Redox regulation of muscle adaptations to contractile activity and aging

Jackson MJ. Redox regulation of muscle adaptations to contractile activity an

In vitro susceptibility of thioredoxins and glutathione to redox modification and aging-related changes in skeletal muscle

AbstractThioredoxins (Trx's) regulate redox signaling and are localized to various cellular compartments. Specific redox-regulated pathways for adaptation of skeletal muscle to contractions are attenuated during aging, but little is known about the roles of Trx's in regulating these pathways. This study investigated the susceptibility of Trx1 and Trx2 in skeletal muscle to oxidation and reduction in vitro and the effects of aging and contractions on Trx1, Trx2, and thioredoxin reductase (TrxR) 1 and 2 contents and nuclear and cytosolic Trx1 and mitochondrial Trx2 redox potentials in vivo. The proportions of cytosolic and nuclear Trx1 and mitochondrial Trx2 in the oxidized or reduced forms were analyzed using redox Western blotting. In myotubes, the mean redox potentials were nuclear Trx1, −251mV; cytosolic Trx1, −242mV; mitochondrial Trx2, −346mV, data supporting the occurrence of differing redox potentials between cell compartments. Exogenous treatment of myoblasts and myotubes with hydrogen peroxide or dithiothreitol modified glutathione redox status and nuclear and cytosolic Trx1, but mitochondrial Trx2 was unchanged. Tibialis anterior muscles from young and old mice were exposed to isometric muscle contractions in vivo. Aging increased muscle contents of Trx1, Trx2, and TrxR2, but neither aging nor endogenous ROS generated during contractions modified Trx redox potentials, although oxidation of glutathione and other thiols occurred. We conclude that glutathione redox couples in skeletal muscle are more susceptible to oxidation than Trx and that Trx proteins are upregulated during aging, but do not appear to modulate redox-regulated adaptations to contractions that fail during aging

Effect of aging in functional redox state of single isolated skeletal muscle fibres

[EN] Skeletal muscle constantly produces reactive oxygen species (ROS). During contractile activity ROS are generated in skeletal muscle fibres. There is considerable support for an involvement of ROS in the process of aging. Several studies indicate that adaptive responses of skeletal muscle that are activated and regulated by ROS are disrupted during aging. The aim of this study was to monitor, in real time, intracellular ROS production in single skeletal muscle fibres from old and young mice and study the effect of contractile activity in these cells. Following evaluate and correlate the potential changes in intracellular ROS production with glutathione redox state and antioxidant enzymatic activities in muscle. Single skeletal muscle fibres were isolated from the Flexor Digitorus Brevis muscle from young (2-4 monthold) and old (26-28 month-old) C57BL/6 mice. Fibres were loaded with DCFH-DA, a fluorophore probe that allows the quantification of intracellular ROS generation by fluorescence microscopy imaging. Contractile activity was induced in fibres by electrical stimulation. Glutathione redox state and activity of antioxidant enzymes were analysed in gastrocnemious muscle. Intracellular basal level of ROS was higher in fibres from old mice. Contractile activity induced increase of ROS generation in fibres from young mice. However, this response was attenuated in fibres from old mice. Glutathione redox state was significant different, in favour of oxidized glutathione, in muscles from old mice. Glutathione peroxidase and catalase activities were significantly augmented in muscles from old mice. In conclusion, the process of aging modifies the basal redox status in skeletal muscle fibres in favour of oxidation and induces adaptation mechanisms of antioxidant defences. These are not able to neutralize the increase of basal oxidation, but they might lead to the attenuation of ROS produced by contractile activity observed in fibres from old mice

Redox proteomic analysis of the gastrocnemius muscle from adult and old mice.

The data provides information in support of the research article, "Differential Cysteine Labeling and Global Label-Free Proteomics Reveals an Altered Metabolic State in Skeletal Muscle Aging", Journal of Proteome Research, 2014, 13 (11), 2008-21 [1]. Raw data is available from ProteomeXchange [2] with identifier PDX001054. The proteome of gastrocnemius muscle from adult and old mice was analyzed by global label-free proteomics and the relative quantification of specific reduced and reversibly oxidized Cysteine (Cys) residues was performed using Skyline [3]. Briefly, reduced Cysteine (Cys) containing peptides was alkylated using N-ethylmalemide (d0-NEM). Samples were desalted and reversibly oxidized Cys residues were reduced using tris(2-carboxyethyl)phosphine (TCEP) and the newly formed reduced Cys residues were labeled with heavy NEM( d5-NEM). Label-free analysis of the global proteome of adult (n=5) and old (n=4) gastrocnemius muscles was performed using Peaks7™ mass spectrometry data analysis software [4]. Relative quantification of Cys containing peptides that were identified as reduced (d(0) NEM labeled) and reversibly oxidized d(5)-NEM labeled was performed using the intensity of their precursor ions in Skyline. Results indicate that muscles from old mice show reduced redox flexibility particularly in proteins involved in the generation of precursor metabolites and energy metabolism, indicating a loss in the flexibility of the redox energy response

Are there functional consequences of a reduction in selenium intake in UK subjects?

