1. Cell Biology
  2. Stem Cells and Regenerative Medicine

The role of Pitx2 and Pitx3 in muscle stem cells gives new insights into P38α MAP kinase and redox regulation of muscle regeneration

  1. Aurore Lhonore  Is a corresponding author
  2. Pierre-Henri Commère
  3. Elisa Negroni
  4. Giorgia Pallafacchina
  5. Bertrand Friguet
  6. Jacques Drouin
  7. Margaret Buckingham
  8. Didier Montarras  Is a corresponding author
  1. CNRS UMR 3738, Institut Pasteur, France
  2. Institut Pasteur, France
  3. Sorbonne Universités, Université Pierre et Marie Curie Université Paris 06, INSERM UMRS974, France
  4. Italian National Research Council (CNR) Neuroscience Institute, Italy
  5. CNRS UMR 8256, INSERM ERL U1164, France
  6. Institut de Recherches Cliniques de Montréal, France
https://doi.org/10.7554/eLife.32991
Share this article
Cite this article
  1. Aurore Lhonore
  2. Pierre-Henri Commère
  3. Elisa Negroni
  4. Giorgia Pallafacchina
  5. Bertrand Friguet
  6. Jacques Drouin
  7. Margaret Buckingham
  8. Didier Montarras
(2018)
The role of Pitx2 and Pitx3 in muscle stem cells gives new insights into P38α MAP kinase and redox regulation of muscle regeneration
eLife 7:e32991.
https://doi.org/10.7554/eLife.32991
  2. Download BibTeX
  3. Download .RIS

Abstract

Skeletal muscle regeneration depends on satellite cells. After injury these muscle stem cells exit quiescence, proliferate and differentiate to regenerate damaged fibres. We show that this progression is accompanied by metabolic changes leading to increased production of reactive oxygen species (ROS). Using Pitx2/3 single and double mutant mice that provide genetic models of deregulated redox states, we demonstrate that moderate overproduction of ROS results in premature differentiation of satellite cells while high levels lead to their senescence and regenerative failure. Using the ROS scavenger, N-Acetyl-Cysteine, (NAC) in primary cultures we show that a physiological increase in ROS is required for satellite cells to exit the cell cycle and initiate differentiation through the redox activation of p38a MAP kinase. Subjecting cultured satellite cells to transient inhibition of P38a MAP kinase in conjunction with NAC treatment leads to their rapid expansion, with striking improvement of their regenerative potential in grafting experiments.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Source data files have been provided for all figures and supplement figures.

The following previously published data sets were used

Article and author information

Author details

  1. Aurore Lhonore

    Department of Developmental and Stem Cell Biology, CNRS UMR 3738, Institut Pasteur, Paris, France
    For correspondence
    alhonore@hotmail.com
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6371-4455

  2. Pierre-Henri Commère

    Platform of Cytometry, Institut Pasteur, Paris, France
    Competing interests
    No competing interests declared.

  3. Elisa Negroni

    Center for Research in Myology, Sorbonne Universités, Université Pierre et Marie Curie Université Paris 06, INSERM UMRS974, Paris, France
    Competing interests
    No competing interests declared.

  4. Giorgia Pallafacchina

    Italian National Research Council (CNR) Neuroscience Institute, Padova, Italy
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9766-5970

  5. Bertrand Friguet

    Biological Adaptation and Aging-IBPS, CNRS UMR 8256, INSERM ERL U1164, Paris, France
    Competing interests
    No competing interests declared.

  6. Jacques Drouin

    Laboratory of Molecular Genetics, Institut de Recherches Cliniques de Montréal, Montréal, France
    Competing interests
    No competing interests declared.

  7. Margaret Buckingham

    Department of Developmental and Stem Cell Biology, CNRS UMR 3738, Institut Pasteur, Paris, France
    Competing interests
    Margaret Buckingham, Reviewing editor, eLife.

  8. Didier Montarras

    Department of Developmental and Stem Cell Biology, CNRS UMR 3738, CNRS UMR 3738, Institut Pasteur, Paris, France
    For correspondence
    didier.montarras@pasteur.fr
    Competing interests
    No competing interests declared.

Funding

Agence Nationale de la Recherche (REGSAT)

  • Margaret Buckingham

AFM-Téléthon

  • Didier Montarras

AFM-Téléthon (Postdoc Fellowship)

  • Aurore Lhonore

Agence Nationale de la Recherche (ANR-10-LABX-73)

  • Margaret Buckingham

Seventh Framework Programme (OptiStem 223098)

  • Margaret Buckingham

Seventh Framework Programme (Marie Curie IRG 248496)

  • Aurore Lhonore

Fondation pour la Recherche Médicale (Postdoc Fellowship)

  • Aurore Lhonore

Fondation pour la Recherche Médicale (Postdoc Fellowship)

  • Giorgia Pallafacchina

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 procedures were approved and conducted in accordance with the Institut Pasteur animal ethics committee (CEEA Institut Pasteur n{degree sign}2013-0017 and APAFIS #2455 2015 1122133311) following the regulations of the Ministry of Agriculture and the European Community guidelines. All surgery were performed under Ketamine /Xylazine anesthesia and and every effort was made to minimize suffering.

