Kyushu University Academic Staff Educational and Research Activities Database
List of Papers
Suzuki Takahiro Last modified date:2022.05.09

Associate Professor / Department of Animal and Marine Bioresource Sciences / Department of Bioresource Sciences / Faculty of Agriculture


Papers
1. Takahiro Suzuki, Aika Mori, Takahiro Maeno, Rio Arimatsu, Emi Ichimura, Yuriko Nishi, Kouga Hisaeda, Yuki Yamaya, Ken Kobayashi, Mako Nakamura, Ryuichi Tatsumi, Koichi Ojima, 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, 10.3390/ijms22094499, 22, 9, 4499-4499, 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..
2. 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, The Journal of Biochemistry, 10.1093/jb/mvab030, 2021.03, Abstract
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..
3. Yusaku Tsugami, Haruka Wakasa, Manabu Kawahara, Atsushi Watanabe, Takahiro Suzuki, Takanori Nishimura, Ken Kobayashi, Adverse effects of LPS on membrane proteins in lactating bovine mammary epithelial cells, Cell and Tissue Research, 10.1007/s00441-020-03344-0, 2021.01.
4. Ken Kobayashi, Yusaku Tsugami, Norihiro Suzuki, Takahiro Suzuki, Takanori Nishimura, Suppressive effects of curcumin on milk production without inflammatory responses in lactating mammary epithelial cells, Phytomedicine, 10.1016/j.phymed.2020.153360, 80, 153360-153360, 2021.01.
5. Norihiro Suzuki, Yusaku Tsugami, Haruka Wakasa, Takahiro Suzuki, Takanori Nishimura, Ken Kobayashi, Menthol from Mentha piperita Suppresses the Milk Production of Lactating Mammary Epithelial Cells In Vivo and In Vitro, Molecular Nutrition & Food Research, 10.1002/mnfr.202000853, 64, 24, 2000853-2000853, 2020.12.
6. Shiori Tsutsui, Haruka Wakasa, Yusaku Tsugami, Takahiro Suzuki, Takanori Nishimura, Ken Kobayashi, Distinct Expression Patterns of Fibrillar Collagen Types I, III, and V in Association with Mammary Gland Remodeling during Pregnancy, Lactation and Weaning, Journal of Mammary Gland Biology and Neoplasia, 10.1007/s10911-020-09457-0, 25, 3, 219-232, 2020.09.
7. Yusaku Tsugami, Norihiro Suzuki, Takahiro Suzuki, Takanori Nishimura, Ken Kobayashi, Regulatory Effects of Soy Isoflavones and Their Metabolites in Milk ProductionviaDifferent Ways in Mice, Journal of Agricultural and Food Chemistry, 10.1021/acs.jafc.0c01288, 68, 21, 5847-5853, 2020.05.
8. Naomi Watanabe, Yusuke Komiya, Yusuke Sato, Yusuke Watanabe, Takahiro Suzuki, Keizo Arihara, Oleic acid up-regulates myosin heavy chain (MyHC) 1 expression and increases mitochondrial mass and maximum respiration in C2C12 myoblasts, Biochemical and Biophysical Research Communications, 10.1016/j.bbrc.2020.02.099, 525, 2, 406-411, 2020.04.
9. Yusaku Tsugami, Norihiro Suzuki, Manabu Kawahara, Takahiro Suzuki, Takanori Nishimura, Ken Kobayashi, Establishment of an in vitro culture model to study milk production and the blood–milk barrier with bovine mammary epithelial cells, Animal Science Journal, 10.1111/asj.13355, 91, 1, 2020.03.
10. Kobayashi, K., Tsugami, Y., Suzuki, N., Suzuki, T., and Nishimura, T., Nicotine directly affects milk production in lactating mammary epithelial cells concurrently with inactivation of STAT5 and glucocorticoid receptor in vitro., Toxicology in Vitro, 10.1016/j.tiv.2019.104741, 63, 104741, 2020.03.