Dietary Se levels in the UK have fallen over the last 20 years and recent surveys indicate that average Se intakes are 30-40 microg/d, which is well below the current UK reference nutrient intake for adult men (75 microg/d) or women (60 microg/d). Functional consequences of this decline have not been recognised, although epidemiological data suggest it may contribute to increased risk of infections and incidence of some cancers. Previous data have indicated that biochemical changes in Se-dependent proteins occur in otherwise healthy UK subjects given small Se supplements. The current studies have focused on the effect of small Se supplements on the immune response since there is evidence of specific interactions between Se intake and viral replication, and since the potential anti-cancer effects of Se may be mediated by non-antioxidant effects of Se such as changes in immune function. Data indicate that subjects given small Se supplements (50 or 100 microg Se/d) have changes in the activity of Se-dependent enzymes and evidence of improved immune function and clearance of an administered live attenuated virus in the form of poliovirus vaccine. Responses of individual subjects to Se supplements are variable, and current work is evaluating potential explanations for this variability, including genetic variability and pre-existing Se status

Repeated bouts of aerobic exercise lead to reductions in skeletal muscle free radical generation and nuclear factor ΚB activation

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/65978/1/jphysiol.2008.155382.pd

Overexpression of HSP70 in mouse skeletal muscle protects against muscle damage and age‐related muscle dysfunction

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/154339/1/fsb2fj030395fje-sup-0001.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/154339/2/fsb2fj030395fje.pd

Reactive oxygen species and loss of muscle fibres during ageing

[ES] Sumario de la revista Neuromuscular Disorders: Especies reactivas de oxígeno y pérdida de fibras musculares durante el envejecimiento

Redox Regulation in Health and Disease Application of redox proteomics to skeletal muscle aging and exercise

Abstract Skeletal muscle represents a physiologically relevant model for the application of redox proteomic techniques to dissect its response to exercise and aging. Contracting skeletal muscles generate ROS (reactive oxygen species) and RNS (reactive nitrogen species) necessary for the regulation of many proteins involved in excitation-contraction coupling. The magnitude and species of ROS/RNS generated by contracting muscles will have downstream effects on specific protein targets and cellular redox signalling. Redox modifications on specific proteins are essential for the adaptive response to exercise and skeletal muscle can develop a dysregulated redox response during aging. In the present article, we discuss how redox proteomics can be applied to identify and quantify the reversible modifications on susceptible cysteine residues within those redox-sensitive proteins, and the integration of oxidative and non-oxidative protein modifications in relation to the functional proteome. Skeletal muscle as a redox model Skeletal muscle represents the largest organ of the human body and comprises approximately 40 % of total body mass in humans. Over the age of 50, there is a decline in skeletal muscle mass in adults. The decline in muscle mass and associated function is referred to as sarcopenia, and can lead to a reduction in the ability to perform daily tasks and loss of independence amongst the elderly Key words: aging, cysteine modification, exercise, redox proteomics, signalling, skeletal muscle. Abbreviations: eNOS, endothelial NOS; ICAT, isotope-coded affinity tag; iTRAQ, isobaric tag for relative and absolute quantification; MRM, multiple reaction monitoring; NOS, nitric oxide synthase; PTM, post-translational modification; RNS, reactive nitrogen species; ROS, reactive oxygen species. 1 To whom correspondence should be addressed (email [email protected]). A number of questions remain as to the source of the signals required for the generation of ROS and RNS, the identification of their specific protein targets (and modifications) and how the redox signals are relayed throughout the muscle fibre. The intensity of muscle contractions, muscle fibre type, and the fitness and the age of the individual may all have an effect on both the levels and type of ROS/RNS generated. Aging muscle has an altered redox response with subsequent physiological and biochemical effects on the cytoskeleton, mitochondria, Ca 2 + signalling and sequestration. Exercise is known to induce the generation of ROS and RNS that results in the activation of a number of transcription factors, including NF-κB (nuclear factor κB), AP-1 (activator protein 1) and HSF-1 (heat-shock factor 1), and induces mitochondrial biogenesis (for a review, see Defining the biochemical pathways and processes within skeletal muscles that are affected by ROS/RNS and how these responses change with age and exercise could help our understanding of biological aging. Moderate levels of ROS/RNS generally act through a reversible thiol-disulfideexchange mechanism on specific cysteine residues and can modify key target proteins by altering both the structur

Involvement of Reactive Oxygen Species (ROS) inskeletal muscle function during ageing: Study in amodel of isolated single skeletal muscle fibre

[ES] Suplemento de la revista Free Radical Biology and Medicine: Involvement of Reactive Oxygen Species (ROS) inskeletal muscle function during ageing: Study in amodel of isolated single skeletal muscle fibre