Calcium homeostasis in myogenic differentiation factor 1 (MyoD)-transformed, virally-transduced, skin-derived equine myotubes
File(s)
Author(s)
Type
Journal Article
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.
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.
Date Issued
2014-08-22
Date Acceptance
2014-07-30
Citation
PLoS ONE, 2014, 9 (8)
ISSN
1932-6203
Publisher
Public Library of Science (PLoS)
Journal / Book Title
PLoS ONE
Volume
9
Issue
8
Copyright Statement
© 2014 Fernandez-Fuente et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which
permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Identifier
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000341230600101&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=1ba7043ffcc86c417c072aa74d649202
Subjects
Science & Technology
Multidisciplinary Sciences
Science & Technology - Other Topics
CENTRAL CORE DISEASE
RECURRENT EXERTIONAL RHABDOMYOLYSIS
RYANODINE RECEPTOR MUTATIONS
RECESSIVE RYR1 MUTATIONS
SKELETAL-MUSCLE FIBERS
CA2+ RELEASE CHANNEL
MALIGNANT HYPERTHERMIA
VENTRICULAR MYOCYTES
DERMAL FIBROBLASTS
DANTROLENE SODIUM
Publication Status
Published
Article Number
e105971
Date Publish Online
2014-08-22