Yuji Ogura

Duration of static stretching influences muscle force production in hamstring muscles.

The purpose of the present study was to investigate whether duration of static stretching could affect the maximal voluntary contraction (MVC).Volunteer male subjects (n = 10) underwent 2 different durations of static stretching of their hamstring muscles in the dominant leg: 30 and 60 seconds. No static stretching condition was used as a control condition. Before and after each stretching trial, hamstring flexibility was measured by a sit and reach test. MVC was then measured using the maximal effort of knee flexion. The hamstring flexibility was significantly increased by 30 and 60 seconds of static stretching (control: 0.5 ± 1.1 cm; 30 seconds: 2.1 ± 1.8 cm; 60 seconds: 3.0 ± 1.6 cm); however, there was no significant difference between 30 and 60 seconds of static stretching conditions. The MVC was significantly lowered with 60 seconds of static stretching compared to the control and 30 seconds of the stretching conditions (control: 287.6 ± 24.0 N; 30 seconds: 281.8 ± 24.2 N; 60 seconds: 262.4 ± 36.2 N). However, there was no significant difference between control and 30 seconds of static stretching conditions. Therefore, it was concluded that the short duration (30 seconds) of static stretching did not have a negative effect on the muscle force production.

Wasting mechanisms in muscular dystrophy.

Muscular dystrophy is a group of more than 30 different clinical genetic disorders that are characterized by progressive skeletal muscle wasting and degeneration. Primary deficiency of specific extracellular matrix, sarcoplasmic, cytoskeletal, or nuclear membrane protein results in several secondary changes such as sarcolemmal instability, calcium influx, fiber necrosis, oxidative stress, inflammatory response, breakdown of extracellular matrix, and eventually fibrosis which leads to loss of ambulance and cardiac and respiratory failure. A number of molecular processes have now been identified which hasten disease progression in human patients and animal models of muscular dystrophy. Accumulating evidence further suggests that aberrant activation of several signaling pathways aggravate pathological cascades in dystrophic muscle. Although replacement of defective gene with wild-type is paramount to cure, management of secondary pathological changes has enormous potential to improving the quality of life and extending lifespan of muscular dystrophy patients. In this article, we have reviewed major cellular and molecular mechanisms leading to muscle wasting in muscular dystrophy. This article is part of a Directed Issue entitled: Molecular basis of muscle wasting.

Regulatory circuitry of TWEAK-Fn14 system and PGC-1α in skeletal muscle atrophy program

Skeletal muscle wasting attributed to inactivity has significant adverse functional consequences. Accumulating evidence suggests that peroxisome proliferator‐activated receptor γ coactivator 1α (PGC‐1a) and TNF‐like weak inducer of apoptosis (TWEAK)‐Fn14 system are key regulators of skeletal muscle mass in various catabolic states. While the activation of TWEAK‐Fn14 signaling causes muscle wasting, PGC‐1α preserves muscle mass in several conditions, including functional denervation and aging. However, it remains unknown whether there is any regulatory interaction between PGC‐1α and TWEAK‐Fn14 system during muscle atrophy. Here we demonstrate that TWEAK significantly reduces the levels of PGC‐1α and mitochondrial content (~50%) in skeletal muscle. Levels of PGC‐1α are significantly increased in skeletal muscle of TWEAK‐knockout (KO) and Fn14‐KO mice compared to wild‐type mice on denervation. Transgenic (Tg) overexpression of PGC‐1α inhibited progressive muscle wasting in TWEAK‐Tg mice. PGC‐1α inhibited the TWEAK‐induced activation of NF‐κB(~50%) and dramatically reduced (‐90%) the expression of atrogenes such as MAFbx and MuRF1. Intriguingly, muscle‐specific overexpression of PGC‐1α also prevented the inducible expression of Fn14 in denervated skeletal muscle. Collectively, our study demonstrates that TWEAK induces muscle atrophy through repressing the levels of PGC‐1a. Overexpression of PGC‐1α not only blocks the TWEAK‐induced atrophy program but also diminishes the expression of Fn14 in denervated skeletal muscle.—Hindi, S. M., Mishra, V., Bhatnagar, S., Tajrishi, M. M., Ogura, Y., Yan, Z., Burkly, L. C., Zheng, T. S., Kumar, A. Regulatory circuitry of TWEAK‐Fn14 system and PGC‐1α in skeletal muscle atrophy program. FASEB J. 28, 1398–1411 (2014). www.fasebj.org

Single bout of running exercise changes LC3-II expression in rat cardiac muscle.

