Differences in transcriptional patterns of extracellular matrix, inflammatory, and myogenic regulatory genes in myofibroblasts, fibroblasts, and muscle precursor cells isolated from old male rat skeletal muscle using a novel cell isolation procedure
Abstract
Aged skeletal muscle displays increased fibrosis and impaired regeneration. While it is not well characterized how skeletal muscle fibroblasts contribute to these phenomena, transforming growth factor-β1 (TGF-β1) and Delta/Notch signaling have been implicated to influence muscle regeneration. In this study, a unique combination of aging phenotypes is identified in differentiating fibroblasts (myofibroblasts), proliferating fibroblasts, and muscle precursor cells (MPCs) that characterize an impaired regenerative potential observed in aged skeletal muscle. Using a novel dual-isolation technique, that isolates fibroblasts and MPCs from the same rat skeletal muscle sample, and cell culture conditions of 5 % O(2) and 5 % CO(2), we report for the first time that myofibroblasts from 32-mo-old skeletal muscle, compared to 3-mo-old, display increased levels of mRNA for the essential extracellular matrix (ECM) genes, collagen 4α1 (83 % increase), collagen 4α2 (98 % increase), and laminin 2 (113 % increase), as well as increased levels of mRNA for the inflammatory markers, interleukin-6 (4.3-fold increase) and tumor necrosis factor α (3.2-fold increase), and TGF-β1 (84 % increase), whose protein controls proliferation and differentiation. Additionally, we demonstrate that proliferating fibroblasts from 32-mo-old skeletal muscle display increased levels of mRNA for the Notch ligand, Delta 1 (≥2.0-fold increase). Together, these findings suggest that increased expression of ECM and inflammatory genes in myofibroblasts from 32-mo-old skeletal muscle may contribute to the fibrogenic phenotype that impairs regeneration in aged skeletal muscle. Furthermore, we believe the novel dual-isolation technique developed here may be useful in studies that investigate communications among MPCs, fibroblasts, and myofibroblasts in skeletal muscle.
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During regeneration of aged skeletal muscle, muscle tissue is replaced with connective tissue and consequently muscle function and muscle quality decline.
“During regeneration of aged skeletal muscle, muscle tissue is replaced with connective tissue and, consequently muscle function and muscle quality decline.”
From Phytoecdysteroids Accelerate Recovery of Skeletal Muscle Function Following in vivo Eccentric Contraction-Induced Injury in Adult and Old Mice (2021) · cites this paper as [66]
One of the major complications with age-related impairment in muscle regeneration after injury is decreased myogenic potential and increased fibrogenesis.
“One of the major complications with the age-related impairment in muscle regeneration after injury is not only the decreased myogenic potential, but also increased fibrogenesis (growth of connective tissue) (66).”
From Phytoecdysteroids Accelerate Recovery of Skeletal Muscle Function Following in vivo Eccentric Contraction-Induced Injury in Adult and Old Mice (2021) · cites this paper as [66]
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The myofibroblast matrix: implications for tissue repair and fibrosis
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- Authors
- Zwetsloot, K. A., Nedergaard, A., Gilpin, L. T., Childs, T. E., & Booth, F. W.
- Affiliations
- Appalachian State University (US)
- University of Copenhagen (DK)
- University of Missouri (US)
- Corresponding author
- Zwetsloot, Kevin A.
- Venue
- Biogerontology
- Published
- 2012-04-26
- Volume
- 13
- Issue
- 4
- Pages
- 383-398
- ISSN
- 1389-5729 (ISSN-L)
- 1573-6768
- OpenAlex
- W2082703856
- Cited by
- 11
- References
- 44
- Citations by year
- 2026: 1 · 2025: 1 · 2022: 2 · 2021: 2 · 2019: 1 · 2017: 1 · 2016: 1 · 2015: 1 · 2012: 1
- FWCI
- 0.26
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Biogerontology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- University of Missouri
- Medical College of Wisconsin
- National Institute on Aging (R01AG018780, R01 AG18780)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Muscle Physiology and Disorders
- Life Sciences › Agricultural and Biological Sciences › Animal Science and Zoology › Meat and Animal Product Quality
- Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Cardiac Fibrosis and Remodeling
- Keywords
- skeletal muscle aging, myofibroblasts, fibroblasts, muscle precursor cells, extracellular matrix genes, type IV collagen, laminin-2, inflammatory genes, interleukin-6, TNF-α, TGF-β1, Delta1, Notch signaling, muscle fibrosis, muscle regeneration, gene expression
- MeSH terms
- Aging, Animals, Cell Separation, Cells, Cultured, Collagen, Extracellular Matrix, Fibroblasts, Inflammation, Laminin, Male, RNA, Messenger, Stem Cells, Transcription, Genetic, Tumor Necrosis Factor-alpha, Interleukin-6, Myogenic Regulatory Factors, Muscle, Skeletal, Models, Animal, Rats, Receptors, Notch, Transforming Growth Factor beta1, Myofibroblasts
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