11. Aogu Kumai, Yusaku Tsugami, Haruka Wakasa, Norihiro Suzuki, Takahiro Suzuki, Takanori Nishimura, Ken Kobayashi, Adverse Effects of Coumestrol and Genistein on Mammary Morphogenesis and Future Milk Production Ability of Mammary Epithelial Cells, Advanced Biosystems, 10.1002/adbi.201900187, 4, 4, 1900187-1900187, 2020.02.
12. Yoshiki Kaihoko, Yusaku Tsugami, Norihiro Suzuki, Takahiro Suzuki, Takanori Nishimura, Ken Kobayashi, Distinct expression patterns of aquaporin 3 and 5 in ductal and alveolar epithelial cells in mouse mammary glands before and after parturition, Cell and Tissue Research, 10.1007/s00441-020-03168-y, 380, 3, 513-526, 2020.01.
13. Kangmin Seo, Takahiro Suzuki, Ken Kobayashi, Takanori Nishimura, Adipocytes suppress differentiation of muscle cells in a co-culture system, Animal Science Journal, 10.1111/asj.13145, 90, 3, 423-434, 2019.03.
14. Matsunaga K, Tsugami Y, Kumai A, Suzuki T, Nishimura T, Kobayashi K, IL-1β directly inhibits milk lipid production in lactating mammary epithelial cells concurrently with enlargement of cytoplasmic lipid droplets., Experimental cell research, 10.1016/j.yexcr.2018.06.038, 370, 2, 365-372, 2018.09.
15. Ojima, K, Ichimura, E, Suzuki, T, Oe, M, Muroya, S, Nishimura, T, HSP90 modulates the myosin replacement rate in myofibrils., American Journal of Physiology-Cell Physiology, 10.1152/ajpcell.00245.2017, 315, 1, C104-C114, 2018.07.
16. Ken Kobayashi, Yusaku Tsugami, Kota Matsunaga, Takahiro Suzuki, Takahiro Nishimura, Moderate High Temperature Condition Induces the Lactation Capacity of Mammary Epithelial Cells Through Control of STAT3 and STAT5 Signaling, Journal of Mammary Gland Biology and Neoplasia, 10.1007/s10911-018-9393-3, 23, 1-2, 75-88, 2018.06.
17. Hideaki Ohtsubo, Yusuke Sato, Yuji Matsuyoshi, Takahiro Suzuki, Wataru Mizunoya, Mako Nakamura, Ryuichi Tatsumi, Yoshihide Ikeuchi, Fluorescence microscopy data on expression of Paired Box Transcription Factor 7 in skeletal muscle of APOBEC2 knockout mice, Data in Brief, 10.1016/j.dib.2018.02.063, 17, 1348-1351, 2018.04, The data presented in this article are related to the research articles entitled “APOBEC2 negatively regulates myoblast differentiation in muscle regeneration” and “Data supporting possible implication of APOBEC2 in self-renewal functions of myogenic stem satellite cells: toward understanding the negative regulation of myoblast differentiation” (Ohtsubo et al., 2017a, 2017b) [1,2]. This article provides in vivo phenotypical data to show that Paired Box Transcription Factor 7 (Pax7)-positive cell number (per myofiber) is significantly lower in APOBEC2 (a member of apoB mRNA editing enzyme, catalytic polypeptide-like family)-knockout muscle than the control wild-type tissue at the same age of 8-wk-old in mice. The emerging results support an essential role for APOBEC2 in the self-renewal functions of myogenic stem satellite cells, namely the re-establishment of quiescent status after activation and proliferation of myoblasts..
