骨骼肌线粒体表型对 H2O2 排放的影响

IF 16.4 1区 化学 Q1 CHEMISTRY, MULTIDISCIPLINARY Accounts of Chemical Research Pub Date : 2024-01-06 DOI:10.1016/j.cbpb.2024.110940
Collins Kamunde , Yashodya Wijayakulathilake , Chidozie Okoye , Nirmala Chinnappareddy , Zahra Kalvani , Pius Tetteh , Michael van den Heuvel , Ravinder Sappal , Don Stevens
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引用次数: 0

摘要

活性氧(ROS)是骨骼肌线粒体信息处理系统在静息和运动时的主要输出。在骨骼肌中,线粒体释放的 ROS 取决于多种因素;然而,纤维类型的特异性差异仍然不明确,部分原因是使用的线粒体来自由混合纤维组成的哺乳动物肌肉。为了阐明纤维类型的特异性差异,我们使用了从虹鳟鱼(Oncorhynchus mykiss)红白骨骼肌中分离出来的线粒体。我们首先确定了在分离的鱼类红骨骼肌和白骨骼肌线粒体(RMM 和 WMM)中测量 ROS(以 H2O2 形式)产生的测定条件,然后比较了不同底物氧化过程中的释放率以及线粒体电子传递系统(ETS)药物调节剂的反应。我们的研究结果表明,RMM 和 WMM 的 H2O2 发射率可通过相同的蛋白质浓度和 Amplex UltraRed 辣根过氧化物酶(AUR-HRP)检测系统的组成进行量化。对于 RMM 和 WMM,蛋白质归一化的 H2O2 发射率在测试的最低蛋白质浓度时最高,之后呈指数下降。然而,H2O2 发射率的绝对值取决于用于将荧光信号转换为 H2O2 的校准曲线,而趋势则取决于归一化策略。我们发现,根据被氧化的底物及其浓度的不同,RMM 和 WMM 的 H2O2 发射率在质量和数量上都存在很大差异。同样,ETS 的药理调节剂也改变了 RMM 和 WMM 中 H2O2 释放的幅度和趋势。虽然相似浓度的底物在 RMM 和 WMM 中能引起最大的释放率,但在数量上却有所不同,因此根据肌肉纤维类型的不同,最大 H2O2 释放率可能需要不同浓度的药理 ETS 调节剂。综上所述,我们的研究表明,与 WMM 相比,RMM 存在生化差异,这种差异会改变底物氧化和对 ETS 调节剂的反应,从而导致特定纤维类型的线粒体 H2O2 发射率。
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Effect of skeletal muscle mitochondrial phenotype on H2O2 emission

Reactive oxygen species (ROS) are a key output of the skeletal muscle mitochondrial information processing system both at rest and during exercise. In skeletal muscle, mitochondrial ROS release depends on multiple factors; however, fiber-type specific differences remain ambiguous in part owing to the use of mitochondria from mammalian muscle that consist of mixed fibers. To elucidate fiber-type specific differences, we used mitochondria isolated from rainbow trout (Oncorhynchus mykiss) red and white skeletal muscles that consist of spatially distinct essentially pure red and white fibers. We first characterized the assay conditions for measuring ROS production (as H2O2) in isolated fish red and white skeletal muscle mitochondria (RMM and WMM) and thereafter compared the rates of emission during oxidation of different substrates and the responses to mitochondrial electron transport system (ETS) pharmacological modulators. Our results showed that H2O2 emission rates by RMM and WMM can be quantified using the same protein concentration and composition of the Amplex UltraRed-horseradish peroxidase (AUR-HRP) detection system. For both RMM and WMM, protein normalized H2O2 emission rates were highest at the lowest protein concentration tested and decreased exponentially thereafter. However, the absolute values of H2O2 emission rates depended on the calibration curves used to convert fluorescent signals to H2O2 while the trends depended on the normalization strategy. We found substantial qualitative and quantitative differences between RMM and WMM in the H2O2 emission rates depending on the substrates being oxidized and their concentrations. Similarly, pharmacological modulators of the ETS altered the magnitudes and trends of the H2O2 emission differently in RMM and WMM. While comparable concentrations of substrates elicited maximal albeit quantitively different emission rates in RMM and WMM, different concentrations of pharmacological ETS modulators may be required for maximal H2O2 emission rates depending on muscle fiber-type. Taken together, our study suggests that biochemical differences exist in RMM compared with WMM that alter substrate oxidation and responses to ETS modulators resulting in fiber-type specific mitochondrial H2O2 emission rates.

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来源期刊
Accounts of Chemical Research
Accounts of Chemical Research 化学-化学综合
CiteScore
31.40
自引率
1.10%
发文量
312
审稿时长
2 months
期刊介绍: Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.
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