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

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

Scopus

PubMed

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

Scopus

PubMed

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

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

Scopus

PubMed

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

Scopus

PubMed

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

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

Scopus

PubMed

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

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

Scopus

PubMed

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

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

Scopus

PubMed

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

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

Scopus

PubMed

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

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

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

Scopus

PubMed

Language：English   Publishing type：Research paper (scientific journal)   Publisher：Asian Australasian Association of Animal Production Societies  

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.

DOI： 10.5713/ajas.19.0682

Scopus

PubMed

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

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.

DOI： 10.1242/bio.048975

Scopus

PubMed

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

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.

DOI： 10.3390/metabo10050188

Scopus

PubMed

Language：Japanese   Publisher：Asian Australasian Association of Animal Production Societies  

DOI： 10.5713/ajas.18.0648

Scopus

PubMed

Language：Japanese  

DOI： 10.1016/j.domaniend.2019.01.002

Scopus

PubMed

Publishing type：Research paper (scientific journal)   Publisher：Oxford University Press (OUP)  

DOI： 10.1093/jas/sky404.783

Language：Japanese  

DOI： 10.1152/ajpcell.00245.2017

Scopus

PubMed

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

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.

DOI： 10.1016/j.meatsci.2017.11.035

Scopus

PubMed

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

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.

DOI： 10.1111/asj.12826

Web of Science

Scopus

PubMed

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

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

DOI： 10.1002/jsfa.8204

Web of Science

Scopus

PubMed

Language：English   Publishing type：Research paper (scientific journal)   Publisher：PUBLIC LIBRARY SCIENCE  

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.

DOI： 10.1371/journal.pone.0162496

Web of Science

Scopus

PubMed

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

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.

DOI： 10.1016/j.meatsci.2016.04.013

Web of Science

Scopus

PubMed

Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL  

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.

DOI： 10.1002/2211-5463.12091

Web of Science

Scopus

PubMed

Language：English   Publishing type：Research paper (scientific journal)   Publisher：BIOMED CENTRAL LTD  

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.

DOI： 10.1186/s40104-016-0068-x

Web of Science

Scopus

PubMed

Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL  

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.

DOI： 10.1111/asj.12381

Web of Science

Scopus

PubMed

Language：English   Publishing type：Research paper (scientific journal)   Publisher：PUBLIC LIBRARY SCIENCE  

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.

DOI： 10.1371/journal.pone.0136475

Web of Science

Scopus

PubMed

Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL  

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.

DOI： 10.1111/asj.12310

Web of Science

Scopus

PubMed

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

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.

DOI： 10.1016/j.euprot.2014.08.001

Scopus

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

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.

DOI： 10.1016/j.meatsci.2014.07.018

Web of Science

Scopus

PubMed

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

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

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.

DOI： 10.1016/j.meatsci.2013.10.021

Web of Science

Scopus

PubMed

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

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

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.

DOI： 10.2527/jas.2012-5371

Scopus

PubMed

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

Language：Japanese   Publisher：The Japanese Society of Swine Science  

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

CiNii Books

Language：English   Publishing type：Research paper (scientific journal)   Publisher：TAYLOR & FRANCIS LTD  

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.

DOI： 10.1271/bbb.120424

Web of Science

Scopus

PubMed

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

Language：Japanese   Publisher：The Japanese Society of Swine Science  

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

CiNii Books

Language：English  

CiNii Books

Other Link： http://agriknowledge.affrc.go.jp/RN/2010831416

Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL  

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.

DOI： 10.1111/j.1740-0929.2011.00954.x

Web of Science

Scopus

PubMed

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

Language：English   Publisher：Warm Regional Society of Animal Science, Japan  

<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

CiNii Books

Other Link： https://jlc.jst.go.jp/DN/JALC/10006074615?from=CiNii

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

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.

DOI： 10.1016/j.meatsci.2011.05.006

Web of Science

Scopus

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

Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL  

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

Web of Science

Scopus

PubMed

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

Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL PUBLISHING, INC  

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.

DOI： 10.1111/j.1740-0929.2010.00823.x

Web of Science

Scopus

PubMed

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

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

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.

DOI： 10.1016/j.meatsci.2010.06.019

Web of Science

Scopus

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

Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL  

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.

DOI： 10.1111/j.1740-0929.2010.00773.x

Web of Science

Scopus

PubMed

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

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

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

Web of Science

Scopus

PubMed

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

Language：English   Publishing type：Research paper (scientific journal)   Publisher：AMER SOC ANIMAL SCIENCE  

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.

DOI： 10.2527/jas.2008-1486

Web of Science

Scopus

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

Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL PUBLISHING, INC  

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.

DOI： 10.1111/j.1740-0929.2008.00613.x

Web of Science

Scopus

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

Language：English   Publishing type：Research paper (scientific journal)   Publisher：TAYLOR & FRANCIS LTD  

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.

DOI： 10.1271/bbb.80390

Web of Science

Scopus

PubMed

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

Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL PUBLISHING, INC  

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

Web of Science

Scopus

PubMed

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

Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL PUBLISHING, INC  

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.

DOI： 10.1111/j.1740-0929.2008.00578.x

Web of Science

Scopus

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

Language：English  

DOI： 10.1016/j.jns.2007.07.011

PubMed

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

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.

DOI： 10.1016/j.meatsci.2007.04.010

Web of Science

Scopus

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

Language：Japanese   Publisher：日本食肉科学会  

CiNii Books

Language：English   Publishing type：Research paper (scientific journal)   Publisher：AMER CHEMICAL SOC  

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.

