Manipulation of environmental oxygen modifies reactive oxygen and nitrogen species generation during myogenesis

McCormick, Rachel, Pearson, Timothy and Vasilaki, Aphrodite (2016) Manipulation of environmental oxygen modifies reactive oxygen and nitrogen species generation during myogenesis. Redox Biology, 8. pp. 243-251. ISSN 2213-2317

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Abstract

Regulated changes in reactive oxygen and nitrogen species (RONS) activities are important in maintaining the normal sequence and development of myogenesis. Both excessive formation and reduction in RONS have been shown to affect muscle differentiation in a negative way. Cultured cells are typically grown in 20% O2 but this is not an appropriate physiological concentration for a number of cell types, including skeletal muscle. The aim was to examine the generation of RONS in cultured skeletal muscle cells under a physiological oxygen concentration condition (6% O2) and determine the effect on muscle myogenesis. Primary mouse satellite cells were grown in 20% or 6% O2 environments and RONS activity was measured at different stages of myogenesis by real-time fluorescent microscopy using fluorescent probes with different specificities i.e. dihydroethidium (DHE), 4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate (DAF-FM DA) and 5-(and-6)-chloromethyl-2′,7′ -dichlorodihydrofluorescein diacetate (CM-DCFH-DA). Data demonstrate that satellite cell proliferation increased when cells were grown in 6% O2 compared with 20% O2. Myoblasts grown in 20% O2 showed an increase in DCF fluorescence and DHE oxidation compared with myoblasts grown at 6% O2. Myotubes grown in 20% O2 also showed an increase in DCF and DAF-FM fluorescence and DHE oxidation compared with myotubes grown in 6% O2. The catalase and MnSOD contents were also increased in myoblasts and myotubes that were maintained in 20% O2 compared with myoblasts and myotubes grown in 6% O2. These data indicate that intracellular RONS activities in myoblasts and myotubes at rest are influenced by changes in environmental oxygen concentration and that the increased ROS may influence myogenesis in a negative manner.

Item Type: Article
Additional Information: Open Access under a Creative Commons Attribution (CC BY) licence
Uncontrolled Keywords: reactive oxygen and nitrogen species,low oxygen concentration,skeletal muscle,antioxidant defence enzymes
Faculty \ School: Faculty of Medicine and Health Sciences > Norwich Medical School
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Depositing User: Pure Connector
Date Deposited: 15 Feb 2016 17:00
Last Modified: 22 Apr 2020 01:04
URI: https://ueaeprints.uea.ac.uk/id/eprint/57091
DOI: 10.1016/j.redox.2016.01.011

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