Macroautophagy (autophagy) is an intracellular catalytic process. We examined the effect of running exercise, which stimulates cardiac work physiologically, on the expression of microtubule-associated protein 1 light chain 3 (LC3)-II, an indicator of autophagy, as well as some autophagy-related proteins in rat cardiac muscle. The left ventricles were taken from rats immediately (0 h), and at 0.5h, 1h or 3h after a single bout of running exercise on a treadmill for 30 min and also from rats in a rest condition. In these samples, we evaluated the level of LC3-II and p62, and the phosphorylation level of mammalian target of rapamycin (mTOR), Akt and AMP-activated protein kinase alpha (AMPKα) by Western blotting. The exercise produced a biphasic change in LC3-II, with an initial decrease observed immediately after the exercise and a subsequent increase 1h thereafter. LC3-II then returned to the rest level at 3h after the exercise. A negative correlation was found between the LC3-II expression and mTOR phosphorylation, which plays a role in inhibiting autophagy. The exercise increased phosphorylation of AMPKα, which stimulates autophagy via suppression of mTOR phosphorylation, immediately after exercise. The level of p62 and phosphorylated Akt was not altered significantly by the exercise. These results suggest for the first time that a single bout of running exercise induces a biphasic change in autophagy in the cardiac muscle. The exercise-induced change in autophagy might be partially mediated by mTOR in the cardiac muscle.

Satellite cell pool enhancement in rat plantaris muscle by endurance training depends on intensity rather than duration

Aim:  Increases in the number of satellite cells are necessary for the maintenance of normal muscle function. Endurance training enhances the satellite cell pool. However, it remains unclear whether exercise intensity or exercise duration is more important to enhance the satellite cell pool. This study examined the effects of different intensity and duration of endurance training on the satellite cell pool in rat skeletal muscle.

Matrix Metalloproteinase-9 Inhibition Improves Proliferation and Engraftment of Myogenic Cells in Dystrophic Muscle of mdx Mice

Duchenne muscular dystrophy (DMD) caused by loss of cytoskeletal protein dystrophin is a devastating disorder of skeletal muscle. Primary deficiency of dystrophin leads to several secondary pathological changes including fiber degeneration and regeneration, extracellular matrix breakdown, inflammation, and fibrosis. Matrix metalloproteinases (MMPs) are a group of extracellular proteases that are involved in tissue remodeling, inflammation, and development of interstitial fibrosis in many disease states. We have recently reported that the inhibition of MMP-9 improves myopathy and augments myofiber regeneration in mdx mice (a mouse model of DMD). However, the mechanisms by which MMP-9 regulates disease progression in mdx mice remain less understood. In this report, we demonstrate that the inhibition of MMP-9 augments the proliferation of satellite cells in dystrophic muscle. MMP-9 inhibition also causes significant reduction in percentage of M1 macrophages with concomitant increase in the proportion of promyogenic M2 macrophages in mdx mice. Moreover, inhibition of MMP-9 increases the expression of Notch ligands and receptors, and Notch target genes in skeletal muscle of mdx mice. Furthermore, our results show that while MMP-9 inhibition augments the expression of components of canonical Wnt signaling, it reduces the expression of genes whose products are involved in activation of non-canonical Wnt signaling in mdx mice. Finally, the inhibition of MMP-9 was found to dramatically improve the engraftment of transplanted myoblasts in skeletal muscle of mdx mice. Collectively, our study suggests that the inhibition of MMP-9 is a promising approach to stimulate myofiber regeneration and improving engraftment of muscle progenitor cells in dystrophic muscle.

