Murine muscle stem cell response to perturbations of the neuromuscular junction are attenuated with aging

  1. Jacqueline Larouche
  2. Mahir Mohiuddin
  3. Jeongmoon J Choi
  4. Peter J Ulintz
  5. Paula M Fraczek
  6. Kaitlyn Sabin
  7. Sethuramasundaram Pitchiaya
  8. Sarah J Kurpiers
  9. Jesus Castor-Macias
  10. Wenxuan Liu
  11. Robert Louis Hastings
  12. Lemuel A Brown
  13. James F Markworth
  14. Kanishka De Silva
  15. Benjamin D Levi
  16. Sofia D Merajver
  17. Gregorio Valdez
  18. Joe V Chakkalakal
  19. Young Jang  Is a corresponding author
  20. Susan Brooks  Is a corresponding author
  21. Carlos A Aguilar  Is a corresponding author
  1. University of Michigan, United States
  2. Georgia Institute of Technology, United States
  3. University of Rochester, United States
  4. Brown University, United States
  5. University of Texas Southwestern, United States
  6. University of Rochester Medical Center, United States

Abstract

During aging and neuromuscular diseases, there is a progressive loss of skeletal muscle volume and function impacting mobility and quality of life. Muscle loss is often associated with denervation and a loss of resident muscle stem cells (satellite cells or MuSCs), however, the relationship between MuSCs and innervation has not been established. Herein, we administered severe neuromuscular trauma to a transgenic murine model that permits MuSC lineage tracing. We show that a subset of MuSCs specifically engraft in a position proximal to the neuromuscular junction (NMJ), the synapse between myofibers and motor neurons, in healthy young adult muscles. In aging and in a mouse model of neuromuscular degeneration (Cu/Zn superoxide dismutase knockout – Sod1-/-), this localized engraftment behavior was reduced. Genetic rescue of motor neurons in Sod1-/- mice reestablished integrity of the NMJ in a manner akin to young muscle and partially restored MuSC ability to engraft into positions proximal to the NMJ. Using single cell RNA-sequencing of MuSCs isolated from aged muscle, we demonstrate that a subset of MuSCs are molecularly distinguishable from MuSCs responding to myofiber injury and share similarity to synaptic myonuclei. Collectively, these data reveal unique features of MuSCs that respond to synaptic perturbations caused by aging and other stressors.

Data availability

Data have been deposited to GEO under accession code GSE165978.

The following data sets were generated
The following previously published data sets were used

Article and author information

Author details

  1. Jacqueline Larouche

    Biomedical Engineering, University of Michigan, Ann Arbor, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9380-3547
  2. Mahir Mohiuddin

    BIomedical Engineering, Georgia Institute of Technology, Atlanta, GA, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Jeongmoon J Choi

    BIomedical Engineering, Georgia Institute of Technology, Atlanta, GA, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Peter J Ulintz

    Biomedical Engineering, University of Michigan, Ann Arbor, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2037-8655
  5. Paula M Fraczek

    Biomedical Engineering, University of Michigan, Ann Arbor, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Kaitlyn Sabin

    Biomedical Engineering, University of Michigan, Ann Arbor, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Sethuramasundaram Pitchiaya

    Biomedical Engineering, University of Michigan, Ann Arbor, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Sarah J Kurpiers

    Biomedical Engineering, University of Michigan, Ann Arbor, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Jesus Castor-Macias

    Biomedical Engineering, University of Michigan, Ann Arbor, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. Wenxuan Liu

    Pharmacology & Physiology, University of Rochester, Rochester, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. Robert Louis Hastings

    10Dept. of Molecular Biology, Cell Biology and Biochemistry, Brown University, Providence, United States
    Competing interests
    The authors declare that no competing interests exist.
  12. Lemuel A Brown

    Biomedical Engineering, University of Michigan, Ann Arbor, United States
    Competing interests
    The authors declare that no competing interests exist.
  13. James F Markworth

    Biomedical Engineering, University of Michigan, Ann Arbor, United States
    Competing interests
    The authors declare that no competing interests exist.
  14. Kanishka De Silva

    Biomedical Engineering, University of Michigan, Ann Arbor, United States
    Competing interests
    The authors declare that no competing interests exist.
  15. Benjamin D Levi

    Dept. of Surgery, University of Texas Southwestern, Dallas, United States
    Competing interests
    The authors declare that no competing interests exist.
  16. Sofia D Merajver

    Internal Medicine-Hematology/Oncology, University of Michigan, Ann Arbor, United States
    Competing interests
    The authors declare that no competing interests exist.
  17. Gregorio Valdez

    Department of Molecular Biology, Cell Biology and Biochemistry, Brown University, Providence, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0375-4532
  18. Joe V Chakkalakal

    Department of Orthopaedics and Rehabilitation, University of Rochester Medical Center, Rochester, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8440-7312
  19. Young Jang

    BIomedical Engineering, Georgia Institute of Technology, Atlanta, GA, United States
    For correspondence
    young.jang@gatech.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9489-2104
  20. Susan Brooks

    Internal Medicine-Hematology/Oncology, University of Michigan, Ann Arbor, United States
    For correspondence
    svbrooks@umich.edu
    Competing interests
    The authors declare that no competing interests exist.
  21. Carlos A Aguilar

    Biomedical Engineering, University of Michigan, Ann Arbor, United States
    For correspondence
    caguilar@umich.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3830-0634

Funding

National Institute on Aging (P01 AG051442)

  • Susan Brooks

Congressionally Directed Medical Research Programs (W81XWH1810653)

