Peroxisome proliferator-activated receptor-γ coactivator 1 α (PGC-1 α) expression and the formation of slow-twitch muscle fibers in porcine and bovine skeletal muscles

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.     

β‐hydroxy‐β‐methyl butyrate promotes leucine metabolism and improves muscle fibre composition in growing pigs

The aim of this study was to investigate the effects of excess leucine (Leu) vs. its metabolites α‐ketoisocaproate (KIC) and β‐hydroxy‐β‐methyl butyrate (HMB) on Leu metabolism, muscle fibre composition and muscle growth in growing pigs. Thirty‐two pigs with a similar initial weight (9.55 ± 0.19 kg) were fed 1 of 4 diets for 45 days: basal diet, basal diet + 1.25% L‐Leu, basal diet + 1.25% KIC‐Ca, basal diet + 0.62% HMB‐Ca. Results indicated that relative to the basal diet and HMB groups, Leu and KIC groups exhibited increased Leu concentrations and decreased concentrations of isoleucine, valine and EAAs in selected muscle (p < 0.05) and had lower mRNA levels of MyHC I and higher expression of MyHC IIx/IIb (p < 0.05), and there was no significant difference between the basal and HMB‐supplemented groups. Moreover, the mRNA expression levels of AMPKα and UCP3 were higher but the myostatin mRNA levels were lower in the soleus muscle of the HMB group than those from other groups (p < 0.05). These findings demonstrated that doubling dietary Leu content exerted growth‐depressing effects in growing pigs; dietary KIC supplementation induced muscular branched‐chain amino acid imbalance and promoted muscle toward a more glycolytic phenotype; while dietary HMB supplementation promoted the generation of more oxidative muscle types and increased muscle growth specially in oxidative skeletal muscle, and these effects of HMB might be associated with the AMPKα‐Sirt1‐PGC‐1α axis and mitochondrial biogenesis.     

Correlation between skeletal muscle fiber type and free amino acid levels in Japanese Black steers

Free amino acids are important components of tastants and flavor precursors in meat. To clarify the correlation between muscle fiber type and free amino acids, we determined the concentrations of various free amino acids and dipeptides in samples of different muscle tissues (n = 21), collected from 26‐month‐old Japanese Black steers (n = 3) at 2 days postmortem. The proportions of the myosin heavy chain (MyHC), slow (MyHC1) and fast (MyHC2) isoforms were determined by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE). The contents of free amino acids and dipeptides were measured by high performance liquid chromatography (HPLC). The MyHC isoform composition varied among the tissue samples. The MyHC1 proportion ranged from 6.9% ± 3.9% to 83.3% ± 16.7%. We confirmed that there was a strong positive correlation between MyHC1 composition and total free amino acid concentrations, including those for two dipeptides. Among the 31 measured free amino acids and dipeptides, 11 showed significant positive correlations and five showed significant negative correlations with MyHC1 composition. These results suggest that a high MyHC1 content induces high free amino acid contents in bovine muscles possibly because of greater oxidative metabolism. This high level of free amino acids could contribute to the intense flavor of meat that is rich in slow‐twitch fibers.     

Satellite cell and myonuclei populations in rat soleus and extensor digitorum longus muscles after maternal nutritional deprivation and realimentation

The manner and rate of nuclear proliferation and accumulation in rat skeletal muscle was investigated using a model of induced permanent growth impairment in rats to gain additional insight into the regulation of skeletal muscle growth. Comparisons were made at 1, 21 and 175 d between control progeny and progeny of dams restricted during gestation and lactation to 50% the daily feed intake of ad libitum fed controls, followed by realimentation after weaning. Estimates of total satellite cells/muscle were lower (P less than .01) in soleus and extensor digitorum longus (EDL) of restricted progeny at 21 and 175 d. Estimates of total satellite cells/muscle increased nearly twofold between 21 and 175 d in the soleus and decreased slightly in EDL in both treatment groups. Satellite cell concentration expressed as a percentage of total muscle nuclei was not different between restricted and control progeny at either 1 or 175 d. However, while satellite cell percentages decreased about 50% between 1 and 21 d in controls, they remained at initial levels in restricted progeny. Incidence of satellite cells/muscle fiber (satellite cell concentration) decreased between 1 and 21 d in control soleus and EDL, but remained unchanged or was slightly higher in soleus and EDL of restricted progeny at 21 d. Incidence of satellite cells continued to decrease between 21 and 175 d in restricted and control EDL and restricted soleus, but was unchanged in control soleus. Incidence of myonuclei/fiber (myonuclei concentration) increased with age (P less than .005), was not affected by growth impairment either before or after realimentation and was higher (P less than .005) in soleus than EDL.(ABSTRACT TRUNCATED AT 250 WORDS)     

