Kyushu University Academic Staff Educational and Research Activities Database
List of Papers
Ryuichi Tatsumi Last modified date:2024.03.01

Professor / Animal & Mairne Bioresource Sciences / Department of Bioresource Sciences / Faculty of Agriculture


Papers
1. Yusuke Komiya, Shugo Iseki, Masaru Ochiai, Yume Takahashi, Issei Yokoyama1, Takahiro Suzuki, Ryuichi Tatsumi, Shoko Sawano, Wataru Mizunoya, Keizo Arihara., Dietary oleic acid intake increases the proportion of type 1 and 2X muscle fibers in mice, Scientific Reports , 10.1038/s41598-023-50464-y, 14, e755, open access, 2024.01, [URL], Skeletal muscle is one of the largest metabolic tissues in mammals and is composed of four different
types of muscle fibers (types 1, 2A, 2X, and 2B); however, type 2B is absent in humans. Given that
slow-twitch fibers are superior to fast-twitch fibers in terms of oxidative metabolism and are rich in
mitochondria, shift of muscle fiber types in direction towards slower fiber types improves metabolic
disorders and endurance capacity. We previously had reported that oleic acid supplementation
increases type 1 fiber formation in C2C12 myotubes; however, its function still remains unclear. This
study aimed to determine the effect of oleic acid on the muscle fiber types and endurance capacity.
An in vivo mouse model was used, and mice were fed a 10% oleic acid diet for 4 weeks. Two different
skeletal muscles, slow soleus muscle with the predominance of slow-twitch fibers and fast extensor
digitorum longus (EDL) muscle with the predominance of fast-twitch fibers, were used. We found
that dietary oleic acid intake improved running endurance and altered fiber type composition of
muscles, the proportion of type 1 and 2X fibers increased in the soleus muscle and type 2X increased
in the EDL muscle. The fiber type shift in the EDL muscle was accompanied by an increased muscle
TAG content. In addition, blood triacylglycerol (TAG) and non-esterified fatty acid levels decreased
during exercise. These changes suggested that lipid utilization as an energy substrate was enhanced
by oleic acid. Increased proliferator-activated receptor γ coactivator-1β protein levels were observed
in the EDL muscle, which potentially enhanced the fiber type transitions towards type 2X and
muscle TAG content. In conclusion, dietary oleic acid intake improved running endurance with the
changes of muscle fiber type shares in mice. This study elucidated a novel functionality of oleic acid in
skeletal muscle fiber types. Further studies are required to elucidate the underlying mechanisms. Our
findings have the potential to contribute to the field of health and sports science through nutritional
approaches, such as the development of supplements aimed at improving muscle function..
2. Alaa Elgaabari, Nana Imatomi, Hirochika Kido, Takashi Nakashima, Shoko Okuda, Yoshitaka Manabe, Shoko Sawan, Wataru Mizunoya, Ryuki Kaneko, Sakiho Tanaka, Takahiro Maeno, Yuji Matsuyoshi, Miyumi Seki, So Kuwakado, Kahona Zushi, Nasibeh Daneshvar, Mako Nakamura, Takahiro Suzuki, Kenji Sunagawa, Judy E. Anderson, Ronald E. Allen, Ryuichi Tatsumi., Age-related nitration/dysfunction of myogenic stem cell activator HGF, Aging Cell, 10.1111/acel.14041, e14041, open access, 2023.11, [URL], Mechanical perturbation triggers activation of resident myogenic stem cells to enter the cell cycle through a cascade of events including hepatocyte growth factor (HGF) release from its extracellular tethering and the subsequent presentation to signaling-receptor c-met. Here, we show that with aging, extracellular HGF undergoes tyrosine-residue (Y) nitration and loses c-met binding, thereby disturbing muscle homeostasis. Biochemical studies demonstrated that nitration/dysfunction is specific to HGF among other major growth factors and is characterized by its locations at Y198 and Y250 in c-met-binding domains. Direct-immunofluorescence microscopy of lower hind limb muscles from three age groups of rat, provided direct in vivo evidence for age-related increases in nitration of ECM-bound HGF, preferentially stained for anti-nitrated Y198 and Y250-HGF mAbs (raised in-house) in fast IIa and IIx myofibers. Overall, findings highlight inhibitory impacts of HGF nitration on myogenic stem cell dynamics, pioneering a cogent discussion for better understanding age-related muscle atrophy and impaired regeneration with fibrosis (including sarcopenia and frailty)..
3. Takahiro Maeno, Rio Arimatsu, Koichi Ojima, Yuki Yamaya, Hikaru Imakyure, Naruha Watanabe, Yusuke Komiya, Ken Kobayashi, Mako Nakamura, Takanori Nishimura, Ryuichi Tatsumi, Takahiro Suzuki, Netrin-4 synthesized in satellite cell-derived myoblasts stimulates autonomous fusion, Experimental Cell Research , 10.1016/j.yexcr.2023.113698, 430, 1, 113698, 2023.09, Satellite cells are indispensable for skeletal muscle regeneration and hypertrophy by forming nascent myofibers (myotubes). They synthesize multi-potent modulator netrins (secreted subtypes: netrin-1, -3, and -4), originally found as classical neural axon guidance molecules. While netrin-1 and -3 have key roles in myogenic differentiation, the physiological significance of netrin-4 is still unclear. This study examined whether netrin-4 regulates myofiber type commitment and myotube formation. Initially, the expression profiles indicated that satellite cells isolated from the extensor digitorum longus muscle (EDL muscle: fast-twitch myofiber-abundant) expressed slightly more netrin-4 than the soleus muscle (slow-type abundant) cells. As netrin-4 knockdown inhibited both slow- and fast-type myotube formation, netrin-4 may not directly regulate myofiber type commitment. However, netrin-4 knockdown in satellite cell-derived myoblasts reduced the myotube fusion index, while exogenous netrin-4 promoted myotube formation, even though netrin-4 expression level was maximum during the initiation stage of myogenic differentiation. Furthermore, netrin-4 knockdown also inhibited MyoD (a master transcriptional factor of myogenesis) and Myomixer (a myoblast fusogenic molecule) expression. These data suggest that satellite cells synthesize netrin-4 during myogenic differentiation initiation to promote their own fusion, stimulating the MyoD–Myomixer signaling axis..
4. Shoko Sawano, Misaki Fukushima, Taiki Akasaka, Mako Nakamura, Ryuichi Tatsumi, Yoshihide Ikeuchi, Wataru Mizunoya, Up- and down-egulated genes after long-term muscle atrophy induced by denervation in mice detected using RNA-Seq, Life , 10.3390/life13051111, 13, 5, e1111, open access, 2023.05, [URL], Skeletal muscle atrophy occurs rapidly as a result of inactivity. Although there are many reports on changes in gene expression during the early phase of muscle atrophy, the patterns of up-and downregulated gene expression after long-term and equilibrated muscle atrophy are poorly understood. In this study, we comprehensively examined the changes in gene expression in long-term denervated mouse muscles using RNA-Seq. The murine right sciatic nerve was denervated, and the mice were housed for five weeks. The cross-sectional areas of the hind limb muscles were measured using an X-ray CT system 35 days after denervation. After 28 d of denervation, the cross-sectional area of the muscle decreased to approximately 65% of that of the intact left muscle and reached a plateau. Gene expression in the soleus and extensor digitorum longus (EDL) muscles on the 36th day was analyzed using RNA-Seq and validated using RT-qPCR. RNA-Seq analysis revealed that three genes—Adora1, E230016M11Rik, and Gm10718—were upregulated and one gene—Gm20515—was downregulated in the soleus muscle; additionally, four genes—Adora1, E230016M11Rik, Pigh, and Gm15557—were upregulated and one gene—Fzd7—was downregulated in the EDL muscle (FDR
5. Ohsawa Y., Ohtsubo H., Munekane A., Ohkubo K., Murakami T., Fujino M., Nishimatsu S.-I., Hagiwara H., Nishimura H., Suzuki T., Tatsumi R., Mizunoya W., Hinohara A., Fukunaga M., Sunada Y., Circulating α-Klotho counteracts transforming growth factor-β-induced sarcopenia, American Journal of Pathology, doi.org/10.1016/j.ajpath.2023.01.009, 193, 5, 591-607, 2023.02.
6. Elgaabari A., Imatomi N., Kido H., Seki M., Tanaka S., Matsuyoshi M., Nakashima T., Sawano S., Mizunoya W., Suzuki T., Nakamura M., Anderson J. E., Tatsumi R., A pilot study on nitration/dysfunction of NK1 segment of myogenic stem cell activator HGF, Biochemistry and Biophysics Reports, 10.1016/j.bbrep.2022.101295, 31, 101295, open access, 2022.06, [URL], Protein tyrosine residue (Y) nitration, a post-translational chemical-modification mode, has been associated with changes in protein activity and function; hence the accumulation of specific nitrated proteins in tissues may be used to monitor the onset and progression of pathological disorders. To verify the possible impact of nitration on postnatal muscle growth and regeneration, a pilot study was designed to examine the nitration/dysfunction of hepatocyte growth factor (HGF), a key ligand that is released from the extracellular tethering and activates myogenic stem satellite cells to enter the cell cycle upon muscle stretch and injury. Exposure of recombinant HGF (a hetero-dimer of α- and β-chains) to peroxynitrite induces Y nitration in HGF α-chain under physiological conditions. Physiological significance of this finding was emphasized by Western blotting that showed the NK1 segment of HGF (including a K1 domain critical for signaling-receptor c-met binding) undergoes nitration with a primary target of Y198. Peroxynitrite treatment abolished HGF-agonistic activity of the NK1 segment, as revealed by in vitro c-met binding and bromodeoxyuridine-incorporation assays. Importantly, direct-immunofluorescence microscopy of rat lower hind-limb muscles from two aged-groups (2-month-old “young” and 12-month-old “retired/adult”) provided in vivo evidence for age-related nitration of extracellular HGF (Y198). Overall, findings provide the insight that HGF/NK1 nitration/dysfunction perturbs myogenic stem cell dynamics and homeostasis; hence NK1 nitration may stimulate progression of muscular disorders and diseases including sarcopenia..
7. Kosuke Tokunaga, Shota Akimoto, Machiko Aiba, Mutsuki Nakagomi, Takahiro Suzuki, Ryuichi Tatsumi, Mako Nakamura, Connexin 43 is localized in gizzard smooth muscle cells during chicken development., The Journal of Poultry Science, doi.org/10.2141/jpsa.0220003, 59, 4, 338-347, published on-line: 25 March 2022, 2022.04.
8. Ryo EBIHARA, Sushil BHANDARI, Seong–Kyu CHOE, Mulan QAHAR, Wataru MIZUNOYA, Ryuichi TATSUMI and Mako NAKAMURA, NUDT7 can promote slow myofiber formation and its Endogenous expression, Journal of the Faculty of Agriculture Kyushu University, doi.org/10.5109/4772335, 67, 1, 1-7, 2022.03, [URL].
9. Takahiro Suzuki, Aika Mori, Takahiro Maeno, Rio Arimatsu, Emi Ichimura, Yuriko Nishi, Kouga Hisaeda, Yuki Yamaya, Ken Kobayashi, Mako Nakamura, Ryuichi Tatsumi, Koichi Ojima and Takanori Nishimura, Abundant Synthesis of Netrin-1 in Satellite Cell-Derived Myoblasts Isolated from EDL Rather Than Soleus Muscle Regulates Fast-Type Myotube Formation, International Journal of Molecular Sciences, 22, e4499-e4499, 2021.04, Resident myogenic stem cells (satellite cells) are attracting attention for their novel roles in
myofiber type regulation. In the myogenic differentiation phase, satellite cells from soleus muscle
(slow fiber-abundant) synthesize and secrete higher levels of semaphorin 3A (Sema3A, a multifunctional
modulator) than those derived from extensor digitorum longus (EDL; fast fiber-abundant),
suggesting the role of Sema3A in forming slow-twitch myofibers. However, the regulatory mechanisms
underlying fast-twitch myotube commitment remain unclear. Herein, we focused on netrin
family members (netrin-1, -3, and -4) that compete with Sema3A in neurogenesis and osteogenesis.
We examined whether netrins affect fast-twitch myotube generation by evaluating their expression in
primary satellite cell cultures. Initially, netrins are upregulated during myogenic differentiation. Next,
we compared the expression levels of netrins and their cell membrane receptors between soleus- and
EDL-derived satellite cells; only netrin-1 showed higher expression in EDL-derived satellite cells than
in soleus-derived satellite cells. We also performed netrin-1 knockdown experiments and additional
experiments with recombinant netrin-1 in differentiated satellite cell-derived myoblasts. Netrin-1
knockdown in myoblasts substantially reduced fast-type myosin heavy chain (MyHC) expression;
exogenous netrin-1 upregulated fast-type MyHC in satellite cells. Thus, netrin-1 synthesized in
EDL-derived satellite cells may promote myofiber type commitment of fast muscles..
