Skeletal muscle is a primary target of SOD1G93A-mediated toxicity

The antioxidant enzyme superoxide dismutase 1 (SOD1) is a critical player of the antioxidative defense whose activity is altered in several chronic diseases, including amyotrophic lateral sclerosis. However, how oxidative insult affects muscle homeostasis remains unclear. This study addresses the role of oxidative stress on muscle homeostasis and function by the generation of a transgenic mouse model expressing a mutant SOD1 gene (SOD1(G93A)) selectively in skeletal muscle. Transgenic mice developed progressive muscle atrophy, associated with a significant reduction in muscle strength, alterations in the contractile apparatus, and mitochondrial dysfunction. The analysis of molecular pathways associated with muscle atrophy revealed that accumulation of oxidative stress served as signaling molecules to initiate autophagy, one of the major intracellular degradation mechanisms. These data demonstrate that skeletal muscle is a primary target of SOD1(G93A) -mediated toxicity and disclose the molecular mechanism whereby oxidative stress triggers muscle atrophy

Training-induced ROS production and adaptations in rat skeletal muscles

Ten male 2-month-old rats were trained to run on a treadmill up to ~55% of their maximal oxygen uptake for 1 h/day, 3 days/week, 14 weeks. Ten age-matched sedentary rats were used as controls. Rats were sacrificed 24 h after the last training session in order to study the effect of such moderate aerobic training on two skeletal muscles differing in fiber type and metabolism: Soleus and Tibialis Anterior. The aim was to evaluate whether moderate endurance training i) induced fiber-type and metabolic adaptations; ii) caused oxidative stress; iii) upregulated anti-oxidant and cytoprotective molecules. A variety of analyses were carried out, ranging from morphometric and histochemical evaluations, RT-PCR analysis of relevant gene expression, western blotting, assessment of plasma membrane and sarcoplasmic reticulum calcium channels and ionic pump activities, quantification of ROS-induced modifications, enzymatic assays. Data showed that the effects of moderate training were similar to that described for intensive training as far as the metabolic and functional adaptations of fast-twitch muscles were concerned, but that also slow-twitch muscles were affected by training in somewhat similar ways. The formation of moderate amounts of ROS was documented even in this model of moderate exercise training. Moreover, ROS thus formed were shown to induce defensive responses but also to be responsible of - or to participate in - the induction of most adaptive training-related modification

Age-Dependent Effects on Functional Aspects in Human Satellite Cells

: In humans aging is a complex process that determines many physical and metabolic alterations correlated to the accumulation of oxidative damage in different tissues. Sarcopenia is an age-related nonpathological condition that includes a progressive loss of mass and strength in skeletal muscle, associated with a decline in the fibers' functional capability. This condition could be correlated to abnormal reactive oxygen species (ROS) accumulation with consequent fiber oxidative damage. This complex situation is not only evident in mature muscle fibers but also in muscle resident satellite cells (involved in fiber damage repairing) in which some functional parameters, at least for that concerns the Ca(2+) homeostasis, seem to be modified. In fact, our data show that there is an age-dependent increase of lipid peroxidation, in cultured myotubes (differentiated and fused satellite cells) after 7 days of in vitro differentiation. In these substrates also the capacity of these cells to produce Ca(2+) transient in response to various stimuli (ATP, caffeine, nicotine, KCl) is, sometimes, drastically modified. In particular, the presence of an age-dependent defective status of excitation-contraction (EC) coupling apparatus is supported by a single cell Ca(2+) analysis obtained from myotubes (derived from aged muscles) in the presence of 40 mM caffeine or 40 mM KCl. The alkaloid presence induces a complete emptying of ryanodine-dependent calcium stores indicating a probable integrity both of SR-terminal cisternae and/or the specific Ca(2+) channel known as RyR1. However, if a sarcolemmal depolarization is induced by the addition of 40 mM KCl in the experimental medium then Ca(2+) release RyR1-dependent can be observed only if Ca(2+) is present in the experimental solution. These results suggest that the EC uncoupling status could be due to the alteration of the interaction between RyR and DHPR. The two receptors are present and functionally active in myotubes from aged donors but they are probably still not in the right localization. These results suggest that during donor's life the satellite cells undergo an aging process similar to the one observed in skeletal muscle tissue, even if they are in a quiescence status for most of the time

Skeletal Muscle Is a Primary Target of SOD1(G93A)-Mediated Toxicity

The antioxidant enzyme superoxide dismutase 1 (SOD1) is a critical player of the antioxidative defense whose activity is altered in several chronic diseases, including amyotrophic lateral sclerosis. However, how oxidative insult affects muscle homeostasis remains unclear. This study addresses the role of oxidative stress on muscle homeostasis and function by the generation of a transgenic mouse model expressing a mutant SOD1 gene (SOD1(G93A)) selectively in skeletal muscle. Transgenic mice developed progressive muscle atrophy, associated with a significant reduction in muscle strength, alterations in the contractile apparatus, and mitochondrial dysfunction. The analysis of molecular pathways associated with muscle atrophy revealed that accumulation of oxidative stress served as signaling molecules to initiate autophagy, one of the major intracellular degradation mechanisms. These data demonstrate that skeletal muscle is a primary target of SOD1(G93A) -mediated toxicity and disclose the molecular mechanism whereby oxidative stress triggers muscle atrophy

Serum S100B Levels in Melanoma

Malignant melanoma is a cancer with increasing incidence worldwide with relevant socioeconomic impact. Despite progress in prevention and early detection, it is one of the most lethal forms of skin cancer. Therefore it is urgent need to identify suitable biomarkers in order to improve early diagnosis, precise staging, and prognosis, as well as for therapy selection and monitoring. In this book chapter, we are focusing on S100B and discuss its clinical relevance in melanoma

Impaired hypertrophy in myoblasts is improved with testosterone administration

We investigated the ability of testosterone (T) to restore differentiation in multiple population doubled (PD) murine myoblasts, previously shown to have a reduced differentiation in monolayer and bioengineered skeletal muscle cultures vs. their parental controls (CON) (Sharples et al., 2011, 2012 [7], [26]). Cells were exposed to low serum conditions in the presence or absence of T (100 nM) ± PI3K inhibitor (LY294002) for 72 h and 7 days (early and late muscle differentiation respectively). Morphological analyses were performed to determine myotube number, diameter (μm) and myonuclear accretion as indices of differentiation and myotube hypertrophy. Changes in gene expression for myogenin, mTOR and myostatin were also performed. Myotube diameter in CON and PD cells increased from 17.32 ± 2.56 μm to 21.02 ± 1.89 μm and 14.58 ± 2.66 μm to 18.29 ± 3.08 μm (P ≤ 0.05) respectively after 72 h of T exposure. The increase was comparable in both PD (+25%) and CON cells (+21%) suggesting a similar intrinsic ability to respond to exogenous T administration. T treatment also significantly increased myonuclear accretion (% of myotubes expressing 5+ nuclei) in both cell types after 7 days exposure (P ≤ 0.05). Addition of PI3K inhibitor (LY294002) in the presence of T attenuated these effects in myotube morphology (in both cell types) suggesting a role for the PI3K pathway in T stimulated hypertrophy. Finally, PD myoblasts showed reduced responsiveness to T stimulated mRNA expression of mTOR vs. CON cells and T also reduced myostatin expression in PD myoblasts only. The present study demonstrates testosterone administration improves hypertrophy in myoblasts that basally display impaired differentiation and hypertrophic capacity vs. their parental controls, the action of testosterone in this model was mediated by PI3K/Akt pathwa