Reviewing Editor

  1. Andrew Brack, University of California, San Francisco, United States

Version history

  1. Received: October 20, 2017
  2. Accepted: August 1, 2018
  3. Accepted Manuscript published: August 14, 2018 (version 1)
  4. Version of Record published: October 16, 2018 (version 2)

Copyright

© 2018, Lhonore 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

  • 2,396
    Page views
  • 514
    Downloads
  • 43
    Citations

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

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. Aurore Lhonore
  2. Pierre-Henri Commère
  3. Elisa Negroni
  4. Giorgia Pallafacchina
  5. Bertrand Friguet
  6. Jacques Drouin
  7. Margaret Buckingham
  8. Didier Montarras
(2018)
The role of Pitx2 and Pitx3 in muscle stem cells gives new insights into P38α MAP kinase and redox regulation of muscle regeneration
eLife 7:e32991.
https://doi.org/10.7554/eLife.32991

Categories and tags

Research organism

Further reading

    1. Cell Biology
    2. Developmental Biology

    Cylicins are a structural component of the sperm calyx being indispensable for male fertility in mice and human

    Simon Schneider, Andjela Kovacevic ... Hubert Schorle
    Research Article

    Cylicins are testis-specific proteins, which are exclusively expressed during spermiogenesis. In mice and humans, two Cylicins, the gonosomal X-linked Cylicin 1 (Cylc1/CYLC1) and the autosomal Cylicin 2 (Cylc2/CYLC2) genes, have been identified. Cylicins are cytoskeletal proteins with an overall positive charge due to lysine-rich repeats. While Cylicins have been localized in the acrosomal region of round spermatids, they resemble a major component of the calyx within the perinuclear theca at the posterior part of mature sperm nuclei. However, the role of Cylicins during spermiogenesis has not yet been investigated. Here, we applied CRISPR/Cas9-mediated gene editing in zygotes to establish Cylc1- and Cylc2-deficient mouse lines as a model to study the function of these proteins. Cylc1 deficiency resulted in male subfertility, whereas Cylc2-/-, Cylc1-/yCylc2+/-, and Cylc1-/yCylc2-/- males were infertile. Phenotypical characterization revealed that loss of Cylicins prevents proper calyx assembly during spermiogenesis. This results in decreased epididymal sperm counts, impaired shedding of excess cytoplasm, and severe structural malformations, ultimately resulting in impaired sperm motility. Furthermore, exome sequencing identified an infertile man with a hemizygous variant in CYLC1 and a heterozygous variant in CYLC2, displaying morphological abnormalities of the sperm including the absence of the acrosome. Thus, our study highlights the relevance and importance of Cylicins for spermiogenic remodeling and male fertility in human and mouse, and provides the basis for further studies on unraveling the complex molecular interactions between perinuclear theca proteins required during spermiogenesis.

    1. Cell Biology
    2. Structural Biology and Molecular Biophysics

    Baited reconstruction with 2D template matching for high-resolution structure determination in vitro and in vivo without template bias

    Bronwyn A Lucas, Benjamin A Himes, Nikolaus Grigorieff
    Research Advance

    Previously we showed that 2D template matching (2DTM) can be used to localize macromolecular complexes in images recorded by cryogenic electron microscopy (cryo-EM) with high precision, even in the presence of noise and cellular background (Lucas et al., 2021; Lucas et al., 2022). Here, we show that once localized, these particles may be averaged together to generate high-resolution 3D reconstructions. However, regions included in the template may suffer from template bias, leading to inflated resolution estimates and making the interpretation of high-resolution features unreliable. We evaluate conditions that minimize template bias while retaining the benefits of high-precision localization, and we show that molecular features not present in the template can be reconstructed at high resolution from targets found by 2DTM, extending prior work at low-resolution. Moreover, we present a quantitative metric for template bias to aid the interpretation of 3D reconstructions calculated with particles localized using high-resolution templates and fine angular sampling.

    1. Cell Biology
    2. Immunology and Inflammation

    Regulation of pDC Fate Determination by Histone Deacetylase 3

    Yijun Zhang, Tao Wu ... Li Wu
    Research Article

    Dendritic cells (DCs), the key antigen-presenting cells, are primary regulators of immune responses. Transcriptional regulation of DC development had been one of the major research interests in DC biology, however, the epigenetic regulatory mechanisms during DC development remains unclear. Here, we report that Histone deacetylase 3 (Hdac3), an important epigenetic regulator, is highly expressed in pDCs, and its deficiency profoundly impaired the development of pDCs. Significant disturbance of homeostasis of hematopoietic progenitors was also observed in HDAC3-deficient mice, manifested by altered cell numbers of these progenitors and defective differentiation potentials for pDCs. Using the in vitro Flt3L supplemented DC culture system, we further demonstrated that HDAC3 was required for the differentiation of pDCs from progenitors at all developmental stages. Mechanistically, HDAC3 deficiency resulted in enhanced expression of cDC1-associated genes, owing to markedly elevated H3K27 acetylation (H3K27ac) at these gene sites in BM pDCs. In contrast, the expression of pDC-associated genes was significantly downregulated, leading to defective pDC differentiation.