10. Alaa Elgaabari, Atsuko Miyawaki-Kuwakado, Kosuke Tomimatsu, Qianmei Wu, Kosuke Tokunaga, Wakana Izumi, Takahiro Suzuki, Ryuichi Tatsumi, Mako Nakamura, Epigenetic effects induced by the ectopic expression of Pax7 in 3T3-L1., Journal of Biochemistry, doi.org/10.1093/jb/mvab030, published on-line: 17 March 2021, 2021.03, Although skeletal muscle cells and adipocytes are derived from the same mesoderm, they do not transdifferentiate in vivo and are strictly distinct at the level of gene expression. To elucidate some of the regulatory mechanisms underlying this strict distinction, Pax7, a myogenic factor, was ectopically expressed in 3T3-L1 adipose progenitor cells to perturb their adipocyte differentiation potential. Transcriptome analysis showed that ectopic expression of Pax7 repressed the expression of some adipocyte genes and induced expression of some skeletal muscle cell genes. We next profiled the epigenomic state altered by Pax7 expression using H3K27ac, an activating histone mark, and H3K27me3, a repressive histone mark, as indicators. Our results show that ectopic expression of Pax7 did not result in the formation of H3K27ac at loci of skeletal muscle-related genes, but instead resulted in the formation of H3K27me3 at adipocyte-related gene loci. These findings suggest that the primary function of ectopic Pax7 expression is the formation of H3K27me3, and muscle gene expression results from secondary regulation..
11. Daneshvar N., Tatsumi R., Peeler J., Anderson J. E., Premature satellite cell activation before injury accelerates myogenesis and disrupts neuromuscular junction maturation in regenerating muscle, American Journal of Physiology - Cell Physiology, 10.1152/ajpcell.00121.2020, 318, 1, C116-C128, 2020.05, Satellite cell (SC) activation, mediated by nitric oxide (NO), is essential to myogenic repair, whereas myotube function requires innervation. Semaphorin (Sema) 3A, a neuro-chemorepellent, is thought to regulate axon guidance to neuromuscular junctions (NMJs) during myotube differentiation. We tested whether “premature” SC activation (SC activation before injury) by a NO donor (isosorbide dinitrate) would disrupt early myogenesis and/or NMJs. Adult muscle was examined during regeneration in two models of injury: myotoxic cardiotoxin (CTX) and traumatic crush (CR) (n = 4 -5/group). Premature SC activation was confirmed by increased DNA synthesis by SCs immediately in pretreated mice after CTX injury. Myotubes grew faster after CTX than after CR; growth was accelerated by pretreatment. NMJ maturation, classified by silver histochemistry (neurites) and acetylcholinesterase (AchE), and α-bungarotoxin staining (Ach receptors, AchRs) were delayed by pretreatment, consistent with a day 6 rise in the denervation marker γ-AchR. With pretreatment, S100B from terminal Schwann cells (TSCs) increased 10- to 20-fold at days 0 and 10 after CTX and doubled 6 days after CR. Premature SC activation disrupted motoneuritogenesis 8 -10 days post-CTX, as pretreatment reduced colocalization of pre- and postsynaptic NMJ features and increased Sema3A-65. Premature SC activation before injury both accelerated myogenic repair and disrupted NMJ remodeling and maturation, possibly by reducing Sema3A neuro-repulsion and altering S100B. This interpretation extends the model of Sema3A-mediated motoneuritogenesis during muscle regeneration. Manipulating the timing and type of Sema3A by brief NO effects on SCs suggests an important role for TSCs and Sema3A-65 processing in axon guidance and NMJ restoration during muscle repair..
12. Han G., Ouchi T., Hirata T., Haraguchi S., Miyazaki T., Arakawa T., Masuhara N., Mizunoya W., Tatsumi R., Tashiro k., Bungo T., Furuse N., Chowdhury V. S., Effects of L-leucine in ovo feeding on thermotolerance, growth and amino acid metabolism under heat stress in broilers., Animal, 10.1017/S1751731120000464, 2020.04.
13. Sawano S., Baba K., Sonoda Y., Wakamatsu J.-I., Tomonaga S., Furuse M., Sato Y., Tatsumi R., Ikeuchi Y., Mizunoya W., Beef extract supplementation promotes myoblast proliferation and myotube growth in C2C12 cells., European Journal of Nutrition, 10.1007/s00394-020-02205-4, 2020.02.
14. Sawano S., Oza K., Murakami T., Nakamura N., Tatsumi R., Mizunoya, W., Effect of gender, rearing, and cooking on the metabolomic profile of porcine muscles., Metabolites, 10.3390/metabo10010010, 10, 1, e10-e10, 2019.12, [URL].
15. Daisuke Mashima, Yoshiaki Oka, Takafumi Gotoh, Shozo Tomonaga, Shoko Sawano, Mako Nakamura, Ryuichi Tatsumi, Wataru Mizunoya, Correlation between skeletal muscle fiber type and free amino acid levels in Japanese Black steers, Animal Science Journal, 10.1111/asj.13185, 90, 4, 604-609, 2019.04, Free amino acids are important components of tastants and flavor precursors in meat. To clarify the correlation between muscle fiber type and free amino acids, we determined the concentrations of various free amino acids and dipeptides in samples of different muscle tissues (n = 21), collected from 26-month-old Japanese Black steers (n = 3) at 2 days postmortem. The proportions of the myosin heavy chain (MyHC), slow (MyHC1) and fast (MyHC2) isoforms were determined by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE). The contents of free amino acids and dipeptides were measured by high performance liquid chromatography (HPLC). The MyHC isoform composition varied among the tissue samples. The MyHC1 proportion ranged from 6.9% ± 3.9% to 83.3% ± 16.7%. We confirmed that there was a strong positive correlation between MyHC1 composition and total free amino acid concentrations, including those for two dipeptides. Among the 31 measured free amino acids and dipeptides, 11 showed significant positive correlations and five showed significant negative correlations with MyHC1 composition. These results suggest that a high MyHC1 content induces high free amino acid contents in bovine muscles possibly because of greater oxidative metabolism. This high level of free amino acids could contribute to the intense flavor of meat that is rich in slow-twitch fibers..
16. Yusuke Komiya, Toshiya Nakamura, Momoko Ishii, Kuniyoshi Shimizu, Eri Hiraki, Fuminori Kawabata, Mako Nakamura, Ryuichi Tatsumi, Yoshihide Ikeuchi, Wataru Mizunoya, Increase in muscle endurance in mice by dietary Yamabushitake mushroom (Hericium erinaceus) possibly via activation of PPARδ, Animal Science Journal, 10.1111/asj.13199, 2019.01, Skeletal muscle fiber is largely classified into two types: type 1 (slow-twitch) and type 2 (fast-twitch) fibers. Meat quality and composition of fiber types are thought to be closely related. Previous research showed that overexpression of constitutively active peroxisome proliferator-activated receptor (PPAR)δ, a nuclear receptor present in skeletal muscle, increased type 1 fibers in mice. In this study, we found that hexane extracts of Yamabushitake mushroom (Hericium erinaceus) showed PPARδ agonistic activity in vitro. Eight-week-old C57BL/6J mice were fed a diet supplemented with 5% (w/w) freeze-dried Yamabushitake mushroom for 24 hr. After the treatment period, the extensor digitorum longus (EDL) muscles were excised. The Yamabushitake-supplemented diet up-regulated the PPARδ target genes Pdk4 and Ucp3 in mouse skeletal muscles in vivo. Furthermore, feeding the Yamabushitake-supplemented diet to mice for 8 weeks resulted in a significant increase in muscle endurance. These results indicate that Yamabushitake mushroom contains PPARδ agonistic ligands and that dietary intake of Yamabushitake mushroom could activate PPARδ in skeletal muscle of mice. Unexpectedly, we observed no significant alterations in composition of muscle fiber types between the mice fed control and Yamabushitake-supplemented diets..
17. Yuji Matsuyoshi, Mariko Akahoshi, Mako Nakamura, Ryuichi Tatsumi, Wataru Mizunoya, Isolation and purification of satellite cells from young rats by percoll density gradient centrifugation, Methods in Molecular Biology, 10.1007/978-1-4939-8897-6_6, 81-93, 2019.01, Satellite cells (SCs) are myogenic stem cells that play an important role in skeletal muscle regeneration and hypertrophy. Primary cultures of SCs are useful to analyze cell functions; however, it is difficult to obtain highly pure SCs from young rats with the conventional procedures. The purpose of this study is to establish a purification method for SC isolation from young rats and quantitatively evaluate the purification procedure employing Percoll, a common research tool to purify cells. We elucidated the purity of SCs collected by Percoll density gradient centrifugation using real-time RT-qPCR and immunocytochemistry for desmin. Percoll treatment increased the purity of SCs isolated from young rats to nearly 90%, which was comparable to that achieved with the conventional method using middle-aged rats..
18. Yuki Yoshida, Arata Tsutaki, Yuki Tamura, Karina Kouzaki, Koichi Sashihara, Shohei Nakashima, Motoyuki Tagashira, Ryuichi Tatsumi, Koichi Nakazato, Dietary apple polyphenols increase skeletal muscle capillaries in Wistar rats, Physiological Reports, 10.14814/phy2.13866, 6, 18, 2018.09, Dietary apple polyphenols (AP) have been shown to exhibit beneficial effects on muscle endurance. Fast-to-slow change in the composition of myosin heavy chains was known as one of the molecular mechanisms. Here, we examined the effects of dietary AP on the capillaries and mitochondria in the rat skeletal muscle to elucidate the mechanisms underlying muscular endurance enhancement. Twenty-four Wistar male rats were divided into three groups, namely, the control group, 0.5% AP group, and 5% AP group (n = 8 in each group). After a feeding period of 4 weeks, rats were dissected, gastrocnemius muscles were removed, and the density of capillaries and levels of mitochondrial proteins were analyzed. Capillary density of the gastrocnemius increased to 17.8% in rats fed with 5% AP as compared to the control rats. No significant change was observed in the mitochondrial content and dynamics (fusion/fission) of regulatory proteins. To investigate the mechanisms underlying the increase in the capillary density, positive (vascular endothelial cell growth factor, VEGF) and negative (thrombosponsin-1, TSP-1) factors of angiogenesis were analyzed. TSP-1 expression significantly decreased in rats fed with 0.5% AP and 5% AP by approximately 25% and 40%, respectively, as compared with the control rats. There were no significant differences in VEGF expression. Thus, dietary AP may increase the muscle capillary density by decreasing TSP-1 expression. We concluded that the increase in the capillary density and the fast-to-slow change in myosin heavy chains by AP feeding are the main causes for muscle endurance enhancement in Wistar rats..
19. Wakana Izumi, Yuko Takuma, Ryo Ebihara, Wataru Mizunoya, Ryuichi Tatsumi, Mako Nakamura, Paired box 7 inhibits differentiation in 3T3-L1 preadipocytes, Animal Science Journal, 10.1111/asj.13050, 89, 8, 1214-1219, 2018.08, Myogenesis is precisely proceeded by myogenic regulatory factors. Myogenic stem cells are activated, proliferated and fused into a multinuclear myofiber. Pax7, paired box 7, one of the earliest markers during myogenesis. It has been reported that Pax7 regulates the muscle marker genes, Myf5 and MyoD toward differentiation. The possible roles of Pax7 in myogenic cells have been well researched. However, it has not yet been clarified if Pax7 itself is able to induce myogenic fate in nonmyogenic lineage cells. In this study, we performed experiments using stably expressed Pax7 in 3T3-L1 preadipocytes to elucidate if Pax7 inhibits adipogenesis. We found that Pax7 represses adipogenic markers and prevents differentiation. These cells showed decreased expression of PDGFRα, PPARγ and Fabp4 and inhibited forming lipid droplets..
20. Sato Y., Ohtsubo H., Nihei N., Kaneko T., Sato Y., Adachi S.-I., Kondo S., Nakamura M., Mizunoya W., Iida H., Tatsumi R., Rada C., Yoshizawa F., Apobec2 deficiency causes mitochondrial defects and mitophagy in skeletal muscle, FASEB JOURNAL, 10.1096/fj.201700493R, 32, 3, 1428-1439, 2018.03.
21. Ohtsubo H., Sato Y., Matsuyoshi Y., Suzuki T., Mizunoya W., Nakamura M., Tatsumi R., Ikeuchi, Y., Fluorescence microscopy data on expression of Paired Box Transcription Factor 7 in skeletal muscle of APOBEC2 knockout mice., Data in Brief, doi.org/10.1016/j.dib.2018.02.063, 17, 1348-1351, 2018.02.
22. Ebihara R., Choe S.-K., Mizunoya W., Tatsumi R., Nakamura M., Possible implication of Nudt7 in C2C12 myoblast differentiation., International Symposium on Agricultural, Food, Environmental and Life Sciences in Asia, 2017 (AFELiSA'17), 2017.11.
23. Judy E. Anderson, Mai-Khoi Q. Do, N. Daneshvar, Takahiro SUZUKI, J. Dort, Wataru Mizunoya, Ryuichi Tatsumi, The role of semaphorin 3A in myogenic regeneration and the formation of functional neuromuscular junctions on new fibers, Biological Reviews, 10.1111/brv.12286, 10.1111/brv.12286, 2017.08.
