皖南花猪骨骼肌AdpR和MyHCmRNA表达的发育性变化

摘要：为探讨皖南花猪骨骼肌组织中脂联素受体（AdpRl、AdpR2）和不同类型肌球蛋白重链（MyHC）mRNA的发育性变化及性别差异,选择0（出生当天）、30、45、90、180日龄的皖南花猪公母各5头,以B～Actin为内标,采用△△Ct相对定量实时荧光PCR方法对背最长肌和半腱肌中AdpRl、AdpR2、MyHCI、MyHC2a、MyHC2b和My-HC2xmRNA进行定量分析。结果显示,背最长肌和半腱肌AdpRl、AdpR2、MyHCl、MyHC2a、MyHC2b和My—HC2XmRNA的表达都有显著或极显著的发育变化规律（P〈0．05或P〈0．01）。总体上AdpRl、AdpR2、My—HC2a、MyHC2b和MyHC2XmRNA在背最长肌显著或极显著高于半腱肌（P〈0．05或P〈0．01）;MyHClmRNA在背最长肌极显著低于半腱肌（P〈0．01）。半腱肌MyHClmRNA在母猪显著大于公猪（P〈0．05）,而MyHC2amRNA在母猪显著小于公猪（P〈0．05）。背最长肌和半腱肌AdpR2mRNA的表达分别与MyHCl正相关（P〈0．05）,半腱肌AdpRlmRNA的表达与MyHC2x正相关（P〈0．05）。结果表明,皖南花猪骨骼肌组织中AdpR和MyHC的基因表达有特定的发育模式和组织特异性,且有一定性别差异。     

Cold exposure increases slow‐type myosin heavy chain 1 (MyHC1) composition of soleus muscle in rats

The aim of this study was to examine the effects of cold exposure on rat skeletal muscle fiber type, according to myosin heavy chain (MyHC) isoform and metabolism‐related factors. Male Wistar rats (7 weeks old) were housed individually at 4 ± 2°C as a cold‐exposed group or at room temperature (22 ± 2°C) as a control group for 4 weeks. We found that cold exposure significantly increased the slow‐type MyHC1 content in the soleus muscle (a typical slow‐type fiber), while the intermediate‐type MyHC2A content was significantly decreased. In contrast to soleus, MyHC composition of extensor digitorum longus (EDL, a typical fast‐type fiber) and gastrocnemius (a mix of slow‐type and fast‐type fibers) muscle did not change from cold exposure. Cold exposure increased mRNA expression of mitochondrial uncoupling protein 3 (UCP3) in both the soleus and EDL. Cold exposure also increased mRNA expression of myoglobin, peroxisome proliferator‐activated receptor gamma coactivator 1α (PGC1α) and forkhead box O1 (FOXO1) in the soleus. Upregulation of UCP3 and PGC1α proteins were observed with Western blotting in the gastrocnemius. Thus, cold exposure increased metabolism‐related factors in all muscle types that were tested, but MyHC isoforms changed only in the soleus.     

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.     

干扰DHX9对牛骨骼肌卫星细胞增殖与成肌分化的影响

为研究天冬氨酸-谷氨酸-丙氨酸-组氨酸盒解旋酶9 (DEAH (Asp-Glu-Ala-His)-box helicase 9,DHX9)对牛骨骼肌细胞增殖与分化的影响,利用已经建立的牛骨骼肌卫星细胞体外成肌分化模型,设计合成DHX9的si-RNA,采用荧光定量PCR和Western blot技术检测DHX9基因在成肌...     

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.     

Dietary thymol supplementation promotes skeletal muscle fibre type switch in longissimus dorsi of finishing pigs

As one of the key points related to meat quality, skeletal muscle fibre type is determined by energy metabolism and genetic factors, but its transformation could be also greatly influenced by many factors. Thymol, the primary effective ingredients of thyme, is well known for its anti-oxidation and anti-inflammatory, while little is known about its effect on skeletal muscle oxidative metabolism and fibre type switch. Therefore, in order to investigate its effects and possibility to be applied in livestock production, 36 150-day-old fattening Pigs were fed with different diet for six-week experiment. As a result, the drip loss ratio of longissimus dorsi (LD) was significantly reduced (p < .05). Oxidative metabolism-related enzyme activity, the mRNA levels and protein expression of COX5B and PGC1α, mRNA level of myosin heavy chain I (MyHC I) and protein level of MyHC IIa were significantly upregulated (p < .05). While compared with control group, the protein expression of MyHC IIb was significantly decreased (p < .05). The result revealed that thymol could promote the oxidative metabolism in the muscle of pigs and improve the meat quality to a certain extent.     

Age-Related Changes in Myosin Light and Heavy Chain Isoforms’ Composition in Gluteus Medius Muscle of Sedentary Akhal-Teke Horses

The primary purpose of the present study was to examine age dependence of gluteus medius muscle phenotypic plasticity with regard to myosin light chain (MyLC) and myosin heavy chain (MyHC) isoform expression of sedentary Akhal-Teke horses. A total of 23 Akhal-Teke horses (11 horses, 1.5 to 2.5-year-old; nine horses, 9 to 20-year-old; and three horses, 21 to 23.5-year-old) were studied. A muscle biopsy of depth 60 mm from the left gluteus medius muscle was obtained. MyHC and MyLC isoform pattern was determined in three age groups. The current study demonstrated that in the 21 to 23.5-year-old age group, the relative content of MyHC I isoform and MyLC 1_slow isoform decreased significantly. The results of the current study indicate that gluteus medius muscle of Akhal-Teke horses consists of a relatively high percentage of MyHC I and IIa isoforms, which together compose about 50% of MyHC. MyHC IIa and IIx isoforms together compose 88% to 94% of MyHC isoforms. Age-related decrease of the relative content of MyHC I and MyLC 1_slow isoforms in gluteus medius muscle starts in Akhal-Teke horses older than 20 years.     