18. Study of dietary fat and skeletal muscle fiber type..
19. Yusaku Tsugami, Kota Matsunaga, Takahiro Suzuki, Takanori Nishimura, Ken Kobayashi, Phytoestrogens Weaken the Blood-Milk Barrier in Lactating Mammary Epithelial Cells by Affecting Tight Junctions and Cell Viability, JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 10.1021/acs.jafc.7b04786, 65, 50, 11118-11124, 2017.12, During lactation, mammary epithelial cells (MECs) form the blood-milk barrier by less-permeable tight junctions (TJs) to prevent the leakage of milk components. Phytoestrogens affect the proliferation, differentiation, and apoptosis of MECs. However, it remains unclear whether phytoestrogens are involved in the blood-milk barrier. Therefore, we investigated the influence of phytoestrogens (coumestrol, genistein, and daidzein) by using an in vitro mouse-MEC-culture model. The results showed that coumestrol and genistein changed the expression of TJ proteins (claudins-3 and -4 and occludin), weakened barrier function, and reduced beta-casein production. Daidzein also weakened barrier function without inhibiting beta-casein production. Additionally, coumestrol and genistein induced apoptosis in MECs. These results indicate that phytoestrogens weaken the blood-milk barrier by directly affecting TJs and the cellular viability of lactating MECs in different ways..
20. Koichi Ojima, Emi Ichimura, Yuya Yasukawa, Mika Oe, Susumu Muroya, Takahiro Suzuki, Jun-ichi Wakamatsu, Takanori Nishimura, Myosin substitution rate is affected by the amount of cytosolic myosin in cultured muscle cells, ANIMAL SCIENCE JOURNAL, 10.1111/asj.12826, 88, 11, 1788-1793, 2017.11, In striated muscles, approximately 300 myosin molecules form a single thick filament in myofibrils. Each myosin is continuously displaced by another myosin to maintain the thick filament structure. Our previous study using a fluorescence recovery after photobleaching (FRAP) technique showed that the myosin replacement rate is decreased by inhibition of protein synthesis, but myosin is still exchangeable. This result prompted us to examine whether myosin in the cytoplasm is involved in myosin replacement in myofibrils. To address this, FRAP was measured in green fluorescent protein (GFP)-tagged myosin heavy chain 3 (Myh3) expressing myotubes that were treated with streptolysin-O (SLO), which forms pores specifically in the plasma membrane to induce leakage of cytoplasmic proteins. Our biochemical data demonstrated that the cytoplasmic myosin content was reduced in SLO-permeabilized semi-intact myotubes. Furthermore, FRAP experiments showed a sluggish substitution rate of GFP-Myh3 in SLO-permeabilized myotubes. Taken together, these results demonstrate that the myosin substitution rate is significantly reduced by a decreased amount of myosin in the cytoplasm and that cytoplasmic myosin contributes to myosin replacement in myofibrils..
21. Yusaku Tsugami, Kota Matsunaga, Takahiro Suzuki, Takanori Nishimura, Ken Kobayashi, Isoflavones and their metabolites influence the milk component synthesis ability of mammary epithelial cells through prolactin/STAT5 signaling, MOLECULAR NUTRITION & FOOD RESEARCH, 10.1002/mnfr.201700156, 61, 10, 1700156, 2017.10, Scope: Isoflavones are a class of polyphonic compounds present in legumes and are called phytoestrogens because of their estrogen-like activity. Estrogen influences the behavior of mammary epithelial cells (MECs) during pregnancy and lactation. In this study, we investigated the direct influences of isoflavones and their metabolites in milk production ability of MECs.
Methods and results: Mouse MECs were cultured with prolactin and dexamethasone (glucocorticoid analog) to induce milk production ability. Subsequently, lactating MECs were treated with each isoflavone. Coumestrol, biochanin A, genistein, and formononetin decreased the intracellular and secreted beta-casein. On the other hand, p-ethylphenol, daidzein, and equol did not significantly influence beta-casein production at any concentration. Coumestrol, biochanin A and genistein down-regulated the mRNA expression of whey acidic protein (WAP), lactoferrin and alpha-lactalbumin. In contrast, p-ethylphenol, daidzein and equol up-regulated beta-casein and/or WAP with alpha-lactalbumin. Furthermore, coumestrol and genistein down-regulated the expression of prolactin receptor and signal transducer and activator of transcription 5 (STAT5) accompanied by a decrease in STAT5 phosphorylation.
