Updated on 2024/05/14

写真a

 
MUROYA Susumu
 
Organization
Research Field in Veterinary Medicine, Agriculture, Fisheries and Veterinary Medicine Area Joint faculty of Veterinary Medicine Department of Animal Science and Welfare Professor
Title
Professor

Research Interests

  • Skeleltal Muscle

  • Meat

  • Muscle fiber type

  • Physiology

  • Bovine

  • exosome

  • Epigenetics

  • Transcriptomics

  • MicroRNA

  • Metabolomics

Research Areas

  • Life Science / Laboratory animal science

  • Life Science / Animal physiological chemistry, physiology and behavioral biology

  • Life Science / Animal production science

Research History

  • Kagoshima University   Joint faculty of Veterinary Medicine   Prof.

    2024.4

  • National Agriculture and Food Research Organization   Institute of Livestock and Grassland Science, NARO

    2021.4

  • National Agriculture and Food Research Organization

    2016.4

  • National Agriculture and Food Research Organization, National Institute of Livestock and Grassland Science

    2011.4 - 2016.3

  • National Agriculture and Food Research Organization, National Institute of Livestock and Grassland Science

    2009.4 - 2009.9

  • National Agriculture and Food Research Organization, National Institute of Livestock and Grassland Science

    2008.3 - 2009.3

  • National Agriculture and Food Research Organization, National Institute of Livestock and Grassland Science   Chief Researcher

    2006.4 - 2011.3

  • 独立行政法人農業技術研究機構 畜産草地研究所   品質開発部   主任研究官

    2001.4 - 2006.3

  • Ministry of Agriculture, Forestry and Fisheries   Researcher

    1996.10 - 2001.3

▼display all

Professional Memberships

  • International Society of Extracellular Vesicles

  • JAPAN SOCIETY FOR BIOSICENCE, BIOTECHNOLOGY, AND AGROCHEMISTRY

  • JAPANESE SOCIETY OF ANIMAL SCIENCE

  • THE MOLECULAR BIOLOGY SOCIETY OF JAPAN

  • American Society of Animal Science

Committee Memberships

  •   Associate Editor - Frontiers in Animal Science  

    2022.5   

  •   68th International Congress of Meat Science and Technology (ICoMST) Organizing committee member  

    2020.4   

  •   日本産肉研究会監事  

    2012.4   

 

Papers

  • Koichi Ojima, Shoji Hata, Fumiko Shinkai-Ouchi, Yasuko Ono, Susumu Muroya .  Calpain-3 not only proteolyzes calpain-1 and -2 but also is a substrate for calpain-1 and -2. .  Journal of biochemistry174 ( 5 ) 421 - 431   2023.7Reviewed International journal

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    Calpain is an intracellular cysteine protease that cleaves its specific substrates in a limited region to modulate cellular function. Calpain-1 (C1) and calpain-2 (C2) are ubiquitously expressed in mammalian cells, but calpain-3 (C3) is a skeletal muscle-specific type. In the course of calpain activation, the N-terminal regions of all three isoforms are clipped off in an intramolecular or intermolecular fashion. C1 proteolyzes C2 to promote further proteolysis, but C2 proteolyzes C1 to suspend C1 proteolysis, indicating the presence of C1-C2 reciprocal proteolysis. However, whether C3 is involved in the calpain proteolysis network is unclear. To address this, we examined whether GFP-tagged C3:C129S (GFP-C3:CS), an inactive protease form of C3, was a substrate for C1 or C2 in HEK cells. Intriguingly, the N-terminal region of C3:CS was cleaved by C1 and C2 at the site identical to that of the C3 autoproteolysis site. Furthermore, the N-terminal clipping of C3:CS by C1 and C2 was observed in mouse skeletal muscle lysates. Meanwhile, C3 preferentially cleaved the N-terminus of C1 over that of C2, and the sizes of these cleaved proteins were identical to their autoproteolysis forms. Our findings suggest an elaborate inter-calpain network to prime and suppress proteolysis of other calpains.

    DOI: 10.1093/jb/mvad057

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  • Susumu Muroya, Konosuke Otomaru, Kazunaga Oshima, Ichiro Oshima, Koichi Ojima, Takafumi Gotoh .  DNA Methylation of Genes Participating in Hepatic Metabolisms and Function in Fetal Calf Liver Is Altered by Maternal Undernutrition during Gestation. .  International journal of molecular sciences24 ( 13 )   2023.6Reviewed International journal

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    This study aimed to elucidate the effects of maternal undernutrition (MUN) on epigenetic modification of hepatic genes in Japanese Black fetal calves during gestation. Using a previously established experimental design feeding the dams with 60% (LN) or 120% (HN) of their global nutritional requirements during the 8.5-month gestational period, DNA methylation in the fetal liver was analyzed with reduced representation bisulfite sequencing (RRBS). The promoters and gene bodies in the LN fetuses were hypomethylated compared to HN fetuses. Pathway analysis showed that the genes with DMR in the exon/intron in the LN group were associated with pathways involved in Cushing syndrome, gastric acid secretion, and aldosterone synthesis and secretion. Promoter hypomethylation in the LN group was frequently observed in genes participating in various signaling pathways (thyroid hormone, Ras/Rap1, PIK3-Akt, cAMP), fatty acid metabolism, and cholesterol metabolism. The promoter hypomethylated genes ALPL and GNAS were upregulated in the LN group, whereas the promoter hypermethylated genes GRB10 and POR were downregulated. The intron/exon hypomethylated genes IGF2, IGF2R, ACAD8, TAT, RARB, PINK1, and SOAT2 were downregulated, whereas the hypermethylated genes IGF2BP2, NOS3, and NR2F1 were upregulated. Collectively, MUN alters the promoter and gene body methylation of genes associated with hepatic metabolisms (energy, cholesterol, mitochondria) and function, suggesting an impact of altered gene methylation on the dysregulation of gene expression in the fetal liver.

    DOI: 10.3390/ijms241310682

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  • Susumu Muroya .  - Invited Review - Postmortem skeletal muscle metabolism of farm animals approached with metabolomics. .  Animal bioscience36 ( 2 ) 374 - 384   2023.2Reviewed International journal

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    Skeletal muscle metabolism regulates homeostatic balance in animals. The metabolic impact persists even after farm animal skeletal muscle is converted to edible meat through postmortem rigor mortis and aging. Muscle metabolites resulting from animal growth and postmortem storage have a significant impact on meat quality, including flavor and color. Metabolomics studies of postmortem muscle aging have identified metabolisms that contain signatures inherent to muscle properties and the altered metabolites by physiological adaptation, with glycolysis as the pivotal metabolism in postmortem aging. Metabolomics has also played a role in mining relevant postmortem metabolisms and pathways, such as the citrate cycle and mitochondrial metabolism. This leads to a deeper understanding of the mechanisms underlying the generation of key compounds that are associated with meat quality. Genetic background, feeding strategy, and muscle type primarily determine skeletal muscle properties in live animals and affect post-mortem muscle metabolism. With comprehensive metabolite detection, metabolomics is also beneficial for exploring biomarker candidates that could be useful to monitor meat production and predict the quality traits. The present review focuses on advances in farm animal muscle metabolomics, especially postmortem muscle metabolism associated with genetic factors and muscle type.

    DOI: 10.5713/ab.22.0370

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  • Susumu Muroya .  An insight into farm animal skeletal muscle metabolism based on a metabolomics approach. .  Meat science195   108995 - 108995   2022.9Reviewed International journal

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    Farm animal skeletal muscle, formed through development, growth, and maturation processes, is converted to edible meat during postmortem rigor mortis and aging. Live and postmortem muscle metabolism is the phenotypic determinant of muscle characteristics and meat quality traits such as flavor and color. Meanwhile, animal muscle metabolism, originally programmed by genetic program, is modulated by feeding and environmental conditions through changes in the biosynthetic network. Metabolomics deepens our understanding of metabolisms underlying skeletal muscle growth, maturation, abnormality, and postmortem meat aging. The metabolomics approach is beneficial to explore biomarkers to monitor meat production processes and products, leading to improvements in livestock productivity and meat quality. One of the recent metabolomics findings in animal muscle could be the impact of mitochondrial activity and energy metabolisms on meat quality. The present review overviews the advances in metabolomics studies of farm animal skeletal muscle, to gain an insight into the muscle metabolisms associated with livestock production and meat quality.

    DOI: 10.1016/j.meatsci.2022.108995

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  • Susumu Muroya, Riko Nomura, Hirotaka Nagai, Koichi Ojima, Kazutsugu Matsukawa .  Metabolomic profiling of postmortem aged muscle in Japanese Brown beef cattle revealed an interbreed difference from Japanese Black beef. .  Animal bioscience36 ( 3 ) 506 - 520   2022.9Reviewed International journal

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    Objective: Japanese Brown (JBR) cattle, especially the Kochi (Tosa) pedigree (JBRT), is a local breed of moderately marbled beef. Despite the increasing demand, the interbreed differences in muscle metabolites from the highly marbled Japanese Black (JBL) beef remain poorly understood. We aimed to determine flavor-related metabolites and postmortem metabolisms characteristic to JBRT beef in comparison with JBL beef. Methods: Lean portions of the longissimus thoracis (LT, loin) muscle from four JBRT cattle were collected at 0, 1, and 14 d postmortem. The muscle metabolomic profiles were analyzed using capillary electrophoresis time-of-flight mass spectrometry. The difference in postmortem metabolisms and aged muscle metabolites were analyzed by statistical and bioinformatic analyses between JBRT (n = 12) and JBL cattle (n = 6). Results: A total of 240 metabolite annotations were obtained from the detected signals of the JBRT muscle samples. Principal component analysis separated the beef samples into three different aging point groups. According to metabolite set enrichment analysis (MSEA), postmortem metabolic changes were associated with the metabolism of pyrimidine, nicotinate and nicotinamide, purine, pyruvate, thiamine, amino sugar, and fatty acid; citric acid cycle; and pentose phosphate pathway as well as various amino acids and mitochondrial fatty acid metabolism. The aged JBRT beef showed higher ultimate pH and lower lactate content than aged JBL beef, suggesting the lower glycolytic activity in postmortem JBRT muscle. JBRT beef was distinguished from JBL beef by significantly different compounds, including choline, amino acids, uridine monophosphate, inosine 5'-monophosphate, fructose 1,6-diphosphate, and betaine, suggesting interbreed differences in the accumulation of nucleotide monophosphate, glutathione metabolism, and phospholipid metabolism. Conclusion: Glycolysis, purine metabolism, fatty acid catabolism, and protein degradation were the most common pathways in beef during postmortem aging. The differentially expressed metabolites and the relevant metabolisms in JBRT beef may contribute to the development of a characteristic flavor.

    DOI: 10.5713/ab.22.0202

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  • Koichi Ojima, Masahiro Kigaki, Emi Ichimura, Takahiro Suzuki, Ken Kobayashi, Susumu Muroya, Takanori Nishimura .  Endogenous slow and fast myosin dynamics in myofibers isolated from mice expressing GFP-Myh7 and Kusabira Orange-Myh1. .  American journal of physiology. Cell physiology323 ( 2 ) C520-C535 - C535   2022.6Reviewed International journal

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    Skeletal muscle consists of slow and fast myofibers in which different myosin isoforms are expressed. Approximately 300 myosins form a single thick filament in the myofibrils, where myosin is continuously exchanged. However, endogenous slow and fast myosin dynamics have not been fully understood. To elucidate those dynamics, here we generated mice expressing green fluorescence protein-tagged slow myosin heavy chain (GFP-Myh7) and Kusabira Orange fluorescence protein-tagged fast myosin heavy chain (KuO-Myh1). First, these mice enabled us to distinguish between GFP- and KuO-myofibers under fluorescence microscopy: GFP-Myh7 and KuO-Myh1 were exclusively expressed in slow myofibers and fast myofibers, respectively. Next, to monitor endogenous myosin dynamics, fluorescence recovery after photobleaching (FRAP) was conducted. The mobile fraction (Mf) of GFP-Myh7 and that of KuO-Myh1 were almost constant values independent of the regions of the myofibers and the muscle portions where the myofibers were isolated. Intriguingly, proteasome inhibitor treatment significantly decreased the Mf in GFP-Myh7 but not in KuO-Myh1 myofibers, indicating that the response to a disturbance in protein turnover depended on muscle fiber type. Taken together, the present results indicated that the mice we generated are promising tools not only for distinguishing between GFP- and KuO-myofibers but also for studying the dynamics of endogenous myosin isoforms by live-cell fluorescence imaging.

    DOI: 10.1152/ajpcell.00415.2021

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  • Susumu Muroya, Yi Zhang, Kounosuke Otomaru, Kazunaga Oshima, Ichiro Oshima, Mitsue Sano, Sanggun Roh, Koichi Ojima, Takafumi Gotoh .  Maternal Nutrient Restriction Disrupts Gene Expression and Metabolites Associated with Urea Cycle, Steroid Synthesis, Glucose Homeostasis, and Glucuronidation in Fetal Calf Liver .  Metabolites12 ( 3 ) 203 - 203   2022.2Reviewed

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    Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:MDPI AG  

    This study aimed to understand the mechanisms underlying the effects of maternal undernutrition (MUN) on liver growth and metabolism in Japanese Black fetal calves (8.5 months in utero) using an approach that integrates metabolomics and transcriptomics. Dams were fed 60% (low-nutrition; LN) or 120% (high-nutrition; HN) of their overall nutritional requirements during gestation. We found that MUN markedly decreased the body and liver weights of the fetuses; metabolomic analysis revealed that aspartate, glycerol, alanine, gluconate 6-phosphate, and ophthalmate levels were decreased, whereas UDP-glucose, UDP-glucuronate, octanoate, and 2-hydroxybutyrate levels were decreased in the LN fetal liver (p ≤ 0.05). According to metabolite set enrichment analysis, the highly different metabolites were associated with metabolisms including the arginine and proline metabolism, nucleotide and sugar metabolism, propanoate metabolism, glutamate metabolism, porphyrin metabolism, and urea cycle. Transcriptomic and qPCR analyses revealed that MUN upregulated QRFPR and downregulated genes associated with the glucose homeostasis (G6PC, PCK1, DPP4), ketogenesis (HMGCS2), glucuronidation (UGT1A1, UGT1A6, UGT2A1), lipid metabolism (ANGPTL4, APOA5, FADS2), cholesterol and steroid homeostasis (FDPS, HSD11B1, HSD17B6), and urea cycle (CPS1, ASS1, ASL, ARG2). These metabolic pathways were extracted as relevant terms in subsequent gene ontology/pathway analyses. Collectively, these results indicate that the citrate cycle was maintained at the expense of activities of the energy metabolism, glucuronidation, steroid hormone homeostasis, and urea cycle in the liver of MUN fetuses.

    DOI: 10.3390/metabo12030203

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  • Emi Ichimura, Koichi Ojima, Susumu Muroya, Ken Kobayashi, Takanori Nishimura .  Thick filament-associated myosin undergoes frequent replacement at the tip of the thick filament. .  FEBS open bio12 ( 4 ) 852 - 863   2022.2Reviewed International journal

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    Myosin plays a fundamental role in muscle contraction. Approximately 300 myosins form a bipolar thick filament, in which myosin is continuously replaced by protein turnover. However, it is unclear how rapidly this process occurs, and whether the myosin exchange rate differs depending on the region of the thick filament. To answer this question, we first measured myosin release and insertion rates over a short period, and monitored myotubes expressing a photoconvertible fluorescence protein-tagged myosin, which enabled us to monitor myosin release and insertion simultaneously. About 20% of myosins were replaced within 10 min, while 70% of myosins were exchanged over 10 h with symmetrical and biphasic alteration of myosin release and insertion rates. Next, a fluorescence pulse-chase assay was conducted to investigate whether myosin is incorporated into specific regions in the thick filament. Newly synthesized myosin was located at the tip of the thick-filament rather than the center in the first 7 min of pulse-chase labeling and was observed in the remainder of the thick filament by 30 min. These results suggest that the myosin replacement rate differs depending on the regions of the thick filament. We concluded that myosin release and insertion occur concurrently, and that myosin is more frequently exchanged at the tip of the thick filament.

    DOI: 10.1002/2211-5463.13379

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  • Yi Zhang, Konosuke Otomaru, Kazunaga Oshima, Yuji Goto, Ichiro Oshima, Susumu Muroya, Mitsue Sano, Sanggun Roh, Takafumi Gotoh .  Maternal Nutrition During Gestation Alters Histochemical Properties, and mRNA and microRNA Expression in Adipose Tissue of Wagyu Fetuses .  Frontiers in Endocrinology12   797680   2022.2Reviewed

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    Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:Frontiers Media {SA}  

    <jats:p>We hypothesized that maternal low or high nutrition would give unique effects to morphological and molecular dynamics in adipose tissue of fetus of fatty breed Wagyu (Japanese Black) cattle which produce highly marbled beef. This study aimed to determine the effects of maternal energy intake in Wagyu cows, during gestation on fetal adipose tissue development, histochemical properties, and gene and microRNA (miRNA) expression. Cows were allocated to one of two nutritional energy groups: 120% (HIGH) or 60% nutritional requirements of (LOW). Fetuses (n = 6 per treatment) were removed from pregnant cows by cesarean section at fetal age 260 ± 8 days and euthanized. Subcutaneous adipose tissue (SAT), thoracic cavity visceral adipose tissue (TVAT), and perirenal adipose tissue (PAT) were collected for analysis. In histochemical analysis, in SAT and PAT, HIGH fetuses had greater diameter of adipocytes than LOW fetuses (P&amp;lt;0.05). Only in SAT, LOW fetuses had more Leptin (LEP) mRNA and tended to have more Peroxisome Proliferator-Activated Receptor gamma (PPARG) CCAAT-enhancer-binding proteins alpha (CEBPA) and Glucose transporter (GLUT) 4 mRNA(P&amp;lt;0.10). In all SAT, TVAT, and PAT, LOW fetuses had higher levels of the brown adipose tissue (BAT) biomarkers Uncoupling Protein (UCP) 1 and PPARG coactivator (PGC) 1α mRNA than HIGH fetuses (P&amp;lt;0.08). Meanwhile, in the other adipose tissue, LOW fetuses had lower PPARG, CEBPA, and Zinc Finger Protein (ZFP) 423 (in TVAT and PAT), FASN (in TVAT), LEP and GLUT4 mRNA (in PAT; P&amp;lt;0.10). In particular, in TVAT and PAT, LOW fetuses exhibited lower expression of WAT biomarkers (PPARG and ZFP423). Differential expression of various miRNAs related to adipogenesis between the LOW and HIGH fetuses was detected in an adipose tissue-specific manner (P&amp;lt;0.10). Based on adipose tissue-specific effects of maternal nutrition, these findings suggested that poor maternal nutrition in Wagyu cattle increased BAT development in SAT, TVAT and PAT, while elevated maternal nutrition stimulated fetal SAT development compared with that of TVAT and PAT.</jats:p>

    DOI: 10.3389/fendo.2021.797680

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  • Yi Zhang, Kounosuke Otomaru, Kazunaga Oshima, Yuji Goto, Ichiro Oshima, Susumu Muroya, Mitsue Sano, Rena Saneshima, Yukiko Nagao, Aoi Kinoshita, Yasuko Okamura, Sanggun Roh, Akira Ohtsuka, Takafumi Gotoh .  Effects of low and high levels of maternal nutrition consumed for the entirety of gestation on the development of muscle, adipose tissue, bone, and the organs of Wagyu cattle fetuses. .  Animal science journal = Nihon chikusan Gakkaiho92 ( 1 ) e13600   2021.12Reviewed International journal

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    This study aimed to investigate the effects of high and low levels of energy intake during the entire gestation period on the skeletal muscle development, organ development, and adipose tissue accumulation in fetuses of Wagyu (Japanese Black) cows, a breed with highly marbled beef. Cows were allocated to a high-nutrition (n = 6) group (fed 120% of the nutritional requirement) or low-nutrition (n = 6) group (fed 60% of the nutritional requirement). The cows were artificially inseminated with semen from the same sire, and the fetuses were removed by cesarean section at 260 ± 8.3 days of fetal age and slaughtered. The whole-body, total muscle, adipose, and bone masses of the fetal half-carcasses were significantly higher in the high-nutrition group than the low-nutrition group (p = 0.0018, 0.009, 0.0004, and 0.0362, respectively). Fifteen of 20 individual muscles, five of six fat depots, nine of 17 organs, and seven of 12 bones that were investigated had significantly higher masses in the high-nutrition group than the low-nutrition group. The crude components and amino acid composition of the longissimus muscle significantly differed between the low- and high-nutrition groups. These data indicate that maternal nutrition during gestation has a marked effect on the muscle, bone, and adipose tissue development of Wagyu cattle fetuses.