Increased expression of NOR‐1 mRNA in the skeletal muscles of cold‐exposed neonatal chicks

Nuclear receptor subfamily 4, group A (NR4A) subgroup orphan receptors are rapidly induced by various physiological stimuli and have been suggested to regulate oxidative metabolism and muscle mass in mammalian skeletal muscle. The results showed that the NR4A subgroup orphan receptor, NOR‐1 (NR4A3), was acutely increased in skeletal muscles of neonatal chicks in response to short‐term cold exposure. The increased NOR‐1 gene expression was concomitant with cold‐induced changes in gene expression of both myostatin and proliferator‐activated receptor‐gamma coactivator‐1 (PGC‐1α), and the increase in skeletal muscle mass. These observations suggest that NOR‐1 might play a role in controlling skeletal muscle growth in cold‐exposed neonatal chicks.     

Possible roles of myostatin and PGC-1α in the increase of skeletal muscle and transformation of fiber type in cold-exposed chicks: Expression of myostatin and PGC-1α in chicks exposed to cold

This study examined the hypothesis that myostatin and PGC-1α are involved in the increase in skeletal muscle mass and transformation of fiber type in cold-exposed chicks. One-week-old chicks were exposed to acute (24 h) or long-term (8 d) cold at 4 °C or kept warm at 30 °C. Acute cold exposure induced a significant increase in the skeletal muscle weight and the ratio of slow- to fast-fiber specific troponin I expression (sTnI/fTnI), accompanied by a significant decrease in lactate dehydrogenase activity. Expression of myostatin mRNA in the muscle was significantly lower in cold-exposed chicks than in the controls, whereas PGC-1α mRNA expression was significantly enhanced. These changes in the gene expression rapidly returned to the levels of the control chicks after the end of cold exposure, whereas the changes in fiber type and enzymatic activity were not resumed within 24 h after removal of cold exposure. On the other hand, long-term exposure to cold resulted in a remarkable increase in skeletal muscle weight, accompanied by a significant increase in the ratio of sTnI/fTnI and the enzymatic activities of cytochrome oxidase and lactate dehydrogenase. However, the expression level of myostatin mRNA in cold-exposed chicks was not different from that in their age-matched control chicks and that of PGC-1α mRNA was significantly lower than in the controls. These results indicate that myostatin and PGC-1α expression in the skeletal muscle rapidly change in response to acute cold, suggesting the possibility that these two genes could be involved in the increase in muscle mass and transformation of fiber type, respectively, at the initial stage of adaptation in cold-exposed chicks.     

Glut4突变调控脂肪重分布及肌纤维重塑的机制研究

旨在探讨Glut4基因突变后骨骼肌能量代谢及肌纤维转化的分子机制。本研究利用CRISPR/Cas9技术构建Glut4^Q177L突变鼠。选取22周龄健康的野生雄鼠和Glut4^Q177L突变雄鼠,即试验分为两组,每组3只,3个重复,进行表型评价和糖耐量、胰岛素耐量试验;荧光定量检测两组小鼠腓肠肌葡萄糖转运蛋白、脂代谢、肌纤维类型相关调控基因的表达差异;Western blot测定腓肠肌AMPK及其磷酸化蛋白含量。结果表明,突变鼠皮下脂肪和附睾脂重量显著低于对照组（P<0.05）;突变鼠糖耐量曲线下面积极显著增加（P<0.01）,表明其糖耐量受损;突变鼠血清中甘油三酯的含量显著下降（P<0.05）。相较于野生型小鼠,Glut4^Q177L突变鼠腓肠肌中Glut4、Glut1和Glut12等多个葡萄糖转运蛋白表达量显著升高（P<0.05）;葡萄糖摄取受限导致调控能量代谢关键基因AMPK在mRNA、蛋白和磷酸化修饰水平均显著升高（P<0.05）,同时促进与脂肪酸摄取与合成代谢相关的CD36和ATGL等基因表达上调（P<0.05）;慢速氧化型肌纤维（I型）和快速氧化型肌纤维（IIa型）相关基因表达极显著升高（P<0.01）,而快速酵解型纤维（IIb型）相关基因表达显著降低（P<0.05）。本研究结果显示,Glut4葡萄糖结合关键位点突变降低了骨骼肌和脂肪葡萄糖摄取能力,通过上调其它转运蛋白增加葡萄糖摄取;激活AMPK信号通路及分泌肌细胞因子调控脂肪分解以满足能量需求;促进线粒体丰富的氧化型肌纤维生成以提高能量利用效率。Glut4突变不仅可以为骨骼肌胰岛素抵抗提供有效的动物模型,还可以为家畜生物育种提供基因编辑参考位点。     