24. Tatsumi, Ryuichi; Suzuki, Takahiro; Do, Mai-Khoi Q.; Ohya, Yuki; Anderson, Judy E.; Shibata, Ayumi; Kawaguchi, Mai; Ohya, Shunpei; Ohtsubo, Hideaki; Mizunoya, Wataru; Sawano, Shoko; Komiya, Yusuke; Ichitsubo, Riho; Ojima, Koichi; Nishimatsu, Shin-ichiro; Nohno, Tsutomu; Ohsawa, Yutaka; Sunada, Yoshihide; Nakamura, Mako; Furuse, Mitsuhiro; Ikeuchi, Yoshihide; Nishimura, Takanori; Yagi, Takeshi; Allen, Ronald E., Slow-Myofiber Commitment by Semaphorin 3A Secreted from Myogenic Stem Cells, STEM CELLS, 10.1002/stem.2639, 35, 7, 1815-1834, 2017.07, Recently, we found that resident myogenic stem satellite cells up-regulate a multi-functional secreted protein, semaphorin 3A (Sema3A), exclusively at the early- differentiation phase in response to muscle injury; however, its physiological significance is still unknown. Here we show that Sema3A impacts slow-twitch fiber generation through a signaling pathway, cell-membrane receptor (neuropilin2-plexinA3) → myogenin-myocyte enhancer factor 2D (MEF2D) → slow myosin heavy chain. This novel axis was found by small interfering RNA (siRNA)-transfection experiments in myoblast cultures, which also revealed an additional element that Sema3A-neuropilin1/plexinA1, A2 may enhance slow-fiber formation by activating signals that inhibit fast-myosin expression. Importantly, satellite cell-specific Sema3A conditional-knockout adult mice (Pax7CreERT2-Sema3Aflox activated by tamoxifen-i.p. injection) provided direct in vivo evidence for the Sema3A-driven program, by showing that slow-fiber generation and muscle endurance were diminished after repair from cardiotoxin-injury of gastrocnemius muscle. Overall, the findings highlight an active role for satellite cell-secreted Sema3A ligand as a key “commitment factor” for the slow-fiber population during muscle regeneration. Results extend our understanding of the myogenic stem-cell strategy that regulates fiber-type differentiation and is responsible for skeletal muscle contractility, energy metabolism, fatigue resistance, and its susceptibility to aging and disease..
25. Wataru Mizunoya, Shinpei Okamoto, Hideo Miyahara, Mariko Akahoshi, Takahiro SUZUKI, Mai-Khoi Q. Do, Hideaki Ohtsubo, Yusuke Komiya, Mulan Qahar, Toshiaki Waga, Koichi Nakazato, Yoshihide Ikeuchi, Judy E. Anderson, Ryuichi Tatsumi, Fast-to-slow shift of muscle fiber-type composition by dietary apple polyphenols in rats: Impact of the low-dose supplementation, Animal Science Journal, 10.1111/asj.12655, 88, 3, 489-499, 10.1111/asj.12655, 2017.04, Our previous studies demonstrated that an 8-week intake of 5% (w/w) apple polyphenol (APP) in the diet improves muscle endurance of young-adult rats. In order to identify a lower limit of the dietary contribution of APP to the effect, the experiments were designed for lower-dose supplementation (8-week feeding of 0.5% APP in AIN-93G diet) to 12-week-old male Sprague-Dawley rats. Results clearly showed that the 0.5% APP diet significantly up-regulates slower myosin-heavy-chain (MyHC) isoform ratios (IIx and IIa relative to total MyHC) and myoglobin expression in lower hind-limb muscles examined (P
26. Hideaki Ohtsubo, Yusuke Sato, Takahiro SUZUKI, Wataru Mizunoya, Mako Nakamura, Ryuichi Tatsumi, Yoshihide Ikeuchi, Data supporting possible implication of APOBEC2 in self-renewal functions of myogenic stem satellite cells: toward understanding the negative regulation of myoblast differentiation, Data in Brief, 10.1016/j.dib.2017.03.051, 10.1016/j.dib.2017.03.051, 2017.04, This article provides in vitro phenotypical data to show that APOBEC2, a member of apoB mRNA editing enzyme, catalytic polypeptide-like family, may implicate in self-renewal functions of myogenic stem satellite cells, namely in the re-establishment of quiescent status after activation and proliferation of myoblasts in single-myofiber culture..
27. Hideaki Ohtsubo, Yusuke Sato, Takahiro SUZUKI, Wataru Mizunoya, Mako Nakamura, Ryuichi Tatsumi, Yoshihide Ikeuchi, APOBEC2 negatively regulates myoblast differentiation in muscle regeneration, International Journal of Biochemistry and Cell Biology , 10.1016/j.biocel.2017.02.005, 85, 91-101, 10.1016/j.biocel.2017.02.005, 2017.03, Recently we found that the deficiency of APOBEC2, a member of apoB mRNA editing enzyme, catalytic polypeptide-like family, leads to a diminished muscle mass and increased myofiber with centrally-located nuclei known as dystrophic phenotypes. APOBEC2 expression is predominant in skeletal and cardiac muscles and elevated exclusively at the early-differentiation phase of wild-type (WT) myoblast cultures; however the physiological significance is still un-known. Here we show that APOBEC2 is a key negative regulator of myoblast differentiation in muscle regeneration. APOBEC2-knockout (A2KO) mice myoblast cultures displayed a normal morphology of primary myotubes along with earlier increase in fusion index and higher expression levels of myosin heavy chain (MyHC), myogenin and its cooperating factor MEF2C than WT myoblasts. Similar response was observable in APOBEC2-knockdown cultures of WT myoblasts that were transfected with the specific siRNA at the differentiation phase (not proliferation phase). Importantly, cardiotoxin-injured A2KO gastrocnemius muscle provided in vivo evidence by showing larger up-regulation of neonatal MyHC and myogenin and hence earlier regeneration of myofiber structures with diminished cross-sectional areas and minimal Feret diameters. Therefore, the findings highlight a promising role for APOBEC2 in normal progression of regenerative myogenesis at the early-differentiation phase upon muscle injury..
28. Yusuke Komiya, Shoko Sawano, Daisuke Mashima, Riho Ichitsubo, Mako Nakamura, Ryuichi Tatsumi, Yoshihide Ikeuchi, Wataru Mizunoya, Mouse soleus (slow) muscle shows greater intramyocellular lipid droplets accumulation than EDL (fast) muscle: Fiber type-specific analysis, Journal of Muscle Research and Cell Motility, 10.1007/s10974-017-9468-6, 10.1007/s10974-017-9468-6, 2017.03, Skeletal muscle is the main tissue of lipid metabolism and accordingly is critical for homeostasis and energy production; however, the determinants of lipid accumulation in skeletal muscle are unknown. Here, we examined whether the soleus muscle (predominantly slow-twitch fibers) has a higher lipid accumulation capacity than that of the extensor digitorum longus (EDL, predominantly fast-twitch fibers) muscle in mice. Soleus and EDL muscles were harvested from male C57BL/6J mice. The mRNA levels of genes involved in fatty acid import and triglyceride synthesis and accumulation were examined in soleus and EDL muscles. The intramyocellular lipid (IMCL) droplets of muscle cross sections and isolated single fibers were visualized by staining with BODIPY493/503, and fiber types were determined by immunofluorescent detection of myosin heavy chain (MyHC) isoforms. We detected higher mRNA expression of genes related to lipid accumulation in the soleus than the EDL. We also observed a marked increase of IMCL in single fibers from the soleus, but not the EDL, after treatment with a high-fat diet plus denervation. Interestingly, greater accumulation of IMCL droplets was observed in type 2A and 2X fibers (MyHC2A- and MyHC2X-positive fibers) than type 1 fibers (MyHC1-positive fibers) in soleus muscles. These results suggest that the soleus contains more IMCL owing to the higher population of type 2A fibers, and the difference in lipid accumulation between the soleus and EDL could depend on fiber type composition..
29. Shoko Sawano, Yusuke Komiya, Riho Ichitsubo, Yasuyuki Ohkawa, Mako Nakamura, Ryuichi Tatsumi, Yoshihide Ikeuchi, Wataru Mizunoya, A One-Step Immunostaining Method to Visualize Rodent Muscle Fiber Type within a Single Specimen, PLoS ONE, 10.1371/journal.pone.0166080, 11, 11, e0166080, 10.1371/journal.pone.0166080, 2016.11.
30. Mulan Qahar, Yuko Takuma, Wataru Mizunoya, Ryuichi Tatsumi, Yoshihide Ikeuchi, Mako Nakamura, Semaphorin 3A promotes activation of Pax7, Myf5, and MyoD through inhibition of emerin expression in activated satellite cells, FEBS openbio, 10.1002/2211-5463.12050, 2016.03.
31. Sang-Gun Roh, Yutaka Suzuki, Takaufmi Gotoh, Ryuichi Tatsumi, Kazuo Katoh, Physiological roles of adipokines, hepatokines, and myokines in ruminants, Asian-Australasian Journal of Animal Sciences, 印刷中, 2015.12, Since the discovery of leptin secreted from adipocytes, specialized tissues and cells secreted the several peptides (or cytokines) that are characterized to negatively and positively regulate the metabolic process. Different types of adipokines, hepatokines, and myokines, which act as cytokines, are secreted from adipose, liver, and muscle tissue, respectively, and have been identified and examined for their physiological roles in humans and disease in animal models. Recently, various studies of these cytokines have been conducted in ruminants, including dairy cattle, beef cattle, sheep, and goat. Interestingly, a few cytokines from these tissues play an important role in the post-parturition, lactation, and fattening (marbling) periods in ruminants. Thus, understanding these hormones is important for improving nutritional management in dairy cows and beef cattle. However, to our knowledge, there have been no reviews of the characteristics of these cytokines in beef and dairy products in ruminants. In particular, lipid and glucose metabolism in adipose tissue, liver tissue, and muscle tissue are very important for energy storage, production, and synthesis, which are regulated by these cytokines in ruminant production. In this review, we summarize the physiological roles of adipokines, hepatokines, and myokines in ruminants. This discussion provides a foundation for animal production in ruminants..
32. Mai-Khoi Q. Do, Naomi Shimizu, Takahiro Suzuki, Hideaki Ohtsubo, Wataru Mizunoya, Mako Nakamura, Shoko Sawano, Mitsuhiro Furuse, Yoshihide Ikeuchi, Judy E. Anderson, Ryuichi Tatsumi, Transmembrane proteoglycans syndecan-2, 4, receptor candidates for the impact of HGF and FGF2 on semaphorin 3A expression in early-differentiated myoblasts, Physiological Reports, 10.14814/phy2.12553, 3, 9, e12553, 2015.09, Regenerative mechanisms that regulate intramuscular motor innervation are thought to reside in the spatiotemporal expression of axon-guidance molecules. Our previous studies proposed an unexplored role of resident myogenic stem cell (satellite cell)-derived myoblasts as a key presenter of a secreted neural chemorepellent semaphorin 3A (Sema3A); hepatocyte growth factor (HGF) and basic fibroblast growth factor (FGF2) triggered its expression exclusively at the early-differentiation phase. In order to advance this concept, the present study described that transmembrane heparan/chondroitin-sulfate proteoglycans syndecan-2, 4 may be the plausible receptor candidates for HGF and FGF2 to signal Sema3A expression. Results showed that mRNA expression of syndecan-2, 4 was abundant (two-magnitude higher than syndecan-1, 3) in early-differentiated myoblasts and their in vitro knock-down diminished the HGF/FGF2-induced expression of Sema3A down to a baseline level. Pre-treatment with heparitinase and chondroitinase ABC decreased the HGF and FGF2 responses, respectively in non-knock-down cultures, supporting a possible model that HGF and FGF2 may bind to heparan and chondroitin sulfate chains of syndecan-2, 4 to signal Sema3A expression. The findings, therefore, extend our understanding that HGF/FGF2-syndecan-2, 4 association may stimulate a burst of Sema3A secretion by myoblasts recruited to the site of muscle injury; this would ensure a coordinated delay in the attachment of motoneuron terminals onto fibers early in muscle regeneration, and thus synchronize the recovery of muscle-fiber integrity and the early resolution of inflammation after injury with reinnervation toward functional recovery..
33. Wataru Mizunoya, Hideo Miyahara, Shinpei Okamoto, Mariko Akahoshi, Takahiro Suzuki, Mai-Khoi Q. Do, Hideaki Ohtsubo, Yusuke Komiya, Mu Lan, Toshiaki Waga, Akira Iwata, Koichi Nakazato, Yoshihide Ikeuchi, Judy E. Anderson, Ryuichi Tatsumi, Improvement of endurance based on muscle fiber-type composition by treatment with dietary apple polyphenols in rats, PLoS ONE, 10.1371/journal.pone.0134303, 10, 7, e0134303, 2015.07, A recent study demonstrated a positive effect of apple polyphenol (APP) intake on muscle endurance of young-adult animals. While an enhancement of lipid metabolism may be responsible, in part, for the improvement, the contributing mechanisms still need clarification. Here we show that an 8-week intake of 5% (w/w) APP in the diet, up-regulates two features related to fiber type: the ratio of myosin heavy chain (MyHC) type IIx/IIb and myoglobin protein expression in plantaris muscle of 9-week-old male Fischer F344 rats compared to pair-fed controls (P
34. Yusuke Komiya, Judy E. Anderson, Mariko Akahoshi, Mako Nakamura, Ryuichi Tatsumi, Yoshihide Ikeuchi, Wataru Mizunoya, Data in support for protocol for rat single muscle-fiber isolation and culture, Data in Brief, 10.1016/j.dib.2015.04.016, 2015.05, .
35. Yusuke Komiya, Judy E. Anderson, Mariko Akahoshi, Mako Nakamura, Ryuichi Tatsumi, Yoshihide Ikeuchi, Wataru Mizunoya, Protocol for rat single muscle-fiber isolation and culture, Analytical Biochemistry, 10.1016/j.ab.2015.03.034, 2015.04, .