    DOI: 10.1111/asj.13600

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  • Emi Ichimura, Koichi Ojima, Susumu Muroya, Takahiro Suzuki, Ken Kobayashi, Takanori Nishimura .  The ubiquitin ligase Ozz decreases the replacement rate of embryonic myosin in myofibrils .  Physiological Reports9 ( 17 ) e15003   2021.9Reviewed International journal

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    Myosin, the most abundant myofibrillar protein in skeletal muscle, functions as a motor protein in muscle contraction. Myosin polymerizes into the thick filaments in the sarcomere where approximately 50% of embryonic myosin (Myh3) are replaced within 3 h (Ojima K, Ichimura E, Yasukawa Y, Wakamatsu J, Nishimura T, Am J Physiol Cell Physiol 309: C669-C679, 2015). The sarcomere structure including the thick filament is maintained by a balance between protein biosynthesis and degradation. However, the involvement of a protein degradation system in the myosin replacement process remains unclear. Here, we show that the muscle-specific ubiquitin ligase Ozz regulates replacement rate of Myh3. To examine the direct effect of Ozz on myosin replacement, eGFP-Myh3 replacement rate was measured in myotubes overexpressing Ozz by fluorescence recovery after photobleaching. Ozz overexpression significantly decreased the replacement rate of eGFP-Myh3 in the myofibrils, whereas it had no effect on other myosin isoforms. It is likely that ectopic Ozz promoted myosin degradation through increment of ubiquitinated myosin, and decreased myosin supply for replacement, thereby reducing myosin replacement rate. Intriguingly, treatment with a proteasome inhibitor MG132 also decreased myosin replacement rate, although MG132 enhanced the accumulation of ubiquitinated myosin in the cytosol where replaceable myosin is pooled, suggesting that ubiquitinated myosin is not replaced by myosin in the myofibril. Collectively, our findings showed that Myh3 replacement rate was reduced in the presence of overexpressed Ozz probably through enhanced ubiquitination and degradation of Myh3 by Ozz.

    DOI: 10.14814/phy2.15003

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  • Susumu Muroya, Yi Zhang, Aoi Kinoshita, Kounosuke Otomaru, Kazunaga Oshima, Yuji Gotoh, Ichiro Oshima, Mitsue Sano, Sanggun Roh, Mika Oe, Koichi Ojima, Takafumi Gotoh .  Maternal Undernutrition during Pregnancy Alters Amino Acid Metabolism and Gene Expression Associated with Energy Metabolism and Angiogenesis in Fetal Calf Muscle .  Metabolites11 ( 9 )   2021.8Reviewed International journal

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    To elucidate the mechanisms underlying maternal undernutrition (MUN)-induced fetal skeletal muscle growth impairment in cattle, the longissimus thoracis muscle of Japanese Black fetal calves at 8.5 months in utero was analyzed by an integrative approach with metabolomics and transcriptomics. The pregnant cows were fed on 60% (low-nutrition, LN) or 120% (high-nutrition, HN) of their overall nutritional requirement during gestation. MUN markedly decreased the bodyweight and muscle weight of the fetus. The levels of amino acids (AAs) and arginine-related metabolites including glutamine, gamma-aminobutyric acid (GABA), and putrescine were higher in the LN group than those in the HN group. Metabolite set enrichment analysis revealed that the highly different metabolites were associated with the metabolic pathways of pyrimidine, glutathione, and AAs such as arginine and glutamate, suggesting that MUN resulted in AA accumulation rather than protein accumulation. The mRNA expression levels of energy metabolism-associated genes, such as PRKAA1, ANGPTL4, APLNR, CPT1B, NOS2, NOS3, UCP2, and glycolytic genes were lower in the LN group than in the HN group. The gene ontology/pathway analysis revealed that the downregulated genes in the LN group were associated with glucose metabolism, angiogenesis, HIF-1 signaling, PI3K-Akt signaling, pentose phosphate, and insulin signaling pathways. Thus, MUN altered the levels of AAs and expression of genes associated with energy expenditure, glucose homeostasis, and angiogenesis in the fetal muscle.

    DOI: 10.3390/metabo11090582

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  • Zhang Yi, Otomaru Kounosuke, Oshima Kazunaga, Goto Yuji, Oshima Ichiro, Muroya Susumu, Sano Mitsue, Saneshima Rena, Nagao Yukiko, Kinoshita Aoi, Okamura Yasuko, Roh Sanggun, Ohtsuka Akira, Gotoh Takafumi .  和牛胎仔の筋肉、脂肪組織、骨および臓器の発生与える全妊娠期における母体の高栄養および低栄養の影響(Effects of low and high levels of maternal nutrition consumed for the entirety of gestation on the development of muscle, adipose tissue, bone, and the organs of Wagyu cattle fetuses) .  Animal Science Journal92 ( 1 ) 1 of 13 - 13 of 13   2021.7和牛胎仔の筋肉、脂肪組織、骨および臓器の発生与える全妊娠期における母体の高栄養および低栄養の影響(Effects of low and high levels of maternal nutrition consumed for the entirety of gestation on the development of muscle, adipose tissue, bone, and the organs of Wagyu cattle fetuses)

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    Language:English   Publisher:John Wiley & Sons Australia, Ltd  

  • Koichi Ojima, Susumu Muroya, Hiromu Wada, Kotaro Ogawa, Mika Oe, Koichi Takimoto, Takanori Nishimura .  Immature adipocyte‐derived exosomes inhibit expression of muscle differentiation markers .  FEBS Open Bio11 ( 3 ) 768 - 781   2021.3Reviewed International journal

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Wiley  

    Exosomes are released from a variety of cells to communicate with recipient cells. Exosomes contain microRNAs (miRNAs), which are noncoding RNAs that suppress target genes. Our previous proteomic study (FEBS Open Bio 2016, 6, 816-826) demonstrated that 3T3-L1 adipocytes secrete exosome components as well as growth factors, inspiring us to investigate what type of miRNA is involved in adipocyte-secreted exosomes and what functions they carry out in recipient cells. Here, we profiled miRNAs in 3T3-L1 adipocyte-secreted exosomes and revealed suppression of muscle differentiation by adipocyte-derived exosomes. Through our microarray analysis, we detected over 300 exosomal miRNAs during adipocyte differentiation. Exosomal miRNAs present during adipocyte differentiation included not only pro-adipogenic miRNAs but also miRNAs associated with muscular dystrophy. Gene ontology analysis predicted that the target genes of miRNAs are associated primarily with transcriptional regulation. To further investigate whether adipocyte-secreted exosomes regulate the expression levels of genes involved in muscle differentiation, we treated cultured myoblasts with adipocyte-derived exosome fractions. Intriguingly, the expression levels of myogenic regulatory factors, Myog and Myf6, and other muscle differentiation markers, myosin heavy-chain 3 and insulin-like growth factor 2, were significantly downregulated in myoblasts treated with adipocyte-derived exosomes. Immature adipocyte-derived exosomes exhibited a stronger suppressive effect than mature adipocyte-derived exosomes. Our results suggest that adipocytes suppress the expression levels of muscle differentiation-associated genes in myoblasts via adipocyte-secreted exosomes containing miRNAs.

    DOI: 10.1002/2211-5463.13100

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    Other Link: https://onlinelibrary.wiley.com/doi/full-xml/10.1002/2211-5463.13100

  • Mika Oe, Koichi Ojima, Susumu Muroya .  Difference in potential DNA methylation impact on gene expression between fast- and slow-type myofibers. .  Physiological genomics53 ( 2 ) 69 - 83   2021.1Reviewed International journal

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    Skeletal muscles are comprised of two major types of myofibers, fast and slow. It is hypothesized that once myofiber type is determined, muscle fiber-type specificity is maintained by an epigenetic mechanism, however, this remains poorly understood. To address this, we conducted a comprehensive CpG methylation analysis with a reduced representation of bisulfite sequencing (RRBS). Using GFP-myh7 mouse, we visually distinguished and separately pooled slow-type and myh7-negative fast-type fibers for analyses. A total of 31,967 and 26,274 CpGs were hypermethylated by ≥10% difference in the fast- and slow-type fibers, respectively. Notably, the number of promoter-hypermethylated genes with down-regulated expression in the slow-type fibers was 3.5 times higher than that in the fast-type fibers. Gene bodies of the fast-type-specific myofibrillar genes Actn3, Tnnt3, Tnni2, Tnnc2, and Tpm1 were hypermethylated in the slow-type fibers, whereas those of the slow-type-specific genes Myh7, Tnnt1, and Tpm3 were hypermethylated in the fast-type fibers. Each of the instances of gene hypermethylation was associated with the respective down-regulated expression. In particular, a relationship between CpG methylation sites and the transcription variant distribution of Tpm1 was observed, suggesting a regulation of Tpm1 alternative promoter usage by gene body CpG methylation. An association of hypermethylation with the regulation of gene expression was also observed in Wdr70, and transcription factors Sim2 and Tbx1. These results suggest not only a myofiber type-specific regulation of gene expression and alternative promoter usage by gene body CpG methylation, but also a dominant effect of promoter-hypermethylation on the gene expressions in slow myofibers.

    DOI: 10.1152/physiolgenomics.00099.2020

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  • Susumu Muroya, Hideki Ogasawara, Kana Nohara, Mika Oe, Koichi Ojima, Masayuki Hojito .  Coordinated alteration of mRNA-microRNA transcriptomes associated with exosomes and fatty acid metabolism in adipose tissue and skeletal muscle in grazing cattle .  Asian-Australasian Journal of Animal Sciences33 ( 11 ) 1824 - 1836   2020.11Reviewed International journal

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    Objective: On the hypothesis that grazing of cattle prompts organs to secrete or internalize circulating microRNAs (c-miRNAs) in parallel with changes in energy metabolism, we aimed to clarify biological events in adipose, skeletal muscle, and liver tissues in grazing Japanese Shorthorn (JSH) steers by a transcriptomic approach. Methods: The subcutaneous fat (SCF), biceps femoris muscle (BFM), and liver in JSH steers after three months of grazing or housing were analyzed using microarray and qPCR, followed by gene ontology (GO) and functional annotation analyses. Results: The results of transcriptomics indicated that SCF was highly responsive to grazing compared to BFM and liver tissues. The 'Exosome', 'Carbohydrate metabolism' and 'Lipid metabolism' were extracted as the relevant GO terms in SCF and BFM, and/or liver from the >1.5-fold-altered mRNAs in grazing steers. The qPCR analyses showed a trend of upregulated gene expression related to exosome secretion and internalization (CHMP4A, VPS4B, VAMP7, CAV1) in the BFM and SCF, as well as upregulation of lipolysis-associated mRNAs (CPT1A, HSL, PLIN, ATGL, FABP4) and most of the microRNAs (miRNAs) in SCF. Moreover, gene expression related to fatty acid uptake and inter-organ signaling (FATP4 and ANGPTL) was upregulated in BFM, suggesting SCF-BFM organ crosstalk for energy metabolism. Meanwhile, expression of plasma exosomal miR-16a, miR-19b, miR-21-5p, and miR-142-5p was reduced. According to bioinformatic analyses, the c-miRNA target genes are associated with the terms 'Endosome', 'Caveola', 'Endocytosis', 'Carbohydrate metabolism', and with pathways related to environmental information processing and the endocrine system. Conclusion: Exosome and fatty acid metabolism-related gene expression was altered in SCF of grazing cattle, which could be regulated by miRNA such as miR-142-5p. These changes occurred coordinately in both the SCF and BFM, suggesting involvement of exosome in the SCF-BFM organ crosstalk to modulate energy metabolism.

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  • Koichi Ojima, Shoji Hata, Fumiko Shinkai-Ouchi, Mika Oe, Susumu Muroya, Hiroyuki Sorimachi, Yasuko Ono .  Developing fluorescence sensor probe to capture activated muscle-specific calpain-3 (CAPN3) in living muscle cells. .  Biology open9 ( 9 )   2020.9Reviewed International journal

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    Calpain-3 (CAPN3) is a muscle-specific type of calpain whose protease activity is triggered by Ca2+ Here, we developed CAPN3 sensor probes (SPs) to detect activated-CAPN3 using a fluorescence/Förster resonance energy transfer (FRET) technique. In our SPs, partial amino acid sequence of calpastatin, endogenous CAPN inhibitor but CAPN3 substrate, is inserted between two different fluorescence proteins that cause FRET. Biochemical and spectral studies revealed that CAPN3 cleaved SPs and changed emission wavelengths of SPs. Importantly, SPs were scarcely cleaved by CAPN1 and CAPN2. Furthermore, our SP successfully captured the activation of endogenous CAPN3 in living myotubes treated with ouabain. Our SPs would become a promising tool to detect the dynamics of CAPN3 protease activity in living cells.

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  • Susumu Muroya, Shuji Ueda, Tomohiko Komatsu, Takuya Miyakawa, Per Ertbjerg .  MEATabolomics: Muscle and Meat Metabolomics in Domestic Animals. .  Metabolites10 ( 5 )   2020.5Reviewed International journal

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    In the past decades, metabolomics has been used to comprehensively understand a variety of food materials for improvement and assessment of food quality. Farm animal skeletal muscles and meat are one of the major targets of metabolomics for the characterization of meat and the exploration of biomarkers in the production system. For identification of potential biomarkers to control meat quality, studies of animal muscles and meat with metabolomics (MEATabolomics) has been conducted in combination with analyses of meat quality traits, focusing on specific factors associated with animal genetic background and sensory scores, or conditions in feeding system and treatments of meat in the processes such as postmortem storage, processing, and hygiene control. Currently, most of MEATabolomics approaches combine separation techniques (gas or liquid chromatography, and capillary electrophoresis)-mass spectrometry (MS) or nuclear magnetic resonance (NMR) approaches with the downstream multivariate analyses, depending on the polarity and/or hydrophobicity of the targeted metabolites. Studies employing these approaches provide useful information to monitor meat quality traits efficiently and to understand the genetic background and production system of animals behind the meat quality. MEATabolomics is expected to improve the knowledge and methodologies in animal breeding and feeding, meat storage and processing, and prediction of meat quality.

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  • Susumu Muroya, Mika Oe, Koichi Ojima, Akira Watanabe .  Metabolomic approach to key metabolites characterizing postmortem aged loin muscle of Japanese Black (Wagyu) cattle .  Asian-Australasian Journal of Animal Sciences32 ( 8 ) 1172 - 1185   2019.8Reviewed

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  • Nakajima I, Kojima M, Oe M, Ojima K, Muroya S, Chikuni K .  Comparing pig breeds with genetically low and high backfat thickness: differences in expression of adiponectin, its receptor, and blood metabolites. .  Domestic animal endocrinology68   54 - 63   2019.7Reviewed

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  • S Muroya, H Ogasawara, K Nohara, M Oe, K Ojima, M Hojito .  PSVII-25 Grazing-induced transcriptomic changes in bovine biceps femoris muscle, subcutaneous fat, and liver mRNAs and plasma exosome microRNAs. .  Journal of Animal Science96 ( suppl_3 ) 356 - 356   2018.12

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  • 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 physiology315 ( 1 ) C104 - C114   2018.7Reviewed

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  • S. Muroya, M. Oe, K. Ojima .  Thiamine accumulation and thiamine triphosphate decline occur in parallel with ATP exhaustion during postmortem aging of pork muscles .  Meat Science137   228 - 234   2018.3Reviewed

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    We aimed to clarify the mechanisms affecting postmortem thiamine and its phosphoester contents in major edible pork muscles, namely the longissimus lumborum (LL) in addition to vastus intermedius (VI). Metabolomic analysis by capillary electrophoresis-time of flight mass spectrometry revealed that the level of thiamine triphosphate (ThTP), approximately 1.8-fold higher in LL than in VI muscle at 0 h postmortem, declined in the first 24 h, resulting in an undetectable level at 168 h postmortem in both muscles. In contrast, the thiamine content in both muscles increased after 24 h postmortem during the aging process. The thiamine accumulation and ThTP decline progressed in parallel with a drastic reduction of the ATP level. The intermuscular differences in pH at 24 h, and expression of thiamine transporter and thiamine pyrophosphokinase might result in delayed thiamine generation in LL. These results suggest that postmortem ATP exhaustion forced ThTP hydrolysis and further depyrophosphorylation of thiamine diphosphate in the porcine muscles, which resulted in thiamine accumulation.

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  • 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 JOURNAL88 ( 11 ) 1788 - 1793   2017.11Reviewed

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    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.

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  • Keisuke Sasaki, Motoki Ooi, Naoto Nagura, Michiyo Motoyama, Takumi Narita, Mika Oe, Ikuyo Nakajima, Tatsuro Hagi, Koichi Ojima, Miho Kobayashi, Masaru Nomura, Susumu Muroya, Takeshi Hayashi, Kyoko Akama, Akira Fujikawa, Hironao Hokiyama, Kuniyuki Kobayashi, Takanori Nishimura .  Classification and characterization of Japanese consumers' beef preferences by external preference mapping .  JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE97 ( 10 ) 3453 - 3462   2017.8Reviewed

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    BACKGROUNDOver the past few decades, beef producers in Japan have improved marbling in their beef products. It was recently reported that marbling is not well correlated with palatability as rated by Japanese consumers. This study sought to identify the consumer segments in Japan that prefer sensory characteristics of beef other than high marbling.
    RESULTSThree Wagyu beef, one Holstein beef and two lean imported beef longissimus samples were subjected to a descriptive sensory test, physicochemical analysis and a consumer (n = 307) preference test. According to consumer classification and external preference mapping, four consumer segments were identified as gradual high-fat likers', moderate-fat and distinctive taste likers', Wagyu likers' and distinctive texture likers'. Although the major trend of Japanese consumers' beef preference was marbling liking', 16.9% of the consumers preferred beef samples that had moderate marbling and distinctive taste. The consumers' attitudes expressed in a questionnaire survey were in good agreement with the preference for marbling among the moderate-fat and distinctive taste likers'.
    CONCLUSIONThese results indicate that moderately marbled beef is a potent category in the Japanese beef market. (c) 2017 Society of Chemical Industry

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  • Susumu Muroya, Masahiro Shibata, Masayuki Hayashi, Mika Oe, Koichi Ojima .  Differences in Circulating microRNAs between Grazing and Grain-Fed Wagyu Cattle Are Associated with Altered Expression of Intramuscular microRNA, the Potential Target PTEN, and Lipogenic Genes .  PLOS ONE11 ( 9 ) e0162496   2016.9Reviewed

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    We aimed to understand the roles of miRNAs in the muscle tissue maturation and those of circulating microRNAs (c-miRNAs) in beef production of Japanese Black (JB) cattle (Wagyu), a breed with genetically background of superior intermuscular fat depot, by comparing different feeding conditions (indoor grain-feeding vs. grazing on pasture). The cattle at 18 months old were assigned to pasture feeding or conventional indoor grain feeding conditions for 5 months. Microarray analysis of c-miRNAs from the plasma extracellular vesicles led to the detection of a total of 202 bovine miRNAs in the plasma, including 15 miRNAs that differed between the feeding conditions. Validation of the microarray results by qPCR showed that the circulating miR-10b level in the grazing cattle was upregulated compared to that of the grain-fed cattle. In contrast, the levels of miR-17-5p, miR-19b, miR-29b, miR-30b-5p, miR-98, miR-142-5p, miR-301a, miR-374b, miR-425-5p, and miR-652 were lower in the grazing cattle than in the grain-fed cattle. Bioinformatic analysis indicated that the predicted target genes of those c-miRNAs were enriched in gene ontology terms associated with blood vessel morphogenesis, plasma membrane, focal adhesion, endocytosis, collagen, ECM-receptor interaction, and phosphorylation. In the grazing cattle, the elevation of miR-10b expression in the plasma was coincident with its elevation in the longissimus lumborum (LL) muscle. Expression of bovine-specific miR-2478, the most plasma-enriched miRNA, tended to be also upregulated in the muscle but not in the plasma. Furthermore, grazing caused the downregulated mRNA expression of predicted miR-10b and/or miR-2478 target genes, such as DNAJB2, PTEN, and SCD1. Thus, the feeding system used for JB cattle affected the c-miRNAs that could be indicators of grain feeding. Among these, miR-10b expression was especially associated with feeding-induced changes and with the expression of the potential target genes responsible for glucose homeo-stasis and intramuscular fat depot in the LL muscle of JB cattle.

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  • M. Oe, K. Ojima, I. Nakajima, K. Chikuni, M. Shibata, S. Muroya .  Distribution of tropomyosin isoforms in different types of single fibers isolated from bovine skeletal muscles .  MEAT SCIENCE118   129 - 132   2016.8Reviewed

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    To clarify the relationship between myosin heavy chain (MyHC) isoforms and tropomyosin (TPM) isoforms in single fibers, 64 single fibers were isolated from each of bovine three muscles (masseter, semispinalis and semitendinosus). mRNA expressions of MyHC and TPM isoforms were analyzed by real-time PCR. All single fibers from the masseter expressed MyHC-slow. The fibers from the semispinalis expressed both MyHC-slow and 2a. The fibers from the semitendinosus expressed MyHC-slow, 2a and 2x. TPM-1 and TPM-2 were co-expressed in 2a and 2x type fibers, and TPM-2 and TPM-3 were co-expressed in slow type fibers. The expression pattern of TPM isoforms in each fiber type was similar between fibers isolated from different muscles. These results suggest that TPM-1 and TPM-3 isoforms correspond to the function of 2a or 2x type fibers and slow type fibers, respectively, with TPM-2 in common. Furthermore, the patterns of MyHC and TPM isoform combinations did not vary among single fibers isolated from the individual muscles examined. (C) 2016 Elsevier Ltd. All rights reserved.