Influence of halothane genotype and body-weight on myosin heavy chain composition in pig muscle as related to meat quality

In an effort to understand the relationship between muscle fiber type, live weight, genotype, and PSE development, enzyme-linked immunosorbent analyses were used to evaluate myosin heavy chain (MyHC) isoform content in the longissimus muscle of pigs differing in halothane gene status (nn, homozygous mutant; Nn, heterozygous; NN, homozygous normal) that were slaughtered at three different weights (100, 120 and 140 kg). Pigs carrying the n gene (Nn and nn) exhibited more IIB MyHC and less slow type I MyHC than those pigs free of the n gene, while NN pigs had greater amounts of IIAX MyHC. The relative abundance of IIB and IIAX MyHC in muscle of all pigs studied was strongly negatively correlated (r=−0.834). Heavier pigs (140 kg) had the greatest amounts of slow and IIA MyHC. Across all genotypes, the relative abundance of IIB MyHC and muscle pH at 45 min postexsanguination (pH45) was negatively correlated (r=−0.418). In addition, the relative amount of slow was positively correlated with pH45 (r=0.386). Because muscle of homozygous nn positive pigs exhibited similar IIB/slow MyHC ratios to that of heterozygous Nn pigs, yet less desirable pH45 values and ultimate meat quality scores argues against a role of MyHC content per se in contributing to PSE development. However, these data do not preclude that those pigs with greater amounts of IIB MyHC are more ‘susceptible’ to adverse pork quality development than those pigs with less IIB MyHC.     

Mitochondrial biogenesis and PGC‐1α gene expression in male broilers from ascites‐susceptible and ‐resistant lines

Ascites is a cardiovascular metabolic disease characterized by accumulation of fluid around the heart and in the abdominal cavity that eventually leads to death. This syndrome is the end‐point result of a series of metabolic incidents that are generally caused by impaired oxygen availability. Mitochondria are the major sites of oxygen consumption, therefore major contributors to oxidative stress. Genetic, metabolic and dietary factors can influence variations in mitochondrial biogenesis (mitochondrial size, number and mass) that might have an effect on oxygen consumption and reactive oxygen species production. This study evaluated the effect of genotype on PGC‐1α mRNA gene expression and mitochondrial biogenesis. These parameters were examined in male broiler chickens at 22 weeks of age from the SUS and RES lines divergently selected for ascites phenotype. From each line, five birds were sampled for right ventricle and breast muscle. Gene expression and mtDNA copy number were assessed by quantitative PCR. Results showed that birds from SUS had significantly higher PGC‐1α mRNA gene (p = .033) and mitochondrial DNA copy number (p = .038) in breast muscle. There was no difference in right ventricle PGC‐1α expression or mitochondrial DNA copy number between the two lines. These findings indicate that mitochondrial biogenesis and PGC‐1α mRNA gene expression differ between male broiler chickens from RES and SUS lines in a tissue‐specific manner.     

Triiodothyronine differentially regulates key metabolic factors in lean and obese cats