36. Shoko Sawano, Takahiro Suzuki, Mai-Khoi Q. Do, Hideaki Ohtsubo, Wataru Mizunoya, Yoshihide Ikeuchi, Ryuichi Tatsumi, Supplementary immunocytochemistry of hepatocyte growth factor production in activated macrophages early in muscle regeneration, Animal Science Journal, 10.1111/asj.12264, 85, 12, 994-1000, 2014.12, Regenerative intramuscular motor-innervation is thought to reside in the spatiotemporal expression of axon-guidance molecules. Our previous studies showed that resident myogenic stem cells, satellite cells, up-regulate a secreted neural-chemorepellent semaphorin 3A (Sema3A) during the early-differentiation period, in response to hepatocyte growth factor (HGF) elevated in injured muscle. However, a paracrine source of the HGF release is still unknown. Very recently, we proposed a possible contribution of anti-inflammatory macrophages (CD206-positive M2) by showing that M2 cells infiltrate predominantly at the early-differentiation phase (3-5 days post-injury) and produce/secrete large amounts of HGF. In understanding this concept, however, there still remains a critical need to examine if phagocytotic pro-inflammatory macrophages (CD86-positive M1), another activated-phenotype still present at the early-differentiation phase concerned, produce HGF upon muscle injury. The current immunocytochemical study demonstrated that the HGF expression is negative for M1 prepared from cardiotoxin-injured tibialis anterior muscle at day-5, in contrast to the intense fluorescent-signal of M2 served as a positive control. This supplementary result advances our understanding of a spatiotemporal burst of HGF secretion from M2 populations (not M1) to impact Sema3A expression, which ensures a coordinated delay in attachment of motoneuron terminals onto damaged and generating fibers during early phase of muscle regeneration..
37. Shohei Sakaguchi, Jun-ichi Shono, Takahiro Suzuki, Shoko Sawano, Judy E. Anderson, Mai-Khoi Q. Do, Hideaki Ohtsubo, Wataru Mizunoya, Mako Nakamura, Mitsuhiro Furuse, Yoshihide Ikeuchi, Ryuichi Tatsumi, Anti-inflammatory macrophages implicate in regenerative moto-neuritogenesis, by promoting myoblast migration and Sema3A expression, Animal Science Congress 2014 of the Asian-Australian Association of Animal Production Societies (AAAP) Grha Sabha Pramana, Universitas Gadjah Mada (UGM), Yogyakarta, Indonesia (November 10-14, 2014), 2014.11, Successful regeneration and remodeling of neuromuscular connections are critical for restoring functional properties of muscle fiber contractility. While the spatiotemporal regulatory mechanisms coordinating these processes (moto-neuritogenesis) with myogenesis itself remain unclear, various neural factors including attractive and repulsive axon-guidance cue ligands may be involved. Our previous studies proposed a heretofore unexplored role of resident myogenic stem cell (satellite cell)-derived myoblasts as a key presenter of a secreted neural chemorepellent semaphorin 3A (Sema3A); hepatocyte growth factor (HGF) triggered its expression exclusively at the early-differentiation phase. In order to verify this concept, the present study was designed to clarify a paracrine source of HGF release. In vitro experiments demonstrated that activated anti-inflammatory macrophages (CD206-positive M2) produce HGF and thereby promote myoblast chemoattraction and Sema3A expression. Media from pro-inflammatory macrophage cultures (M1) did not show any significant effect. M2 also enhanced the expression of myoblast-differentiation markers in culture, and infiltrated predominantly at the early-differentiation phase (3-5 d post-injury); M2 were confirmed to produce HGF as monitored by in vivo/ex vivo immunocytochemistry of CD11b/CD206/HGF-positive cells and by HGF in situ hybridization of cardiotoxin- or crush-injured tibialis anterior muscle, respectively. Findings, therefore, encourage the idea that M2 contribute to spatiotemporal up-regulation of extracellular Sema3A concentrations by producing HGF that, in turn, stimulates a burst of Sema3A secretion by myoblasts that are recruited to site of injury. This model may ensure a coordinated delay in re-attachment of motoneuron terminals onto damaged fibers early in muscle regeneration, and thus synchronize the recovery of muscle-fiber integrity after injury.
(Key Words: Mouse muscle, Activated macrophages, Hepatocyte growth factor (HGF), Semaphorin 3A (Sema3A), Regenerative moto-neuritogenesis).
38. Mulan, Nakamura, M., Suzuki, T., Mizunoya, W., Ojima, K., Tatsumi, R. and Ikeuchi, Y., Functional interaction between Sema3A and Pax7 in proliferation and differentiation of myoblasts, International Symposium on Agriculture, Forestry, Environment and Life Sciences in Asia, 2014 (AFELiSA'14) Kangwon National University, Chuncheon, Gangwon, Korea (October 28-31, 2014), 2014.10.
39. Wataru Mizunoya, Ayumi Tashima, Yusuke Sato, Ryuichi Tatsumi, Yoshihide Ikeuchi, The growth-promoting activity of egg white proteins in the C2C12 myoblast cell line, Animal Science Journal, 10.1111/asj.12257 , 86, 2, 194-199, 2014.09, .
40. Tatsumi, R.,* Sakaguchi, S.,* Shono, J.,* Suzuki, T.,* Sawano, S., Anderson, J.E., Do, M.-K.Q., Ohtsubo, H., Mizunoya, W., Nakamura, M., Furuse, M., and Ikeuchi, Y. (*equal contributors), M2 macrophages may implicate in regenerative moto-neuritogenesis, by promoting myoblast migration and Sema3A expression, 2014 FASEB Science Research Conference on “Skeletal Muscle Satellite and Stem Cells” Steamboat Springs, CO, USA (July 20-25, 2014), 2014.07, Successful regeneration and remodeling of neuromuscular connections are critical for restoring functional properties of muscle fiber contractility. While the spatiotemporal regulatory mechanisms coordinating these processes (moto-neuritogenesis) with myogenesis itself remain unclear, various neural factors including attractive and repulsive axon-guidance cue ligands and their membrane receptors may be involved. Our previous studies proposed a heretofore unexplored role of resident myogenic stem cell (satellite cell)-derived myoblasts as a key presenter of a secreted neural chemorepellent semaphorin 3A (Sema3A); hepatocyte growth factor (HGF) triggered its expression exclusively at the early-differentiation phase (Tatsumi et al., 2009; Do et al., 2011, 2012; Sato et al., 2013; Suzuki et al., 2013). In order to verify this concept, the present study was designed to clarify a paracrine source of HGF release. In vitro experiments demonstrated that activated anti-inflammatory macrophages (CD206-positive M2) produce HGF and thereby promote myoblast chemoattraction and Sema3A expression. Media from pro-inflammatory macrophage cultures (CD86-positive M1) did not show any significant effect. M2 infiltrated predominantly at the early-differentiation phase (3-5 d post-injury) and enhanced the expression of myoblast-differentiation markers; M2 were confirmed to produce HGF as monitored by in vivo/ex vivo immunocytochemistry of CD11b/CD206/HGF-positive cells and by HGF in situ hybridization of cardiotoxin- or crush-injured tibialis anterior muscle, respectively. Findings, therefore, encourage the idea that M2 contribute to spatiotemporal up-regulation of extracellular Sema3A concentrations by producing HGF that, in turn, stimulates a burst of Sema3A secretion by myoblasts that are recruited to site of injury. This regulatory axis of “M2 macrophages → satellite cell-derived myoblasts → intramuscular motoneuron terminals” may ensure a coordinated delay in attachment of motoneuron terminals onto regenerating/generating (new) fibers early in muscle regeneration, and thus synchronize the recovery of muscle-fiber integrity after injury..
41. Ohtsubo, H., Sato, Y., Suzuki, T., Mizunoya, W., Nakamura, M., Tatsumi, R., and Ikeuchi, Y. , APOBEC2 deficiency up-regulates myoblast differentiation, 2014 FASEB Science Research Conference on “Skeletal Muscle Satellite and Stem Cells” Steamboat Springs, CO, USA (July 20-25, 2014), 2014.07.
42. Suzuki, T., Ohya, Y., Ojima, K., Mizunoya, W., Sawano, S., Ohtsubo, H., Nishimatsu, S., Anderson, J. E., Do, M.-K. Q., Nakamura, M., Furuse, M., Ikeuchi, Y., Nohno, T., and Tatsumi, R., Sema3A secreted from satellite cells promotes slow-twitch fiber generation, 2014 FASEB Science Research Conference on “Skeletal Muscle Satellite and Stem Cells” Steamboat Springs, CO, USA (July 20-25, 2014), 2014.07.
43. Ryuichi Tatsumi, Investigating muscle regeneration: The secret of Sema3A, International Innovation, 139, 89-91, 2014.06, Sema3A is a multifunctional molecule involved in several significant processes, but studies have so far neglected its potential impact in muscle biology. Research from Kyushu University in Japan is beginning to unravel its important role and results to date are promising..
44. Shohei Sakaguchi, Jun-ichi Shono, Takahiro Suzuki, Shoko Sawano, Judy E. Anderson, Mai-Khoi Q. Do, Hideaki Ohtsubo, Wataru Mizunoya, Yusuke Sato, Mako Nakamura, Mitsuhiro Furuse, Koji Yamada, Yoshihide Ikeuchi, Ryuichi A Tatsumi, Implication of anti-inflammatory macrophages in regenerative moto-neuritogenesis: promotion of myoblast migration and neural chemorepellent semaphorin 3A expression in injured muscle, International Journal of Biochemistry and Cell Biology, 10.1016/j.biocel.2014.05.032, 54, 272-285, 2014.06, Regenerative mechanisms that regulate intramuscular motor innervation are thought to reside in the spatiotemporal expression of axon-guidance molecules. Our previous studies proposed a heretofore unexplored role of resident myogenic stem cell (satellite cell)-derived myoblasts as a key presenter of a secreted neural chemorepellent semaphorin 3A (Sema3A); hepatocyte growth factor (HGF) triggered its expression exclusively at the early-differentiation phase. In order to verify this concept, the present study was designed to clarify a paracrine source of HGF release. In vitro experiments demonstrated that activated anti-inflammatory macrophages (CD206-positive M2) produce HGF and thereby promote myoblast chemoattraction and Sema3A expression. Media from pro-inflammatory macrophage cultures (M1) did not show any significant effect. M2 also enhanced the expression of myoblast-differentiation markers in culture, and infiltrated predominantly at the early-differentiation phase (3-5 days post-injury); M2 were confirmed to produce HGF as monitored by in vivo/ex vivo immunocytochemistry of CD11b/CD206/HGF-positive cells and by HGF in situ hybridization of cardiotoxin- or crush-injured tibialis anterior muscle, respectively. These studies advance our understanding of the stage-specific activation of Sema3A expression signaling. Findings, therefore, encourage the idea that M2 contribute to spatiotemporal up-regulation of extracellular Sema3A concentrations by producing HGF that, in turn, stimulates a burst of Sema3A secretion by myoblasts that are recruited to site of injury. This model may ensure a coordinated delay in re-attachment of motoneuron terminals onto damaged fibers early in muscle regeneration, and thus synchronize the recovery of muscle-fiber integrity and the early resolution of inflammation after injury..
45. Wataru Mizunoya, Yohei Iwamoto, Yusuke Sato, Ryuichi A Tatsumi, Yoshihide Ikeuchi, Cold Exposure Increases Slow-Type Myosin Heavy Chain 1 (MyHC1) Composition of Soleus Muscle in Wistar Rats, Animal Science Journal, 10.1111/asj.12143., 85, 3, 293-304, 2014.03, .
46. Jun-ichi Shono, Shohei Sakaguchi, Takahiro Suzuki, Mai-Khoi Q. Do, Wataru Mizunoya, Mako Nakamura, Yusuke Sato, Mitsuhiro Furuse, Koji Yamada, Yoshihide Ikeuchi, Ryuichi A Tatsumi, Preliminary Time-Course Study of Antiinflammatory Macrophage Infiltration in Crush-Injured Skeletal Muscle, Animal Science Journal, 10.1111/asj.12105, 84, 11, 744-750, 2013.11, Muscle damage induces massive macrophage infiltration of the injury site, in which activated proinflammatory and antiinflammatory phenotypes (currently classified as M1 and M2, respectively) have been documented as distinct functional populations predominant at different times after the conventional acute injury by intramuscular injection of snake venoms (cardiotoxin, notexin) or chemicals (bupivacaine hydrochloride, barium chloride). The present study employed a muscle-crush injury model that may reflect the more physiologic damage and repair processes initiated by contusing a gastrocnemius muscle in lower hind-limb of adult mice with hemostat forceps, and examined the time-course invasion of M1 and M2 macrophages during muscle regeneration by immunocytochemistry of CD197 and CD206 marker proteins. CD197-positive M1 macrophages were observed exclusively at 1-4 days after crush followed by the alternative prevalence of CD206-positive M2 at 7-days of myogenic differentiation time-point characterized by increasing levels of myogenin mRNA expression. Preliminary PCR analysis showed that M2 may produce hepatocyte growth factor (HGF) in culture, providing additional benefit to understanding that M2 populations actively promote regenerative myogenesis (muscle fiber repair) and moto-neuritogenesis (re-attachment of motoneuron terminals onto damaged fibers) through their time-specific infiltration and release of the growth factor at the injury site early in muscle regeneration. .
