Tohru Takemasa

Oblique Alignment of Stress Fibers in Cells Reduces the Mechanical Stress in Cyclically Deforming Fields

The stress fiber (bundles of actin filaments) is one of the most prominent cytoskeletal components that contributes to the maintenance of cell architecture. It has generally been believed that upon cyclic stretching, both cells and their stress fibers become perpendicularly aligned to the direction of stretching. However, using our newly developed stretching device, we have recently found the contrary evidence that stress fibers in endothelial cells rapidly become rearranged at a specific oblique angle relative to the direction of stretching [Takemasa, T., K. Sugimoto, K. Yamashita: Exp. Cell Res. 230, 407-410 (1997)]. In light of this finding, we attempted to establish the explanation for such a phenomenon. First, we investigated the effects of possible modulators on the angle of the stress fibers; those were, modification of the stretching program, dependency of extracellular matrix types, and their reproducibility in other cell species. However, it seemed that the orientation was solely depending on the stretching amplitude applied. Next, we analyzed alterations in stress fiber length during loading tests using two kinds of deforming experiment systems. It was thus revealed that stress fibers aligned at a particular angle so as to minimize their length alterations in cyclic deforming fields. Rearrangement of the stress fibers at this angle probably occurs as a result of avoiding compressive stress and may be interpreted as a way of reducing the mechanical stress to which they are subjected during the deformation. This hypothesis well explains the reason not only for the survival of the stress fibers at a particular oblique angle, but also for the reduced numbers of stress fibers found at the other angles on cyclic deforming fields.

Muscle type-specific response of PGC-1α and oxidative enzymes during voluntary wheel running in mouse skeletal muscle

Aim:  It is generally accepted that endurance exercise increases the expression of peroxisome proliferator‐activated receptor γ coactivator‐1α (PGC‐1α), which governs the expression of oxidative metabolic enzymes. A previous report demonstrated that the regulation of mitochondrial protein expression in skeletal muscles in response to cold exposure depends on muscle fibre type. Cold exposure and endurance exercise are both metabolic challenges that require adjustments in mitochondrial energy metabolism, we hypothesized that the exercise‐induced increase in oxidative enzymes and PGC‐1α expression is higher in fast‐type than in slow‐type muscle.

Adaptation of macrophages to exercise training improves innate immunity.

The effects of 3-week exercise training on the functions of peritoneal macrophages from BALB/c mice were investigated. Lipopolysaccharide (LPS)-stimulated nitric oxide (NO) and proinflammatory cytokine production in macrophages from trained mice was markedly higher than those from control mice. Meanwhile, exercise training decreased the steady state level of beta(2)-adrenergic receptor (beta(2)AR) mRNA in macrophages. Overexpression of beta(2)AR in the macrophage cell line RAW264 by transfecting with beta(2)AR cDNA suppressed NO synthase (NOS) II expression but dose not influenced proinflammatory cytokine expression. When expression of transfected beta(2)AR in RAWar cells was downregulated by a tetracycline repressor-regulated mammalian expression system, NOS II mRNA expression was significantly increased; this suggested that the changes in the beta(2)AR expression level in macrophages associated with exercise training play a role in the regulation of NO production following LPS stimulation. These findings indicate that exercise training improves macrophage innate immune function in a beta(2)AR-dependent and -independent manner.

Reduction of ribosome biogenesis with activation of the mTOR pathway in denervated atrophic muscle.

Mammalian target of rapamycin (mTOR) pathway positively regulates the cell growth through ribosome biogenesis in many cell type. In general, myostatin is understood to repress skeletal muscle hypertrophy through inhibition of mTOR pathway and myogenesis. However, these relationships have not been clarified in skeletal muscle undergoing atrophy. Here, we observed a significant decrease of skeletal muscle mass at 2 weeks after denervation. Unexpectedly, however, mTOR pathway and the expression of genes related to myogenesis were markedly increased, and that of myostatin was decreased. However, de novo ribosomal RNA synthesis and the levels of ribosomal RNAs were dramatically decreased in denervated muscle. These results indicate that ribosome biogenesis is strongly controlled by factors other than the mTOR pathway in denervated atrophic muscle. Finally, we assessed rRNA transcription factors expression and observed that TAFIa was the only factor decreased. TAFIa might be a one of the limiting factor for rRNA synthesis in denervated muscle. J. Cell. Physiol. 227: 1569–1576, 2012.

Wnt Protein-mediated Satellite Cell Conversion in Adult and Aged Mice Following Voluntary Wheel Running

Background: Satellite cells are activated in response to muscle injury or mechanical stimuli. Results: Running-induced up-regulation of Wnt/β-catenin signaling activates Myf5 and MyoD transcription. Conclusion: Voluntary wheel running induces satellite cell activation by direct regulation of Wnt/β-catenin signaling. Significance: Focusing on Wnt signaling may help to understand the function and intrinsic ability of satellite cells in adult myogenesis. Muscle represents an abundant, accessible, and replenishable source of adult stem cells. Skeletal muscle-derived stem cells, called satellite cells, play essential roles in regeneration after muscle injury in adult skeletal muscle. Although the molecular mechanism of muscle regeneration process after an injury has been extensively investigated, the regulation of satellite cells under steady state during the adult stage, including the reaction to exercise stimuli, is relatively unknown. Here, we show that voluntary wheel running exercise, which is a low stress exercise, converts satellite cells to the activated state due to accelerated Wnt signaling. Our analysis showed that up-regulated canonical Wnt/β-catenin signaling directly modulated chromatin structures of both MyoD and Myf5 genes, resulting in increases in the mRNA expression of Myf5 and MyoD and the number of proliferative Pax7+Myf5+ and Pax7+ MyoD+ cells in skeletal muscle. The effect of Wnt signaling on the activation of satellite cells, rather than Wnt-mediated fibrosis, was observed in both adult and aged mice. The association of β-catenin, T-cell factor, and lymphoid enhancer transcription factors of multiple T-cell factor/lymphoid enhancer factor regulatory elements, conserved in mouse, rat, and human species, with the promoters of both the Myf5 and MyoD genes drives the de novo myogenesis in satellite cells even in aged muscle. These results indicate that exercise-stimulated extracellular Wnts play a critical role in the regulation of satellite cells in adult and aged skeletal muscle.