The effect of a 2-week administration of 75microg triiodothyronine (T3) on substrate oxidation, heat production, non-esterified fatty acids, and leptin was evaluated in eight lean (three females and five males) and eight obese (five females and three males) age-matched adult neutered cats. In addition, using real-time RT-PCR, expression of muscle and adipose tissue uncoupling proteins (UCP2 and UCP3), deiodinase 1 and 2 (D1; D2), and peroxisome proliferator-activated receptor (PPAR) alpha and gamma and peroxisome-proliferator-activator receptor-gamma co-activator 1alpha (PGC1) was examined. Compared to lean cats, obese cats had increased NEFA, leptin, UCP2, and D1mRNA in muscle and UCP3mRNA levels in fat, but lower heat production, and fat PPARs and PGC1. T3 administration increased thermogenesis and NEFA in lean and obese cats, and adipose tissue PPARgamma in lean cats. It also increased muscle D1 in lean and D2 in obese cats. The increase in muscle D2 was interpreted to be reflective of the reduced serum total T4 concentration following T3 suppression of the pituitary. No effect was seen on leptin, or UCP2 and 3. This shows that T3 regulates thermogenesis but not through changes in uncoupling protein expression. It also indicates that PPARs have an important role in the pathogenesis of obesity in cats.     

Increase in muscle endurance in mice by dietary Yamabushitake mushroom (Hericium erinaceus) possibly via activation of PPARδ

Skeletal muscle fiber is largely classified into two types: type 1 (slow‐twitch) and type 2 (fast‐twitch) fibers. Meat quality and composition of fiber types are thought to be closely related. Previous research showed that overexpression of constitutively active peroxisome proliferator‐activated receptor (PPAR)δ, a nuclear receptor present in skeletal muscle, increased type 1 fibers in mice. In this study, we found that hexane extracts of Yamabushitake mushroom (Hericium erinaceus) showed PPARδ agonistic activity in vitro. Eight‐week‐old C57BL/6J mice were fed a diet supplemented with 5% (w/w) freeze‐dried Yamabushitake mushroom for 24 hr. After the treatment period, the extensor digitorum longus (EDL) muscles were excised. The Yamabushitake‐supplemented diet up‐regulated the PPARδ target genes Pdk4 and Ucp3 in mouse skeletal muscles in vivo. Furthermore, feeding the Yamabushitake‐supplemented diet to mice for 8 weeks resulted in a significant increase in muscle endurance. These results indicate that Yamabushitake mushroom contains PPARδ agonistic ligands and that dietary intake of Yamabushitake mushroom could activate PPARδ in skeletal muscle of mice. Unexpectedly, we observed no significant alterations in composition of muscle fiber types between the mice fed control and Yamabushitake‐supplemented diets.     

肌球蛋白重链基因与肉品质的关系

肌纤维是构成肌肉的基本单位,肌纤维的特性直接影响猪肉品质.猪的肌纤维根据肌球蛋白重链(MyHC)的多态性可分为1、2a、2b和2x 4种类型,在代谢上分别与慢速氧化型、快速氧化型、快速酵解型和中间类型相对应.肌纤维的生成在分子水平上受到肌细胞生成素基因的精确调控,肌纤维的类型在生长过程中不断发生转化,并受遗传、生长、性别和营养等多因素的影响.     

Correlation between skeletal muscle fiber type and responses of a taste sensing system in various beef samples

The difference of muscle fiber type composition affects several parameters related to meat quality; however, the relationship between muscle fiber types and meat taste is unclear. To elucidate this relationship, we determined the taste of various beef samples using a taste sensor (INSENT SA402B) and analyzed its correlation with different muscle fiber type composition. We used 22 kinds of beef samples and measured nine tastes, including the relative and change of membrane potential caused by adsorption (CPA) values, using six sensors (GL1, CT0, CA0, AAE, C00, and AE1). The taste sensor analysis indicated positive value outputs for the relative C00, AAE, and GL1 values as well as for the CPA value of AAE, which corresponded to bitterness, umami, sweetness, and richness, respectively. We found significant positive correlations of the myosin heavy chain 1 (MyHC1) composition with umami taste, and with richness. This result suggests that high levels of slow MyHC1 can induce strong umami taste and richness in beef. We expect that our results will contribute to the elucidation of the relationship between muscle fiber types and meat palatability.     