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  • Koichi Ojima, Mika Oe, Ikuyo Nakajima, Susumu Muroya, Takanori Nishimura .  Dynamics of protein secretion during adipocyte differentiation .  FEBS OPEN BIO6 ( 8 ) 816 - 826   2016.8Reviewed

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    The major functions of adipocytes include both lipid storage and the production of secretory factors. However, the type of proteins released from mouse 3T3-L1 cells during adipocyte differentiation remains poorly understood. We examined the dynamics of secreted proteins during adipocyte differentiation using mass spectrometry (MS) combined with an iTRAQ((R)) labeling method that enables the simultaneous analysis of relative protein expression levels. A total of 215 proteins were identified and quantified from approximately 10000 MS/MS spectra. Of these, approximately 38% were categorized as secreted proteins based on gene ontology classification. Adipokine secretion levels were increased with the progression of differentiation. By contrast, levels of fibril collagen components, such as subunits of type I and III collagens, were decreased during differentiation. Basement membrane components attained their peak levels at day 4 when small lipid droplets accumulated in differentiated 3T3-L1 cells. Simultaneously, peak levels of collagen microfibril components that comprise type V and VI collagen subunits were also observed. Our data demonstrated that extracellular matrix components were predominantly released during the early and middle stages of adipocyte differentiation, with a subsequent increase in the secretion of adipokines. This suggests that 3T3-L1 cells secrete adipokines after their ECM is constructed during adipocyte differentiation.

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  • Susumu Muroya, Tatsuro Hagi, Ataru Kimura, Hisashi Aso, Masatoshi Matsuzaki, Masaru Nomura .  Lactogenic hormones alter cellular and extracellular microRNA expression in bovine mammary epithelial cell culture .  JOURNAL OF ANIMAL SCIENCE AND BIOTECHNOLOGY7 ( 1 ) 8   2016.2Reviewed

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    Background: Bovine milk contains not only a variety of nutritional ingredients but also microRNAs (miRNAs) that are thought to be secreted by the bovine mammary epithelial cells (BMECs). The objective of this study was to elucidate the production of milk-related miRNAs in BMECs under the influence of lactogenic hormones.
    Results: According to a microarray result of milk exosomal miRNAs prior to cellular analyses, a total of 257 miRNAs were detected in a Holstein cow milk. Of these, 18 major miRNAs of interest in the milk were selected for an expression analysis in BMEC culture that was treated with or without dexamethasone, insulin, and prolactin (DIP) to induce a lactogenic differentiation. Quantitative polymerase chain reaction (qPCR) results showed that the expressions of miR-21-5p (P = 0.005), miR-26a (P = 0.016), and miR-320a (P = 0.011) were lower in the DIP-treated cells than in the untreated cells. In contrast, the expression of miR-339a (P = 0.017) in the cell culture medium were lower in the DIP-treated culture than in the untreated culture. Intriguingly, the miR-148a expression in cell culture medium was elevated by DIP treatment of BMEC culture (P = 0.018). The medium-to-cell expression ratios of miR103 (P = 0.025), miR-148a (P &lt; 0.001), and miR-223 (P = 0.013) were elevated in the DIP-treated BMECs, suggesting that the lactogenic differentiation-induced secretion of these three miRNAs in BMECs. A bioinformatic analysis showed that the miRNAs down-regulated in the BMECs were associated with the suppression of genes related to transcriptional regulation, protein phosphorylation, and tube development.
    Conclusion: The results suggest that the miRNAs changed by lactogenic hormones are associated with milk protein synthesis, and mammary gland development and maturation. The elevated miR-148a level in DIP-treated BMECs may be associated with its increase in milk during the lactation period of cows.

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  • Akihiko Horikawa, Hideki Ogasawara, Kaito Okada, Misato Kobayashi, Susumu Muroya, Masayuki Hojito .  Grazing-induced changes in muscle microRNA-206 and-208b expression in association with myogenic gene expression in cattle .  ANIMAL SCIENCE JOURNAL86 ( 11 ) 952 - 960   2015.11Reviewed

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    To investigate the roles of microRNAs (miRNAs) in muscle type conversion, the effects of 4 months of grazing on the expression levels of miRNAs and mRNAs associated with skeletal muscle development were analyzed by quantitative RT-PCR using the Biceps femoris muscle of Japanese Shorthorn cattle. After 4 months of grazing, the expression of muscle fiber type-associated miR-208b was higher in the grazed cattle than in the housed. In concordance with the pattern in miR-208b expression, the expression of MyoD, a myogenic regulatory factor associated with the shifting of muscle property to the fast type, was lower in the grazed cattle after 4 months of grazing than in the housed cattle. In addition, the expression of MyHC-2x (a fast type) was higher in the housed cattle than in the grazed, after 4 months of grazing. During the grazing period, miR-206 expression decreased in the housed cattle, whereas expression in the grazed cattle did not change, but rather remained higher than that of the housed cattle even at 3 months after the grazing ended. These miRNAs including miR-206 persisting with muscles of grazed cattle may be associated with regulation of muscle gene expression during skeletal muscle adaptation to grazing.

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  • Susumu Muroya, Hideki Ogasawara, Masayuki Hojito .  Grazing Affects Exosomal Circulating MicroRNAs in Cattle .  PLOS ONE10 ( 8 ) e0136475   2015.8Reviewed

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    Circulating microRNAs (c-miRNAs) are associated with physiological adaptation to acute and chronic aerobic exercise in humans. To investigate the potential effect of grazing movement on miRNA circulation in cattle, here we profiled miRNA expression in centrifugally prepared exosomes from the plasma of both grazing and housed Japanese Shorthorn cattle. Microarray analysis of the c-miRNAs resulted in detection of a total of 231 bovine exosomal miRNAs in the plasma, with a constant expression level of let-7g across the duration and cattle groups. Expression of muscle-specific miRNAs such as miR-1, miR-133a, miR-206, miR-208a/b, and miR-499 were undetectable, suggesting the mildness of grazing movement as exercise. According to validation by quantitative RT-PCR, the circulating miR-150 level in the grazing cattle normalized by the endogenous let-7g level was down-regulated after 2 and 4 months of grazing (P &lt; 0.05), and then its levels in housed and grazing cattle equalized when the grazing cattle were returned to a housed situation. Likewise, the levels of miR-19b, miR-148a, miR-221, miR-223, miR-320a, miR-361, and miR-486 were temporarily lowered in the cattle at 1 and/or 2 month of grazing compared to those of the housed cattle (P &lt; 0.05). In contrast, the miR-451 level was up-regulated in the grazing cattle at 2 months of grazing (P = 0.044). The elevation of miR-451 level in the plasma was coincident with that in the biceps femoris muscle of the grazing cattle (P = 0.008), which suggests the secretion or intake of miR-451 between skeletal muscle cells and circulation during grazing. These results revealed that exosomal c-miRNAs in cattle were affected by grazing, suggesting their usefulness as molecular grazing markers and functions in physiological adaptation of grazing cattle associated with endocytosis, focal adhesion, axon guidance, and a variety of intracellular signaling, as predicted by bioinformatic analysis.

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  • Koichi Ojima, Mika Oe, Ikuyo Nakajima, Masahiro Shibata, Susumu Muroya, Koichi Chikuni, Akihito Hattori, Takanori Nishimura .  The importance of subfragment 2 and C-terminus of myosin heavy chain for thick filament assembly in skeletal muscle cells .  ANIMAL SCIENCE JOURNAL86 ( 4 ) 459 - 467   2015.4Reviewed

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    In skeletal muscle cells, myofibrillar proteins are highly organized into sarcomeres in which thick filaments interdigitate with thin filaments to generate contractile force. The size of thick filaments, which consist mainly of myosin molecules, is strictly controlled. However, little is known about the mechanisms by which myosin molecules assemble into thick filaments. Here, we assessed the ability of each domain of myosin heavy chain (Myh) to form thick filaments. We showed that exogenously expressed subfragment 2 (S2)+light meromyosin (LMM) of Myh was efficiently incorporated into thick filaments in muscle cells, although neither solely expressed S2 nor LMM targeted to thick filaments properly. In nonmuscle COS7 cells, S2+LMM formed more enlarged filaments/speckles than LMM. These results suggest that Myh filament formation is induced by S2 accompanying LMM. We further examined the effects of Myh C-terminus on thick filament assembly. C-terminal deletion mutants were incorporated not into entire thick filaments but rather into restricted regions of thick filaments. Our findings suggest that the elongation of myosin filaments to form thick filaments is regulated by S2 as well as C-terminus of LMM.

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  • Koichi Ojima, Mika Oe, Ikuyo Nakajima, Masahiro Shibata, Koichi Chikuni, Susumu Muroya, Takanori Nishimura .  Proteomic analysis of secreted proteins from skeletal muscle cells during differentiation .  EuPA Open Proteomics5   1 - 9   2014.12Reviewed

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    Myokines are muscle-secreted factors to regulate cellular functions. However, it remains elusive what type of myokine is released during muscle differentiation. Here we evaluated the dynamics of myokines. More than 400 proteins were detected in conditioned medium and approximately 8% of them were categorized as myokines. The levels of myokines which promote myotube formation, vascularization or neurogenesis peaked during early differentiation, whereas myokines contributing to repellent activity against nerve cells or suppression of adipogenesis decreased after differentiation. Our findings suggest that muscle cells secrete different types of myokines at different developmental stages to communicate with various types of cells.

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  • Susumu Muroya, Mika Oe, Ikuyo Nakajima, Koichi Ojima, Koichi Chikuni .  CE-TOF MS-based metabolomic profiling revealed characteristic metabolic pathways in postmortem porcine fast and slow type muscles .  MEAT SCIENCE98 ( 4 ) 726 - 735   2014.12Reviewed

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    To determine key compounds and metabolic pathways associated with meat quality, we profiled metabolites in postmortem porcine longissimus lumborum (LL) and vastus intermedius (VI) muscles with different aging times by global metabolomics using capillary electrophoresis-time of flight mass spectrometry. Loading analyses of the principal component analysis showed that hydrophilic amino acids and beta-alanine-related compounds contributed to the muscle type positively and negatively, respectively, whereas glycolytic and ATP degradation products contributed to aging time. At 168 h postmortem, LL samples were characterized by abundance of combinations of amino acids, dipeptides, and glycolytic products, whereas the VI samples were characterized by abundance of both sulfur-containing compounds and amino acids. The AMP and inosine contents in the VI were approx. 10 times higher than those in the LL at 4 h postmortem, suggesting different rates of inosine 5'-monophosphate (IMP) accumulation by adenylate kinase 7 and 5'-nucleotidase, and subsequent different production levels of IMP and hypoxanthine between these two porcine muscles. (C) 2014 Elsevier Ltd. All rights reserved.

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  • Keisuke Sasaki, Michiyo Motoyama, Takumi Narita, Tatsuro Hagi, Koichi Ojima, Mika Oe, Ikuyo Nakajima, Katsuhiro Kitsunai, Yosuke Saito, Hikari Hatori, Susumu Muroya, Masaru Nomura, Yuji Miyaguchi, Koichi Chikuni .  Characterization and classification of Japanese consumer perceptions for beef tenderness using descriptive texture characteristics assessed by a trained sensory panel .  MEAT SCIENCE96 ( 2 ) 994 - 1002   2014.2Reviewed

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    Meat tenderness is an important characteristic in terms of consumer preference and satisfaction. However, each consumer may have his/her own criteria to judge meat tenderness, because consumers are neither selected nor trained like an expert sensory panel. This study aimed to characterize consumer tenderness using descriptive texture profiles such as chewiness and hardness assessed by a trained panel. Longissimus muscles cooked at four different end-point temperatures were subjected to a trained sensory panel (n = 18) and consumer (n = 107) tenderness tests. Multiple regression analysis showed that consumer tenderness was characterized as 'low-chewiness and low hardness texture.' Subsequently, consumers were divided into two groups by cluster analysis according to tenderness perceptions in each participant, and the two groups were characterized as 'tenderness is mainly low-chewiness' and 'tenderness is mainly low-hardness' for tenderness perception, respectively. These results demonstrate objective characteristics and variability of consumer meat tenderness, and provide new information regarding the evaluation and management of meat tenderness for meat manufacturers. (C) 2013 Elsevier Ltd. All rights reserved.

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  • S. Muroya, M. Taniguchi, M. Shibata, M. Oe, K. Ojima, I. Nakajima, K. Chikuni .  Profiling of differentially expressed microRNA and the bioinformatic target gene analyses in bovine fast- and slow-type muscles by massively parallel sequencing .  Journal of Animal Science91 ( 1 ) 90 - 103   2013.1Reviewed

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    MicroRNA (miRNA) are highly conserved, noncoding small RNA involved in post-transcriptional gene regulation in a variety of biological processes. To elucidate roles of miRNA in bovine muscle type specification and maintenance, we sought to determine differentially expressed miRNA between semitendinosus (STD) and masseter (MS) muscles from 3 Japanese black cattle by massively parallel sequencing. Differential gene expression of myosin heavy chain (MyHC) isoforms confirmed that STD and MS were MyHC-2x-and MyHC-1-abundant muscles, respectively. In total, 192 known miRNA and 20 potential new bovine miRNA were obtained from the sequencing. The differentially expressed miRNA with more than 2-fold difference in each muscle were identified. In particular, miR-196a and miR-885 were exclusively expressed in STD muscle, which was validated by quantitative reverse transcription-PCR (P = 0.045 and P &lt
    0.001, respectively), whereas a slow type-directing miR-208b was highly expressed in MS compared with STD (false discovery rate &lt
    0.05). In addition, 16 potential novel miRNA were mapped and confirmed for their precursor structures by computational analyses. The results of functional annotation combined with in silico target analysis showed that the predicted target genes of miR-196a/b and miR-885 enriched gene ontology (GO) terms related to skeletal system development and regulation of transcription, respectively. Moreover, GO terms enriched from predicted targets miRNA suggested that STD-abundant and MS-abundant-miRNA were associated with embryonic body planning and organ/ tissue pattern formation, respectively. The present results revealed that the differentially expressed miRNA between the STD and MS muscles may play key roles to determine muscle type-specific tissue formation and maintenance in cattle thorough attenuating putative target genes involved in different developmental events. © 2013 American Society of Animal Science. All rights reserved.

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  • Koichi CHIKUNI, Keisuke SASAKI, Michiyo MOTOYAMA, Ikuyo NAKAJIMA, Koichi OJIMA, Mika OE, Susumu MUROYA .  ブタ肉中のイノシン酸含量におよぼす筋肉型の影響 .  Nihon Yoton Gakkaishi50 ( 1 ) 8 - 14   2013Reviewed

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    Nucleotides are important components of flavor-precursor complex of meat. To clarify the effects of muscle type on the nucleotide contents, the concentration of nucleotides were determined from porcine muscles. The <I>masseter, diaphragm, semispinalis, psoas major, longissimus thoracis</I>, and <I>semitendinosus </I>muscles were sampled at 7 day <I>post-mortem</I> from 80 and 110 kg pigs (four animals in each weight group). The contents of myosin heavy chain (MyHC) slow isoform determined with ELISA were 56.1, 51.5, 39.7, 24.0, 10.2 and 7.8% in <I>diaphragm, semispinalis, masseter, psoas major, longissimus thoracis </I>and <I>semitendinosus </I>muscles, respectively. The contents of inosine 5´-monophosphate (IMP) were significantly higher in the fast type muscles than that in the slow type muscles. The IMP contents were 5.28, 4.99, 4.83, 3.28, 2.28 and 2.07 μmol/g in the <I>longissimus thoracis, psoas major, semitendinosus, semispinalis masseter </I>and <I>diaphragm </I>muscles, respectively. The total content of ATP degradation products was higher in the fast type muscles, and ultimate pH was higher in the slow type muscles. These results suggested that muscle types affected the IMP contents in porcine muscles through the ATP degradation procedures <i>post-mortem</i>.

    DOI: 10.5938/youton.50.8

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  • Keisuke Sasaki, Masayuki Hayashi, Takumi Narita, Michiyo Motoyama, Mika Oe, Koichi Ojima, Ikuyo Nakajima, Susumu Muroya, Koichi Chikuni, Katsuhiro Aikawa, Yasuyuki Ide, Naoto Nakanishi, Nobuaki Suzuki, Shigeru Shioya, Akio Takenaka .  Radiocesium Distribution in the Tissues of Japanese Black Beef Heifers Fed Fallout-Contaminated Roughage Due to the Fukushima Daiichi Nuclear Power Station Accident .  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY76 ( 8 ) 1596 - 1599   2012.8Reviewed

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    This study examined the accumulation and tissue distribution of radioactive cesium nuclides in Japanese Black beef heifers raised on roughage contaminated with radioactive fallout due to the accident at the Fukushima Daiichi Nuclear Power Station on March 2011. Radiocesium feeding increased both Cs-134 and Cs-137 levels in all tissues tested. The kidney had the highest level and subcutaneous adipose had the lowest of radioactive cesium in the tissues. Different radioactive cesium levels were not found among parts of the muscles. These results indicate that radiocesium accumulated highly in the kidney and homogenously in the skeletal muscles in the heifers.

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  • Seiichi IRIE, Koichi OJIMA, Mika OE, Susumu MUROYA, Koichi CHIKUNI, Ikuyo NAKAJIMA .  ブタ皮下脂肪前駆細胞株の分化に対するオクタン酸の濃度及び添加時期の影響 .  Nihon Yoton Gakkaishi49 ( 4 ) 150 - 159   2012Reviewed

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    To control the amount of adipose tissue is one of the greatest concern in pig industries. In this study, we investigated the effect of varying doses and times of adding octanoate on differentiation of pig preadipocytes under culture conditions including oleate. A clonal cell line of preadipocytes established from pig subcutaneous tissue (PSPA) were exposed to 1 μM∼5 mM of octanoate in addition to adipocyte inducing factors such as insulin, dexamethasone, biotin, pantothenate, and oleate. At the concentration of more than 1 mM octanoate, number of cells significantly decreased while triglyceride (TG) accumulation increased in a dose-dependent manner. Gene expression levels by RT-PCR analysis showed that the key adipogenic transcription factors, i.e. peroxisome proliferators-activated receptor (PPAR) γ2 and CCAAT enhancer-binding protein (C/EBP) α were not affected by the addition of octanoate. However, mRNA expression patterns of proliferation and differentiation markers, such as proliferating cell nuclear antigen (PCNA), adipocyte-specific fatty acid binding protein (aP2), adiponectin and perilipin 1 were consistent with the results of cell number and TG contents. Adding octanoate at various times after cell confluence showed that octanoate should be supplemented throughout the differentiation to obtain more lipid laden-adipocytes, because TG amounts were lower in cells treated with octanoate only during the first, mid or last 4 days than the cells fully treated with octanoate during 10 days culture. Instead of low TG content, short term octanoate-treated cells had high cell numbers, indicating that they contiuned cell-cycle progression. Overall, these results suggest that function of octanoate in adipocyte differentiation is reversible and its dose of more than 1 mM would be useful to increase lipids of PSPA cells even under oleate-added condition.

    DOI: 10.5938/youton.49.150

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  • Shibata M, Matsumoto K, Hikino Y, Oe M, Ojima K, Nakajima I, Muroya S, Chikuni K, Yamamoto N .  Effect of Grass Hay Feeding on Meat Production, Carcass Characteristics, and Meat Quality in Japanese Black Steers. .  Bulletin of NARO Western Region Agricultural Research Center11 ( 11 ) 15 - 25   2012Reviewed

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  • Susumu Muroya, Kate E. Neath, Ikuyo Nakajima, Mika Oe, Masahiro Shibata, Koichi Ojima, Koichi Chikuni .  Differences in mRNA expression of calpains, calpastatin isoforms and calpain/calpastatin ratios among bovine skeletal muscles .  ANIMAL SCIENCE JOURNAL83 ( 3 ) 252 - 259   2012Reviewed

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    Messenger RNA (mRNA) expression of calpain-1 (mu-calpain), -2 (m-calpain), -3 (p94), small subunit (calpain-4; 28 kDa), and three types of calpastatin (CSTN) isoform were investigated for 10 skeletal muscles of Holstein cattle by real-time and/or semi-quantitative reverse transcription polymerase chain reaction. Noticeably, effect of muscle type was observed on 28 kDa expression (P &lt; 0.001) with a tendency of higher 28 kDa expression in myosin heavy chain (MyHC)-2x-rich muscles compared to MyHC-slow-rich muscles. The CSTN-I and III expression in Longissimus thoracis (LT) showed the lowest value among the muscles tested. Moreover, 28 kDa/CSTN-I ratio was higher in the diaphragm (DP), psoas major (PM), and LT than those in the lingual muscles (TN), masseter (MS) and pectoralis (PP) (P &lt; 0.05). Calpain-1/CSTN I, calpain-2/CSTN I in LT and PM were higher than that in TN (P &lt; 0.05). Calpain-3/CSTN-I and -III in LT and/or PM showed higher values than that in TN (P &lt; 0.05). These results indicated that the calpain and CSTN expressions are regulated by muscle type, suggesting especially by muscle fiber type. Calpains/CSTN-I ratios, especially 28 kDa/CSTN-I, may account for higher extent of post mortem proteolysis previously observed in LT and PM muscles.