Tetrahymena 14-nm filament-forming protein has citrate synthase activity.

The Tetrahymena 14-nm filament-forming protein (49K protein) is a structural protein involved in oral morphogenesis and in pronuclear behavior during conjugation. Cloning the 49K protein gene from a Tetrahymena thermophila cDNA library, we found that its primary structure exhibits a high sequence identity (51.5%) with porcine heart citrate synthase and retains functional domains. The 49K protein actually possesses citrate synthase activity, and is detected in mitochondria. These results suggest that the 49K protein has dual functions as both a respiratory enzyme and a structural protein in the cytoskeleton.

Effect of training and detraining on monocarboxylate transporter (MCT) 1 and MCT4 in Thoroughbred horses

The aim of this study was to investigate the effects of training and detraining on the monocarboxylate transporter (MCT) 1 and MCT4 levels in the gluteus medius muscle of Thoroughbred horses. Twelve Thoroughbred horses were used for the analysis. For 18 weeks, all the horses underwent high‐intensity training (HIT), with running at 90–110% maximal oxygen consumption ( ) for 3 min, 5 days week−1. Thereafter, the horses either underwent detraining for 6 weeks by either 3 min of moderate‐intensity training (MIT) at 70% , 5 days week−1 (HIT‐MIT group) or stall rest (HIT‐SR group). The horses underwent an incremental exercise test, was measured and resting muscle samples were obtained from the middle gluteus muscle at 0, 18 and 24 weeks. The content of MCT1 and MCT4 proteins increased after 18 weeks of HIT. At the end of this period, an increase was noted in the citrate synthase activity, while phosphofructokinase activity remained unchanged. After 6 weeks of detraining, all these indexes returned to the pretraining levels in the HIT‐SR group. However, in the HIT‐MIT group, the increase in the MCT1 protein content and citrate synthase activity was maintained after 6 weeks of MIT, while the MCT4 protein content decreased to the pretraining value. These results suggest that the content of MCT1 and MCT4 proteins increases after HIT in Thoroughbred horses. In addition, the increase in the MCT1 protein content and oxidative capacity induced by HIT can be maintained by MIT of 70% , but the increase in the MCT4 protein content cannot be maintained by MIT.

Acute exercise induces biphasic increase in respiratory mRNA in skeletal muscle.

Peroxisome proliferator-activated receptor gamma coactivator-1alpha (PGC-1alpha) promotes the expression of oxidative enzymes in skeletal muscle. We hypothesized that activation of the p38 MAPK (mitogen-activated protein kinase) in response to exercise was associated with exercise-induced PGC-1alpha and respiratory enzymes expression and aimed to demonstrate this under the physiological level. We subjected mice to a single bout of treadmill running and found that the exercise induced a biphasic increase in the expression of respiratory enzymes mRNA. The second phase of the increase was accompanied by an increase in PGC-1alpha protein, but the other was not. Administration of SB203580 (SB), an inhibitor of p38 MAPK, suppressed the increase in PGC-1alpha expression and respiratory enzymes mRNA in both phases. These data suggest that p38 MAPK is associated with the exercise-induced expression of PGC-1alpha and biphasic increase in respiratory enzyme mRNAs in mouse skeletal muscle under physiological conditions.

Acute exercise increases expression of extracellular superoxide dismutase in skeletal muscle and the aorta.

Abstract Exercise dramatically increases oxygen consumption and causes oxidative stress. Superoxide dismutase (SOD) is important in the first-line defence mechanisms against oxidative stress. To investigate the effect of acute exercise on the expression of SOD, we examined the expression of mRNA for three SOD isozymes, in mice run on a treadmill to exhaustion. Six hours after exercise, the expression of extracellular SOD (EC-SOD) mRNA increased significantly in skeletal muscle and persisted for 24 h, whereas no change was observed for cytoplasmic and mitochondrial SOD mRNA. Moreover, acute exercise also induced EC-SOD mRNA in the aorta. These results suggest that a single bout of exercise is enough to augment the expression EC-SOD mRNA in skeletal muscle and the aorta, and may partly explain the beneficial effect of exercise.

The primary structure of Tetrahymena profilin

The cDNA of Tetrahymena profilin was cloned and sequenced. The deduced product has a molecular mass of 16,785 Da which is the largest among profilins known so far. Tetrahymena profilin shows higher homologies with lower eukaryotic profilins than with mammalian profilins. Although the homologies with mammalian and lower eukaryotic profilins are only 20-29% which is the lowest one among lower eukaryotic profilins, the N- and C-terminal regions of Tetrahymena profilin are considerably conserved as those in other profilins, suggesting that these regions are responsible for the essential properties common to profilins.