47. Wataru Mizunoya, Yohei Iwamoto, Bungo Shirouchi, MASAO SATO, Yusuke Komiya, Farzaneh R. Razin, Ryuichi A Tatsumi, Yusuke Sato, Mako Nakamura, Yoshihide Ikeuchi, Dietary fat influences the expression of contractile and metabolic genes in rat skeletal muscle, PLoS ONE, 10.1371/journal.pone.0080152, 8, e80152, 2013.11, .
48. Takahiro Suzuki, Koichi Ojima, Mai-Khoi Q. Do, Minako Hara, Wataru Mizunoya, Mako Nakamura, Yoshihide Ikeuchi, Judy E. Anderson, Ryuichi Tatsumi, Semaphorin 3A secreted from myogenic stem cells promotes slow-twitch muscle fiber generation, 2013 EMBO Workshop on “Semaphorin Function and Mechanism in Action”, Cemay-la-Ville, France (October 29-31, 2013), 2013.10, Semaphorin 3A (Sema3A), a class 3 vertebrate-secreted semaphorin originally characterized as a potent neural chemorepellent, is now recognized to play crucial roles in angiogenesis, organogenesis, osteoclastogenesis and immune responses as a multi-functional modulator. Recently, we found that resident myogenic stem cells, satellite cells, up-regulate Sema3A expression and secretion exclusively at the early-myogenic differentiation phase in response to in vivo crush injury and hepatocyte growth factor treatment in the primary cultures (Tatsumi et al., 2009); however, its physiological significance in muscle regeneration is still unknown. Here we show that Sema3A impacts slow-type myosin heavy chain (slow-MyHC) expression in mouse satellite cell cultures. Sema3A-siRNA transfection significantly reduced expression of slow-MyHC as well as the muscle-specific transcription factor myogenin, and the down-stream mediators MEF2 as revealed by qPCR and western blotting analyses. Total MyHC expression level was unchanged, likely due to compensatory up-regulation of fast-MyHC during the 72-hr transfection period. Similar responses (except for fast-MyHC) were also observed in myogenin-knockdown cultures. In addition, reduced myogenin expression induced by immunoneutralization of the receptor neuropilin1 (Npn1) was rescued by co-addition of Sema3A protein. These results therefore indicate that Sema3A may activate a slow-MyHC expression-signaling axis consisting of Npn1, myogenin and MEF2. This model is supported by our comparative observations that satellite cells from soleus muscle (abundant in slow fibers) showed higher expression of Sema3A, myogenin and a co-receptor plexinA2 than those from EDL (Suzuki et al. 2013). Overall, the findings highlight a heretofore unexplored and active role for satellite cell-derived Sema3A as a key modulator of slow fiber generation during muscle regeneration, and advance our understanding of the multi-functional contributions of Sema3A..
49. Ryuichi Tatsumi, Judy E. Anderson, Ronald E. Allen, Muscle regeneration dynamics mediated by resident myogenic stem cells: a possible implication in moto-neuritogenesis and fiber-type regulation, Tucson Symposium 2013 on “The Biology of Muscle Growth and Repair”, 2013.09, Skeletal muscle regeneration and work-induced hypertrophy are initiated by mechanical insult or other perturbation and one of the earliest events is triggering the activation (re-entry to cell cycle from G0) of quiescent resident myogenic stem cells, satellite cells. Recent studies of satellite cells in culture and in vivo addressed the possible dual-roles of hepatocyte growth factor (HGF) in the stretch-induced activation and the re-establishing quiescence. In the proposed scenario, the time-coordinated increase in extracellular HGF is a key modulator for the two contrary pathways having low and high thresholds to impact activation and the counterpart quiescence of satellite cells, respectively. Moreover, the role of HGF in muscle repair may not be restricted to myogenesis; we demonstrated that HGF up-regulates expression of secreted axon-guidance molecule semaphorin 3A (Sema3A) in satellite cells at early-differentiation phase in primary cultures and in vivo. The results encourage a possible implication of satellite cells in the spatiotemporal regulation of extracellular Sema3A concentrations, which potentially mediates restoration or remodeling of nerve-muscle connections in muscle regeneration in synchrony with recovery of muscle-fiber integrity and types (fast and slow). Very recently, we found in satellite cell cultures that the Sema3A ligand impacts slow-twitch fiber generation through a muscle-specific transcription factor myogenin-dependent pathway; Sema3A-siRNA transfection significantly reduced expression of slow-type myosin heavy chain (slow-MyHC) as well as myogenin and its co-mediator MEF2. Total MyHC protein expression level was likely unchanged, due to compensatory up-regulation of fast-MyHC and similar responses (except for fast-MyHC) were also observed in myogenin-knockdown cultures. Overall our results highlight the “programmed mechano-biology dynamics” that successful muscle regeneration, comprised of satellite cell-driving myogenesis, intramuscular moto-neuritogenesis and fiber-type regulation (survival), may be a programmed sequence of events that respond to a mechanical perturbation in a synchronous, HGF-dependent and time-coordinated manner..
50. Wataru Mizunoya, Shoko Sawano, Yohei Iwamoto, Yusuke Sato, Ryuichi A Tatsumi, Yoshihide Ikeuchi, Effect of 48-h Food Deprivation on the Expressions of Myosin Heavy Chain Isoforms and Fiber Type-Related Factors in Rats, Journal of Nutrition Science and Vitaminology, org/10.3177/jnsv.59.289, 59, 4, 289-298, 2013.08, The primary aim of this study was to examine the effects of 48-h food deprivation on rat skeletal muscle fiber type, according to myosin heavy-chain (MyHC) isoform composition and some metabolism-related factors in both slow-type dominant and fast-type dominant muscle tissues. Male Wistar rats (7 wk old) were treated with 48-h food deprivation or ad libitum feeding as control. After the treatment, the soleus muscle (slow-type dominant) and the extensor digitorum longus (EDL, fast-type dominant) were excised. We found that 48-h food deprivation did not affect MyHC composition in either the soleus or EDL, compared with fed rats by electrophoretic separation of MyHC isoforms. However, 48-h food deprivation significantly increased the mRNA expression of fast-type MyHC2B in the EDL muscle. Moreover, food deprivation increased fatty acid metabolism, as shown by elevated levels of related serum energy substrates and mRNA expression of mitochondrial uncoupling protein (UCP) 3 and lipoprotein lipase (LPL) in both the soleus and EDL. UCP3 and LPL are generally expressed at higher levels in slow-type fibers. Furthermore, we found that food deprivation significantly decreased the protein amounts of PGC1α and phosphorylated FOXO1, which are known as skeletal muscle fiber type regulators. In conclusion, 48-h food deprivation increased mRNA expression of fast-type MyHC isoform and oxidative metabolism-related factors in EDL, whereas MyHC composition at the protein level did not change in either the soleus or EDL..
51. Wataru Mizunoya, Koichiro Ohnuki, Kento Baba, Hideo Miyahara, Naomi Shimizu, Kuniko Tabata, Takako Kino, Yusuke Sato, Ryuichi A Tatsumi, Yoshihide Ikeuchi, Effect of Dietary Fat Type on Anxiety-Like and Depression-Like Behavior in Mice, SpringerPlus , 10.1186/2193-1801-2-165, 2, 165, 2013.04, Dietary fat plays an important role in higher brain functions. We aimed to assess the short and long term intake of three different types of dietary fat (soybean oil, lard, and fish oil) on anxiety-like and depression-like behavior in mice. For the short term intake assessment, a behavioral test battery for anxiety and depression was carried out for a 3-day feeding period. For the long term intake assessment, a behavioral test battery began after the 4-week feeding period. During the short term intake, the time spent in the open arms of the elevated plus-maze was the longest in the fish oil fed group, followed by the soybean oil and lard-fed groups. The elevated plus-maze is a common animal model to assess anxiety, in which an increased time spent in the open arms indicates an anxiolytic effect. The difference between the fish oil-fed group and lard-fed group was statistically significant (p
52. Yusuke Sato, Mai-Khoi Q. Do, Takahiro Suzuki, Hideaki Ohtsubo, Wataru Mizunoya, Mako Nakamura, Mitsuhiro Furuse, Yoshihide Ikeuchi, Ryuichi A Tatsumi, Satellite Cells Produce Neural Chemorepellent Semaphorin 3A upon Muscle Injury, Animal Science Journal, 10.1111/asj.12104, 84, 185-189, Listed on ScienceAlerts.com, 2013.02, Regenerative mechanisms that regulate intramuscular motor innervation including configuration of the neuromuscular connections are thought to reside in the spatiotemporal expression of axon-guidance molecules. Our previous studies proposed a heretofore unexplored role of satellite cells as a key source of a secreted neural chemorepellent semaphorin 3A (Sema3A) expression. In order to verify this concept, there is still a critical need to provide direct evidence to show the up-regulation of Sema3A protein in satellite cells in vivo upon muscle injury. The present study employed a Sema3A/MyoD double-immunohistochemical staining for cryo-sections prepared from cardiotoxin (CTX)-injected gastrocnemius muscle of adult mouse lower hind-limb. Results clearly demonstrated that Sema3A expression was up-regulated in MyoD-positive satellite cells at 4-12 days post-injury period, the time that corresponds to the cell differentiation phase characterized by increasing myogenin mRNA expression. This direct proof encourages a possible implication of satellite cells in the spatiotemporal regulation of extracellular Sema3A concentrations, which potentially ensures coordinating a delay in neurite sprouting and re-attachment of motoneuron terminals onto damaged muscle fibers early in muscle regeneration in synchrony with recovery of muscle-fiber integrity..
53. Takahiro Suzuk, Mai-Khoi Q. Do, Yusuke Sato, Koichi Ojima, Minako Hara, Wataru Mizunoya, Mako Nakamura, Mitsuhiro Furuse, Yoshihide Ikeuchi, Judy E. Anderson, Ryuichi A Tatsumi, Comparative Analysis of Semaphorin 3A in Soleus and EDL Muscle Satellite Cells In Vitro toward Understanding Its Role in Modulating Myogenin Expression, International Journal of Biochemistry and Cell Biology, 45, 2, 476-482, Listed on ScienceAlerts.com, 2013.02, Resident myogenic stem cells, satellite cells, up-regulate a secreted multi-functional modulator, semaphorin 3A (Sema3A), exclusively at the early-differentiation phase in response to muscle-crush injury and treatment with hepatocyte growth factor (HGF) or basic fibroblast growth factor (FGF2). Here, we add evidence that the Sema3A expression and secretion induced by the growth factors is significantly higher in primary cultures from adult rat soleus than from the fast-twitch extensor digitorum longus (EDL) muscle. The higher Sema3A response, revealed by quantitative PCR and western blotting of cell lysates and conditioned media, may account for the higher myogenin expression of soleus muscle satellite cells early in differentiation since addition of recombinant Sema3A stimulates myogenin expression in cultures. These experiments also showed that mRNA expression of plexin A2, which together with neuropilins, constitutes Sema3A composite-receptors, was higher in satellite cells from soleus than EDL with no difference in plexin A1 and A3 and neuropilin-1 and 2 levels. These comparative studies, therefore, highlight a possible Sema3A-plexin A2-myogenin signaling axis that may ensure promoting early differentiation by soleus muscle satellite cells. .
54. Mai-Khoi Q. Do, Takahiro Suzuki, Borjigin Gerelt, Yusuke Sato, Wataru Mizunoya, Mako Nakamura, Yoshihide Ikeuchi, Judy E. Anderson, and Ryuichi Tatsumi, Time-Coordinated Prevalence of Extracellular HGF, FGF2 and TGF-β3 in Crush-Injured Skeletal Muscle, Animal Science Journal, 83, 712-717, 2012.10, Successful regeneration and remodeling of neuromuscular junctions are critical for restoring functional capacities and properties of skeletal muscle after damage, and axon-guidance molecules may be involved in the signaling that regulates such restoration. Recently, we found that early-differentiated satellite cells up-regulate a secreted neural chemorepellent Sema3A upon in vivo muscle-crush injury. The study also revealed that Sema3A expression is up-regulated in primary satellite-cell cultures in response to hepatocyte growth factor (HGF) and basic fibroblast growth factor (FGF2) and prevented by transforming growth factor (TGF)-β2, 3 (Tatsumi et al. 2009. Am J Physiol Cell Physiol; Do et al. 2011. Am J Physiol Cell Physiol). In order to verify the physiological significance of this regulation in vitro, the present study was designed to estimate the time-course of extracellular HGF, FGF2 and TGF-β3 concentrations after crush-injury of gastrocnemius muscle in the rat lower hind-limb, using a combination of a non-homogenization/non-spin extraction of extracellular wound fluids and ECL-western blotting analyses. Results clearly demonstrated that active HGF and FGF2 are prevalent in 2-8 days post-crush, whereas active TGF-β3 increases after 12 days, providing a better understanding of the time-coordinated levels of HGF, FGF2, and TGF-β3 that drive regulation of Sema3A expression during regenerative intramuscular moto-neuritogenesis..