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  • KAMIYA Mitusru, MUROYA Susumu, KAMIYA Yuko, KAMADA Toshihiko, TANAKA Masahito .  <b>Effects of High Ambient Temperature on Plasma 3-methylhistidine Concentration and Expression of Muscle Calpain and Calpastatin mRNA in High-yielding Lactating Holstein Co</b><b>ws </b> .  Nihon Souchi Gakkai Kyushu Shibukaiho55 ( 1 ) 49 - 53   2012

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    <tt>The effect of high ambient temperature on myofibrillar proteolysis in high-yielding lactating cows was investigated. In a 2 x 2 crossover design, four multiparous lactating Holstein cows were maintained in a chamber under conditions of constant moderate ambient temperature (MT) or high ambient temperature (HT). Dry matter intake (P < 0.10) and milk yield (P < 0.10), and milk protein yield (P < 0.05) were lower in HT than MT. Urinary 3-methylhistidine excretion were 1.87 mmol/day in MT and 1.92 mmol/day in HT, but no statistically significant difference was found. Plasma 3-methylhistidine concentrations were higher (P < 0.05) in HT (6.9 nmol/ml) than MT (4.1 nmol/ml), indicating that high ambient temperature increased the myofibrillar proteolysis. The expression of muscle calpain SS (small subunit (28kDa)), -1, and -3, and calpastatin mRNA in HT were increased 1.4, 1.1, 1.6, and 1.4-fold of MT, respectively, but no statistically significant difference was found. Meanwhile, the correlation between the level of plasma 3-methylhistidine and the level of muscle calpain 3 mRNA was significant (P < 0.05) and its correlation coefficient was the highest in calpain family. In conclusion, high ambient temperature affected the plasma index of myofibrillar proteolysis related to the expression of muscle calpain 3 </tt><tt>and calpastatin mRNA in high-yielding lactating dairy cows. </tt>

    DOI: 10.11461/jwaras.55.49

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  • Masahiro Shibata, Kazunori Matsumoto, Yasuko Hikino, Mika Oe, Koichi Ojima, Ikuyo Nakajima, Susumu Muroya, Koichi Chikuni .  Influence of different feeding systems on the growth performance and muscle development of Japanese Black steers .  MEAT SCIENCE89 ( 4 ) 451 - 456   2011.12Reviewed

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    This study investigated the growth performance and gene expression for muscle development between grass hay-fed (GH) and concentrate-fed (CT) steers. Daily gain and energy intake during the fattening period of the GH group were lower than those of the CT group. Analysis of C/EBP alpha, PPAR gamma 2, myosin heavy chain (MHC), and myostatin gene expressions was performed by real-time PCR. Expressions of C/EBP alpha and myostatin in semitendinosus and longissimus lumborum (LL) muscles were higher in the CT group than in the GH group at the end of fattening. In LL muscle, MHC expression at the end of fattening was greater in the GH group than in the CT group. These results suggest that regulation of adipogenesis and myogenesis by the expression of genes involved in muscle development might have occurred in the skeletal muscle of the GH group by the feeding of grass hay and/or because of the low energy intakes, (C) 2011 Elsevier Ltd. All rights reserved.

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  • Ikuyo Nakajima, Mika Oe, Koichi Ojima, Susumu Muroya, Masahiro Shibata, Koichi Chikuni .  Cellularity of developing subcutaneous adipose tissue in Landrace and Meishan pigs: Adipocyte size differences between two breeds .  ANIMAL SCIENCE JOURNAL82 ( 1 ) 144 - 149   2011Reviewed

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    Experiments were designed to compare the adipocyte cellularity of subcutaneous adipose tissue between growing Landrace (low backfat) and Meishan (high backfat) pigs at 1 week, 3 weeks, 6 weeks, 3 months and 5 months of age. As pigs aged, body weight and backfat thickness of both breeds significantly increased. When compared at equal ages, backfat thickness adjusted to equal body weight was greater for Meishan pigs. The mean diameter of fat cell size also increased with age, and by 6 weeks adipocytes from both outer and inner layers of subcutaneous adipose tissue were larger in Meishan pigs. At 5 months, approximately 80% of the adipose tissue mass in Meishan pigs was attributable to adipocytes measuring 95-165 mu m in diameter, whereas adipocytes of 75-145 mu m comprised most of the tissue mass in the Landrace. Although the contribution of smaller adipocytes (25-45 mu m) to the tissue volume was negligible, both breeds showed a biphasic diameter distribution at all ages, suggesting that adipocyte hyperplasia is still active. Our results demonstrate that cellularity differences exist between the subcutaneous adipose tissues of Landrace and Meishan pigs, and adipocyte hypertrophy is the most overwhelming contributor to the greater backfat deposition for Meishan pigs.

    DOI: 10.1111/j.1740-0929.2010.00810.x

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  • Mika Oe, Mayumi Ohnishi-Kameyama, Ikuyo Nakajima, Susumu Muroya, Masahiro Shibata, Koichi Ojima, Shiro Kushibiki, Koichi Chikuni .  Proteome analysis of whole and water-soluble proteins in masseter and semitendinosus muscles of Holstein cows .  ANIMAL SCIENCE JOURNAL82 ( 1 ) 181 - 186   2011Reviewed

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    To assess both quantitative and qualitative differences between the slow- and fast-type muscles, masseter (slow) and semitendinosus (fast) from four Holstein cows were analyzed by two-dimensional difference gel electrophoresis (2D DIGE) and mass spectrometry. The proteome analysis identified 27 spots as 20 proteins in the whole protein fraction extracted with 8 mol/L urea solution, and 16 spots were identified as 11 proteins in the water-soluble protein fraction. Two slow-type myofibrillar proteins (myosin light chain-1 slow-b and myosin light chain-2 slow), and aconitase-2 mitochondria were present at higher levels in the masseter muscle (P &lt; 0.05). Four fast-type myofibrillar proteins (myosin light chain-1 fast, myosin light chain-2 fast, myosin light chain-3 fast and tropomyosin-1), and three enzymes of glycolytic pathway (enolase-3, aldolase-A and triosephosphate isomerase), were present at higher levels in the semitendinosus muscle (P &lt; 0.05). Our proteome analysis showed that the composition of sarcoplasmic proteins as well as myofibrillar proteins was clearly different between slow- and fast-type muscles.

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  • Susumu Muroya, Per Ertbjerg, Luigi Pomponio, Mette Christensen .  Desmin and troponin T are degraded faster in type IIb muscle fibers than in type I fibers during postmortem aging of porcine muscle .  MEAT SCIENCE86 ( 3 ) 764 - 769   2010.11Reviewed

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    A novel approach was applied in this study to directly evaluate the effect of muscle fiber type on postmortem protein degradation. Porcine muscle fibers were isolated from longissimus muscle at day 1, 3, and 6 postmortem. Fibers were sorted by immunochemical myosin heavy chain isoform typing. Western blot analysis of fibers pooled separately into type I or IIb showed that the relative amounts of 39- and 50-kDa desmin degradation fragments at day 6, and 28- to 31-kDa fragments of troponin T fast type isoform (fTnT) at day 1 and 6 postmortem were higher in type IIb than in type I fibers. At day 6 troponin T slow type isoform (sTnT) was less degraded than fTnT in type I fibers. These results indicated greater rate and extent of proteolysis in type IIb than in type I fibers and higher susceptibility of fTnT to proteolysis than that of sTnT isoform. (C) 2010 The American Meat Science Association. Published by Elsevier Ltd. All rights reserved.

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  • Koichi Chikuni, Mika Oe, Keisuke Sasaki, Masahiro Shibata, Ikuyo Nakajima, Koichi Ojima, Susumu Muroya .  Effects of muscle type on beef taste-traits assessed by an electric sensing system .  ANIMAL SCIENCE JOURNAL81 ( 5 ) 600 - 605   2010Reviewed

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    To assess the role of muscle fiber type in beef taste-traits, we analyzed cooked meats from bovine masseter, diaphragm, psoas major, longissimus thoracis, and semitendinosus muscles with an electric taste sensing system (INSENT SA402B). The system is composed of five taste sensors of polymer membranes fixing different lipids. The sensors, CT0, CA0, AAE, C00 and AE1 are designed to respond to the individual tastes of salty, sour, umami, bitter and astringent, respectively. The system found significant differences in the converted outputs of CA0 (cvCA0), C00 (cvC00) and AE1 (cvAE1) among the bovine muscles. The slow-type muscles (masseter and diaphragm) showed lower cvCA0, higher cvC00, and higher cvAE1 than did the fast-type muscles (psoas major, longissimus thoracis, and semitendinosus). Lactic acid content was different among muscle types and was highly related to the cvCA0 output and pH. carbonyl compounds and free fatty acids were higher in the slow-type muscles. Free fatty acids were major components causing the difference in the C00 output among the muscle types. Iron content was also different among the muscle types and related to the cvC00 and cvAE1 outputs. These results suggested that the muscle fiber type affects the beef taste characteristics.

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  • Susumu Muroya, Mika Oe, Ikuyo Nakajima, Masahiro Shibata, Koichi Chikuni .  A non-destructive method to monitor changes in a troponin T peptide in beef drip with a monoclonal antibody .  MEAT SCIENCE83 ( 1 ) 155 - 160   2009.9Reviewed

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    To simplify the monitoring of postmortem beef aging, we established a system to detect a troponin T (TnT) peptide fragment in bovine muscle drip (natural exudates) with an original monoclonal antibody. The antibody was raised against a synthetic peptide APPPPAEVPEVHEEVH corresponding to the N-terminal region of bovine fast-type TnT. In a competitive enzyme-linked immunosorbent assay (ELISA), our antibody detected the standard peptide dose-dependently. According to the monitoring examination with a competitive ELISA during 22 days postmortem, the concentration of the peptide in both the drip and trichloroacetic acid extracts from the longissimus muscle (n = 4) significantly increased in parallel, up to 10 nmol/ml and 16.4 nmol/g at day 14 postmortem, respectively. These events were accompanied by an increase in the conventional 30 kDa fragment in western blot analysis and a decrease in the Warner-Bratzler shear force value of the beef from 5.0 to 2.4 N/cm(2). The peptide detection system using drips with the antibody has advantages applicable to a non-destructive, simple, quick, and on-site monitoring method, such as immunochromatography. (C) 2009 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.meatsci.2009.04.012

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  • M. Shibata, K. Matsumoto, M. Oe, M. Ohnishi-Kameyama, K. Ojima, I. Nakajima, S. Muroya, K. Chikuni .  Differential expression of the skeletal muscle proteome in grazed cattle .  JOURNAL OF ANIMAL SCIENCE87 ( 8 ) 2700 - 2708   2009.8Reviewed

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    The objective of this study was to investigate the differences in the muscle proteome of grass-fed and grain-fed cattle. Eight Japanese Black Cattle 10 mo of age were separated randomly into 2 groups: 1) grazing (grass-fed) and 2) concentrate (grain-fed) groups. All cattle were first housed individually in a stall barn and fed a combination of concentrate ad libitum and Italian ryegrass hay until 21 mo of age. After this control period, the 4 grass-fed cattle were placed on outdoor pasture, whereas the other 4 grain-fed cattle continued on the concentrate diet. The cattle were slaughtered at 27 mo of age, and tissues from the semitendinosus muscle were obtained for use in proteome analysis. Differential expression of muscle proteins in the 2 groups was carried out using 2-dimensional gel electrophoresis (2DE) and Western blot analyses, with subsequent mass spectrometry. Approximately 200 individual protein spots were detected and compared in each group using 2DE, of which 20 and 9 spots, respectively, showed differences in the spot intensity for the sarcoplasmic fraction and myofibrillar fraction. In the grazing group, the relative intensity of spots was significantly greater for adenylate kinase 1 and myoglobin in the sarcoplasmic fraction, and for slow-twitch myosin light chain 2 in the myofibrillar fraction (P &lt; 0.05), than the concentrate group. The relative spot intensity of several glycolytic enzymes was significantly greater in the grazing group, such as beta-enolase 3, fructose-1,6-bis-phosphate aldolase A, triosephosphate isomerase, and heat shock 27 kDa protein (P &lt; 0.05). Moreover, significantly greater slow twitch of troponin T, troponin I, and myosin heavy chain of semitendinosus muscle was detected in the grazing group than in the concentrate group using Western blot analysis (P &lt; 0.05). Several previous reports have described that the slow-twitch muscle contents affect elements of nutrition, flavor, and food texture of meat. This study revealed muscle fiber type conversion to slow-twitch tissues from fast-twitch tissues occurring with change in the energy metabolic enzyme when cattle were grazed in the latter fattening period. Although analyses of the influence on elements of nutrition, flavor, and food texture were not done for this study, these results show that slow-twitch converted muscle resulting from the grazing of cattle might modify several meat characteristics.

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  • Mika Oe, Ikuyo Nakajima, Susumu Muroya, Masahiro Shibata, Koich Chikuni .  Relationships between tropomyosin and myosin heavy chain isoforms in bovine skeletal muscle .  ANIMAL SCIENCE JOURNAL80 ( 2 ) 193 - 197   2009.4Reviewed

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    The composition of tropomyosin (TPM) and myosin heavy chain (MyHC) isoforms was analyzed in 10 physiologically different bovine muscles (masseter, diaphragm, tongue, semispinalis, pectoralis profundus, biceps femoris, psoas major, semimembranosus, longissimus thoracis and semitendinosus) to clarify the relationships between TPM and MyHC isoforms in different muscle fiber types. The content of TPM1 and TPM3 was different in muscles according to their function in muscle contraction, although the content of TPM2 was constantly about 50% of the total TPM in all muscles. The content of TPM1 was higher in semimembranosus, longissimus thoracis and semitendinosus, while that of TPM3 was higher in masseter and diaphragm. The high positive correlation between MyHC-slow content and TPM3 content (r = 0.92) suggested a coexpression of TPM3 and MyHC-slow isoforms in a muscle fiber. MyHC-slow and TPM3 were expressed at the same level in masseter and diaphragm, whereas there was more TPM3 than MyHC-slow in tongue and semispinalis, so it appears that the excess TPM3 in tongue and semispinalis is expressed with other MyHC isoforms. MyHC-2a was the only fast type isoform expressed in tongue and semispinalis. Therefore, the excess TPM3 was composed of myofibrils with MyHC-2a. The results suggested that a fiber expressing MyHC-2a would be regulated delicately by changing the TPM isoform types.

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  • Keisuke Sasaki, Koichi Chikuni, Ikuyo Nakajima, Mika Oe, Michiyo Motoyama, Susumu Muroya .  Changes in Thiamin Contents in Porcine Muscles and Liver during Growth .  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY73 ( 1 ) 177 - 179   2009.1Reviewed

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    Changes in the thiamin contents in three types of porcine muscle and porcine liver during growth were investigated. The muscular thiamin content was lower at the newborn stage than at fetal stage, and increased after the weaning period. The liver thiamin content, however, remained unchanged from the fetal stage to 5 months old. The changes in thiamin contents were different between Landrace and Meishan pigs.

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  • Susumu Muroya, Kouichi Watanabe, Shinichiro Hayashi, Masato Miyake, Shigeru Konashi, Youichi Sato, Manabu Takahashi, Shigeki Kawahata, Yoshisato Yoshikawa, Hisashi Aso, Koichi Chikuni, Takahiro Yamaguchi .  Muscle type-specific effect of myostatin deficiency on myogenic regulatory factor expression in adult double-muscled Japanese Shorthorn cattle .  ANIMAL SCIENCE JOURNAL80 ( 6 ) 678 - 685   2009Reviewed

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    To clarify muscle type-specific effect of myostatin on myogenic regulatory factors (MRFs), we examined mRNA expression of MRFs in five skeletal muscles of normal (NM) and myostatin-deficient double-muscled (DM) adult Japanese Shorthorn cattle by quantitative reverse-transcribed PCR. Among the four MRFs, namely, Myf5, MyoD, myogenin, and MRF4, MyoD expression was different among the muscles of the DM cattle (P &lt; 0.01) but not of the NM cattle. Meanwhile, MyoD expression was significantly elevated only in masseter (MS) muscle in the DM cattle due to the myostatin deficiency (P &lt; 0.05). Myf5 and MRF4 expression in semitendinosus (ST) was higher in the DM than in the NM cattle (P &lt; 0.05). According to analysis of myosin heavy chain (MyHC) isoform expression, more MyHC-2x and -2a and less -slow isoforms were expressed in the longissimus and ST muscles compared to the MS muscle in both cattle (P &lt; 0.05), but no significant difference in MyHC expression was observed between the NM and DM cattle. Taken together, myostatin has influences on Myf5 and MRF4 expression in faster-type muscles and on MyoD expression in slower-type muscles, suggesting a possible muscle type-specific effect of myostatin in skeletal muscle growth and maintenance.

    DOI: 10.1111/j.1740-0929.2009.00684.x

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  • Koichi Chikuni, Atsushi Horiuchi, Hanako Ide, Masatoshi Shibata, Takeshi Hayashi, Ikuyo Nakajima, Mika Oe, Susumu Muroya .  Nucleotide sequence polymorphisms of beta1-, beta2-, and beta3-adrenergic receptor genes on Jinhua, Meishan, Duroc and Landrace pigs .  ANIMAL SCIENCE JOURNAL79 ( 6 ) 665 - 672   2008.12Reviewed

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    The full amino acid coding sequences of adrenergic receptor genes beta1, beta2, and beta3 (ADRB1, ADRB2, and ADRB3)were determined for Jinhua, Meishan, Duroc and Landrace pigs. Non-synonymous substitution of Arg458Pro was found in the porcine ADRB1 gene, resulting in a 469 amino acid sequence. Continuous substitutions of Asn29Asp and Glu30Gln were found in the porcine ADRB2 gene, resulting in a 418 amino acid sequence. Additionally, a Lys30 polymorphism of the ADRB2 gene was found in the Jinhua pigs. There were three non-synonymous substitutions of Asn24Thr, Arg264Gln and Asn398Asp on the porcine ADRB3 gene. A thymine insertion in the ADRB3 gene, resulting in a protein with two fewer amino acids, was found in the Jinhua and Meishan pigs. To assess the effect of ADRB polymorphisms on porcine subcutaneous fat layer thickness, we calculated the genetic frequency of the variants in fatty and lean groups, each consisting of 24 pigs that were crossbreds of Duroc and Jinhua pigs. The effect of the ADRB3 gene polymorphism was not evaluated, because there was insufficient variation on the ADRB3 gene in the examined groups. Although Fisher&apos;s exact test showed no significant difference in the frequency of ADRB1 and ADBR2 variants between the two groups, the Arg458 variant of ADRB1 was higher (P = 0.11) in the lean group, and pigs in that group had a thinner fat layer than did those with the Pro458 variant. These results imply a possibility of ADRB1 polymorphism as a minor factor in porcine fat layer thickness. The Asp29 variant of ADRB2 was higher in the lean group (P = 0.11), and the Glu30 variant was higher in the fatty group (P = 0.15), but the Asp29 variant was found only in the Chinese pigs. Thus, the effect of ADRB2 polymorphisms was not clear in this study.

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  • Ichiba M, Nakamura M, Kusumoto A, Mizuno E, Kurano Y, Matsuda M, Kato M, Agemura A, Tomemori Y, Muroya S, Nakabeppu Y, Sano A .  Clinical and molecular genetic assessment of a chorea-acanthocytosis pedigree. .  Journal of the neurological sciences263 ( 1-2 ) 124 - 32   2007.12Clinical and molecular genetic assessment of a chorea-acanthocytosis pedigree.

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  • K. E. Neath, A. N. Del Barrio, R. M. Lapitan, J. R. V. Herrera, L. C. Cruz, T. Fujihara, S. Muroya, K. Chikuni, M. Hirabayashi, Y. Kanai .  Protease activity higher in postmortem water buffalo meat than Brahman beef .  MEAT SCIENCE77 ( 3 ) 389 - 396   2007.11Reviewed

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    We previously demonstrated that postmortem water buffalo meat had higher tenderness than Brahman beef. In order to explain this difference in tenderness, the objective of the current study was to investigate the protease activity in these two meats. Five female crossbred water buffalo (Philippine Carabao x Bulgarian Murrah) and five female crossbred cattle (Brahman x Philippine Native) were slaughtered at 30 months of age, followed by immediate sampling of Longissimus thoracis muscle for measurement of protease activity. Results showed that buffalo meat had significantly higher protease activity compared to beef (P &lt; 0.05). Furthermore, calpain inhibitor 1, a specific inhibitor of calpains 1 and 2, was the most effective inhibitor of protease activity. There was no difference in calpastatin activity, and no major differences were observed in calpains 1, 2, and calpastatin expression by Western blotting. This study suggests that higher calpain activity in early postmortem buffalo meat was responsible for the increased tenderness of water buffalo meat compared to beef. (c) 2007 Elsevier Ltd. All rights reserved.