Conclusion: Isoflavones and their metabolites influence the milk production ability of MECs through different interactions with prolactin/STAT5 signaling. Simultaneous intake of multiple isoflavones by consumption of legumes may induce promotive or adverse effects on lactating MECs..
22. Judy E. Anderson, Mai-Khoi Q. Do, Nasibeh Daneshvar, Takahiro Suzuki, Junio Dort, Wataru Mizunoya, Ryuichi Tatsumi, The role of semaphorin3A in myogenic regeneration and the formation of functional neuromuscular junctions on new fibres, BIOLOGICAL REVIEWS, 10.1111/brv.12286, 92, 3, 1389-1405, 2017.08, Current research on skeletal muscle injury and regeneration highlights the crucial role of nerve-muscle interaction in the restoration of innervation during that process. Activities of muscle satellite or stem cells, recognized as the 'currency' of myogenic repair, have a pivotal role in these events, as shown by ongoing research. More recent investigation of myogenic signalling events reveals intriguing roles for semaphorin3A (Sema3A), secreted by activated satellite cells, in the muscle environment during development and regeneration. For example, Sema3A makes important contributions to regulating the formation of blood vessels, balancing bone formation and bone remodelling, and inflammation, and was recently implicated in the establishment of fibre-type distribution through effects on myosin heavy chain gene expression. This review highlights the active or potential contributions of satellite-cell-derived Sema3A to regulation of the processes of motor neurite ingrowth into a regenerating muscle bed. Successful restoration of functional innervation during muscle repair is essential; this review emphasizes the integrative role of satellite-cell biology in the progressive coordination of adaptive cellular and tissue responses during the injury-repair process in voluntary muscle..
23. Tatsumi R, Suzuki T, Do MQ, Ohya Y, Anderson JE, Shibata A, Kawaguchi M, Ohya S, Ohtsubo H, Mizunoya W, Sawano S, Komiya Y, Ichitsubo R, Ojima K, Nishimatsu SI, Nohno T, Ohsawa Y, Sunada Y, Nakamura M, Furuse M, Ikeuchi Y, Nishimura T, Yagi T, Allen RE, Slow-Myofiber Commitment by Semaphorin 3A Secreted from Myogenic Stem Cells., Stem cells, 10.1002/stem.2639, 35, 7, 1815-1834, 2017.07.
24. 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, 12, 269-273, 2017.06, This paper 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..
25. 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 & CELL BIOLOGY, 10.1016/j.bioce1.2017.02.005, 85, 91-101, 2017.04, 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. (C) 2017 Elsevier Ltd. All rights reserved..
26. 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, 2017.03, 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<0.05). There was a trend to increased fatigue resistance detected from measurements of relative isometric plantar-flexion force torque generated by a stimulus train delivered to the tibial nerve (F(98, 1372)=1.246, P=0.0574). Importantly, there was no significant difference in the animal body-phenotypes or locomotor activity shown as total moving distance in light and dark periods. Therefore, the present study encourages the notion that even low APP-intake may increase the proportions of fatigue-resistant myofibers, and has promise as a strategy for modifying performance in human sports and improving function in age-related muscle atrophy..
27. Kobayashi, K, Oyama, S, Kuki, C, Tsugami, Y, Matsunaga, K, Suzuki, T, Nishimura, T, Distinct roles of prolactin, epidermal growth factor, and glucocorticoids in β-casein secretion pathway in lactating mammary epithelial cells., Molecular and Cellular Endocrinology, 10.1016/j.mce.2016.11.006, 440, C, 16-24, 2017.01.
