Molecular Mechanisms of Stress-induced Reactive Oxygen Species Formation in Skeletal Muscle
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ndltd-OhioLink-oai-etd.ohiolink.edu-osu10388538942021-08-03T05:47:34Z Molecular Mechanisms of Stress-induced Reactive Oxygen Species Formation in Skeletal Muscle Zuo, Li fluorescence cytochrome c diaphragm superoxide mitochondria lipoxygenase laser scan confocal microscopy <p>Reactive oxygen species (ROS) play an important role in many biological systems. Skeletal muscles have been shown to generate considerable ROS in resting and in contracting conditions. In this study, I tested the hypothesis that increased ROS production in skeletal muscles is also associated with exposure to two other conditions of stress which are common in normal skeletal muscle during exercise, namely heat stress and hypoxia.</p> <p>There is no previous direct evidence that ROS are produced during these stresses, particularly in skeletal muscle, but they may play important roles in normal contractile and cell signaling responses. Two assays for superoxide (O<sub>2</sub><sup>•-</sup>) formation were used in rodent diaphragm, the cytochrome c assay for extracellular O<sub>2</sub><sup>•-</sup> release and the hydroethidine oxidation for intracellular O<sub>2</sub><sup>•-</sup> formation. The results demonstrate the following: 1) Markedly increased intra- and extracellular ROS formation was observed, particularly O<sub>2</sub><sup>•-</sup>, at temperatures known to be physiologically relevant to exercise (i.e. 42°C). 2) The process of O<sub>2</sub><sup>•-</sup>, release (extracellular formation) is not directly related to mitochondria, NADPH oxidase, or anion channels, though these are normally believed to be involved in either O<sub>2</sub><sup>•-</sup> generation or the exit pathway of O<sub>2</sub><sup>•-</sup> through membranes. 3) Upstream pathways of arachidonic acid (AA) metabolism, both phospholipase A<sub>2</sub> and nitric oxide synthase are associated with O<sub>2</sub><sup>•-</sup> release. 4) Downstream pathways of AA metabolism are also involved. Though blockage of either cyclooxygenase or cytochrome P450-dependent enzymes does not cause any inhibition of O<sub>2</sub><sup>•-</sup> release, blockage of lipoxygenase (LOX) results in near elimination of the signal. This suggests that O<sub>2</sub><sup>•-</sup> release is dependent on AA metabolism through the LOX pathway. However, confocal measurements of intracellular O<sub>2</sub><sup>•-</sup> formation suggest that intracellular ROS are produced by a separate mechanism. 5) Tissue fluorometry techniques, using a fluorescence probe sensitive to hydrogen peroxide, showed that acute hypoxia also induces ROS formation in skeletal muscle.</p> <p>These findings provide insight into understanding the potential role of ROS in physiologic and pathophysiologic responses to heat exposure and acute hypoxia in skeletal muscle.</p> 2002-12-20 English text The Ohio State University / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=osu1038853894 http://rave.ohiolink.edu/etdc/view?acc_num=osu1038853894 unrestricted This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws. |
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language |
English |
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topic |
fluorescence cytochrome c diaphragm superoxide mitochondria lipoxygenase laser scan confocal microscopy |
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fluorescence cytochrome c diaphragm superoxide mitochondria lipoxygenase laser scan confocal microscopy Zuo, Li Molecular Mechanisms of Stress-induced Reactive Oxygen Species Formation in Skeletal Muscle |
author |
Zuo, Li |
author_facet |
Zuo, Li |
author_sort |
Zuo, Li |
title |
Molecular Mechanisms of Stress-induced Reactive Oxygen Species Formation in Skeletal Muscle |
title_short |
Molecular Mechanisms of Stress-induced Reactive Oxygen Species Formation in Skeletal Muscle |
title_full |
Molecular Mechanisms of Stress-induced Reactive Oxygen Species Formation in Skeletal Muscle |
title_fullStr |
Molecular Mechanisms of Stress-induced Reactive Oxygen Species Formation in Skeletal Muscle |
title_full_unstemmed |
Molecular Mechanisms of Stress-induced Reactive Oxygen Species Formation in Skeletal Muscle |
title_sort |
molecular mechanisms of stress-induced reactive oxygen species formation in skeletal muscle |
publisher |
The Ohio State University / OhioLINK |
publishDate |
2002 |
url |
http://rave.ohiolink.edu/etdc/view?acc_num=osu1038853894 |
work_keys_str_mv |
AT zuoli molecularmechanismsofstressinducedreactiveoxygenspeciesformationinskeletalmuscle |
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1719425378138718208 |