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  • 室谷 進 .  食肉の付加価値向上をめざした筋タンパク質トロポニンTの基礎研究 .  食肉の科学48 ( 1 ) 20 - 28   2007.6

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  • Susumu Muroya, Mayumi Ohnishi-Kameyama, Mika Oe, Ikuyo Nakajima, Masahiro Shibata, Koichi Chikuni .  Double phosphorylation of the myosin regulatory light chain during rigor mortis of bovine longissimus muscle .  JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY55 ( 10 ) 3998 - 4004   2007.5Reviewed

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    To investigate changes in myosin light chains (MyLCs) during postmortem aging of the bovine longissimus muscle, we performed two-dimensional gel electrophoresis followed by identification with matrix-assisted laser desorption ionization time-of-flight mass spectrometry. The results of fluorescent differential gel electrophoresis showed that two spots of the myosin regulatory light chain (MyLC2) at pI values of 4.6 and 4.7 shifted toward those at pI values of 4.5 and 4.6, respectively, by 24 h postmortem when rigor mortis was completed. Meanwhile, the MyLC1 and MyLC3 spots did not change during the 14 days postmortem. Phosphoprotein-specific staining of the gels demonstrated that the MyLC2 proteins at pI values of 4.5 and 4.6 were phosphorylated. Furthermore, possible N-terminal region peptides containing one and two phosphoserine residues were detected in each mass spectrum of the MyLC2 spots at pI values of 4.5 and 4.6, respectively. These results demonstrated that MyLC2 became doubly phosphorylated during rigor formation of the bovine longissimus, suggesting involvement of the MyLC2 phosphorylation in the progress of beef rigor mortis.

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  • M. Oe, M. Ohnishi-Kameyama, I. Nakajima, S. Muroya, K. Chikuni .  Muscle type specific expression of tropomyosin isoforms in bovine skeletal muscles .  MEAT SCIENCE75 ( 4 ) 558 - 563   2007.4Reviewed

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    Nucleotide sequences encoding an entire coding region for bovine tropomyosin (TPM) isoforms were determined. Three TPM isoforms, TPM1, TPM2 and TPM3, were expressed in bovine skeletal muscles, and exhibited a 93.3%, 99.6% and 100% amino acid homology to the human sequence, respectively. Based on the sequences, the composition of TPM isoforms was analyzed on cDNA and protein levels from five physiologically different muscles (masseter, diaphragm, psoas major, longissimus thoracis and semitendinosus) using RTPCR and proteome analyses. Although the content of TPM2 was constantly about 50% of the total TPM in all muscles, the contents of TPM1 and TPM3 were different in muscles according to their function in muscle contraction. In masseter, the content of TPM3 cDNA was about 50% and higher than that of other muscles. In longissimus thoracis and semitendinosus, the contents of TPM1 cDNA were 29.6% and 31.7%, respectively, which were comparatively higher than that of other muscles. The result suggests that the TPM dimer consists of the TPM2 subunit regularly and TPM1 or TPM3 depending on whether the muscle is fast or slow type, respectively. (c) 2006 Elsevier Ltd. All rights reserved.

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  • K. E. Neath, A. N. Del Barrio, R. M. Lapitan, J. R. V. Herrera, L. C. Cruz, T. Fujihara, S. Muroya, K. Chikuni, M. Hirabayashi, Y. Kanai .  Difference in tenderness and pH decline between water buffalo meat and beef during postmortem aging .  MEAT SCIENCE75 ( 3 ) 499 - 505   2007.3Reviewed

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    The objective of this research was to determine the difference in tenderness and some characteristics of water buffalo meat and beef during postmortem aging. Five female crossbred water-buffalo (Philippine Carabao x Bulgarian Murrah) and five female crossbred cattle (Brahman x Philippine Native), were finished on the same diet for 6 months and slaughtered at 30 months of age. The muscle pH was measured at 40 min, 3 It, 7 h, 24 h, and 48 h postmortem. Longissimus thoracis (LT) and semimembranosus (SM) muscles were excised at 2 d postmortem, and shear force was measured at 2, 4, 7, and 14 d postmortem. Glycogen and lactate concentrations were determined from 0, 2, and 4 d LT samples, and myosin heavy chain type of buffalo and cattle LT was determined by ELISA methods. Myofibrillar protein degradation was also observed by SDS-PAGE and Western blotting of fast-type troponin T. Results showed that the buffalo meat had significantly lower shear force values compared to beef for LT and SM muscles, which was supported by a difference in troponin T degradation. Postmortem pH decline of buffalo meat was significantly slower than that of beef, which was confirmed by lactic acid concentrations, but was not explained by glycogen content. In addition, there was no significant difference in the ratio of slow to fast type muscle fibers in buffalo and cattle, indicating that myosin heavy chain type was not responsible for the difference in pH decline and tenderness between the buffalo meat and beef. This study demonstrated that the tenderness of water buffalo meat was superior to that of Brahman beef, which may have been due to the difference in pH decline and the subsequent effect on muscle protease activity. (c) 2006 Elsevier Ltd. All rights reserved.

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  • Susumu Muroya, Mayumi Ohnishi-Kameyama, Mika Oe, Ikuyo Nakajima, Koichi Chikuni .  Postmortem changes in bovine troponin T isoforms on two-dimensional electrophoretic gel analyzed using mass spectrometry and western blotting: The limited fragmentation into basic polypeptides .  MEAT SCIENCE75 ( 3 ) 506 - 514   2007.3Reviewed

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    To comprehend postmortem changes in troponin T (TnT), whole beef proteins were developed on a two-dimensional electrophoretic gel. Multiple TnT-related spots were identified by both western blotting and MALDI-TOF MS utilizing bovine TnT isoform mRNA sequences. More than 10 TnT fast-type isoform spots (pI 5.7-9.6 &lt;) and the two slow-type isoform spots (pI 5.6-5.7) were observed at slaughter. All the isoforms were degraded exclusively into basic spots (pI 9.6 &lt;) at day 14 postmortem. Some TnT-related phosphorylated spots present at slaughter had disappeared by day 14, suggesting that the phosphorylated N-terminal region was cut off during beef aging. The intact isoforms and the fragments were identified by the MS with sequence coverage of 20.8-62.7%, and two of the fragments included the cutting site peptide of a conventional 30 kDa or of a slow TnT-derived fragment. These results revealed that all of the TnT isoforms are cut exclusively in the glutamic acid-rich amino-terminal region during postmortem aging. (c) 2006 Elsevier Ltd. All rights reserved.

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  • Mizuno E, Nakamura M, Agemura A, Kusumoto A, Ichiba M, Kurano Y, Muroya S, Sano A .  Brain-specific transcript variants of 5' and 3' ends of mouse VPS13A and VPS13C. .  Biochemical and biophysical research communications353 ( 4 ) 902 - 7   2007.2Brain-specific transcript variants of 5' and 3' ends of mouse VPS13A and VPS13C.

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  • K. E. Neath, A. N. Del Barrio, R. M. Lapitan, J. R. V. Herrera, L. C. Cruz, T. Fujihara, S. Muroya, K. Chikuni, M. Hirabayashi, Y. Kanai .  Relation of postmortem protease activity to tenderness in buffalo meat and Brahman beef .  ITALIAN JOURNAL OF ANIMAL SCIENCE6 ( SUPPL. 2 ) 1175 - 1177   2007Reviewed

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    We previously showed that meat from crossbred water buffalo had significantly higher tenderness than beef from crossbred Brahman cattle of the same age, gender, and diet. Extensive studies on meat tenderness have indicated that proteases degrade muscle fibre proteins during postmortem storage, leading to weakening of the myofibrillar structure and an increase in tenderness. Thus, we investigated the difference in protease activity immediately postmortem, in order to explain the difference in tenderness between buffalo meat and beef. Five female crossbred water-buffalo (Philippine Carabao x Bulgarian Murrah) and five female crossbred cattle (Brahman x Philippine Native) were slaughtered at 30 months of age, and Longissimus thoracis muscle was sampled immediately post-slaughter. Protease activity at different pH levels and the effect of various inhibitors on protease activity were examined Results showed that buffalo meat had significantly higher protease activity compared to beef, and calpain inhibitor 1 was the most effective inhibitor. As calpain inhibitor 1 is a specific inhibitor of calpain 1 and 2, the results suggest that higher calpain activity in buffalo meat was responsible for the higher tenderness of buffalo meat compared to Brahman beef.

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  • Shibata M, Matsumoto K, Aikawa K, Muramoto T, Muroya S, Oe M, Nakajima I, Chikuni K, Fujimura S, Kadowaki M .  Myostatin and Adipogenic Transcription Factor Expressions During Skeletal Muscular Growth of Japanese Black Cattle .  Proceedings of 52nd International Congress Meat Science and Technology ( CD )   2006.8Myostatin and Adipogenic Transcription Factor Expressions During Skeletal Muscular Growth of Japanese Black CattleReviewed

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  • S Kitamura, S Muroya, Nakajima, I, K Chikuni, T Nishimura .  Amino acid sequences of porcine fast and slow troponin T isoforms .  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY70 ( 3 ) 726 - 728   2006.3Reviewed

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    In this study, 10 troponin T isoforms from adult porcine skeletal muscle messenger RNA were clarified. These were eight fast- and two slow-type isoforms. Fast-type isoforms had three and two variable exons in the N-terminal and the C-terminal region respectively. Slowtype isoforms had one variable exon in the N-terminal region.

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  • S Muroya, Nakajima, I, M Oe, K Chikuni .  Difference in postmortem degradation pattern among troponin T isoforms expressed in bovine longissimus, diaphragm, and masseter muscles .  MEAT SCIENCE72 ( 2 ) 245 - 251   2006.2Reviewed

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    The postmortem degradation of troponin T (TnT) in bovine longissimus (LT), diaphragm (DP), and masseter (MS) was analyzed. A 28.3 kDa (conventional 30 kDa) fragment of fast-type TnT isoforms showed the highest content in both LT and DP, where a 35.4 kDa isoform had the highest expression among the other fast isoforms. Meanwhile, a 26.0 kDa fragment was found to be the most highly produced among the fast TnT fragments in MS, where the expression of 36.5 and 32.8 kDa isoforms was higher than that of 35.4 and 34.8 kDa isoforms. Thus, the compositions of both the intact TnT isoform proteins and the postmortem fragments differed among the muscles examined, indicating that each TnT isoform degrades into a specific fragment in each muscle. Among the muscles, the LT muscle showed a high extent of TnT degradation and the highest expression of fast TnT isoforms containing a taste-related peptide sequence. (c) 2005 Elsevier Ltd. All rights reserved.

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  • Susumu MUROYA .  筋タンパク質トロポニンTの多様性と食肉における「機能」 .  Kagaku To Seibutsu44 ( 12 ) 831 - 840   2006Reviewed

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    DOI: 10.1271/kagakutoseibutsu1962.44.831

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  • S Muroya, Nakajima, I, M Oe, K Chikuni .  Effect of phase limited inhibition of MyoD expression on the terminal differentiation of bovine myoblasts: No alteration of Myf5 or myogenin expression .  DEVELOPMENT GROWTH & DIFFERENTIATION47 ( 7 ) 483 - 492   2005.9Reviewed

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    To investigate the roles played by MyoD in the terminal differentiation of satellite cell-derived myoblasts, the effect of antisense inhibition of MyoD expression was examined in bovine adult myoblast culture, in which inhibition treatment was limited to the terminal differentiation phase. MyoD antisense oligonucleotide DNA (AS-mD) suppressed the formation of multinucleated myotubes in the cell culture. Myotube formation was suppressed even when AS-mD treatment was limited to the period preceding the onset of myotube formation. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis revealed that treatment with AS-mD suppressed the expression of myosin heavy chain embryonic isoform and troponin T isoforms at 4 days after the induction of differentiation. AS-mD also suppressed the expression of MRF4, but did not alter the expression of either Myf5 or myogenin, in contrast to previous results using mouse cells possessing MyoD(-/-) genetic background. These findings suggest that MyoD controls myogenesis but not Myf5 or myogenin mRNA expression during the terminal differentiation phase. Furthermore, among the alpha 4, alpha 5, alpha 6, and alpha 7 integrins, alpha 4, alpha 5, and alpha 7 integrin expression was suppressed by AS-mD treatment, in parallel with the suppression of myotube formation, which suggests that MyoD controls myotube formation by regulating the expression of alpha 4, alpha 5, and alpha 7 integrins.

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  • Koichi Chikuni, Ikuyo Nakajima, Mika Oe, Susumu Muroya .  Peroxisome proliferator-activated receptor-γ coactivator 1 α(PGC-1 α) expression and the formation of slow-twitch muscle fibers in porcine and bovine skeletal muscles .  Animal Science Journal76 ( 4 ) 375 - 380   2005.8Reviewed

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    The peroxisome proliferator-activated receptor-γ coactivator-1 α(PGC-1 α) induces mitochondria biogenesis in skeletal muscles. To determine the relationships between PGC-1 α and the muscle fiber types, the expression levels of PGC-1 α were analyzed in porcine and bovine skeletal muscles. As a first step, the nucleotide sequences of the porcine and bovine PGC-1 α were determined. The porcine and bovine PGC-1 α cDNA encoded 796 amino acid sequences and showed 95.1 % identity between the two species. The expression levels of the PGC-1 α mRNA were analyzed in the same 10 skeletal muscles from four pigs and three cattle. The contents of porcine and bovine PGC-1 α were higher in the tongue, masseter and diaphragm, and lower in the Biceps femoris, semimembranosus, Longissimus thoracis and semitendinosus muscles. The contents of myosin heavy chain slow-type protein (MyHC-slow) were also determined in the same muscles by ELISA. The analysis of MyHC-slow showed results similar to those for the PGC-1 α contents in all of the muscles except for the tongue. The content of MyHC-slow in the tongue was the lowest among the porcine muscles, and moderate among the bovine muscles. The results suggest that PGC-1 α relates to the development of oxidative muscle fibers, but is not the principal factor in determining type I fiber content.

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  • MUROYA Susumu .  Significance of troponin T in live and postmortem muscles of meat animals .  The journal of animal genetics32 ( 2 ) 141 - 151   2005.6

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    DOI: 10.5924/abgri2000.32.2_141

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  • Kitamura S., Muroya S., Tanabe S., Okumura T., Chikuni K., Nishimura T. .  Mechanism of production of troponin T fragments during postmortem aging of porcine muscle .  Journal of Agricultural and Food Chemistry53 ( 10 ) 4178 - 4181   2005.5

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    Troponin T (TnT) is one of the myofibrillar proteins that is easily degraded during postmortem aging of pork. In this study, we determined the N-terminal amino acid sequences of TnT degradation fragments produced during postmortem aging and by m-calpain hydrolysis. The N-terminal amino acid sequences of TnT fragments produced during postmortem aging were EVHEPEEKPRPKLTAP, EKPRPKLTAPKIPEG, and APKIPEGEKVDF. On the other hand, the N-terminal amino acid sequences of TnT fragments produced by the action of m-calpain were APPPPAEV, EVHEPEEK, and APK. These sequences of degradation fragments could be mapped on fast type TnT isoform 2. The peptide bonds of His37-Glu38 and Thr51-Ala52 in fTnT2 were cleaved during postmortem aging as well as by the calpain hydrolysis; therefore, calpain was concluded to have an important role in TnT degradation during postmortem aging. It was also found that the sourness-suppressing peptide APPPPAEVHEVHEEVH (Okumura et al. Biosci. Biotechnol. Biochem. 2004, 68, 1657-1662) derived from TnT degradation could be produced by the action of calpains on Glu21-Ala22 and His37-Glu38 sites. © 2005 American Chemical Society.

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  • SI Kitamura, S Muroya, S Tanabe, T Okumura, K Chikuni, T Nishimura .  Mechanism of production of troponin T fragments during postmortem aging of porcine muscle .  JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY53 ( 10 ) 4178 - 4181   2005.5Reviewed

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    Troponin T (TnT) is one of the myofibrillar proteins that is easily degraded during postmortem aging of pork. In this study, we determined the N-terminal amino acid sequences of TnT degradation fragments produced during postmortem aging and by m-calpain hydrolysis. The N-terminal amino acid sequences of TnT fragments produced during postmortem aging were EVHEPEEKPRPKLTAP, EKPRPKLTAPKIPEG, and APKIPEGEKVDF. On the other hand, the N-terminal amino acid sequences of TnT fragments produced by the action of m-calpain were APPPPAEV, EVHEPEEK, and APK These sequences of degradation fragments could be mapped on fast type TnT isoform 2. The peptide bonds of His(37)-Glu(38) and Thr(51)-Ala(52) in fTnT2 were cleaved during postmortem aging as well as by the calpain hydrolysis; therefore, calpain was concluded to have an important role in TnT degradation during postmortem aging. It was also found that the sourness-suppressing peptide APPPPAEVHEVHEEVH (Okumura et al. Biosci. BiotechnoL Biochem. 2004, 68,1657-1662) derived from TnT degradation could be produced by the action of calpains on Glu(21)-Ala(22) and His(37)-Glu(38) sites.

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  • K Chikuni, S Muroya, Nakajima, I .  Absence of the functional myosin heavy chain 2b isoform in equine skeletal muscles .  ZOOLOGICAL SCIENCE21 ( 5 ) 589 - 596   2004.5Reviewed

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    Nucleotide sequences which included the full coding region for three types of myosin heavy chain (MyHC) isoforms were determined from equine skeletal muscles. The deduced amino acid sequences were 1937, 1938, and 1935 residues for the MyHC-2a, -2x, and -slow, respectively. No MyHC-2b isoform was amplified from the equine muscle cDNA except for one pseudogene fragment. One nucleotide was inserted in the coding region of the equine pseudogene product, a minute amount of which was expressed in the skeletal muscle. The 596 bp sequence of the equine MyHC pseudogene was categorized into the MyHC-2b genes on the phylogenetic tree of the mammalian MyHC genes. These results suggest that an ancestral MyHC-2b gene had lost its function and changed to a pseudogene during the course of horse history. The MyHC genes in some ungulates were analyzed through the PCR amplifications using the MyHC isoform-specific primers to confirm the presence of the MyHC-2b and -2x genes. The exon coding the 3' untranslated region of the MyHC-2x was successfully amplified from the all ungulates examined; however, that of the MyHC-2b gene was amplified only from horses, pigs and lesser mouse deer. The PCR analyses from rhinoceros, sika deer, moose, giraffes, water buffalo, bovine, Japanese serow and sheep genes implied the absence of the MyHC-2b-specific sequence in their genomes. These results suggest that the MyHC-2b gene independently lost its function in some ungulate species.

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  • S Muroya, S Kitamura, S Tanabe, T Nishimura, Nakajima, I, K Chikuni .  N-terminal amino acid sequences of troponin T fragments, including 30 kDa one, produced during postmortem aging of bovine longissimus muscle .  MEAT SCIENCE67 ( 1 ) 19 - 24   2004.5Reviewed

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    We have determined the amino (N)-terminal amino acid (AA) sequences of five troponin T (TnT) fragments produced during postmortem aging of bovine longissimus muscle. Western blot analysis showed that 32.1, 28.8, 27, and 25.8 kDa anti-fast-type TnT (fTnT)-positive fragments and a 31 kDa anti-slow-type TnT (sTnT)-positive fragment were present at 14 d postmortem. The N-terminal AA sequences of the 32.1, 28.8 (conventional 30 kDa), 27, and 25.8 kDa fragments were APPPPAEV, EVHEPEEK, EKPRPRLT, and APKIPEGE, respectively, and they were mapped to the N-terminal region of bovine fTnT isoforms. The N-terminal sequences of the 31 kDa fragment, EAPEEPEP, were mapped to the sTnT isoforms. These findings indicate that the two isoform types of fTnT predominantly expressed in the longissimus muscle are cleaved specifically at GlU(21)-Ala(22) and Glu(15)-Ala(16), His(37)-Glu(38) and His(31)-Glu(32), Glu(43)-Glu(44) and Glu(37)-Glu(38), and/or Thr(51)-Ala(52) and Thr(45)-Ala(46), respectively, and that a sTnT isoform is cleaved specifically at Glu(23)-Glu(24). (C) 2003 Elsevier Ltd. All rights reserved.