28. 辰巳隆一, 鈴木貴弘, 大屋雄暉, ド クイ マイコイ, 大坪秀明, 川口舞, アンダーソン ジュディー, 水野谷航, 小宮佑介, チャハラ モクラン, 尾嶋孝一, 澤野祥子, 中村真子, 古瀬充宏, 池内義秀, アレン ロナルド, 巳隆一, 鈴木貴弘, 大屋雄暉, ド クイ マイコイは等しい, Experiments reveal a novel mechanism to regulate myofiber types and its activation by functional food ingredients., 栄養生理研究会報, 60, 2, 69-77, 2016.07.
29. Naoki Katase, Kumiko Terada, Takahiro Suzuki, Shin-ichiro Nishimatsu, Tsutomu Nohno, miR-487b, miR-3963 and miR-6412 delay myogenic differentiation in mouse myoblast-derived C2C12 cells (vol 16, 13, 2015), BMC CELL BIOLOGY, 10.1186/s12860-016-0083-y, 17, 1, 9, 2016.03.
30. 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-e12553, 2015.09.
31. 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 < 0.05). Results were demonstrated by our SDS-PAGE system specialized for MyHC isoform separation and western blotting of whole muscles. Animal-growth profiles (food intake, body-weight gain, and internal-organ weights) did not differ between the control and 5% APP-fed animals (n = 9/group). Findings may account for the increase in fatigue resistance of lower hind limb muscles, as evidenced by a slower decline in the maximum isometric planter-flexion torque generated by a 100-s train of electrical stimulation of the tibial nerve. Additionally, the fatigue resistance was lower after 8 weeks of a 0.5% APP diet than after 5% APP, supporting an APP-dose dependency of the shift in fiber-type composition. Therefore, the present study highlights a promising contribution of dietary APP intake to increasing endurance based on fiber-type composition in rat muscle. Results may help in developing a novel strategy for application in animal sciences, and human sports and age-related health sciences..
32. 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. However, in understanding this concept 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 the early phase of muscle regeneration..
33. 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 Tatsumi, Implication of anti-inflammatory macrophages in regenerative moto-neuritogenesis: Promotion of myoblast migration and neural chemorepellent semaphorin 3A expression in injured muscle, The International Journal of Biochemistry & Cell Biology, 10.1016/j.biocel.2014.05.032, 54, 272-285, 2014.09.
34. Autonomous determination of muscle fiber-types by semaphorin 3A secreted from resident myogenic stem cells..
35. Jun-ichi Shono, Shohei Sakaguchi, Takahiro Suzuki, Mai-Khoi Q. Do, Wataru Mizunoya, Mako Nakamura, Yusuke Sato, Mitsuhiro Furuse, Koji Yamada, Yoshihide Ikeuchi, Ryuichi 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 pro-inflammatory and anti-inflammatory 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 better reflect the physiologic damage and repair processes initiated by contusing a gastrocnemius muscle in the 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, characterized by increasing levels of myogenin messenger RNA 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 growth factor at the injury site early in muscle regeneration. © 2013 Japanese Society of Animal Science..
36. Takahiro Suzuki, Mai-Khoi Q. Do, Yusuke Sato, Koichi Ojima, Minako Hara, Wataru Mizunoya, Mako Nakamura, Mitsuhiro Furuse, Yoshihide Ikeuchi, Judy E. Anderson, Ryuichi Tatsumi, Comparative analysis of semaphorin 3A in soleus and EDL muscle satellite cells in vitro toward understanding its role in modulating myogenin expression, The International Journal of Biochemistry & Cell Biology, 10.1016/j.biocel.2012.10.003, 45, 2, 476-482, 2013.02.
37. Yusuke SATO, Mai-Khoi Q. DO, Takahiro SUZUKI, Hideaki OHTSUBO, Wataru MIZUNOYA, Mako NAKAMURA, Mitsuhiro FURUSE, Yoshihide IKEUCHI, Ryuichi TATSUMI, Satellite cells produce neural chemorepellent semaphorin 3A upon muscle injury, Animal Science Journal, 10.1111/asj.12014, 84, 2, 185-189, 2013.02.