Atypical masticatory muscle myositis in three cavalier King Charles spaniel littermates

This case report describes a novel manifestation of the immune-mediated disease, masticatory muscle myositis. Clinical signs, including difficulty in opening the mouth (trismus), were seen in three of four 12-week-old cavalier King Charles spaniel littermates. Diagnosis was established by 2M immunohistochemistry, supported by characteristic histopathological changes in affected temporal muscle. Treatment using corticosteroids at immune-modifying doses resulted in resolution of clinical signs in all the affected animals. Masticatory muscle myositis should be considered as a differential diagnosis in groups of young dogs with clinical signs of myositis localised to the head.     

Muscle type‐specific effect of myostatin deficiency on myogenic regulatory factor expression in adult double‐muscled Japanese Shorthorn cattle

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 < 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 < 0.05). Myf5 and MRF4 expression in semitendinosus (ST) was higher in the DM than in the NM cattle (P < 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 < 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.     

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。     

Differential response of cull cow muscles to the hypertrophic actions of ractopamine-hydrogen chloride

Ractopamine-HCl (RAC) is a beta-adrenergic agonist with variable effects on cattle performance and carcass variables. Cull cows fed RAC (200 mg . head(-1) . d(-1)) demonstrate an increased size of type I and II muscle fibers that does not translate into a larger ribeye area. The objective of this study was to examine the dose-dependent effects of RAC on cull cow muscle morphometrics. Eighty-eight cull beef cows representing 2 breed types (n = 44 each) were fed 0, 100, 200, and 300 mg . head(-1) . d(-1) of RAC for the last 28 d of a 54-d feeding period. On d 54, cows were slaughtered, and samples of the LM and semimembranosus muscle (SM) from 16 randomly selected carcasses (n = 4 per treatment) were taken for measurement of beta (2)-adrenergic receptors and type I, IIA, and IIX myosin heavy chain (MyHC) gene expression. Twenty-four hours postmortem, LM, SM, infraspinatus (INF), and vastus lateralis samples from 40 randomly selected carcasses (n = 10 per treatment) were obtained and frozen for immunohistochemical analysis. Muscle fiber cross-sectional area and diameter, MyHC isoform expression, and fiber-associated nuclei numbers were measured. Ractopamine dosage exhibited differential effects on muscle morphometrics and MyHC gene expression. Muscle fiber cross-sectional area and diameter were increased (P < 0.05) by RAC in INF type I and IIA fibers and SM type IIA fibers. Ractopamine increased (P < 0.05) MyHC type IIX mRNA and tended to increase (P < 0.10) beta(2)-adrenergic receptors in the SM; a change in mRNA abundance was not detected for either gene in the LM. Treatment with RAC decreased (P < 0.05) fiber-associated nuclei numbers in the INF, vastus lateralis, and LM but did not affect (P > 0.05) MyHC or beta-adrenergic receptor expression. These results indicate that cull cow feeding programs may consider supplementing RAC as a means of adding value to cuts within the chuck, such as the INF.     

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.     

饲粮能氨水平对不同品种猪肉质性状及相关基因表达的影响

本试验旨在研究饲粮能氮水平对不同品种猪肉质性状以及肌纤维类型和细胞因子相关基因表达的影响.选取巴马香猪和长白猪共96头(48头/品种),随机分为4组(每组24头猪),分别为长白猪饲喂NRC饲粮组、长白猪饲喂我国地方猪标准(GB)饲粮组、巴马香猪饲喂NRC饲粮组、巴马香猪饲喂GB饲粮组.试验从5周龄开始,至出栏时结束.分别于保育期、生长期和肥育期末,每组选择8头体况相近猪只屠宰,采集肌肉样品分析.结果表明:生长阶段显著影响了试验猪背最长肌熟肉率和肌球蛋白重链(MyHC)Ⅱx、MyHCⅡ易及白细胞介素-15(IL-15)mRNA表达水平(P<0.05),且随生长阶段呈增加趋势;与长白猪相比,巴马香猪的背最长肌熟肉率、MyHC Ⅰ mRNA表达水平显著升高(P<0.05),而滴水损失显著降低(P<0.05);与NRC饲粮相比,GB饲粮显著提高了生长期长白猪和巴马香猪的pH45 min(P<0.05),显著上调了保育期和肥育期猪背最长肌MyHC Ⅰ mRNA表达水平(P<0.05);与GB饲粮相比,NRC饲粮显著上调了生长期猪背最长肌MyHC Ⅰ mRNA表达水平(P<0.05).由此可见,品种和生长阶段均可显著影响肉质性状及相关基因的表达,饲粮能氮水平则主要通过与生长阶段产生互作效应影响肉质性状及肌纤维类型相关基因的表达.     

Relationships between tropomyosin and myosin heavy chain isoforms in bovine skeletal muscle

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.     

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.