55. Tatsumi, R.,* Hara, M.,* Tabata, K.,* Suzuki, T., Do, M.-K. Q., Mizunoya, W., Nakamura, M., Ikeuchi, Y., Anderson, J. E., and Allen, R. E. (*equal contributors), Mechano-Sensing Calcium-Influx Machinery that Instigates Skeletal Muscle Satellite Cell Activation, 2012 FASEB Science Research Conference on “Skeletal Muscle Satellite & Stem Cells", Lucca, Italy, 2012.08, When skeletal muscle is stretched or injured, satellite cells, resident myogenic stem cells positioned beneath the basal lamina of mature muscle fibers, are activated to enter the cell cycle. This signaling pathway is a cascade of events including calcium-calmodulin formation, nitric oxide (NO) radical production by NO synthase, matrix metalloproteinase activation, release of hepatocyte growth factor (HGF) from the extracellular matrix, and presentation of HGF to the receptor c-met, as demonstrated by assays of primary cultures and in vivo experiments. Here, we add evidence that two ion channels, the mechano-sensitive cation channel (MS-channel) and the long-lasting-type voltage-gated calcium-ion channel (L-VGC-channel), mediate the influx of extracellular calcium ions in response to cyclic stretch in satellite-cell cultures. When applied to 1-hr stretch cultures with individual inhibitors for MS- and L-VGC-channels (GsMTx-4 and nifedipine, respectively) or with a less specific inhibitor for MS-channels (gadolinium chloride, Gd), satellite cell activation and upstream HGF release were abolished, as revealed by bromodeoxyuridine-incorporation assays and western blotting of conditioned media, respectively. The inhibition was dose-dependent with a maximum at 0.1 μM (GsMTx-4), 10 μM (nifedipine) or 100 μM (Gd) and cancelled by addition of HGF to the culture media; a potent inhibitor for transient-type VGC-channels (NNC55-0396, 100 μM) did not show any significant inhibitory effect. The stretch response was also abolished when calcium-chelator EGTA (1.8 mM) was added to the medium, indicating the significance of extracellular free calcium ions in our present activation model. Finally, cation/calcium channel dependencies were further documented by calcium-imaging analyses on stretched cells; results clearly demonstrated that calcium ion influx was abolished by GsMTx-4, nifedipine and EGTA. Therefore, these results provide an additional insight that calcium ions may flow in through L-VGC-channels by possible coupling with adjacent MS-channel gating that promotes the local depolarization of cell membranes to impact the satellite cell activation cascade..
56. Suzuki, T., Ojima, K., Do, M.Q., Hara, M., Mizunoya, W., Nakamura, M., Ikeuchi, Y., Anderson, J.E. and Tatsumi, R., Semaphorin 3A regulates the early differentiation of satellite cells, 2012 FASEB Science Research Conference on “Skeletal Muscle Satellite & Stem Cells", Lucca, Italy, 2012.08, Semaphorin 3A (Sema3A), a class 3 vertebrate-secreted semaphorin, is a potent axon-guidance molecule for sensory, sympathetic and motor neurons. Recently, we found that resident myogenic stem cells, satellite cells, up-regulate Sema3A expression and secretion exclusively at the early-myogenic differentiation phase in response to in vivo crush injury and HGF/FGF2 treatments in the primary cultures, suggesting possible implication of satellite cells in regenerative motoneuritogenesis including sprouting and attachment of motoneuron terminals onto damaged muscle fibers in synchrony with recovery of muscle-fiber structures (Tatsumi et al., Am. J. Physiol. Cell Physiol. 2009; Do et al., Am. J. Physiol. Cell Physiol. 2011). Here, we show that Sema3A also mediates post-natal myogenesis by stimulating the early differentiation of satellite cells. When recombinant Sema3A (R&D Systems) was added to satellite cell cultures for 24 hr from 48-hr post-plating, myogenin message (an early differentiation marker) was up-regulated in a dose-dependent manner with a maximum at 10 ng/ml. Sema3A-specific knock-down by RNAi technique (about 60-75% reduction efficiency) remarkably down-regulated myogenin expression at message and protein levels at the early-differentiation stage, however showed no significant effect on MyoD expression at the same phase and myosin heavy chain expression and myotube formation at the later stage. Immunofluorescence analysis revealed the presence of Sema3A membrane-receptor neuropilin1 (Npn1) at 48-hr post-plating, which is the early-differentiation time-point which siRNA was transfected to cells; immunoneutralization of Npn1 activity also reduced the myogenin expression, which can be rescued competitively by co-addition of Sema3A protein, to a level equivalent to control cultures without anti-Npn1 antibody and Sema3A. These results indicate that satellite cell-secreted Sema3A may bind to the receptor Npn1 to generate myogenin expression signaling that stimulates early differentiation of satellite cells in autocrine and/or paracrine fashions. This topic may be supported potentially by our comparative observations that rat soleus muscle satellite cells showed higher expression activities of Sema3A, myogenin, and plexinA2 (a signaling co-receptor protein for Sema3A) than EDL muscle cells at the early-differentiation stage. Overall, the data highlight again a heretofore unexplored and active role for Sema3A as a key regulator of the early myogenic differentiation of satellite cells during muscle regeneration, therefore providing a better understanding of multi-functional contributions of Sema3A..
57. Do, M.Q., Shimizu, N., Suzuki, T., Mizunoya, W., Nakamura, M., Ikeuchi, Y., Anderson, J.E. and Tatsumi, R., Heparan/chondroitin sulfate chains may mediate HGF/FGF2-induced up-regulation of neural chemorepellent Sema3A in satellite cell cultures, 2012 FASEB Science Research Conference on “Skeletal Muscle Satellite & Stem Cells", Lucca, Italy, 2012.08, Semaphorin 3A (Sema3A, also referred to as SemaIII, SemD and collapsin), a class 3 vertebrate-secreted semaphorin, is a potent axon-guidance cue for sensory, sympathetic and motor axons. Recently, we found that satellite cells up- or down-regulate Sema3A in time-dependent responses to in vivo muscle crush-injury and in vitro treatment with recombinant HGF, FGF2 or TGF-βs. Such responses imply that satellite cells are involved in regenerative motoneuritogenesis, including sprouting and re-attachment of motoneuron terminals onto damaged muscle fibers. In order to explore the mechanism of HGF/FGF2-induced Sema3A up-regulation, the present study was designed to investigate possible membrane receptors that are present on satellite cells and could be involved in that signaling pathway. First, we tested whether c-met and FGFR1, high-affinity receptors of HGF and FGF2 respectively, are responsible for Sema3A up-regulation. Treatment with anti-c-met and anti-FGFR1 neutralizing antibodies did not diminish Sema3A up-regulation, indicating that c-met and FGFR1 may not mediate the response. In addition to such high-affinity receptors, HGF and FGF2 also bind with lower affinity to glycosaminoglycan (GAG) chains of proteoglycans - an important component of extracellular matrix- for a variety of biological functions. We therefore hypothesized that the association between HGF/FGF2 and GAG chains might mediate the Sema3A up-regulation. To test the hypothesis, satellite cell cultures were pretreated with GAG-degrading enzymes before the addition of HGF or FGF2, and the level of Sema3A expression was quantified by real-time qPCR. Results showed that treatment with heparitinase for partial removal of heparan sulfate chains significantly decreased induction of Sema3A up-regulation by HGF, but not FGF2. Similarly, treatment with chondroitinase ABC for partial removal of chondroitin sulfate chains significantly diminished the Sema3A up-regulation induced by FGF2, but not HGF. These results suggest that HGF and FGF2 bind to receptors carrying heparan and/or chondroitin sulfate chains; syndecans or other transmembrane-type proteoglycans appear to be plausible receptor candidates for signaling via growth factors to up-regulate Sema3A in satellite cells in culture. .
58. Komiya, Y., Mizunoya, W., Anderson, J.E., Goto, T., Takahashi, N., Kawada, T., Sato, Y., Nakamura, M., Tatsumi, R., and Ikeuchi, Y., Change of skeletal muscle fiber types by n-3 poly unsaturated fatty acid, 2012 FASEB Science Research Conference on “Skeletal Muscle Satellite & Stem Cells", Lucca, Italy, 2012.08, Skeletal muscle fibers are classified as type I (red/oxidative/slow) and type II (white/glycolytic/fast) based on color, metabolism and contractile properties. Type I fibers are especially mitochondria-rich and use oxidative metabolism for energy production. Thus, an increase of type I fibers can counteract obesity and reduce fatigue. In a recent study, the nuclear receptor peroxisome proliferator-activated receptor (PPAR) δ attracted attention as a factor regulating muscle fiber type. Over-expression of activated PPARδ caused a shift to slow-type fibers (Wang et al. 2004, PLoS Biol., 2: 1532-1539) and polyunsaturated fatty acid (PUFA) has the ligand activity of PPARδ (Forman et al. 1996, Ann. N.Y.Acad.Sci., 804: 266-275). Recently, we found in rat skeletal muscle that fiber type distribution was slower in vivo after feeding with fish oil. To elucidate this phenomenon, we hypothesized that the change to slow muscle with fish oil results from fish oil PUFA activating the receptor activity of PPARδ. Here, we tested the hypothesis using a rat muscle fiber culture assay. Since muscle fiber-type transformation generally occurs in mature fibers, this culture model has the advantage that fibers can express mature MyHC isoforms while differentiated cell lines such as C2C12 and L6 cannot. Rat fiber cultures were established according to the mouse protocol (Wozniak and Anderson 2005, Biochem.Cell Biol., 83: 674-676) and over 90% muscle fibers were alive until 7 days of culture. When the PPARδ−specific agonist (GW501516) was added to fibers cultured for 7 days, transcript expression of MyHC1and 2A was up-regulated at day 1, lipoprotein lipase message was up-regulated at day 3, and pyruvate dehydrogenase kinase (PDK) 4 and uncoupling protein 3 were up-regulated at day 7. Using luciferase reporter assays, PPARδ ligand activity was measured for several kinds of fatty acid. Results showed that, eicosapentaenoic acid (EPA), abundant in fish oil, had higher ligand activity to PPARδ compared with other fatty acids. When EPA was added to fiber cultures for 24 hours, PDK4 mRNA was up-regulated. In addition, when both EPA and PPARδ-specific antagonist (GSK0660) were added to fiber cultures, the up-regulation of PDK4 mRNA by addition of EPA was reseted to the control level. In conclusion, EPA up-regulates a protein characteristic of slow-type muscle via PPARδ activation and may transform muscle fiber type into slow type. .
59. Sato, Y., Ohtsubo, H., Kaneko, T., Mizunoya, W., Nakamura, M., Tatsumi, R., Iida, H., Ikeuchi, Y., Yoshizawa, F., Sugahara, K., Neuberger, M.S. and Rada, C., APOBEC2 deficiency causes increased autophagy and abnormal mitochondria in skeletal muscle, 2012 FASEB Science Research Conference on “Skeletal Muscle Satellite & Stem Cells", Lucca, Italy, 2012.08, APOBEC2 is one a member of AID/APOBEC cytidine deaminase family with putative DNA/RNA editing enzyme. Loss of APOBEC2 leads to a shift in muscle fiber-type, diminished body mass and myofiber with centrally-located nuclei although the molecular mechanisms are unknown. APOBEC2 is localized in z-bands of sarcomere in mouse muscle. Its deficiency did not affect the sarcomere structure although skeletal muscles of APOBEC2 KO (A2-/-) mice had degenerating fibers and rimmed vacuoles (RVs). To test whether the defect of APOBEC2 affected protein degradation system, we investigated the expression level of autophagy-related genes (Map1lc3b, Park2, Pink1 and p62) and ubiquitin ligases (Trim63, Fbxo32). The expression levels of autophagy-related genes were increased although those of ubiquitin ligases were not changed. Immunohistochemical analysis showed accumulated LC3 and p62 in RVs of A2-/- myofiber. Subsequent electron microscopy showed enlarged mitochondria and mitochondria engulfed by autophagosomes in A2-/- muscle. In addition, we found that activated LC3-Ⅱ and poly-ubiquitin were increased and localized to mitochondria of A2-/- muscle. In this study, we demonstrated that APOBEC2 deficiency caused enlarged mitochondria and increased autophagy (especially mitophagy). APOBEC2 may be unknown autophagy-related gene in skeletal muscle..
60. Minako Hara, Kuniko Tabata, Takahiro Suzuki, Mai-Khoi Q. Do, Wataru Mizunoya, Mako Nakamura, Shotaro Nishimura, Shoji Tabata, Yoshihide Ikeuchi, Kenji Sunagawa, Judy E. Anderson, Ronald E. Allen, and Ryuichi Tatsumi, Calcium Influx through a Possible Coupling of Cation Channels Impacts Skeletal Muscle Satellite Cell Activation in Response to Mechanical Stretch, American Journal of Physiology-Cell Physiology, 2012年3月26日掲載受理、セットで先行公開, 2012.03.
61. Do, M.-K. Q., Sato, Y., Shimizu, N., Suzuki, T., Shono, J.-I., Mizunoya, W., Nakamura, M., Ikeuchi, Y., Anderson, J. E., and Tatsumi, R., Growth Factor Regulation of Neural Chemorepellent Sema3A Expression in Satellite Cell Cultures, American Journal of Physiology-Cell Physiology, 301, C1270-C1279, Listed on the March edition of Global Medical Discovery Series, 2011.11, 筋肥大・再生において筋幹細胞(衛星細胞)が合成・分泌する神経軸索ガイダンス因子Sema3Aによって運動神経末端の再接着(神経支配の回復)が制御されていることをはじめて見出した他、このSema3Aの発現がHGF/FGF2/TGF-beta2,3によって時系列的に調節されていることを示した。.
62. Tatsumi, R., Mizunoya, W., Ikeuchi, Y., Hattori, A., Anderson, J. E., and Allen, R. E., Mechano-Biology of Resident Myogenic Stem Cells: Implication in Postnatal Myogenesis and Intramuscular Motoneuritogenesis, Animal Science Congress 2010 of the Asian-Australian Association of Animal Production Societies (AAAP), National Pingtung University of Science and Technology (NPUST), Pingtung, Taiwan , 2010.08.