38. Do, M-KQ, Suzuki, T, Gerelt, B, Sato, Y, Mizunoya, W, Nakamura, M, Ikeuchi, Y, Anderson, JE, Tatsumi, R, Time-coordinated prevalence of extracellular HGF, FGF2 and TGFβ3 in crush-injured skeletal muscle., Animal Science Journal, 10.1111/j.1740-0929.2012.01057.x, 83, 10, 712-717, 2012.10.
39. 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, 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, 10.1152/ajpcell.00068.2012, 302, 12, C1741-C1750, 2012.06, Hara M, Tabata K, Suzuki T, Do MQ, Mizunoya W, Nakamura M, Nishimura S, Tabata S, Ikeuchi Y, Sunagawa K, Anderson JE, Allen RE, Tatsumi R. Calcium influx through a possible coupling of cation channels impacts skeletal muscle satellite cell activation in response to mechanical stretch. Am J Physiol Cell Physiol 302: C1741-C1750, 2012. First published March 28, 2012; doi:10.1152/ajpcell.00068.2012.-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 mechanosensitive 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-h stretch cultures with individual inhibitors for MS and L-VGC channels (GsMTx-4 and nifedipine, respectively) or with a less specific inhibitor (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 mu M (GsMTx-4), 10 mu M (nifedipine), or 100 mu M (Gd) and canceled by addition of HGF to the culture media; a potent inhibitor for transient-type VGC channels (NNC55-0396, 100 mu M) 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 initiate the satellite cell activation cascade..
40. Mai-Khoi Q. Do, Yusuke Sato, Naomi Shimizu, Takahiro Suzuki, Jun-ichi Shono, Wataru Mizunoya, Mako Nakamura, Yoshihide Ikeuchi, Judy E. Anderson, Ryuichi Tatsumi, Growth factor regulation of neural chemorepellent Sema3A expression in satellite cell cultures, AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY, 10.1152/ajpcell.00257.2011, 301, 5, C1270-C1279, 2011.11, Do MKQ, Sato Y, Shimizu N, Suzuki T, Shono J, Mizunoya W, Nakamura M, Ikeuchi Y, Anderson JE, Tatsumi R. Growth factor regulation of neural chemorepellent Sema3A expression in satellite cell cultures. Am J Physiol Cell Physiol 301: C1270-C1279, 2011. First published August 24, 2011; doi: 10.1152/ajpcell.00257.2011.-Successful regeneration and remodeling of the intramuscular motoneuron network and neuromuscular connections are critical for restoring skeletal muscle function and physiological properties. The regulatory signals of such coordination remain unclear, although axon-guidance molecules may be involved. Recently, satellite cells, resident myogenic stem cells positioned beneath the basal lamina and at high density at the myoneural junction regions of mature fibers, were shown to upregulate a secreted neural chemorepellent semaphorin 3A (Sema3A) in response to in vivo muscle-crush injury. The initial report on that expression centered on the observation that hepatocyte growth factor (HGF), an essential cue in muscle fiber growth and regeneration, remarkably upregulates Sema3A expression in early differentiated satellite cells in vitro [Tatsumi et al., Am J Physiol Cell Physiol 297: C238-C252, 2009]. Here, we address regulatory effects of basic fibroblast growth factor (FGF2) and transforming growth factor (TGF)-beta s on Sema3A expression in satellite cell cultures. When treated with FGF2, Sema3A message and protein were upregulated as revealed by reverse transcription-polymerase chain reaction and immunochemical studies. Sema3A upregulation by FGF2 was dose dependent with a maximum (8- to 1-fold relative to the control) at 2.5 ng/ml (150 pM) and occurred exclusively at the early differentiation stage. The response was highly comparable in dose response and timing to effects of HGF treatment, without any additive or synergistic effect from treatment with a combination of both potent upregulators. In contrast, TGF-beta 2 and -beta 3 potently decreased basal Sema3A expression; the maximum effect was at very low concentrations (40 and 8 pM, respectively) and completely cancelled the activities of FGF2 and HGF to upregulate Sema3A. These results therefore encourage the prospect that a time-coordinated increase in HGF, FGF2, and TGF-beta ligands and their receptors promotes a programmed strategy for Sema3A expression that guarantees successful intramuscular motor reinnervation by delaying sprouting and reattachment of motoneuron terminals onto damaged muscle fibers early in regeneration pending restoration of muscle fiber contractile integrity..