Elevation of body temperature is an essential factor for exercise-increased extracellular heat shock protein 72 level in rat plasma

This study examined whether the exercise-increased extracellular heat shock protein 72 (eHsp72) levels in rats was associated with body temperature elevation during exercise. In all, 26 female Sprague-Dawley rats (3 mo old) were assigned randomly to control (CON; n = 8), exercise under warm temperature (WEx; n = 9), or exercise under cold temperature (CEx; n = 9). The WEx and CEx were trained at 25 degrees C or 4 degrees C, respectively, for nine days using a treadmill. Before and immediately after the final exercise bout, the colonic temperatures were measured as an index of body temperature. The animals were subsequently anesthetized, and blood samples were collected and centrifuged. Plasma samples were obtained to assess their eHsp72 levels. Only the colonic temperature in WEx was increased significantly (P < 0.05) by exercise. The eHsp72 level in WEx was significantly higher (P < 0.05) than that of either the CON or CEx. However, no significant difference was found between CON and CEx. Regression analyses revealed that the eHsp72 level increased as a function of the body temperature. In another experiment, the eHsp72 level of animals with body temperature that was passively elevated through similar kinetics to those of the exercise was studied. Results of this experiment showed that mere body temperature elevation was insufficient to induce eHsp72 responses. Collectively, our results suggest that body temperature elevation during exercise is important for induction of exercise-increased eHsp72. In addition, the possible role of body temperature elevation is displayed when the exercise stressor is combined with it.

Blood flow restriction by low compressive force prevents disuse muscular weakness

Repetitive blood flow restriction prevents muscular atrophy and weakness induced by chronic unloading. However, it was unclear which external compressive force for blood flow restriction was optimal to prevent muscular dysfunction. The present study was intended to investigate the effects of repeated muscle blood flow restriction at low pressure on muscular weakness induced by immobilization without weight bearing. Using casts, the left ankles of 11 healthy males were immobilized for 2 weeks. Subjects were instructed to walk using crutches with no weight bearing during the period. Subjects were divided randomly into two groups: a restriction of blood flow (RBF) group (application of external compressive force of 50 mm Hg) and a control (CON) group (no intervention). We measured changes in the muscle strength of the knee extensor-flexor and ankle plantar flexor. The percent changes in knee extensor torque at 60°/s under eccentric contraction in the RBF group were significantly smaller than in the CON group (-12.5±10.7% and -30.1±10.9%, p<0.05). The percent changes in knee flexor torque when performing an eccentric contraction at 60°/s, an isometric contraction, or a concentric contraction at both 60 and 300°/s in the RBF group were significantly smaller than those in the CON group (p<0.05). In conclusion, our results show that repetitive restriction of blood flow with 50 mm Hg cuff pressure to the lower extremity reduces muscular weakness induced by chronic unloading.

Heat stress activates the Akt/mTOR signalling pathway in rat skeletal muscle.

It is well known that various stimuli, such as mechanical stress and nutrients, induce muscle hypertrophy thorough the Akt/mTOR signalling pathway, which is a key mediator of protein synthesis and hypertrophy in skeletal muscle. It was recently reported that heat stress also induces an increase in muscle weight and muscle protein content. In addition, heat stress enhances Akt/mTOR signalling after one bout of resistance exercise. However, it remains unclear whether increased temperature itself stimulates the Akt/mTOR signalling pathway.

TWEAK/Fn14 Signaling Axis Mediates Skeletal Muscle Atrophy and Metabolic Dysfunction

Tumor necrosis factor-like weak inducer of apoptosis (TWEAK) through binding to its receptor fibroblast growth factor inducible 14 (Fn14) has been shown to regulate many cellular responses including proliferation, differentiation, apoptosis, inflammation, and fibrosis, under both physiological and pathological conditions. Emerging evidence suggests that TWEAK is also a major muscle wasting cytokine. TWEAK activates nuclear factor-κB signaling and proteolytic pathways such as ubiquitin–proteasome system, autophagy, and caspases to induce muscle proteolysis in cultured myotubes. Fn14 is dormant or expressed in minimal amounts in normal healthy muscle. However, specific atrophic conditions, such as denervation, immobilization, and starvation stimulate the expression of Fn14 leading to activation of TWEAK/Fn14 signaling and eventually skeletal muscle atrophy. TWEAK also causes slow- to fast-type fiber transition in skeletal muscle. Furthermore, recent studies suggest that TWEAK diminishes mitochondrial content and represses skeletal muscle oxidative phosphorylation capacity. TWEAK mediates these effects through affecting the expression of a number of genes and microRNAs. In this review article, we have discussed the recent advancements toward understanding the role and mechanisms of action of TWEAK/Fn14 signaling in skeletal muscle with particular reference to different models of atrophy and oxidative metabolism.