  • Benjamin D Levi

Congressionally Directed Medical Research Programs (W81XWH2010795)

  • Benjamin D Levi

Breast Cancer Research Foundation

  • Peter J Ulintz
  • Sofia D Merajver

National Science Foundation (DGE 1256260)

  • Jacqueline Larouche

National Institute on Aging (R01 AG051456)

  • Joe V Chakkalakal

National Institute of Arthritis and Musculoskeletal and Skin Diseases (P30 AR069620)

  • Susan Brooks
  • Carlos A Aguilar

National Institute of Arthritis and Musculoskeletal and Skin Diseases (R01 AR071379)

  • Benjamin D Levi

National Institute of Arthritis and Musculoskeletal and Skin Diseases (R61 AR078072)

  • Benjamin D Levi

3M Foundation

  • Carlos A Aguilar

American Federation for Aging Research

  • Carlos A Aguilar

National Institute on Aging (P30 AG024824)

  • Susan Brooks
  • Carlos A Aguilar

Congressionally Directed Medical Research Programs (W81XWH2010336)

  • Young Jang
  • Carlos A Aguilar

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 (IACUC protocol #: PRO00008428, PRO00006689) of the University of Michigan.

Reviewing Editor

  1. Shahragim Tajbakhsh, Institut Pasteur, France

Publication history

  1. Preprint posted: May 29, 2020 (view preprint)
  2. Received: January 21, 2021
  3. Accepted: July 28, 2021
  4. Accepted Manuscript published: July 29, 2021 (version 1)
  5. Version of Record published: August 12, 2021 (version 2)

Copyright

© 2021, Larouche 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

  • 2,103
    Page views
  • 289
    Downloads
  • 4
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, 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. Jacqueline Larouche
  2. Mahir Mohiuddin
  3. Jeongmoon J Choi
  4. Peter J Ulintz
  5. Paula M Fraczek
  6. Kaitlyn Sabin
  7. Sethuramasundaram Pitchiaya
  8. Sarah J Kurpiers
  9. Jesus Castor-Macias
  10. Wenxuan Liu
  11. Robert Louis Hastings
  12. Lemuel A Brown
  13. James F Markworth
  14. Kanishka De Silva
  15. Benjamin D Levi
  16. Sofia D Merajver
  17. Gregorio Valdez
  18. Joe V Chakkalakal
  19. Young Jang
  20. Susan Brooks
  21. Carlos A Aguilar
(2021)
Murine muscle stem cell response to perturbations of the neuromuscular junction are attenuated with aging
eLife 10:e66749.
https://doi.org/10.7554/eLife.66749

Further reading

    1. Stem Cells and Regenerative Medicine
    Han Xiao et al.
    Research Article Updated

    Proper mechanical stimulation can improve rotator cuff enthesis injury repair. However, the underlying mechanism of mechanical stimulation promoting injury repair is still unknown. In this study, we found that Prrx1+ cell was essential for murine rotator cuff enthesis development identified by single-cell RNA sequence and involved in the injury repair. Proper mechanical stimulation could promote the migration of Prrx1+ cells to enhance enthesis injury repair. Meantime, TGF-β signaling and primary cilia played an essential role in mediating mechanical stimulation signaling transmission. Proper mechanical stimulation enhanced the release of active TGF-β1 to promote migration of Prrx1+ cells. Inhibition of TGF-β signaling eliminated the stimulatory effect of mechanical stimulation on Prrx1+ cell migration and enthesis injury repair. In addition, knockdown of Pallidin to inhibit TGF-βR2 translocation to the primary cilia or deletion of Ift88 in Prrx1+ cells also restrained the mechanics-induced Prrx1+ cells migration. These findings suggested that mechanical stimulation could increase the release of active TGF-β1 and enhance the mobilization of Prrx1+ cells to promote enthesis injury repair via ciliary TGF-β signaling.

    1. Neuroscience
    2. Stem Cells and Regenerative Medicine
    Weizhao Chen et al.
    Short Report

    Lineage reprograming of resident glial cells to dopaminergic neurons (DAns) is an attractive prospect of the cell-replacement therapy for Parkinson's disease (PD). However, it is unclear whether repressing polypyrimidine tract binding protein 1 (PTBP1) could efficiently convert astrocyte to DAns in the substantia nigra and striatum. Although reporter-positive DAns were observed in both groups after delivering the adeno-associated virus (AAV) expressing a reporter with shRNA or CRISPR-CasRx to repress astroglial PTBP1, the possibility of AAV leaking into endogenous DAns could not be excluded without using a reliable lineage-tracing method. By adopting stringent lineage-tracing strategy, two other studies showed that either knockdown or genetic deletion of quiescent astroglial PTBP1 fails to obtain induced DAns under physiological condition. However, the role of reactive astrocytes might be underestimated because upon brain injury, reactive astrocyte can acquire certain stem cell hallmarks that may facilitate the lineage conversion process. Therefore, whether reactive astrocytes could be genuinely converted to DAns after PTBP1 repression in a PD model needs further validation. In this study, we used Aldh1l1-CreERT2-mediated specific astrocyte-lineage-tracing method to investigate whether reactive astrocytes can be converted to DAns in a 6-hydroxydopamine (6-OHDA) mouse model of PD. However, we found that no astrocyte-originated DAn was generated after effective and persistent knockdown of astroglial PTBP1 either in the substantia nigra or in striatum, while AAV 'leakage' to nearby neurons was easily observed. Our results confirmed that repressing PTBP1 does not convert astrocytes to DAns, regardless of physiological or PD-related pathological conditions.