41. Takahiro SUZUKI, Hideyuki TAKAISHI, Tomowa SAKATA, Mai-Khoi Q. DO, Minako HARA, Akiko SATO, Wataru MIZUNOYA, Takanori NISHIMURA, Akihito HATTORI, Yoshihide IKEUCHI, Ryuichi TATSUMI, In vitromeasurement of post-natal changes in proliferating satellite cell frequency during rat muscle growth, Animal Science Journal, 10.1111/j.1740-0929.2009.00734.x, 81, 2, 245-251, 2010.04.
42. Ryuichi Tatsumi, Yoriko Sankoda, Judy E. Anderson, Yusuke Sato, Wataru Mizunoya, Naomi Shimizu, Takahiro Suzuki, Michiko Yamada, Robert P. Rhoads, Yoshihide Ikeuchi, Ronald E. Allen, Possible implication of satellite cells in regenerative motoneuritogenesis: HGF upregulates neural chemorepellent Sema3A during myogenic differentiation, AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY, 10.1152/ajpcell.00161.2009, 297, 2, C238-C252, 2009.08, Tatsumi R, Sankoda Y, Anderson JE, Sato Y, Mizunoya W, Shimizu N, Suzuki T, Yamada M, Rhoads RP Jr, Ikeuchi Y, Allen RE. Possible implication of satellite cells in regenerative motoneuritogenesis: HGF upregulates neural chemorepellent Sema3A during myogenic differentiation. Am J Physiol Cell Physiol 297: C238-C252, 2009. First published June 10, 2009; doi: 10.1152/ajpcell.00161.2009.-Regenerative coordination and remodeling of the intramuscular motoneuron network and neuromuscular connections are critical for restoring skeletal muscle function and physiological properties. The regulatory mechanisms of such coordination remain unclear, although both attractive and repulsive axon guidance molecules may be involved in the signaling pathway. Here we show that expression of a neural secreted chemorepellent semaphorin 3A (Sema3A) is remarkably upregulated in satellite cells of resident myogenic stem cells that are positioned beneath the basal lamina of mature muscle fibers, when treated with hepatocyte growth factor (HGF), established as an essential cue in muscle fiber growth and regeneration. When satellite cells were treated with HGF in primary cultures of cells or muscle fibers, Sema3A message and protein were upregulated as revealed by reverse transcription-polymerase chain reaction and immunochemical studies. Other growth factors had no inductive effect except for a slight effect of epidermal growth factor treatment. Sema3A upregulation was HGF dose dependent with a maximum (about 7-to 8-fold units relative to the control) at 10-25 ng/ml and occurred exclusively at the early-differentiation stage, as characterized by the level of myogenin expression and proliferation (bromodeoxyuridine incorporation) of the cells. Neutralizing antibody to the HGF-specific receptor, c-met, did not abolish the HGF response, indicating that c-met may not mediate the Sema3A expression signaling. Finally, in vivo Sema3A was upregulated in the differentiation phase of satellite cells isolated from muscle regenerating following crush injury. Overall, the data highlight a heretofore unexplored and active role for satellite cells as a key source of Sema3A expression triggered by HGF, hence suggesting that regenerative activity toward motor innervation may importantly reside in satellite cells and could be a crucial contributor during postnatal myogenesis..