63. Tatsumi, R.,* Do, M.-K. Q.,* Shimizu, N.,* Sankoda, Y.,* Anderson, J. E., Sato, Y., Suzuki, T., Mizunoya, W., Ikeuchi, Y., and Allen, R. E. (*equal contributors), Possible Implication of Satellite Cells in Regenerative Motoneuritogenesis: HGF and FGF2 Upregulate Neural Chemorepellent Sema3A Expression, Theme 3: Tissue Mechanics, Track 3.6 Muscle Mechanics and Motor Control, at the 6th World Congress on Biomechanics (WCB) 2010, in conjunction with the 14th International Conference on Biomedical Engineering (ICBME) 2010 and the 5th Asian Pacific Conference on Biomechanics (APBiomech) 2010, Suntec Convention and Exhibition Center, Singapore , 2010.08, 筋肥大・再生の過程で、筋幹細胞(衛星細胞)が肝細胞増殖因子HGF依存的に神経軸索ガイダンス因子Sema3Aを合成・分泌することを見出し、衛星細胞が運動神経末端の再接着制御に能動的に関与している可能性を提起した。このアイデアが評価され、掲載雑誌のEditorial Focusに選定された。.
64. Tatsumi, R., Anderson, J. E. and Allen, R. E.., Mechano-Biology on Stretch-Induced Activation of Resident Myogenic Stem Cells, Symposium on “Muscle Injury” section at the 6th World Congress on Biomechanics (WCB) 2010, in conjunction with the 14th International Conference on Biomedical Engineering (ICBME) 2010 and the 5th Asian Pacific Conference on Biomechanics (APBiomech) 2010, Suntec Convention and Exhibition Center, Singapore , 2010.08, In undamaged postnatal muscle fibers with normal contraction and relaxation activities, quiescent satellite cells of resident
myogenic stem cells are interposed between the overlying external lamina and the sarcolemma of a subjacent mature
muscle fiber. When muscle is injured, exercised, overused or mechanically stretched, these cells are activated to enter the
cell proliferation cycle, divide, differentiate, and fuse with the adjacent muscle fiber, and are responsible for regeneration
and work-induced hypertrophy of muscle fibers. Therefore, a mechanism must exist to translate mechanical changes in
muscle tissue into chemical signals that can activate satellite cells. Recent studies of satellite cells or single muscle fibers in
culture and in vivo demonstrated the essential role of hepatocyte growth factor (HGF) and nitric oxide (NO) radical in the
activation pathway. These experiments have also reported that mechanically stretching satellite cells or living skeletal
muscles triggers the activation by rapid release of HGF from its extracellular tethering and the subsequent presentation to
the receptor c-met. HGF release has been shown to rely on calcium-calmodulin formation and NO radical production in
satellite cells and/or muscle fibers in response to the mechanical perturbation, and depend on the subsequent
up-regulation of matrix metalloproteinase (MMP) activity. These results indicate that the activation mechanism is a cascade
of events including calcium ion influx, calcium-calmodulin formation, NO synthase activation, NO radical production, MMP
activation, HGF release and binding to c-met. Better understanding of ‘mechano-biology’ on the satellite cell activation is
essential for designing procedures that could enhance muscle growth and repair activities in meat-animal agriculture and
also in neuromuscular disease and aging in humans..
65. Sato, Y., Probst, H. C., Mizunoya, W., Tatsumi, R., Rada, C., Neuberger M. S., and Ikeuchi, Y., Skeletal Muscle Fiber-Type Change, Diminished Body Mass and Myopathy in APOBEC2 KO Mouse, 2010 FASEB Summer Research Conference on “Skeletal Muscle Satellite & Stem Cells", Carefree, AZ, US, 2010.07.
66. Do, M.-K. Q.,* Shimizu, N.,* Sato, Y., Suzuki, T., Tatsumi, R., Mizunoya, W., Ikeuchi, Y., Anderson, J. E., and Allen, R. E. (*equal contributors), Possible Implication of Satellite Cells in Regenerative Motoneuritogenesis: Temporal Coordination of HGF/FGF-2/TGF-beta may Regulate Neural Chemorepellent Sema3A expression, 2010 FASEB Summer Research Conference on “Skeletal Muscle Satellite & Stem Cells", Carefree, AZ, US, 2010.07, 筋肥大・再生の過程で、筋幹細胞(衛星細胞)が肝細胞増殖因子HGF依存的に神経軸索ガイダンス因子Sema3Aを合成・分泌することを見出し、衛星細胞が運動神経末端の再接着制御に能動的に関与している可能性を提起した。このアイデアが評価され、掲載雑誌のEditorial Focusに選定された。.
67. Suzuki, T., Takaishi, H., Sakata, T., Do, M. Q., Hara, M., Sato, A., Mizunoya, W., Nishimura, T, Hattori, A., Ikeuchi, Y., and Tatsumi, R., In Vitro Measurement of Postnatal Changes in Proliferating Satellite Cell Frequency during Rat Muscle Growth, Animal Science Journal, 81, 2, 245-251, 2010.04.
68. Sato, Y., Probst, H. C., Tatsumi, R., Ikeuchi, Y., Neuberger, M. S., and Rada, C., Deficiency in APOBEC2 Leads to a Shift in Muscle Fiber Type, Diminished Body Mass and Myopathy, Journal of Biological Chemistry, 285, 7111-7118 , 2010.03.
69. Yamada, M., Tatsumi, R., Yamanouchi, K., Hosoyama, T., Shiratsuchi, S., Sato, A., Mizunoya, W., Ikeuchi, Y., Furuse, M., and Allen, R. E. , High Concentrations of HGF Inhibit Skeletal Muscle Satellite Cell Proliferation In Vitro By Inducing Expression of Myostatin: A Possible Mechanism for Re-Establishing Satellite Cell Quiescence In Vivo. , American Journal of Physiology-Cell Physiology, 298, C465–C476 , Editorial Focusに選定, 2010.03, 骨格筋の肥大・再生は筋幹細胞である衛星細胞の増殖活性に依存しており、これは衛星細胞の活性化と休止化のバランスによって制御されている。これまでに、肝細胞増殖因子HGFによる衛星細胞の活性化機構を明らかにしてきたが、これと逆反応の休止化もまたHGFによって誘導されることを見出した。物理刺激をトリガーにして、HGF濃度依存的に衛星細胞が活性化・増殖した後に自立的に休止化するという、独創的な時系列進行モデルを提起した。.
70. Ryuichi A Tatsumi, Mechano-Biology of Skeletal Muscle Hypertrophy and Regeneration: Possible Mechanism of Stretch-Induced Activation of Resident Myogenic Stem Cells, Animal Science Journal, 81, 11-20, invited review, 2010.01, 筋幹細胞である衛星細胞が物理刺激を引き金として活性化・増殖し筋肥大・再生に寄与する分子メカニズムを解明した一連の研究を、Animal Science Jouranlの招待総説論文として発表した。.
71. Ryuichi Tatsumi, Yoriko Sankoda, Judy E. Anderson, Yusuke Sato, Wataru Mizunoya, Naomi Shimizu, Takahiro Suzuki, Michiko Yamada, Robert P. Rhoads, Jr., Yoshihide Ikeuchi, and Ronald E. Allen, Possible Implication of Satellite Cells in Regenerative Motoneuritogenesis: HGF Up-Regulates Neural Chemorepellent Sema3A during Myogenic Differentiation, American Journal of Physiology-Cell Physiology, 297, C238-C252, Editorial Focusに選定, 2009.08.
72. Sato, Y., Shimizu, M., Mizunoya, W., Wariishi, H., Tatsumi, R., Buchman, V. L., and Ikeuchi, Y. , Differential Expression of Sarcoplasmic and Myofibrillar Proteins of Rat Soleus Muscle during Denervation Atrophy, Bioscience, Biotechnology and Biochemistry, 73, 1748-1756, 2009年度BBB論文賞を受賞, 2009.06.
73. Sato, Y., Shimizu, M., Mizunoya, W., Tatsumi, R., Probst, H., Rada, C., Neuberger, M. S., and Ikeuchi, Y., Skeletal Muscle Fiber Type Change and Myopathy in APOBEC-2 KO Mouse, A Joint meeting of Frontiers in Myogenesis and Skeletal Muscle Satellite and Stem Cells, 2009.05.
74. Yamada, M., Tatsumi, R., Mizunoya, W., Ikeuchi, Y., and Allen, R. E., HGF Induces Myostatin Expression and the Subsequent Satellite Cell Quiescence in a Neuropilin-1 Dependent Manner, A Joint meeting of Frontiers in Myogenesis and Skeletal Muscle Satellite and Stem Cells, 2009.05.
75. Tatsumi, R., Sankoda, Y., Anderson, J. E., Sato, Y., Mizunoya, W., Shimizu, N., Suzuki, T., Yamada, M., Rhoads, R. P., Jr., Ikeuchi, Y., and Allen, R. E. , A Possible Role for Satellite Cells in Regenerative Motoneurogenesis: HGF Up-Regulates Neural Chemorepellent Sema3A Expression, A Joint meeting of Frontiers in Myogenesis and Skeletal Muscle Satellite and Stem Cells, 2009.05.
76. Tatsumi, R., Wuollet, A. L., Tabata, K., Nishimura, S., Tabata, S., Mizunoya, W., Ikeuchi, Y., and Allen, R. E., A Role for Calcium-Calmodulin in Regulating Nitric Oxide Production during Skeletal Muscle Satellite Cell Activation, American Journal of Physiology-Cell Physiology, 296, C922-C929, 2009.03.
77. Yamada, M., Shiratsuchi, S.-I., Tatsumi, R., Mizunoya, W., Ikeuchi, Y., and Allen, R. E., A Receptor Neuropilin-1 Mediates HGF-Induced Muscle Satellite Cell Quiescence, The American Society for Cell Biology 48th Annual Meeting, 2008.12.
78. Yamada, M., Sankoda, Y., Tatsumi, R., Mizunoya, W., Ikeuchi, Y., Sunagawa, K., and Allen, R. E., Matrix Metalloproteinase-2 Mediates Stretch-Induced Activation of Skeletal Muscle Satellite Cells in A Nitric Oxide Dependent Manner, International Journal of Biochemistry and Cell Biology, 40, 2183-2191., 2008.07.
79. Mizunoya, W., Wakamatsu, J.-I., Tatsumi, R., and Ikeuchi, Y., Protocol for High-Resolution Separation of Rodent Myosin Heavy Chain Isoforms in a Mini-Gel Electrophoresis System, Analytical Biochemistry, 377, 111-113, 2008.06.
80. Sultana, A., Nakanishi, A., Roy, B.C., Mizunoya, W., Tatsumi, R., Ito, T., Tabata, S., Rashid, H., Katayama, S., and Ikeuchi, Y., Quality Improvement of Frozen and Chilled Beef Biceps Femoris with the Application of Salt-Bicarbonate Solution., Asian-Australasian Journal of Animal Sciences, 21, 903-911, 2008.05.
81. Tatsumi, R. and Allen, R.E., Mechano-Biology of Resident Myogenic Stem Cells: Molecular Mechanism of Stretch-Induced Activation of Satellite Cells., Animal Science Journal, 79, 279-290; invited review, 2008.05, 筋幹細胞である衛星細胞が物理刺激を引き金として活性化・増殖し筋肥大・再生に寄与する分子メカニズムを解明した一連の研究を、招待総説論文に紹介した。本論文は2008年に掲載され、その後の論文引用回数などが評価され2011年度の優秀論文賞を受賞した。.
82. Yamada, M., Sankoda, Y., Tatsumi, R., Mizunoya, W., Ikeuchi, Y., and Allen, R. E., Matrix Metalloproteinase-2 Is Involved In Mechanical Stretch-Induced Activation of Skeletal Muscle Satellite Cells., 2007 FASEB Summer Research Conference on “Skeletal Muscle Satellite & Stem Cells, 2007.07.
83. Sato, Y., Shimizu, M., Mizunoya, W., Wariishi, H., Tatsumi, R., and Ikeuchi, Y. , Differential Expression of Sarcoplasmic and Myofibrillar Proteins of Rat Soleus Muscle during Denervation Atrophy., Adult Skeletal Muscle Symposium: Growth, Function and Motility , 2007.06.
84. Mizunoya, Y., Okamoto, S., Wakamatsu, J.-I., Sonoda, Y., Sekiguchi, T., Waga, T., Tatsumi, R., and Ikeuchi, Y. , Food Components Do Affect Skeletal Muscle Mass and Fiber Types: Effects of Beef Extract and Apple Polyphenol. , Adult Skeletal Muscle Symposium: Growth, Function and Motility , 2007.06.
85. Mori, S., Yokoyama, A., Iguchi, R., Yamamoto, S., Suzuki, A., Mizunoya, W., Tatsumi, R., Yoshioka, K., and Ikeuchi, Y., Effect of High Pressure Treatment on Cytoplasmic 5'-Nucleotidase from Rabbit Skeletal Muscle., Journal of Food Biochemistry, 31 (3), 314-327, 2007.06.
86. Mori, S., Uchida, A., Yamamoto, S., Sultana, A., Tatsumi, R., Mizunoya, W., Suzuki, A., and Ikeuchi, Y., Effect of High Pressure on the Accumulation of IMP and on the Stability of AMP Deaminase in Rabbit Skeletal Muscle., Journal of Food Biochemistry, 31 (3), 328-342, 2007.06.