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  • K Chikuni, S Muroya, Nakajima, I .  Myosin heavy chain isoforms expressed in bovine skeletal muscles .  MEAT SCIENCE67 ( 1 ) 87 - 94   2004.5Reviewed

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    Nucleotide sequences including the full coding region for three types of myosin heavy chain (MyHC) isofoms were determined from bovine adult skeletal muscles. The deduced amino acid sequences were 1940, 1938, and 1935 residues for the MyHC-2a, -2x, and -slow, respectively. Like other mammalian MyHC isoforms, the bovine MyHC isoforms had homologous sequences except for substitutions concentrated on the loop 1, loop 2, and light chain binding regions. RT-PCR amplifications showed that the adult bovine skeletal muscles expressed the MyHC-2a, -2x, and -slow isoforms but no -2b isoform. The absence of the MyHC-2b isoform and substitutions on the loop2 region could explain some differences in meat quality between beef and pork. (C) 2003 Elsevier Ltd. All rights reserved.

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  • Nakajima, I, S Muroya, K Chikuni .  Growth arrest by octanoate is required for porcine preadipocyte differentiation .  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS309 ( 3 ) 702 - 708   2003.9Reviewed

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    A preadipocyte clonal line has been established from porcine subcutaneous tissue. This line, designated PSPA, showed a fibroblastic phenotype and kept on growing under a preadipose condition even after reaching confluence. When confluent cultures were stimulated with insulin, dexamethasone, biotin, pantothenate, and octanoate, growth was arrested, and the cells exhibited a marked increase in lipogenesis. However, adipose conversion was not induced upon exposure of PSPA cells to a standard hormonal mixture of mouse 3T3-L1 cells, and they continued dividing as did the preadipocytes in growth-medium. By serially omitting each individual adipogenic agent from the PSPA differentiation medium, it was determined that octanoate was one of the most essential but the only factor able to induce growth arrest. Octanoate supplementation to 3T3-L1 medium increased the triglyceride accumulation of PSPA cells accompanied by growth arrest. Both RT-PCR and Western blot analysis supported the idea of octanoate as a potential agent with the antiproliferative activity requisite for porcine preadipocytes to enter terminal differentiation. (C) 2003 Elsevier Inc. All rights reserved.

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  • S Muroya, Nakajima, I, K Chikuni .  Amino acid sequences of multiple fast and slow troponin T isoforms expressed in adult bovine skeletal muscles .  JOURNAL OF ANIMAL SCIENCE81 ( 5 ) 1185 - 1192   2003.5Reviewed

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    Multiple nucleotide sequences of complementary DNA (cDNA) of bovine troponin T (TnT) isoforms expressed in the adult skeletal muscles were determined to facilitate the elucidation of the TnT degradation progress during postmortem aging of muscles. Fresh muscle samples were excised from the lingual, masseter, pectoralis, diaphragm, psoas major, longissimus thoracis, spinnalis, semitendinosus, semimembranosus, and biceps femoris muscles of three Holstein cows within 1 h of slaughter. Complementary DNA fragments of fast and slow TnT isoforms expressed in each muscle were amplified by reverse-transcribed PCR. Consequently, four major fragments of fast TnT and two fragments of slow TnT, all of which contained the complete coding region, were obtained. The sequence determination of these fragments revealed that at least eight and two isoforms were generated by the alternative splicing from bovine fast and slow TnT messenger RNA, respectively. In the fast TnT isoforms, five small variable exons were observed; three of these five exons were in the amino (N)-terminal region. The calculated molecular weight of fast and slow TnT isoforms ranged from 29,816 to 32,125 and from 30,166 to 31,284, respectively. The deduced amino acid sequences revealed that the N-terminal region of all the TnT isoforms was extremely glutamic acid-rich. Reverse-transcribed PCR analysis revealed that expression of each of these isoforms was distributed in a fast or slow muscle-specific manner. Given that TnT degradation has been reported to accompany a decrease in glutamic acid content in the conventional 30-kDa degradation product, the sequence data suggested that the 30-kDa fragment seem to be generated by the proteolytic removal of the glutamic acid-rich N-terminal ends. The multiplicity of TnT isoforms may result in a complicated pattern of TnT degradation on SDS-PAGE gel during beef aging.

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  • Susumu MUROYA, Ikuyo NAKAJIMA, Koichi CHIKUNI .  Sequential expression of myogenic regulatory factors in bovine skeletal muscle and the satellite cell culture .  Animal Science Journal73 ( 5 ) 375 - 381   2002.10Reviewed

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    Language:English   Publisher:Wiley-Blackwell  

    DOI: 10.1046/j.1344-3941.2002.00052.x

    CiNii Books

  • Koichi CHIKUNI, Susumu MUROYA, Ryo-ichi TANABE, Ikuyo NAKAJIMA .  Comparative sequence analysis of four myosin heavy chain isoforms expressed in porcine skeletal muscles: Sequencing and characterization of the porcine myosin heavy chain slow isoform .  Animal Science Journal73 ( 4 ) 257 - 262   2002.8Reviewed

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    Language:English   Publisher:Wiley-Blackwell  

    DOI: 10.1046/j.1344-3941.2002.00035.x

    CiNii Books

  • S Muroya, Nakajima, I, K Chikuni .  Related expression of MyoD and Myf5 with myosin heavy chain isoform types in bovine adult skeletal muscles .  ZOOLOGICAL SCIENCE19 ( 7 ) 755 - 761   2002.7Reviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ZOOLOGICAL SOC JAPAN  

    Skeletal muscles are characterized as fast and slow muscles, according to the expression pattern of myosin heavy chain (MyHC) isoforms in the muscle fibers. To investigate the relationships between MyHC isoforms and myogenic regulatory factors (MRFs) including MyoD, Myf5, myogenin, and MRF4 in adult skeletal muscles, expressions of these MRFs in the ten muscles of three cows were analyzed by a semi-quantitative RT-PCR. The results showed that MyoD expression was significantly lower in the lingual muscles (TN), masseter (MS) and diaphragm (DP), which lack MyHC-2x (fast glycolytic) expression and abound with MyHC-slow (slow oxidative) and/or MyHC-2a (fast oxidative), than it was in the pectoralis (PP), psoas major (PM), longissimus thoracis (LT), spinnalls (SP), semitendinosus (ST), semimembranosus (SM), and biceps femoris (BF). In contrast, the Myf5 expression in TN, MS, and DIP was significantly higher than in PM, LT, ST, SM, and BF No significant difference was observed in myogenin and MRF4 expression among the muscles tested. The results suggest that MyoD and Myf5 influence the MyHC isoform expression, although the effects are not decisive in specifying the phenotypes of adult muscles.

    DOI: 10.2108/zsj.19.755

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    PubMed

    Other Link: http://orcid.org/0000-0002-2376-9352

  • Nakajima, I, S Muroya, R Tanabe, K Chikuni .  Extracellular matrix development during differentiation into adipocytes with a unique increase in type V and VI collagen .  BIOLOGY OF THE CELL94 ( 3 ) 197 - 203   2002.6Reviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER  

    In order to study how adipose conversion affects the extracellular environment, levels of extracellular matrix (ECM) proteins during differentiation were analyzed by I-125-labeled antibody binding to each specific primary antibody. When confluent bovine intramuscular preadipocytes (BIP) were stimulated with adipogenic medium, there was a significant accretion on the cell surface of type I-VI collagens, laminin and fibronectin, compared with undifferentiated cells. The deposition amount of ECM proteins had reached near maximal levels at an early stage of differentiation and lasted throughout the culture. However, the increasing manners were not all the same in these eight proteins. Type V and type VI collagen tended to show a transient decline after the rapid rise at the beginning of stimulation, and fibronectin instead, subsequently decreased. Further analysis by immunocytochemical staining showed that remodeling occurred in type V and VI collagen matrices during this period; extensive fibrillar networks seen at 10 d after stimulation were quite unlike that formed earlier. These specific increases and development of matrix during adipocyte differentiation imply some significance for organizing fat lobules in each ECM proteins, especially type V and VI collagens. (C) 2002 Editions scientifiques et medicales Elsevier SAS. All rights reserved.

    DOI: 10.1016/S0248-4900(02)01189-9

    DOI: 10.1016/s0248-4900(02)01189-9

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  • Nakajima, I, S Muroya, R Tanabe, K Chikuni .  Positive effect of collagen V and VI on triglyceride accumulation during differentiation in cultures of bovine intramuscular adipocytes .  DIFFERENTIATION70 ( 2-3 ) 84 - 91   2002.5Reviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:BLACKWELL VERLAG GMBH  

    Ethyl-3,4-dihydroxybenzoate (EDHB), a specific inhibitor of collagen synthesis, was used to study the role of collagen in the differentiation of bovine intramuscular preadipocytes (BIP). Triglyceride (TG) accumulation levels of BIP cells were dose-dependently inhibited by EDHB and were reduced to 50% at a 0.1 mM concentration. EDHB addition prevented the accretion of collagens (types I-VI) on the cell surface, which generally increases during adipose conversion. Western blotting and immunofluorescence studies showed in detail that triple-helical conformation of procollagen molecules was drastically interrupted by EDHB, and as a result, their matrix assembly was not performed in the extracellular space of adipocytes. Particularly, the development of collagen types IV, V and VI during differentiation was severely damaged. When exogenous collagens were supplied to make up for the lack of endogenous products, cultured EDHB-treated cells on type V and VI collagen-coated dishes were the only ones among six collagens to accumulate more TG, although their TG content did not reach that of normal adipocytes. This result implies the importance and the active role of collagens V and VI for adipogenesis. However, these findings also indicate that collagen newly synthesized and organized by the adipocyte itself during differentiation is still necessary for the growth of adipose tissue.

    DOI: 10.1046/j.1432-0436.2002.700203.x

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    Other Link: http://orcid.org/0000-0002-2376-9352

  • Koichi CHIKUNI, Keisuke SASAKI, Tadasu EMORI, Fumiyuki IWAKI, Fumio TANI, Ikuyo NAKAJIMA, Susumu MUROYA, Mitsuru MITSUMOTO .  豚肉風味関連物質の含量に対する加熱処理の影響 .  Nihon Yoton Gakkaishi39 ( 3 ) 191   2002Reviewed

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    Publisher:The Japanese Society of Swine Science  

    DOI: 10.5938/youton.39.191

  • S Muroya, Nakajima, I, K Chikuni .  Bovine skeletal muscle cells predominantly express a vascular cell adhesion molecule-1 seven-Ig domain splice form .  ZOOLOGICAL SCIENCE18 ( 6 ) 797 - 805   2001.8Reviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ZOOLOGICAL SOC JAPAN  

    Vascular cell adhesion molecule-1 (VCAM-1), which has several alternatively splicing variants, plays a role in myotube formation. To investigate which form functions in myogenesis, we analyzed VCAM-1 mRNA expression in bovine skeletal muscle cells. We detected the expression of two VCAM-1 splice forms in the muscle tissue and in the primary satellite cell culture. The longer form was predominantly expressed at the muscle and during myotube formation of the cells. The nucleotide sequences of the two forms were determined by cDNA direct sequencing. The sequence data showed that the predominant form in skeletal muscle was a full-length VCAM-1 (VCAM-7D) that consists of seven immunoglobulin-like (Ig) domains, and the minor form was a novel six-domain form that lacks the seventh Ig domain. Compared to this, bovine pulmonary artery endothelial cells also express a variant which lacks domain 7, but VCAM-7D was not detected by RT-PCR in the culture. No VCAM-1 expression was detected in bovine kidney epithelial cell, lymph node epithelial cell, or leukemic B-lymphocyte culture even under stimulation by tumor necrosis factor-alpha. These data suggest that the splicing of the VCAM-1 gene alternatively varies depending on the cell type where it is expressed, and that VCAM-7D plays a predominant role in myotube formation.

    DOI: 10.2108/zsj.18.797

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    Other Link: http://orcid.org/0000-0002-2376-9352

  • Uramura K, Funahashi H, Muroya S, Shioda S, Takigawa M, Yada T .  Orexin-a activates phospholipase C- and protein kinase C-mediated Ca2+ signaling in dopamine neurons of the ventral tegmental area. .  Neuroreport12 ( 9 ) 1885 - 9   2001.7Orexin-a activates phospholipase C- and protein kinase C-mediated Ca2+ signaling in dopamine neurons of the ventral tegmental area.

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    DOI: 10.1097/00001756-200107030-00024

    PubMed

  • K Chikuni, R Tanabe, S Muroya, Nakajima, I .  Differences in molecular structure among the porcine myosin heavy chain-2a,-2x, and-2b isoforms .  MEAT SCIENCE57 ( 3 ) 311 - 317   2001.3Reviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCI LTD  

    Full coding regions for fast type myosin heavy chain (MyHC) isoforms were sequenced from a porcine skeletal muscle to analyze sequence diversity relating to the contractile properties of muscle fibers. An approximately 6-kb fragment for each MyHC was amplified through RT-PCR using isoform type-specific primers, which were designed in the 5' and 3' non-coding regions of the porcine MyHCs. The lengths of deduced amino acid sequences were 1939, 1939, and 1937 for the porcine MyHC-2a,-2x, and-2b, respectively. The entire amino acid sequences were highly conserved among the three MyHCs, except for the 50/20 k junction region (loop 2) which would weakly bind actin molecules. The porcine MyHC-2b possessed different amino acids from MyHC-2a and-2x, in loop1 and ELC binding region. The sequence data suggested the diversity of contractile properties among the porcine MyHC isoforms. (C) 2000 Elsevier Science Ltd. All rights reserved.

    DOI: 10.1016/S0309-1740(00)00107-8

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    Other Link: http://orcid.org/0000-0002-2376-9352

  • Ryo-ichi TANABE, Tetsuya MURAKAMI, Takahiro KAWAHARA, Rinko YAMASHIRO, Mitsuru MITSUMOTO, Susumu MUROYA, Ikuyo NAKAJIMA, Koichi CHIKUNI .  Composition of Myosin Heavy Chain Isoforms in Relation to Meat Texture in Duroc, Landrace and Meishan Pigs .  Nihon Chikusan Gakkaiho72 ( 3 ) 230 - 237   2001Reviewed

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    Publisher:Japanese Society of Animal Science  

    DOI: 10.2508/chikusan.72.230

  • Susumu MUROYA, Ikuyo NAKAJIMA, Koichi CHIKUNI .  Nucleotide Sequencing of Bovine α4 Integrin cDNA, and its Expression in the Primarily-cultured Skeletal Muscle Satellite Cells .  Nihon Chikusan Gakkaiho72 ( 6 ) 505 - 512   2001Reviewed

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    Publisher:Japanese Society of Animal Science  

    DOI: 10.2508/chikusan.72.505

  • Uramura K, Yada T, Muroya S, Takigawa M .  Ca2+ oscillations in response to methamphetamine in dopamine neurons of the ventral tegmental area in rats subchronically treated with this drug. .  Annals of the New York Academy of Sciences914   316 - 22   2000.9Ca2+ oscillations in response to methamphetamine in dopamine neurons of the ventral tegmental area in rats subchronically treated with this drug.

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  • Uramura K, Yada T, Muroya S, Shioda S, Shiratani T, Takigawa M .  Methamphetamine induces cytosolic Ca2+ oscillations in the VTA dopamine neurons. .  Neuroreport11 ( 5 ) 1057 - 61   2000.4Methamphetamine induces cytosolic Ca2+ oscillations in the VTA dopamine neurons.

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    DOI: 10.1097/00001756-200004070-00031

    PubMed

  • S Muroya, R Tanabe, Nakajima, I, K Chikuni .  Molecular characteristics and site specific distribution of the pigment of the Silky fowl .  JOURNAL OF VETERINARY MEDICAL SCIENCE62 ( 4 ) 391 - 395   2000.4Reviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPAN SOC VET SCI  

    Silky fowl, a breed of chicken, is hyperpigmented in its various internal tissues. The pigment was extracted from various tissues of two strains of Silky fowl to determine its molecular structure and internal distribution. Analysis by infrared spectroscopy showed two spectrum patterns of the pigment in Silky fowl; one is from ovary and testis, the other is from periosteum and feather. The difference between the two spectra is possibly due to the sulfur contents of melanin. Especially, the spectra of the pigments from feather and periosteum shared the characteristics of synthesized melanin spectrum in common, which indicates that the melanocytes dispersed in these tissues were functionally the same. According to our quantitative analysis, the tissues examined were classified significantly in the order of the pigment content (p&lt;0.05): periosteum &gt; gonads (ovary or testis) = trachea greater than or equal to heart, liver, gizzard, cecum, muscles (Pectoralis and Supracoracoideus) and skin. In addition, the specific regions of embryonic neural crest derived cells, such as cardiac artery and various parts of cephalic tissues, were found to be locally hyperpigmented. These data suggest that hyperpigmentation (fibromelanosis) in Silky fowl chicken occurs in a tissue- and organ-specific manner, which is strongly related to neural crest cell development. It is hypothesized that neural crest cells of the bird, containing melanocyte progenitors, acquire unusual ability to differentiate into melanocytes excessively, and to extend the distribution of their descendant along the destinations of neural crest derivatives.

    DOI: 10.1292/jvms.62.391

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  • Muroya S, Yada T, Shioda S, Takigawa M .  Glucose-sensitive neurons in the rat arcuate nucleus contain neuropeptide Y. .  Neuroscience letters264 ( 1-3 ) 113 - 6   1999.4Glucose-sensitive neurons in the rat arcuate nucleus contain neuropeptide Y.

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    DOI: 10.1016/s0304-3940(99)00185-8

    PubMed

  • R Tanabe, S Muroya, K Chikuni .  Expression of myosin heavy chain isoforms in porcine muscles determined by multiplex PCR .  JOURNAL OF FOOD SCIENCE64 ( 2 ) 222 - 225   1999.3Reviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:INST FOOD TECHNOLOGISTS  

    A cDNA sequence of 5'-terminal region of myosin heavy chain (MHC) isoform 2x was determined. Based on this and published sequences, the procedure of reverse transcription-polymerase chain reaction (RT-PCR) employing a multiplex PCR technique was developed to analyze the expression patterns of MHC isoforms in porcine muscles, All four MHC isoforms in postnatal porcine muscles could be analyzed by this method and showed different expression patterns according to the porcine muscle types. The procedure developed in this study would be very useful to elucidate the relationships between meat quality and MHC isoform composition.

    DOI: 10.1111/j.1365-2621.1999.tb15869.x

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    Other Link: http://orcid.org/0000-0002-2376-9352

  • T Matsunaga, K Chikuni, R Tanabe, S Muroya, K Shibata, J Yamada, Y Shinmura .  A quick and simple method for the identification of meat species and meat products by PCR assay .  MEAT SCIENCE51 ( 2 ) 143 - 148   1999.2Reviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCI LTD  

    The polymerase chain reaction (PCR) was applied to identify six meats (cattle, pig, chicken, sheep, goat and horse) as raw materials for products. By mixing seven primers in appropriate ratios, species-specific DNA fragments could be identified by only one multiplex PCR. A forward primer was designed on a conserved DNA sequence in the mitochondrial cytochrome b gene, and reverse primers on species-specific DNA sequences for each species. PCR primers were designed to give different length fragments from the six meats. The products showed species-specific DNA fragments of 157, 227, 274, 331, 398 and 439 bp from goat, chicken, cattle, sheep, pig and horse meats, respectively. Identification is possible by electrophoresis of PCR products. Cattle, pig, chicken, sheep and goat fragments were amplified from cooked meat heated at 100 or 120 degrees C for 30 min, but horse DNA fragments could not be detected from the 120 degrees C sample. Detection limits of the DNA samples were 0.25 ng for all meats. (C) 1998 Elsevier Science Ltd. All rights reserved.

    DOI: 10.1016/S0309-1740(98)00112-0

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    Other Link: http://orcid.org/0000-0002-2376-9352

  • R Tanabe, S Muroya, K Chikuni .  Sequencing of the 2a, 2x, and slow isoforms of the bovine myosin heavy chain and the different expression among muscles .  MAMMALIAN GENOME9 ( 12 ) 1056 - 1058   1998.12Reviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPRINGER VERLAG  

    DOI: 10.1007/s003359900924

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    Other Link: http://orcid.org/0000-0002-2376-9352

  • T Matsunaga, K Chikuni, R Tanabe, S Muroya, H Nakai, K Shibata, J Yamada, Y Shinmura .  Determination of mitochondrial cytochrome B gene sequence for Red deer (Cervus elaphus) and the differentiation of closely related deer meats .  MEAT SCIENCE49 ( 4 ) 379 - 385   1998.8Reviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCI LTD  

    The cytochrome b gene sequence for red deer was determined using the Dye Terminator Cycle Sequencing method and used for identification of deer meat in meat and meat products. Red deer showed a similarity of 94.1, 84.0, 81.1, 85.5 and 85.6% to sika deer (Cervus nippon), bovine, pigs, sheep and goats, respectively. To differentiate the deer meat, oligonucleotide primers RD-1 (5'-TCATCGCAGCACTCGCTATAGTACACT-3'), RD-2(5'-ATCTCCAAGTAGGTCTGGTGCGAATAA-3') were designed for the region of the cytochrome b gene of red neer. The PCR amplified 194 bp fragments from red and sika deer, but no fragments from bovine, pig, chicken, sheep, goat, horse and rabbit DNA. Although cooking the meats reduced the PCR products, red deer could still be detected in meat heated at 120 degrees C. To discriminate between red and sika deer, these PCR products were digested by a restriction enzyme (EcoRI,BamHI,ScaI) and analyzed by 4% agarose gel electrophoresis. As a result, the red deer fragment was digested by EcoRI to 67/127 bp fragments but not by BamHI and Seal. The sika deer fragment was digested to 48/146 bp and 49/145 bp fragments with the two other enzymes, and thus it is possible to differentiate between the two kinds of deer from the digestion pattern of restriction enzymes, (C) 1998 Elsevier Science Ltd. All rights reserved.

