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PloS one2014; 9(8); e105971; doi: 10.1371/journal.pone.0105971

Calcium homeostasis in myogenic differentiation factor 1 (MyoD)-transformed, virally-transduced, skin-derived equine myotubes.

Abstract: Dysfunctional skeletal muscle calcium homeostasis plays a central role in the pathophysiology of several human and animal skeletal muscle disorders, in particular, genetic disorders associated with ryanodine receptor 1 (RYR1) mutations, such as malignant hyperthermia, central core disease, multiminicore disease and certain centronuclear myopathies. In addition, aberrant skeletal muscle calcium handling is believed to play a pivotal role in the highly prevalent disorder of Thoroughbred racehorses, known as Recurrent Exertional Rhabdomyolysis. Traditionally, such defects were studied in human and equine subjects by examining the contractile responses of biopsied muscle strips exposed to caffeine, a potent RYR1 agonist. However, this test is not widely available and, due to its invasive nature, is potentially less suitable for valuable animals in training or in the human paediatric setting. Furthermore, increasingly, RYR1 gene polymorphisms (of unknown pathogenicity and significance) are being identified through next generation sequencing projects. Consequently, we have investigated a less invasive test that can be used to study calcium homeostasis in cultured, skin-derived fibroblasts that are converted to the muscle lineage by viral transduction with a MyoD (myogenic differentiation 1) transgene. Similar models have been utilised to examine calcium homeostasis in human patient cells, however, to date, there has been no detailed assessment of the cells' calcium homeostasis, and in particular, the responses to agonists and antagonists of RYR1. Here we describe experiments conducted to assess calcium handling of the cells and examine responses to treatment with dantrolene, a drug commonly used for prophylaxis of recurrent exertional rhabdomyolysis in horses and malignant hyperthermia in humans.
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Publication Date: 2014-08-22 PubMed ID: 25148524PubMed Central: PMC4141859DOI: 10.1371/journal.pone.0105971Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research paper discusses an alternative test method for studying calcium homeostasis in muscle cells. This is done through transforming skin-derived fibroblasts into muscle lineage by viral transduction with a MyoD gene. The method is seen as less invasive, and hence, potentially more suitable in situations where conventional testing is unsuitable, such as in valuable racehorses or in human pediatric settings.

Understanding Calcium Homeostasis and Muscle Disorders

  • Skeletal muscle calcium homeostasis is key when it comes to muscle disorders in both humans and animals.
  • Specially, disorders associated with ryanodine receptor 1 (RYR1) mutations such as malignant hyperthermia, central core disease, multiminicore disease, certain centronuclear myopathies, and Recurrent Exertional Rhabdomyolysis in Thoroughbred racehorses.
  • Traditionally, defects in this were studied through testing the contractile responses of muscle biopsies to caffeine, a potent RYR1 agonist.

Limitations of Traditional Testing

  • Due to its invasive nature, this method is often unsuitable for use on valuable animals in training, or in a human pediatric setting.
  • In addition, the conventional test is not widely available.
  • Certain RYR1 gene polymorphisms, found through next generation sequencing, are of uncertain pathogenicity, adding another layer of complexity to this existing testing method.

Alternative Testing Method

  • Given these issues, the researchers explored a less invasive test.
  • This involved studying calcium homeostasis in skin-derived fibroblasts that had been converted to muscle lineage through viral transduction with a MyoD gene.
  • This alternative test method, while not new, has hitherto not been thoroughly examined in terms of its assessment of the cells’ calcium homeostasis.

Experimental Process and Results

  • The researchers conducted experiments to assess how these transformed cells handled calcium.
  • They specifically examined how these cells responded to agonists and antagonists of RYR1.
  • One treatment examined was dantrolene, a drug commonly used for preventing recurring exertional rhabdomyolysis in racehorses and malignant hyperthermia in humans.

Cite This Article

APA
Fernandez-Fuente M, Terracciano CM, Martin-Duque P, Brown SC, Vassaux G, Piercy RJ. (2014). Calcium homeostasis in myogenic differentiation factor 1 (MyoD)-transformed, virally-transduced, skin-derived equine myotubes. PLoS One, 9(8), e105971. https://doi.org/10.1371/journal.pone.0105971

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 9
Issue: 8
Pages: e105971

Researcher Affiliations

Fernandez-Fuente, Marta
  • Comparative Neuromuscular Diseases Laboratory, Department of Clinical Sciences and Services, Royal Veterinary College, London, United Kingdom.
Terracciano, Cesare M
  • Laboratory of Cell Electrophysiology, Imperial College London, Myocardial Function, National Heart and Lung Institute, Hammersmith Hospital, London, United Kingdom.
Martin-Duque, Pilar
  • Universidad Francisco de Vitoria, Facultad de Ciencias Biosanitarias: Pozuelo de Alarcón (Madrid), Madrid, Spain.
Brown, Susan C
  • Comparative Biomedical Sciences, Royal Veterinary College, London, United Kingdom.
Vassaux, Georges
  • Laboratoire TIRO, UMRE 4320, iBEB, DSV, Commissariat a' l'Energie Atomique, Nice, France.
Piercy, Richard J
  • Comparative Neuromuscular Diseases Laboratory, Department of Clinical Sciences and Services, Royal Veterinary College, London, United Kingdom.

MeSH Terms

  • Animals
  • Caffeine / pharmacology
  • Calcium / metabolism
  • Calcium Channel Agonists / pharmacology
  • Calcium Channel Blockers / pharmacology
  • Cells, Cultured
  • Dantrolene / pharmacology
  • Dose-Response Relationship, Drug
  • HEK293 Cells
  • Homeostasis / drug effects
  • Horses
  • Humans
  • Muscle Fibers, Skeletal / drug effects
  • Muscle Fibers, Skeletal / metabolism
  • MyoD Protein / genetics
  • MyoD Protein / metabolism
  • Skin / cytology
  • Thapsigargin / pharmacology
  • Transduction, Genetic
  • Transgenes

Conflict of Interest Statement

Competing Interests: The authors have declared that no competing interests exist.

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Citations

This article has been cited 1 times.
  1. Li X, Zhao X, Qin Z, Li J, Sun B, Liu L. Regulation of calcium homeostasis in endoplasmic reticulum-mitochondria crosstalk: implications for skeletal muscle atrophy. Cell Commun Signal 2025 Jan 9;23(1):17.
    doi: 10.1186/s12964-024-02014-wpubmed: 39789595google scholar: lookup
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