DOI： 10.1021/jf063200o

Web of Science

Scopus

PubMed

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

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

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.

DOI： 10.1016/j.meatsci.2006.09.003

Web of Science

Scopus

PubMed

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

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

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.

DOI： 10.1016/j.meatsci.2006.08.016

Web of Science

Scopus

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

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

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.

DOI： 10.1016/j.meatsci.2006.08.012

Web of Science

Scopus

PubMed

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

Language：English  

DOI： 10.1016/j.bbrc.2006.12.122

PubMed

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

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.

DOI： 10.4081/ijas.2007.s2.1175

Web of Science

Scopus

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

Language：English   Publishing type：Research paper (international conference proceedings)  

Language：English   Publishing type：Research paper (scientific journal)   Publisher：TAYLOR & FRANCIS LTD  

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.

DOI： 10.1271/bbb.70.726

Web of Science

Scopus

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

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

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.

DOI： 10.1016/j.meatsci.2005.07.008

Web of Science

Scopus

PubMed

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

Language：Japanese   Publisher：Japan Society for Bioscience, Biotechnology, and Agrochemistry  

DOI： 10.1271/kagakutoseibutsu1962.44.831

CiNii Books

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

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.

DOI： 10.1111/j.1440-169X.2005.00822.x

DOI： 10.1111/j.1440-169x.2005.00822.x

Web of Science

Scopus

PubMed

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

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.

DOI： 10.1111/j.1740-0929.2005.00278.x

Scopus

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

Language：Japanese   Publisher：Japanese Society of Animal Breeding and Genetics  

DOI： 10.5924/abgri2000.32.2_141

CiNii Books

Language：Japanese   Publisher：Journal of Agricultural and Food Chemistry  

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.

DOI： 10.1021/jf047974l

Scopus

Language：English   Publishing type：Research paper (scientific journal)   Publisher：AMER CHEMICAL SOC  

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.

DOI： 10.1021/jf0479741

Web of Science

Scopus

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

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

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.

DOI： 10.2108/zsj.21.589

Web of Science

Scopus

PubMed

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

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

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.

DOI： 10.1016/j.meatsci.2003.08.018

Web of Science

Scopus

PubMed

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

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

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.

DOI： 10.1016/j.meatsci.2003.09.011

Web of Science

Scopus

PubMed

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

Language：English   Publishing type：Research paper (scientific journal)   Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE  

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.

DOI： 10.1016/j.bbrc.2003.08.057

Web of Science

Scopus

PubMed

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

Language：English   Publishing type：Research paper (scientific journal)   Publisher：AMER SOC ANIMAL SCIENCE  

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.

DOI： 10.2527/2003.8151185x

Web of Science

Scopus

PubMed

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

Language：English   Publisher：Wiley-Blackwell  

DOI： 10.1046/j.1344-3941.2002.00052.x

CiNii Books

Language：English   Publisher：Wiley-Blackwell  

DOI： 10.1046/j.1344-3941.2002.00035.x

CiNii Books

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

Web of Science

Scopus

PubMed

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

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

Web of Science

Scopus

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

Web of Science

Scopus

PubMed

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

Publisher：The Japanese Society of Swine Science  

DOI： 10.5938/youton.39.191

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

Web of Science

Scopus

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

Language：English  

DOI： 10.1097/00001756-200107030-00024

PubMed

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

Web of Science

Scopus

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

Publisher：Japanese Society of Animal Science  

DOI： 10.2508/chikusan.72.230

Publisher：Japanese Society of Animal Science  

DOI： 10.2508/chikusan.72.505

Language：English  

DOI： 10.1111/j.1749-6632.2000.tb05207.x

PubMed

Language：English  

DOI： 10.1097/00001756-200004070-00031

PubMed

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

Web of Science

Scopus

Language：English  

DOI： 10.1016/s0304-3940(99)00185-8

PubMed

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

Web of Science

Scopus

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

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

Web of Science

Scopus

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

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

DOI： 10.1007/s003359900924

Web of Science

Scopus

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

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

Web of Science

Scopus

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

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

Scopus

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.

Web of Science

Scopus

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

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

Scopus

Publisher：Japanese Society of Animal Science  

DOI： 10.2508/chikusan.68.297

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

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：English   Publisher：(公社)日本畜産学会  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：English   Publisher：ELSEVIER SCI LTD  

DOI： 10.1016/j.meatsci.2015.01.006

Web of Science

Scopus

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：AMER SOC CELL BIOLOGY  

Web of Science

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：AMER SOC CELL BIOLOGY  

Web of Science

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：AMER SOC CELL BIOLOGY  

Web of Science

Language：Japanese  

J-GLOBAL

Language：Japanese   Publisher：日本食肉研究会  

CiNii Books

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：AMER SOC CELL BIOLOGY  

Web of Science

Language：English  

Language：English  

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：AMER SOC CELL BIOLOGY  

Web of Science

Language：Japanese  

J-GLOBAL

Language：Japanese   Publisher：畜産技術協会  

CiNii Books

Language：Japanese  

J-GLOBAL

Language：English  

Language：English  

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese   Publisher：畜産技術協会  

CiNii Books

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese   Publisher：The Japanese Society for Connective Tissue  

CiNii Books

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

J-GLOBAL

Language：Japanese  

Presentation type：Symposium, workshop panel (nominated)  

Language：Japanese  

Language：Japanese  

Presentation type：Oral presentation (invited, special)  

Presentation type：Symposium, workshop panel (nominated)