87. Tabata, S., Takemura, Y., Kobayashi, M., Ikeda, M., Nishimura, S., Sato, Y., Tatsumi, R., Ikeuchi, Y., and Iwamoto, H., Mechanism of Ca2+ Increase in Myoblasts Derived from Chicken Embryos., Journal of Electron Microscopy, 55(5):265-271, 2006.12.
88. Tatsumi, R., Yamada, M., Katsuki, Y., Okamoto, S., Ishizaki, J., Muzunoya, W., Ikeuchi, Y., Hattori, A., Shimokawa, H., and Allen, R. E., Low-pH Preparation of Skeletal Muscle Satellite Cells Can be Used to Study Activation In Vitro., International Journal of Biochemistry and Cell Biology, 38, 1678-1685, 2006.08.
89. Tatsumi, R., Liu, X., Pulido, A., Morales, M., Sakata, T., Dail, S., Hattori, A., Ikeuchi, Y., and Allen, R. E., Satellite Cell Activation in Stretched Skeletal Muscle and The Role of Nitric Oxide and Hepatocyte Growth Factor., American Journal of Physiology-Cell Physiology, 290, 1487-1494, 2006.06.
90. Yamada, M., Tatsumi, R., Kikuiri, T., Okamoto, S., Nonoshita, S., Mizunoya, W., Ikeuchi, Y., Shimokawa, H., Sunagawa, K., and Allen, R. E., Matrix Metalloproteinases are Involved in Mechanical Stretch-Induced Activation of Skeletal Muscle Satellite Cells., Muscle & Nerve, 34, 313-319, 2006.04.
91. Sakata, T., Tatsumi, R., Yamada, M., Shiratsuchi, S., Okamoto, S., Mizunoya, W., Hattori, A., and Ikeuchi, Y., Preliminary Experiments on Mechanical Stretch-Induced Activation of Skeletal Muscle Satellite Cells In Vivo., Animal Science Journal, 77, 518-525, 2006.04.
92. Yoshihara, H., Wakamatsu, J.-I., Kawabata, F., Mori, S., Haruno, H., Hayashi, T., Sekiguchi, T., Mizunoya, M., Tatsumi, R., Ito, T., and Ikeuchi1, Y., Beef Extract Supplementation Increases Leg Muscle Mass and Modifies Skeletal Muscle Fiber Types in Rats., Journal of Nutritional Science and Vitaminology, 52, 183-193, 2006.04.
93. Yamada, M., Tatsumi, R., Okamoto, S., Nonoshita, S., Mizunoya, W., Ikeuchi, Y., and Allen, R. E., Mechanical Stretch-Induced Activation of Skeletal Muscle Satellite Cells Requires Matrix Metalloproteinase Activity., FRONTIERS IN MYOGENESIS, the meeting of the Society for Muscle Biology, 2006.04.
94. Ryuichi Tatsumi, Keitaro Yamanouchi, Toru Hosoyama, Sei-ichi Shiratsuchi, Michiko Yamada, Jun-ichiro Wakamatsu, Wataru Mizunoya, Yoshihide Ikeuchi, and Ronald E. Allen, A Possible Mechanism of Muscle Satellite Cell Quiescence: HGF Induces Myostatin Expression and Secretion., FASEB Summer Research Conference 2005, 4rd International Conference on Skeletal Muscle & Stem Cells, 2005.06.
95. Tatsumi, R. and Allen, R.E., Active Hepatocyte Growth Factor Is Present in Skeletal Muscle Extracellular Matrix, Muscle & Nerve, 10.1002/mus.20114, 30, 5, 654-658, 30, 654-658, 2004.11.
96. Mendias, C., Tatsumi, R., and Allen, R.E., Role of Cyclooxygenase-1 and -2 in Satellite Cell Proliferation, Differentiatio, and Fusion, Muscle & Nerve, 10.1002/mus.20102, 30, 4, 497-500, 30, 497-500, 2004.10.
97. Tatsumi, R., Mitsuhashi, K., Ashida, K., Haruno, A., Hattori, A., Ikeuchi, Y., and Allen, R.E., Comparative Analysis of Mechanical Stretch-Induced Activation Activity of Back and Leg Muscle Satellite Cells In Vitro, Animal Science Journal, 75, 345-351, 2004.08.
98. Mori, S., Haruno, A., Hayashi, T., Haga, S., Iida, S., Tatsumi, R., Ito, T., and Ikeuchi, Y., Rheological Properties of Emulsion-Type Sausage Prepared from Deer Meat, The West Japan Journal of Animal Science, 47, 87-91, 2004.07.
99. Liaw, K.J.-F., Mori, S., Hashimoto, M., Sugimitsu, M., Hayashi, T., Yamanoue, M., Tatsumi, R., Ikeuchi, Y., and Ito, T., Chemical Cross-Linking of Actin and Myosin Subfragment-1 in Rigor Complex, Journal of the Faculty of Agriculture Kyushu University, 49, 1, 111-118, 49, 111-118, 2004.04.
100. Ito, Y., Toki, S., Omori, T., Ide, H., Tatsumi, R., Wakamatsu, J.-I., Nishimura, T., and Hattori, A., Physicochemical Properties of Water-Soluble Myofibrikkar Proteins Prepared from Chicken Breast Muscle, Animal Science Joural, 75、59-65, 2004.01.
101. Ito, Y., Tatsumi, R., Wakamatsu J.-I., Nishimura, T., and Hattori, A., The Solubilization of Myofibrillar Proteins of Vertebrate Skeletal Muscle in Water, Animal Science Journal, 74, 417-425, 2003.12.
102. Tatsumi, R., and Takahashi, K., Structural Changes in Titin and Nebulin Filaments Specific to Calcium Ions at 0.1 mM: Factors of Meat Tenderization during Postmortem Aging, Journal of Food Science, 68, 756-760, 2003.12.
103. Hattori, A., Fujimoto, S., Nishimura, T., Tatsumi, R., Wakamatsu, J., and Ito, T., Localization of Paratropomyosin in Chicken Skeletal Muscle Myofibrils and Its Behavior during Postmortem Storage of Muscles Revealed by Immunoelectron Microscopy
Journal of Muscle Foods 14, 1-10., Journal of Muscle Foods, 14, 1-10, 2003.12.
104. Tatsumi, R. and Allen, R.E., Satellite Cell Activation in Response to Muscle Stretch and Damage, FASEB Summer Research Conference 2003, 3rd International Conference on Muscle Satellite & Stem Cells, 2003.07.
105. Allen, R.E. and Tatsumi, R., Satellite Cell Activation in Response to Muscle Damage, Molecular Biology of Muscle Development and Regeneration, 2003.05.
106. Tatsumi, R., Hattori, A., Ikeuchi, Y., Anderson, J. E., and Allen, R. E., Release of Hepatocyte Growth Factor from Mechanical Stretched Skeletal Muscle Satellite Cells and the Role of pH and Nitric Oxide, Molecular Biology of the Cell, 10.1091/mbc.E02-01-0062, 13, 8, 2909-2918, 13, 2909-2918, 2002.12.
107. Tatsumi, R., Hattori, A., Allen, R. E., Ikeuchi Y., and Ito, T., Mechanical Stretch-Induced Activation of Skeletal Muscle Satellite Cells Is Dependent on Nitric Oxide Production, Animal Science Journal, 73, 235-239, 2002.12.
108. Ikeuchi, Y., Suzuki, A., Oota, T., Hagiwara, K., Tatsumi, R., Ito, T., and Balny, C., Fluorescence Study of the High Pressure-Induced Denaturation of Skeletal Muscle Actin, European Journal of Biochemistry, 10.1046/j.0014-2956.2001.02664.x, 269, 1, 364-371, 269, 364-371, 2002.12.
109. Ogawa, T., Hayashi, T., Nodake, K., Shiraishi, S., Mori, S., Tatsumi, R., Ikeuchi, Y., and Ito, T., A Novel Muscle Cell Growth Factor, S-Myotrophin, Promotes the Expression of Fast Myosin in Myotubes Developed from C2C12 Cells, 48th International Congress of Meat Science and Technology, 2002.08.
110. Tatsumi, R., Maeda, K., Hattori, A., and Takahashi, K., Calcium Binding to an Elastic Portion of Connectin/Titin Filaments, Journal of Muscle Research and Cell Motility, 22, 149-162, 2001.12.
111. Tatsumi, R., Sheehan, S. M., Iwasaki, H., Hattori, A., and Allen, R. E., Mechanical Stretch Induces Activation of Skeletal Muscle Satellite Cells In Vitro, Experimental Cell Research, 267, 107-114, 2001.12.
112. Tatsumi, R., Sakata, T., Sheehan, S.M., Hattori, A., and Allen, R.E., Mechanical Stretch-Induced Activation of Skeletal Muscle Satellite Cells: the Possible Role of HGF in the Mechanism, FASEB Summer Research Conference 2000, 2nd International Conference on Muscle Satellite & Stem Cells, 2001.07.
113. Sheehan, S. M., Tatsumi, R., Temm-Grove, C.J., and Allen, R. E., HGF is an Autocrine Growth Factor for Skeletal Muscle Satellite Cells In Vitro, Muscle & Nerve, 23, 239-245, 2000.12.
114. Tatsumi, R., Sheehan, S.M., Iwasaki, H., Hattori, A., and Allen, R.E., Mechanical Stretch Induces Activation of Skeletal Muscle Satellite Cells In Vitro., 2000 Keystone Symposia on the Molecular Biology of Muscle Development and Disease, 2000.05.
115. Tatsumi, R., Maeda, K., and Takahashi, K., Calcium-Induced Splitting of Connectin/Titin Filaments: Localization and Characterization of Calcium-Binding Sites, 45th International Congress of Meat Science and Technology, 1999.08.
116. Ito, Y., Tatsumi, R., Nishimura, T., and Hattori, A., Solubilization of Myofibrillar Proteins of Chicken Skeletal Muscle in Water, 45th International Congress of Meat Science and Technology, 1999.08.
117. Tatsumi, R., Hattori, A., and Takahashi, K., Deterioration of Connectin/Titin and Nebulin Filaments by an Excess of Protease Inhibitors, Bioscience, Biotechnology and Biochemistry, 62, 927-934, 1998.12.
118. Tatsumi, R., Anderson, J. E., Nevoret, C. J., Halevy, O., and Allen, R. E., HGF/SF is Present in Normal Adult Skeletal Muscle and Is Capable of Activating Satellite Cells, Developmental Biology, 194, 114-128., 1998.12.
119. Allen, R.E. and Tatsumi, R., Satellite Cell Activation and the Role of Hepatocyte Growth Factor/Scatter Factor, Post-natal Myogenesis: Satellite Cells In Action, 1998.08.
120. Tatsumi, R., Shimada, K., and Hattori, A., Fluorescence Detection of Calcium-Binding Proteins with Quinoline Ca-Indicator Quin2, Analytical Biochemistry, 254, 126-131, 1997.01.
121. Tatsumi, R., Hattori, A., and Takahashi, K., Splitting of Connectin/Titin Filaments into β-Connectin/T2 and a 1,200-kDa Subfragment by 0.1 mM Calcium Ions, Advances in Biophysics, 33, 65-77, 1996.01.
122. Hattori, A., Wakamatsu, J.-I., Ishii, T., Kuwahara, K., and Tatsumi, R., Characterization of a Novel 550-kDa Protein in Skeletal Muscle of Chick Embryo, Biochimica et Biophysica Acta, 1245, 201-206, 1995.01.
123. Hattori, A., Wakamatsu, J.-I., Ishii, T., Kuwahara, K., and Tatsumi, R., A Novel 550-kDa Protein in Skeletal Muscle of Chick Embryo: Purification and Localization, Biochimica et Biophysica Acta, 1245, 191-200, 1995.01.
124. Hattori, A., Ishii, T., Tatsumi, R., and Takahashi, K., Changes in the Molecular Types of Connectin and Nebulin during Development of Chicken Skeletal Muscle, Biochimica et Biophysica Acta, 1244, 179-184, 1995.01.
125. Tatsumi, R. and Hattori, A., Detection of Giant Myofibrillar Proteins Connectin and Nebulin by Electrophoresis in 2% Polyacrylamide Slab Gels Strengthened with Agarose, Analytical Biochemistry, 224, 28-31, 1995.01.
126. Tanabe, R., Tatsumi, R., and Takahashi, K., Purification and Characterization of the 1,200-kDa Subfragment of Connectin Filaments Produced by 0.1 mM Calcium Ions, Journal of Biochemistry, 115, 351-355, 1994.01.
127. Tatsumi, R., Hattori, A., and Takahashi, K., Substructure of Nebulin Filaments: Localization and Characterization of Subfragments Produced by 0.1 mM CaCl2, Journal of Biochemistry, 113, 797-804, 1993.01.
128. Tatsumi, R., Hattori, A., and Takahashi, K., Purification and Characterization of Nebulin Subfragments Produced by 0.1 mM CaCl2, Journal of Biochemistry, 112, 780-785, 1992.01.
129. Tatsumi, R. and Takahashi, K., Calcium-Induced Fragmentation of Skeletal Muscle Nebulin Filaments, Journal of Biochemistry, 112, 775-779, 1992.01.
130. Takahashi, K., Hattori, A., Tatsumi, R., and Takai, K., Calcium-Induced Splitting of Connectin Filaments into β-Connectin and a 1,200-kDa Subfragment, Journal of Biochemistry, 111, 778-782, 1992.01.