    DOI: 10.1016/S0309-1740(97)00145-9

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    Other Link: http://orcid.org/0000-0002-2376-9352

  • Chikuni K., Fukumoto Y., Tanabe R., Muroya S., Ozawa S. .  A simple method for genotyping the bovine growth hormone gene .  Animal Genetics28 ( 3 ) 230 - 232   1997.6

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    Language:Japanese   Publisher:Animal Genetics  

    An allele-specific polymerase chain reaction (PCR) amplification method was developed to determine the genotypes at the bovine growth hormone locus that result from two nucleotide substitutions in exon 5 of the gene. This method was a multiplex PCR (ASM-PCR) employing a common primer pair and two allele-specific reverse primers. The common primer pair was designed to amplify a target region containing two substitution points from the three variants of the bovine growth hormone gene. The allele-specific primers were designed to be mismatched with other genotypes at the 3' end of oligonucleotides. When the common and allele-specific reverse primers competed with each other, the shorter allele-specific fragments were amplified preferentially. Consequently, the PCR products of the variant-specific fragments were 347, 483 and 656 bp for allales A, B and C, respectively, of the bovine growth hormone gene. Genotypes of the bovine growth hormone gene were easily identified by agarose gel electrophoresis of PCR products. The results suggested that this multiplex PCR method would be useful for identification of genetic variants caused by point mutations.

    DOI: 10.1111/j.1365-2052.1997.00095.x

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  • R Tanabe, S Muroya, Nakajima, I, K Chikuni, H Nakai .  Skeletal muscle connectin primary structures as related to animal species and muscle type .  JOURNAL OF FOOD SCIENCE62 ( 3 ) 451 - &   1997.5Reviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:INST FOOD TECHNOLOGISTS  

    Differences in molecular weights and partial amino acid sequences of connectin(titin) were determined for cattle, pig and chicken skeletal muscles. Peptide mapping analysis results differed according to animal species. Amino acid sequences deduced from partial nucleotide sequences of connectin also differed according to animal species at immunoglobulin-like (Ig) and fibronectin type 3 (FN3) domains. In chicken, the molecular weight of connectin from leg muscles was higher than that from pectoral muscles. Differences in meat texture and conditioning may relate to connectin and extent of its breakdown.

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    Other Link: http://orcid.org/0000-0002-2376-9352

  • Tanabe R., Muroya S., Nakajima I., Chikuni K., Nakai H. .  Skeletal muscle connectin primary structures as related to animal species and muscle type .  Journal of Food Science62 ( 3 ) 451 - 453   1997.5

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    Language:Japanese   Publisher:Journal of Food Science  

    Differences in molecular weights and partial amino acid sequences of connectin(titin) were determined for cattle, pig and chicken skeletal muscles. Peptide mapping analysis results differed according to animal species. Amino acid sequences deduced from partial nucleotide sequences of connectin also differed according to animal species at immunoglobulin-like (Ig) and fibronectin type 3 (FN3) domains. In chicken, the molecular weight of connectin from leg muscles was higher than that from pectoral muscles. Differences in meat texture and conditioning may relate to connectin and extent of its breakdown.

    DOI: 10.1111/j.1365-2621.1997.tb04404.x

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  • Susumu MUROYA, Fuminori TERADA, Shigeru SHIOYA .  Influence of heat stress on distribution of nitrogen in milk .  Nihon Chikusan Gakkaiho68 ( 3 ) 297 - 300   1997Reviewed

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    Publisher:Japanese Society of Animal Science  

    DOI: 10.2508/chikusan.68.297

    Other Link: http://orcid.org/0000-0002-2376-9352

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    佐々木啓介, 渡邊源哉, 前田恵助, 本山三知代, 中島郁世, 青沼達也, 本間文佳, 尾花尚明, 小平貴都子, 齋藤薫, 渡邉美のり, 小林美穂, 成田卓美, 尾嶋孝一, 萩達朗, 野村将, 大江美香, 室谷進, 松本和典

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    日本畜産学会大会講演要旨   124th   161   2018.3

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    日本畜産学会大会講演要旨   124th   188   2018.3

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    日本畜産学会大会講演要旨   124th   188   2018.3

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    日本養豚学会大会講演要旨   108th   19   2018.3

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    日本畜産学会大会講演要旨   123rd   99   2017.9

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    日本畜産学会大会講演要旨   122nd   174   2017.3

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    日本畜産学会大会講演要旨   122nd   172   2017.3

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    食品試験研究成果情報   ( 28 )   76‐77   2016.3

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    日本畜産学会大会講演要旨   121st   211   2016.3

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    日本畜産学会大会講演要旨   121st   213   2016.3

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    農研機構畜産研究部門成果情報(Web)   2016   2016

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    日本養豚学会大会講演要旨   103rd   11   2015.10

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    日本畜産学会大会講演要旨   120th   106   2015.9

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  • 3T3‐L1細胞が分化過程で分泌する因子のプロテオミクス解析

    尾嶋孝一, 大江美香, 中島郁世, 室谷進, 西邑隆徳

    日本畜産学会大会講演要旨   120th   92   2015.9

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    室谷進, 小笠原英毅, 寶示戸雅之

    日本畜産学会大会講演要旨   120th   89   2015.9

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  • Characterization and classification of Japanese consumer perceptions for beef tenderness using descriptive texture characteristics assessed by a trained sensory panel (vol 96, pg 994, 2014)

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    MEAT SCIENCE   103   104 - 104   2015.5

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    日本畜産学会大会講演要旨   119th   233   2015.3

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    日本畜産学会大会講演要旨   119th   235   2015.3

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    日本畜産学会大会講演要旨   119th   186   2015.3

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    日本畜産学会大会講演要旨   119th   221   2015.3

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    日本畜産学会大会講演要旨   119th   140   2015.3

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    日本養豚学会大会講演要旨   102nd   8   2015.3

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    MOLECULAR BIOLOGY OF THE CELL   26   2015

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  • Dynamics of secreted proteins from skeletal muscle cells during muscle differentiation.

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    MOLECULAR BIOLOGY OF THE CELL   25   2014.12

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    日本養豚学会大会講演要旨   101st   29   2014.10

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    日本養豚学会大会講演要旨   101st   31   2014.10

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  • ウシ半腱様筋および咬筋のmicroRNA発現プロファイル

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    日本畜産学会大会講演要旨   118th   172   2014.3

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    日本畜産学会大会講演要旨   118th   209   2014.3

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    日本養豚学会大会講演要旨   99th   12   2013.10

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    日本畜産学会大会講演要旨   117th   87   2013.9

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    日本畜産学会大会講演要旨   116th   109   2013.3

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    日本畜産学会大会講演要旨   116th   181   2013.3

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    日本畜産学会大会講演要旨   116th   181   2013.3

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    日本畜産学会大会講演要旨   116th   207   2013.3

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  • 粗飼料多給による家畜飼養技術と効率的畜産物生産技術の開発 第3章 粗飼料多給による良質畜産物の効率的生産技術とそのための品質評価法開発 1 網羅的蛋白質解析技術を利用した牛赤肉品質決定因子の解明と評価手法の開発

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  • Effects of phosphorylation of muscle-specific calpain-3 on its proteolytic activity.

    K. Ojima, Y. Ono, S. Hata, M. Oe, I. Nakajima, S. Muroya, K. Chikuni, H. Sorimachi

    MOLECULAR BIOLOGY OF THE CELL   24   2013

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  • 背脂肪厚の異なる梅山豚種とランドレース種におけるアディポネクチンと血液成分の違い

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    日本養豚学会大会講演要旨   97th   28   2012.10

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  • コペンハーゲン大学における在外研究 : シングルファイバーを巡る熟成研究とデンマークの食肉研究者たち

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    食肉の科学   53 ( 1 )   27 - 30   2012.6

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    日本畜産学会大会講演要旨   115th   242   2012.3

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  • ウシ単一筋線維中のトロポミオシンアイソフォーム発現解析

    大江美香, 千国幸一, 尾嶋孝一, 中島郁世, 室谷進

    日本畜産学会大会講演要旨   115th   232   2012.3

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  • 牛肉熟成中のドリップにおけるトロポニンTペプチド量の経時変化

    室谷進, 大江美香, 中島郁世, 柴田昌宏, 尾嶋孝一, 千国幸一

    日本畜産学会大会講演要旨   115th   248   2012.3

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  • 骨格筋特異的に発現するカルパイン3はNa<sup>+</sup>により活性化する

    尾嶋孝一, 小野弥子, 中島郁世, 大江美香, 室谷進, 反町洋之

    日本畜産学会大会講演要旨   115th   232   2012.3

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  • The impact of each domain of myosin heavy chain molecule on forming thick filaments in muscle and non-muscle cells.

    K. Ojima, I. Nakajima, M. Oe, S. Muroya, T. Nishimura, A. Hattori

    MOLECULAR BIOLOGY OF THE CELL   23   2012

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    Web of Science

  • Effect of grazing in the latter fattening period on the nutrient content and gene expression in steer muscle. Reviewed

    Shibata M, Matsumoto K, Hikino Y, Muroya S, Oe M, Nakajima I, Ojima K, Chikuni K

    57th International Congress of Meat Science and Technology Proceedings   P-172   2011.8

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  • Fatty acids modulate adipocyte growth and development in pig: an approach from cell culture study Reviewed

    Nakajima I, Oe M, Ojima K, Muroya S, Shibata M, Chikuni K

    57th International Congress of Meat Science and Technology Proceedings   P-422   2011.8

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  • いくつかの「エコフィード」利用豚肉に対する消費者の嗜好性とその多様性

    佐々木啓介, 本山三知代, 成田卓美, 萩達朗, 尾嶋孝一, 大江美香, 野村将, 千国幸一, 橘内克弘, 室谷進, 中島郁世, 芦原茜, 大森英之, 田島清, 勝俣昌也, 川島知之

    日本養豚学会誌   48 ( 2 )   80   2011.6

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  • 脂肪酸がブタの脂肪細胞分化に及ぼす影響

    中島郁世, 大江美香, 尾嶋孝一, 室谷進, 柴田昌宏, 千国幸一

    日本養豚学会誌   48 ( 2 )   104   2011.6

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  • 放牧仕上げ肥育において,プロテオーム解析から筋肉の遅筋化が認められる

    柴田昌宏, 松本和典, 大江美香, 亀山眞由美, 尾嶋孝一, 中島郁世, 室谷進, 千国幸一

    近畿中国四国農業研究成果情報(CD-ROM)   2010   CHIKUSANKUSACHISUISHIMBUKAI,KENKYU.SANKO,1   2011.6

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  • Structural functions of skeletal muscle-specific calpain in Ca2+ efflux from the sarcoplasmic reticulum

    K. Ojima, Y. Ono, C. Ottenheijm, S. Hata, H. Suzuki, M. Oe, I. Nakajima, S. Muroya, H. Granzier, H. Sorimachi

    MOLECULAR BIOLOGY OF THE CELL   22   2011

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  • Effects of grass-fed and grain-fed diets on expression of genes for muscle development in Japanese Black steers Reviewed

    Shibata M, Matsumoto K, Hikino Y, Oe M, Ojima K, Nakajima I, Muroya S, Chikuni K

    56th International Congress of Meat Science and Technology Proceedings   C0009   2010.8

  • 屠畜後のブタ最長筋IIb型筋線雄ではタンパク質がI型筋線維より速く分解される

    室谷進, ERTBJERG P, POMPONIO L, CHRISTENSEN M

    日本畜産学会大会講演要旨   112th   96   2010.3

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  • 研究所だより シングルファイバーに憧れて--コペンハーゲン大学 生命科学部 食品科学部門 食肉科学グループ

    室谷 進

    畜産技術   ( 658 )   21 - 22,図巻頭1p   2010.3

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  • 粗飼料多給牛肉における骨格筋形成関連遺伝子の発現

    柴田昌宏, 曳野泰子, 松本和典, 大江美香, 中島郁世, 尾嶋孝一, 室谷進, 千国幸一

    日本畜産学会大会講演要旨   111th   65   2009.9

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  • Cellular changes in subcutaneous adipose tissue of Landrace and Meishan pigs during growth Reviewed

    Nakajima I, Oe M, Ojima K, Muroya S, Shibata M, Chikuni K

    55th International Congress of Meat Science and Technology Proceedings   PE1.11   2009.8

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  • Effects of Grass-Fed and Grain-Fed on Carcass Composition and Meat Quality of Japanese Black Cattle Reviewed

    Shibata M, Matsumoto K, Oe M, Ojima K, Nakajima I, Muroya S, Chikuni K

    55th International Congress of Meat Science and Technology Proceedings   PE1.07   2009.8

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  • Comparison of Whole, Soluble and Mitochondrial Protein Profiles between Fast and Slow Bovine Muscles Reviewed

    Oe M, Takeda K, Ohnishi-Kameyama M, Nakajima I, Muroya S, Shibata M, Ojima K, Chikuni K

    55th International Congress of Meat Science and Technology Proceedings   PE1.12   2009.8

  • 牛筋肉におけるリパーゼの発現量と筋線維型の関係

    千国幸一, 大江美香, 柴田昌宏, 松本和典, 中島郁世, 尾嶋孝一, 室谷進

    日本畜産学会大会講演要旨   110th   141   2009.3

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  • 自給粗飼料多給肥育がウシ屠体成績におよぼす影響

    柴田昌宏, 松本和典, 大江美香, 中島郁世, 尾嶋孝一, 室谷進, 千国幸一

    日本畜産学会大会講演要旨   110th   142   2009.3

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  • 梅山豚種とランドレース種の皮下脂肪における脂肪細胞分化マーカーの発育に伴う発現変化

    中島郁世, 大江美香, 室谷進, 柴田昌宏, 尾嶋孝一, 千国幸一

    日本養豚学会大会講演要旨   90th   16   2008.10

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  • 品種の違う牛サーロイン肉に対する味認識装置の応答

    千国幸一, 大江美香, 佐々木啓介, 柴田昌宏, 中島郁世, 室谷進

    日本畜産学会大会講演要旨   109th   72   2008.3

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  • ウシ胎子期骨格筋のプロテオーム解析

    柴田昌宏, 松本和典, 大江美香, 亀山眞由美, 中島郁世, 室谷進, 千国幸一

    日本畜産学会大会講演要旨   109th   138   2008.3

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  • 26.西洋交雑豚及び梅山豚由来脂肪前駆細胞の増殖能の違い

    中島郁世, 大江美香, 室谷進, 柴田昌宏, 千国幸一

    日本養豚学会大会講演要旨   89th   26   2008.3

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  • ウシ培養筋芽細胞における筋管形成因子の探索

    柴田昌宏, 松本和典, 大江美香, 中島郁世, 室谷進, 千国幸一

    生化学   1P-1270   2008

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  • 牛肉の品質研究のためのプロテオミクス--トロポニンTを中心として

    室谷 進

    畜産技術   ( 631 )   2 - 7   2007.12

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  • 梅山豚種とランドレース種における脂質代謝関連成分の成長に伴う変化

    千国幸一, 中島郁世, 大江美香, 室谷進

    日本養豚学会大会講演要旨   88th   12   2007.10

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  • ミオシン調節軽鎖(MyLC2)タンパク質はウシ胸最長筋の死後硬直時に二重にリン酸化される

    室谷進, 亀山眞由美, 大江美香, 中島郁世, 柴田昌宏, 千国幸一

    日本畜産学会大会講演要旨   108th   87   2007.9

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  • ウシ半腱様筋および咬筋より抽出した水溶性蛋白質のプロテオーム解析

    大江美香, 亀山眞由美, 柴田昌宏, 中島郁世, 室谷進, 千国幸一

    日本畜産学会大会講演要旨   108th   88   2007.9

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  • 牛筋肉部位の違いに対する味認識装置の応答

    千国幸一, 大江美香, 佐々木啓介, 柴田昌宏, 中島郁世, 室谷進

    日本畜産学会大会講演要旨   108th   83   2007.9

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  • 肥育後期に放牧を行った黒毛和種骨格筋のプロテオーム解析

    柴田昌宏, 松本和典, 大江美香, 亀山眞由美, 中島郁世, 室谷進, 千国幸一

    日本畜産学会大会講演要旨   108th   88   2007.9

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  • 牛肉のプロテオーム解析データベース

    大江美香, 安藤幹男, 室谷進, 中島郁世, 柴田昌宏, 千国幸一

    畜産草地研究成果情報   ( 6 )   13 - 14   2007.9

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  • 日本短角種DM牛の骨格筋における筋分化調節因子とミオシン重鎖の発現

    室谷進, 渡辺康一, 小梨茂, 佐藤洋一, 高橋学, 川畑茂樹, 吉川恵郷, 麻生久, 千国幸一, 山口高弘

    日本畜産学会大会講演要旨   107th   101   2007.3

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  • 脂肪細胞と前駆細胞における細胞外マトリックス関連遺伝子の発現比較解析

    中島郁世, 大江美香, 室谷進, 柴田昌宏, 千国幸一

    日本結合組織学会学術大会抄録集   39th   131   2007

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  • ブタのアドレナリンβ受容体遺伝子の塩基配列多型

    千国幸一, 中島郁世, 大江美香, 室谷進

    日本畜産学会大会講演要旨   106th   69   2006.3

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  • ウシ骨格筋におけるトロポミオシンアイソフォーム構成とミオシン重鎖アイソフォーム構成の比較

    大江美香, 中島郁世, 室谷進, 千国幸一

    日本畜産学会大会講演要旨   106th   160   2006.3

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  • 梅山豚由来皮下脂肪前駆細胞株(MSPA)の樹立

    中島郁世, 大江美香, 室谷進, 千国幸一

    日本畜産学会大会講演要旨   106th   137   2006.3

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  • 牛肉の熟成に伴い変動する水溶性画分成分の解析

    北村慎一, 村上恭彦, 室谷進, 千国幸一, 西村敏英

    日本畜産学会大会講演要旨   106th   159   2006.3

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  • ブタの骨格筋タンパク質ミオシン重鎖2b型に特異的な抗体の作成

    千国幸一, 大江美香, 中島郁世, 室谷進

    日本養豚学会大会講演要旨   84th   10   2005.10

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  • 日本短角種DM牛の育成期における骨格筋の性状

    渡邊康一, 小梨茂, 鈴木暁之, 佐藤洋一, 大池裕治, 谷藤隆志, 吉川恵郷, 高橋学, 川畑茂樹, 室谷進, 千国幸一, 麻生久, 山口高弘

    東北畜産学会報   55 ( 2 )   36   2005.8

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  • ブタ骨格筋における筋転写調節因子とミオシン重鎖アイソフォーム構成の関係

    千国幸一, 室谷進, 中島郁世, 大江美香

    日本畜産学会大会講演要旨   105th   79   2005.8

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  • ブタの脂肪細胞分化に伴うコラーゲンの変化と分化への関与

    中島郁世, 大江美香, 室谷進, 千国幸一

    日本畜産学会大会講演要旨   105th   90   2005.8

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  • ウシ骨格筋に発現するトロポミオシンアイソフォームの多様性

    大江美香, 亀山真由美, 室谷進, 中島郁世, 千国幸一

    日本畜産学会大会講演要旨   105th   8   2005.8

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  • ウシ骨格筋におけるカルパイン系構成因子の発現解析

    室谷進, NEATH Kate, 中島郁世, 大江美香, 千国幸一

    日本畜産学会大会講演要旨   105th   80   2005.8

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  • 牛肉熟成中のトロポニンTの分解様式にみられる筋肉間差

    室谷進, 中島郁世, 大江美香, 千国幸一

    日本畜産学会大会講演要旨   104th   179   2005.3

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  • ブタ皮下脂肪前駆細胞株(PSPA)の樹立

    中島郁世, 室谷進, 千国幸一

    畜産草地研究成果情報   ( 3 )   33 - 34   2004.8

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  • 筋肉構成因子の発現は筋分化調節因子MyoDに支配され,Myf5に影響されない

    室谷進, 千国幸一, 中島郁世

    畜産草地研究成果情報   ( 3 )   31 - 32   2004.8

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  • ブタの脂肪細胞分化にデキサメタゾンとオクタン酸が必要な理由

    中島郁世, 室谷進, 千国幸一

    日本養豚学会大会講演要旨   81th   26   2004.3

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  • 牛肉熟成中に生成するトロポニンT分解産物の構造解析

