding

National Institutes of Health (R01AR055973)

  • Ronen Schweitzer

Shriners Hospitals for Children (SHC 5410-POR-14)

  • Ronen Schweitzer

National Institutes of Health (R01DC014160)

  • John V Brigande

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 (IP00000717) of the Oregon Health & Science University.

Copyright

© 2020, Tan 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

  • 2,982
    views
  • 506
    downloads
  • 69
    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. Guak-Kim Tan
  2. Brian A Pryce
  3. Anna Stabio
  4. John V Brigande
  5. ChaoJie Wang
  6. Zheng Xia
  7. Sara F Tufa
  8. Douglas R Keene
  9. Ronen Schweitzer
(2020)
TGFβ signaling is critical for maintenance of the tendon cell fate
eLife 9:e52695.
https://doi.org/10.7554/eLife.52695

Share this article

https://doi.org/10.7554/eLife.52695

Further reading

    1. Cell Biology
    Affiong Ika Oqua, Kin Chao ... Alejandra Tomas
    Research Article

    G protein-coupled receptors (GPCRs) are integral membrane proteins which closely interact with their plasma membrane lipid microenvironment. Cholesterol is a lipid enriched at the plasma membrane with pivotal roles in the control of membrane fluidity and maintenance of membrane microarchitecture, directly impacting on GPCR stability, dynamics, and function. Cholesterol extraction from pancreatic beta cells has previously been shown to disrupt the internalisation, clustering, and cAMP responses of the glucagon-like peptide-1 receptor (GLP-1R), a class B1 GPCR with key roles in the control of blood glucose levels via the potentiation of insulin secretion in beta cells and weight reduction via the modulation of brain appetite control centres. Here, we unveil the detrimental effect of a high cholesterol diet on GLP-1R-dependent glucoregulation in vivo, and the improvement in GLP-1R function that a reduction in cholesterol synthesis using simvastatin exerts in pancreatic islets. We next identify and map sites of cholesterol high occupancy and residence time on active vs inactive GLP-1Rs using coarse-grained molecular dynamics (cgMD) simulations, followed by a screen of key residues selected from these sites and detailed analyses of the effects of mutating one of these, Val229, to alanine on GLP-1R-cholesterol interactions, plasma membrane behaviours, clustering, trafficking and signalling in INS-1 832/3 rat pancreatic beta cells and primary mouse islets, unveiling an improved insulin secretion profile for the V229A mutant receptor. This study (1) highlights the role of cholesterol in regulating GLP-1R responses in vivo; (2) provides a detailed map of GLP-1R - cholesterol binding sites in model membranes; (3) validates their functional relevance in beta cells; and (4) highlights their potential as locations for the rational design of novel allosteric modulators with the capacity to fine-tune GLP-1R responses.

    1. Cell Biology
    2. Immunology and Inflammation
    Alejandro Rosell, Agata Adelajda Krygowska ... Esther Castellano Sanchez
    Research Article

    Macrophages are crucial in the body’s inflammatory response, with tightly regulated functions for optimal immune system performance. Our study reveals that the RAS–p110α signalling pathway, known for its involvement in various biological processes and tumourigenesis, regulates two vital aspects of the inflammatory response in macrophages: the initial monocyte movement and later-stage lysosomal function. Disrupting this pathway, either in a mouse model or through drug intervention, hampers the inflammatory response, leading to delayed resolution and the development of more severe acute inflammatory reactions in live models. This discovery uncovers a previously unknown role of the p110α isoform in immune regulation within macrophages, offering insight into the complex mechanisms governing their function during inflammation and opening new avenues for modulating inflammatory responses.