共轭亚油酸对体外培养的猪骨骼肌肌纤维类型组成的影响

旨在研究共轭亚油酸(conjugated linoleic acid,CLA)对体外培养的猪骨骼肌肌纤维类型组成的影响规律。以体外培养的原代猪骨骼肌卫星细胞为材料,在卫星细胞向肌纤维转化时添加不同水平CLA(0、50、100、150、200μg.mL-1),处理后第4、8和12天,分别采用相对定量RT-PCR测定肌纤维中MyHCⅠ、MyHC 2a、MyHC 2b和MyHC 2x 4种MyHC的基因表达。结果表明,肌纤维类型的组成随培养时间的延长发生显著变化,从第4到12天,MyHC2b型肌纤维比例显著上升,而其余3种类型的肌纤维比例均显著下降。添加50μg.mL-1CLA对肌纤维类型组成无显著影响。添加100μg.mL-1CLA主要影响第12天的肌纤维类型组成,而添加150~200μg.ml-1CLA则可显著改变第4~12天的肌纤维类型组成,即显著提高MyHC I和MyHC 2a型肌纤维比例,显著降低MyHC2x和MyHC2b型肌纤维比例。以上结果提示,添加CLA可使肌纤维类型组成发生变化,且该作用与添加水平和处理时间密切相关。CLA对肌纤维类型组成的影响主要表现为提高MyHCI和MyHC2a型肌纤维比例,降低MyHC 2b和MyHC 2x型肌纤维比例,这在一定程度上可解释CLA提高猪肉品质的原因。     

运动对苏尼特羊肌纤维组织学特性和肉品质的影响及其调控机理

本试验以6月龄苏尼特羊为研究对象,采用肌球蛋白重链(MyHC)分子分型法、ATP酶染色法分析肌纤维组织学特性,并对肉品质指标、能量代谢相关基因mRNA相对表达量、代谢酶活性进行测定及分析,以探究运动对苏尼特羊肌纤维特性和肉品质的影响及其调控机理.选取体况良好、平均体重为(19.77±3.81)kg的3月龄苏尼特羊14只...     

Maternal dietary linoleic acid supplementation promotes muscle fibre type transformation in suckling piglets

As meat quality is basically dependent on muscle fibre characteristics, it is important to know how muscle fibres are regulated and transformed. This study aimed to investigate the effect of maternal dietary supplementation on muscle fibre types using 3% saturated fatty acid (palmitic acid, PA) or 3% unsaturated fatty acid (linoleic acid, LA) from 80 days of gestation to the weaning of offspring (25 days post‐natal). The results indicated that higher mRNA levels of MyHCI type genes were found in the soleus muscles of piglets that suckled from LA‐supplemented sows than from PA‐supplemented sows. In addition, LA treatment increased the gene expression of the type I muscle fibre marker troponin I (p < 0.01), suggesting that LA promoted muscle fibre type transformation to type I fibres. Moreover, PGC‐1α (p < 0.01) and MEF2c (p < 0.05) mRNA levels were higher in the piglets from the LA treatment group than in those from the PA treatment group. Furthermore, LA supplementation also significantly increased AMP‐activated protein kinase (AMPK) mRNA levels (p < 0.05), which is an upstream regulator of PGC‐1α. Collectively, these findings demonstrated that maternal dietary LA supplementation promoted muscle fibre transformation to type I fibre and that this process may be mediated through an AMPK‐dependent pathway.     

Effect of show jumping training on the development of locomotory muscle in young horses

OBJECTIVE: To investigate whether training for show jumping that is commenced early after birth affects the characteristics of equine locomotory muscle. ANIMALS: 19 Dutch Warmblood horses. PROCEDURES: Horses were assigned to a trained or not trained (control) group. After weaning, training (free jumping [2 d/wk] that was alternated with a 20-minute period of exercise in a mechanical rotating walker [3 d/wk]) was started and continued until horses were 3 years old. Fiber type composition (determined from myosin heavy chain [MyHC] content), fiber area, diffusion index (area supplied by 1 capillary), citrate synthase activity, and Na(+),K(+)-ATPase content were assessed in gluteus medius muscle specimens collected at 0.5, 1, 2, and 3 years. RESULTS: Developmental changes included an increase in MyHC fiber type IIa and a decrease in type IIad; increases in fiber area, diffusion index, and citrate synthase activity; and a decrease in Na(+),K(+)-ATPase content. The MyHC fiber type I and type IId were detected in high and low proportions, respectively. Training increased Na(+),K(+)-ATPase content, but did not affect other variables. CONCLUSIONS AND CLINICAL RELEVANCE: In horses, show jumping training at an early age resulted in increased Na(+),K(+)-ATPase content of the deep portions of the gluteus medius muscle. The lack of training effects on the other muscle characteristics can partly be explained by the fact that an appropriate (aerobic) fiber type composition was already established at training commencement. These data also suggested that the developmental changes in equine muscle represent sufficient adaptation to meet the demands of this specific training.