    室谷進, 北村慎一, 田辺創一, 西村敏英, 中島郁世, 千国幸一

    日本畜産学会大会講演要旨   103rd   160   2004.3

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  • 豚筋肉トロボニンTの一次構造解析

    北村慎一, 室谷進, 田辺創一, 奥村朋之, 千国幸一, 西村敏英

    日本農芸化学会大会講演要旨集   2004   64   2004.3

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  • ブタの脂肪細胞分化に必要な因子

    中島郁世, 室谷進, 千国幸一

    日本養豚学会大会講演要旨   80th   21   2003.10

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  • MyoDおよびMyf5アンチセンスオリゴDNAの添加によるウシ培養筋衛星細胞の分化制御

    室谷進, 中島郁世, 千国幸一

    日本畜産学会大会講演要旨   102nd   58   2003.9

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  • ウシ骨格筋の形成機構に関する研究

    室谷進

    日本畜産学会大会講演要旨   101st   6   2003.3

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  • ウシ骨格筋に発現するトロポニンTのアミノ酸配列の決定

    室谷進, 中島郁世, 千国幸一

    日本畜産学会大会講演要旨   101st   227   2003.3

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  • ウシ,ブタの骨格筋に存在するPGC‐1αの塩基配列

    千国幸一, 室谷進, 中島郁世

    日本畜産学会大会講演要旨   101st   203   2003.3

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  • EPC‐1遺伝子発現の分布と老化にともなう変動

    宮下範和, 久保康明, 中島郁世, 室谷進, 高橋ひとみ, 下司雅也, 千国幸一, 岡野彰

    日本畜産学会大会講演要旨   101st   208   2003.3

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  • 豚肉熟成中に生じる30kDaフラグメントの解析

    北村慎一, 田辺創一, 奥村朋之, 室谷進, 千国幸一, 西村敏英

    日本畜産学会大会講演要旨   101st   157   2003.3

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  • ブタ脂肪細胞株とマウス脂肪細胞株の比較

    中島郁世, 室谷進, 千国幸一

    日本養豚学会大会講演要旨   77th   16   2002.3

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  • ウシ骨格筋におけるミオシン重鎖アイソフォームと筋転写因子の発現の関係

    室谷進, 中島郁世, 千国幸一

    日本畜産学会大会講演要旨   100th   232   2002.3

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  • ブタ皮下脂肪細胞株の樹立

    中島郁世, 室谷進, 千国幸一

    日本養豚学会大会講演要旨   76th   19   2001.10

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  • ウシα4インテグリンcDNAの塩基配列の決定

    室谷進, 中島郁世, 千国幸一

    日本畜産学会大会講演要旨   99th   69   2001.8

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  • ウシの骨格筋で発現しているミオシン重鎖アイソフォームの特徴

    千国幸一, 中島郁世, 室谷進

    日本畜産学会大会講演要旨   99th   77   2001.8

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  • ブタ骨格筋で発現しているミオシン重鎖アイソフォームの配列比較 (農林水産省畜産試験場S)

    千国幸一, 田辺亮一, 中島郁世, 室谷進

    畜産研究成果情報   ( 15 )   55 - 56   2001.3

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  • ウシおよびブタの培養筋衛星細胞における筋分化転写因子の検出法とその発現様式 (農林水産省畜産試験場S)

    室谷進, 中島郁世, 千国幸一

    畜産研究成果情報   ( 15 )   57 - 58   2001.3

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    Language:Japanese  

    J-GLOBAL

  • ブタ遅筋型ミオシン重鎖アイソフォームの機能領域

    千国幸一, 室谷進, 中島郁世

    日本畜産学会大会講演要旨   98th   158   2001.3

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    Language:Japanese  

    J-GLOBAL

  • 脂肪細胞分化に伴うV型,VI型コラーゲンの特異的変化

    中島郁世, 室谷進, 千国幸一

    Connective Tissue   32 ( 2 )   121 - 121   2000.5

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    Language:Japanese   Publisher:The Japanese Society for Connective Tissue  

    CiNii Books

    J-GLOBAL

  • 家畜筋衛星細胞の筋管形成におけるmyogeninとVLA‐4の役割

    室谷進, 中島郁世, 千国幸一

    日本畜産学会大会講演要旨   97th   146   2000.3

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    Language:Japanese  

    J-GLOBAL

  • ブタ骨格筋における,ミオシン重鎖アイソフォーム発現様式解析法

    田辺亮一, 千国幸一, 中島郁世, 室谷進

    畜産研究成果情報   ( 13 )   31 - 32   1999.11

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    Language:Japanese  

    J-GLOBAL

  • 脂肪組織形成における細胞外基質の重要性

    中島郁世, 室谷進, 田辺亮一, 千国幸一

    畜産研究成果情報   ( 13 )   35 - 36   1999.11

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    Language:Japanese  

    J-GLOBAL

  • 食肉および食肉製品の簡易な肉種鑑別法

    千国幸一, 田辺亮一, 室谷進

    畜産研究成果情報   ( 13 )   33 - 34   1999.11

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    Language:Japanese  

    J-GLOBAL

  • 筋分化を誘導する無血清培地においてブタ筋衛星細胞は筋管を形成しない

    室谷進, 田辺亮一, 中島郁世, 千国幸一

    日本畜産学会大会講演要旨   96th   98   1999.9

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    Language:Japanese  

    J-GLOBAL

  • Analysis methods for expression pattern of myosin heavy chain isoforms in pig skeletal muscle. (Ministry of Agriculture, Forestry and Fisheries, National Food Research Institute S).

    田辺亮一, 千国幸一, 中島郁世, 室谷進

    食品研究成果情報   ( 11 )   66 - 67   1999.6

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    Language:Japanese  

    J-GLOBAL

  • Simple discrimination methods for meat species in meat and meat products. (Ministry of Agriculture, Forestry and Fisheries, National Food Research Institute S ).

    千国幸一, 田辺亮一, 室谷進

    食品研究成果情報   ( 11 )   12 - 13   1999.6

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    Language:Japanese  

    J-GLOBAL

  • Expressional patterns of myosin heavy chain isoforms, and meat characteristics.

    田辺亮一, 村上徹哉, 河原貴裕, 山城倫子, 中島郁世, 室谷進, 千国幸一

    食肉の科学   40 ( 1 )   170 - 171   1999.5

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    Language:Japanese  

    J-GLOBAL

  • ミオシン重鎖アイソフォーム発現様式と肉質

    田辺亮一, 村上徹哉, 河原貴裕, 山城倫子, 中島郁世, 室谷進, 千国幸一

    日本食肉研究会総会提出議案及び大会講演要旨   40th   16 - 18   1999.3

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    Language:Japanese  

    J-GLOBAL

  • ブタの品種による,ミオシン重鎖アイソフォーム発現様式と肉質の違い

    田辺亮一, 村上徹哉, 河原貴裕, 山城倫子, 中島郁世, 室谷進, 千国幸一

    日本畜産学会大会講演要旨   95th   147   1999.3

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    Language:Japanese  

    J-GLOBAL

  • Sequence comparison of myosin heavy chain isoform cDNA5’ ends of cattle and pig.

    田辺亮一, 室谷進, 千国幸一

    日本畜産学会大会講演要旨   94th   273   1998.3

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    Language:Japanese  

    J-GLOBAL

  • On the effect of hot environment on body fat mobilization of holstein milk cow by the epinephrine load.

    勝俣昌也, 室谷進, 高木匡, 塩谷繁, 寺田文典

    日本畜産学会大会講演要旨   94th   121   1998.3

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    Language:Japanese  

    J-GLOBAL

  • Difference of proportion of fast and slow muscle type myosin heavy chain isoform based on variety of pig and skeletal muscle.

    村上徹哉, 田辺亮一, 室谷進, 千園幸一

    日本畜産学会大会講演要旨   94th   273   1998.3

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    Language:Japanese  

    J-GLOBAL

  • Culture of the neural crest cells of silky fowl.

    室谷進, 千国幸一, 田辺亮一

    日本畜産学会大会講演要旨   94th   95   1998.3

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    Language:Japanese  

    J-GLOBAL

  • Change and structure of the muscle protein connectin under the meat maturing. ( Ministry of Agriculture, Forestry and Fisheries, Natl. Inst. of Animal Industry S ).

    田辺亮一, 室谷進, 千国幸一

    畜産研究成果情報   ( 11 )   51 - 52   1997.8

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    Language:Japanese  

    J-GLOBAL

  • Comparison of the structure of a desmin molecule head region of a chicken, a pig, and cattle.

    千国幸一, 田辺亮一, 室谷進

    日本畜産学会大会講演要旨   93rd   163   1997.8

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    Language:Japanese  

    J-GLOBAL

  • Comparison structure of a myosin heavy chain of cattle and a pig.

    田辺亮一, 室谷進, 千国幸一

    日本畜産学会大会講演要旨   93rd   163   1997.8

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    Language:Japanese  

    J-GLOBAL

  • Ripening change of protein connectin which influences meat elasticity. ( Ministry of Agriculture, Forestry and Fisheries, National Food Research Institute S ).

    田辺亮一, 室谷進, 千国幸一

    食品研究成果情報   ( 9 )   36 - 37   1997.8

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    Language:Japanese  

    J-GLOBAL

  • Simplified discriminating method of the genotype using multiplex PCR.

    千国幸一, 福本泰之, 田なべ亮一, 室谷進, 小沢忍

    日本畜産学会大会講演要旨   92nd   221   1997.3

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    Language:Japanese  

    J-GLOBAL

  • Distribution of black pigment in body of silky fowl.

    室谷進, 千国幸一, 田辺亮一

    日本畜産学会大会講演要旨   92nd   152   1997.3

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    Language:Japanese  

    J-GLOBAL

  • Polymorphism of somatotropin in Mishima Island cattles.

    福本泰之, 千国幸一, 田辺亮一, 室谷進, 篠田稔彦, 阪田昭次, 石川豊, 細井栄嗣, 小沢忍

    日本畜産学会大会講演要旨   92nd   249   1997.3

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    Language:Japanese  

    J-GLOBAL

  • Difference in existence proportion of pig myosin heavy chain isoform by the type of skeletal muscle.

    田辺亮一, 室谷進, 千国幸一

    日本畜産学会大会講演要旨   92nd   116   1997.3

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    Language:Japanese  

    J-GLOBAL

  • Analysis of pig myosin isoform by electrophoresis.

    田辺亮一, 室谷進, 村田勝巳, 福本泰之, 千国幸一

    日本養豚学会大会講演要旨   66th   22   1996.10

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    Language:Japanese  

    J-GLOBAL

  • Effect of NDF content in feed on milk production and milk constituents during hot time.

    塩谷繁, 寺田文典, 高木匡, 室谷進, 岩間裕子

    日本畜産学会大会講演要旨   91st   90   1996.3

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    Language:Japanese  

    J-GLOBAL

  • Simple discriminating method of the quality meat and meat products.

    千国幸一, 松永孝光, 田なベ亮一, 室谷進

    日本畜産学会大会講演要旨   91st   161   1996.3

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    Language:Japanese  

    J-GLOBAL

  • Effects of heat and fish meal on nitrogen compounds compositions in milk.

    室谷進, 寺田文典, 塩谷繁

    日本畜産学会大会講演要旨   91st   92   1996.3

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    Language:Japanese  

    J-GLOBAL

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Presentations

  • 尾嶋 孝一, 秦 勝志, 大内 史子, 小野 弥子, 室谷 進   骨格筋特異的に発現するカルパイン3は組織普遍的なカルパイン1および2を基質とする  

    日本畜産学会大会講演要旨集  2023.9  (公社)日本畜産学会

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    Language:Japanese  

  • 室谷 進   赤身肉の肉質と熟成   Invited

    日本産肉研究会第8回学術集会  2011.8 

  • MUROYA Susumu   Association of extracellular vesicle microRNAs with feeding conditions in beef cattle   Invited International conference

    2017 ISEV Workshop on Diet, Environment and Extracellular Vesicles  2017.1  International Society of Extracellular Vesicles

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    Language:English   Presentation type:Oral presentation (general)  

    Venue:La Trobe University, Melbourne, Australia  

  • 尾嶋 孝一, 秦 勝志, 室谷 進, 小野 弥子   組織普遍的に発現するCAPNはCAPN3を部分切断する  

    日本畜産学会大会講演要旨集  2022.9  (公社)日本畜産学会

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    Language:Japanese  

  • 室谷 進   筋衛星細胞の筋管形成に伴う筋分化転写因子の発現   Invited

    第98回日本畜産学会大会講演要旨集シンポジウム  2001.3 

  • 市村 恵美, 尾嶋 孝一, 室谷 進, 小林 謙, 西邑 隆徳   筋原線維内の太いフィラメントにおけるミオシン分子の置換パターン  

    日本畜産学会大会講演要旨集  2021.9  (公社)日本畜産学会

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    Language:Japanese  

  • Susumu Muroya   Metabolomic profiling in beef focusing on changes during postmortem aging   Invited

    Symposium in JRA Livestock Promotion Program  2022.3 

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    Presentation type:Symposium, workshop panel (nominated)  

  • 室谷 進, Y Zhang, 乙丸 孝之介, 大島 一修, 大島 一郎, 佐野 光枝, 盧 尚健, 尾嶋 孝一, 後藤 貴文   母牛の低栄養は胎子の肝臓における糖代謝,尿素回路,ステロイド代謝に影響を及ぼす  

    日本畜産学会大会講演要旨集  2023.9  (公社)日本畜産学会

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  • 西野 大地, 室谷 進, 後藤 貴文   妊娠期の栄養環境がウシ胎仔の胸最長筋におけるDNAメチル化に及ぼす影響  

    日本畜産学会大会講演要旨集  2023.9  (公社)日本畜産学会

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    Language:Japanese  

  • Susumu MUROYA   Farm Animal Muscle Metabolism Approached with Omics Technologies.   Invited

    2022.8 

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    Presentation type:Oral presentation (invited, special)  

  • 室谷 進   10年先にあってほしい牛肉とその研究開発を考える   Invited

    日本産肉研究会第29回学術集会シンポジウム「持続可能な畜産とは何か?」  2022.3 

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    Presentation type:Symposium, workshop panel (nominated)  

  • 室谷 進   ダブルマッスル牛骨格筋における筋転写因子およびミオシン重鎖の発現   Invited

    ミオスタチンワークショップ「高品質赤肉を効率的に生産する肉用牛資源の造成」-骨格筋形成抑制因子、ミオスタチンの作用機構と産肉への応用―  2006.1 

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Intellectual Property

  • ブタ脂肪前駆細胞株とその分化誘導方法

    中島 郁世, 室谷 進, 千国 幸一

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    Applicant:独立行政法人農業技術研究機構

    Application no:特願2001-316258  Date applied:2001.10

    Announcement no:特開2003-116528  Date announced:2003.4

    J-GLOBAL

  • 遺伝子型の判定方法

    千國 幸一, 田邉 亮一, 室谷 進

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    Applicant:農林水産省畜産試験場長

    Application no:特願平8-312654  Date applied:1996.11

    Announcement no:特開平10-136985  Date announced:1998.5

    Patent/Registration no:特許第3005668号  Date issued:1999.11

    J-GLOBAL

  • 遺伝子型の判定方法

    千國 幸一, 田邉 亮一, 室谷 進

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    Applicant:農林水産省畜産試験場長

    Application no:特願平8-312654  Date applied:1996.11

    Announcement no:特開平10-136985  Date announced:1998.5

    J-GLOBAL

Awards

  • 16th AB/CAPI Outstanding Research Award (Best Paper)

    2022.3   Animal Bioscience office   Metabolomic approach to key metabolites characterizing postmortem aged loin muscle of Japanese Black (Wagyu) cattle

    Susumu Muroya

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    Award type:Honored in official journal of a scientific society, scientific journal 

  • 日本農学進歩賞

    2006.11   公益財団法人農学会   ウシ骨格筋の形成機構とその熟成に関する分子生物学的研究

    室谷 進

  • 日本畜産学会奨励賞

    2003.3   日本畜産学会  

    室谷 進

Research Projects

  • The role of histone modification in bovine skeletal muscle altered by nutrition during fetal life

    Grant number:22K05963  2022.4 - 2025.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)

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    Grant amount:\4160000 ( Direct Cost: \3200000 、 Indirect Cost:\960000 )

  • 代謝刷り込みによる牛放牧肥育技術開発事業

    2021.4 - 2024.3

    令和3年度日本中央競馬会畜産振興事業 

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    Authorship:Coinvestigator(s) 

  • 褐毛和種高知系の肉質・ブランド力強化事業

    2021.4 - 2024.3

    令和3年度日本中央競馬会畜産振興事業 

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    Authorship:Coinvestigator(s) 

  • 豚特有のチアミン三リン酸蓄積機構の解明

    2018.6 - 2019.3

    伊藤記念財団  研究助成事業 

    室谷 進

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    Authorship:Principal investigator  Grant type:Competitive

  • 黒毛和種牛の代謝プログラミングにおけるmicroRNAの役割の解明

    2018.4 - 2021.3

    日本学術振興会  科学研究費補助金 

    室谷 進

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    Authorship:Principal investigator  Grant type:Competitive

  • 国産和牛肉の新たな差別化のための評価指標及び育種手法の開発

    2016.4 - 2020.3

    農研機構生物系特定産業技術研究支援センター  革新的技術開発・緊急展開事業(うち先導プロジェクト) 

    小林栄治

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    Grant type:Competitive

  • 胎児期と初期成長期の代謝プログラミングによる和牛肥育期間の大幅な短縮技術の開発

    2016.4 - 2018.3

    農研機構生物系特定産業技術研究支援センター  革新的技術開発・緊急展開事業(うち先導プロジェクト) 

    後藤貴文

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    Grant type:Competitive

  • Roles of circulating microRNAs in skeletal muscle gene expression in grazing cattle

    Grant number:15K14849  2015.4 - 2017.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research  Grant-in-Aid for Challenging Exploratory Research

    Hayashi Masayuki

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    Grant amount:\3900000 ( Direct Cost: \3000000 、 Indirect Cost:\900000 )

    MicroRNAs are around 22 nt-long small RNAs. MicroRNAs have roles on RNA degradation or inhibition of translation of the target genes. MicroRNAs are present in several solid tissues and body fluids, including blood in which they are packed in small extracellular vesicles, called exosomes. To explore miRNAs characterizing feeding condition, we compared blood microRNAs between pastured and concentrate-fed Japanese Black cattle. We consequently determined differently expressed microRNAs, including miR-10b and miR-17-5p, between the feeding conditions. Furthermore, gene expression including DNAJB2 and PTEN, potential targets of those miRNAs, were different between the feeding conditions. We conclude that difference in feeding condition between pasture and high concentrate feeding affected blood microRNAs and skeletal muscle gene expression in Japanese Black cattle.

  • 速筋に発現するmicroRNAの細胞局在性と機能の解明

    2015.4

    日本学術振興会  科学研究費補助金 

    室谷 進

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    Authorship:Principal investigator  Grant type:Competitive

  • 牛乳中microRNAの新規な食品機能性の探索

    2013.4

    日本学術振興会  科学研究費補助金 

    室谷 進

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    Authorship:Principal investigator  Grant type:Competitive

  • メタボロミクスによる豚肉品質関連物質の網羅的変動解析

    2011.6

    財団法人旗影会  研究助成(一般助成) 

    室谷 進

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    Authorship:Principal investigator  Grant type:Competitive

  • Taste-forming mechanisms in single muscle fibers

    Grant number:23580380  2011 - 2013

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    CHIKUNI Koichi, MUROYA Susumu

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    Grant amount:\4810000 ( Direct Cost: \3700000 、 Indirect Cost:\1110000 )

    To clarify the effects of muscle fiber type on the nucleotide content, the concentration of nucleotides were determined from bovine muscles, porcine muscles and porcine single muscle fibers. The concentration of inosine 5'-monophosphate (IMP) was significantly lower in the slow type muscles than that in the fast type muscles, and ultimate pH was higher in the slow type muscles. The results suggested that the higher pH condition accelerated ATP degradation process and then decreased the IMP concentration in the slow type muscles. The IMP concentration of an each single muscle fiber was determined in the fibers separated from porcine psoas major muscles.Although slow and fast type muscle fibers were separated from the same psoas major muscle, there was no significant difference in IMP concentration between the slow and fast muscle fibers. The equilibration of pH in a muscle post-mortem may explain the conflict of the results from muscles and single muscle fibers.

  • 機能性小分子RNAの網羅的解析による筋線維型調節機構の解明

    2010.4

    日本学術振興会  科学研究費補助金 

    室谷 進

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    Authorship:Principal investigator  Grant type:Competitive

  • 筋線維型が食肉のやわらかさに与える影響の解明

    2009.4

    日本学術振興会  研究者交流(特定国派遣研究者:長期) 

    室谷 進

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    Authorship:Principal investigator  Grant type:Competitive

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Teaching Experience

  • 畜産物利用学(肉)

    2011.3
    -
    2022.3
    Institution:筑波大学

  • 畜産物特別講義Ⅰ

    2007.4
    -
    2008.3
    Institution:麻布大学

  • 畜産物特別講義Ⅱ

    2007.4
    -
    2008.3
    Institution:麻布大学