Early and 24 h post-mortem thigh (ilio tibialis) muscle metabolism and meat quality in two genetic types of turkeys and their reciprocal crosses,raised under semi-confined conditions

1. The experiment was conducted in order to determine the effect of the direction of turkey crossing on quality traits of their thigh muscle.

2. In total, 1358 turkeys of slow- (SG) and fast-growing (FG) lines as well as SF crosses (SG × FG) and FS crosses (FG × SG) were reared with access to free range to 21 weeks of age in the case of males and 15 weeks of age in the case of hens.

3. After slaughter of 15 males and 15 hens from each genetic line, their thigh muscles were examined and the following traits were measured after 5-min, 45-min, 2-h and 24-h post-mortem: temperature, pH, glycogen content (G), lactate content (L) and electrical conductivity (EC). Quality attributes were evaluated based on chemical composition, water holding capacity, cooking loss (%) and colour.

4. The rate of post-mortem changes in temperature, G, L and pH in muscles differed among the 4 lines of turkeys, with the highest metabolic rate determined for muscles of SG turkeys, followed by muscles of SF, FS and FG birds. A more beneficial muscle water holding capacity of both sexes of turkeys and better results of cooking loss in male muscles were found in SG and SF turkeys. The thigh muscles of the crosses were characterised by a higher content of protein and a lower content of fat compared to the muscles of FG turkeys, and in the case of the males also by a higher protein content compared to the muscles of SG birds.

5. Owing to the faster post-mortem metabolism, better water holding capacity, lower cooking loss and fat content at a similar content of protein, the thigh muscles of SF crosses raised in the free range system represent a better quality of meat for consumers compared to the thigh muscles of FS turkeys.     

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.     

Identification of Differentially Expressed Genes and Lipid Metabolism Signaling Pathways between Muscle and Fat Tissues in Broiler Chickens

In this study, signaling pathways and key differentially expressed genes (DEGs) involved in lipid metabolism in muscle and fat tissues were investigated. Muscle and abdominal fat tissues were obtained from 35-day-old female broilers for RNA sequencing. DEGs between muscle and fat tissues were identified. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses of DEGs were performed. A total of 6130 DEGs were identified to be significantly enriched in 365 GO terms, most of which were involved in biological processes, cellular components, and molecular functions in muscle and fat tissues. Three important lipid signaling pathways (pyruvate metabolism, the insulin signaling pathway, and the adipocytokine signaling pathway) were identified among the fat and muscle tissues of broilers. The key common DEGs in these pathways included phosphoenolpyruvate carboxykinase 2 (PCK2), acetyl-CoA carboxylase 1 alpha and beta (ACACA and ACACB), and the mitogen-activated protein kinase (AMPK) gene family. Hence, our findings revealed the pathways and key genes and gene families involved in the regulation of fat deposition in the muscle and fat tissues of broilers.     

m6A甲基化在鸡肌肉生长发育中的表达研究

试验旨在研究RNA m6A修饰相关基因去甲基化酶Alk B同源蛋白5（Alk B homologue 5，ALKBH5）、去甲基化酶肥胖相关蛋白（fat mass and obesity-associated protein，FTO）、甲基转移酶样蛋白3（methyltransferase like 3，METTL3）、甲基转移酶样蛋白14（methyltransferase like 14，METTL14）和成肾细胞瘤1-结合蛋白（Wilms’tumor 1-associating protein，WTAP）在鸡骨骼肌发育过程中的表达，分析其与骨骼肌m6A甲基化水平的相关性。首先，利用实时荧光定量PCR技术检测m6A甲基化相关基因在金茅花鸡12（E12）、14（E14）、16（E16）、18（E18）胚龄和1日龄腿肌和胸肌组织中mRNA表达水平，以及其在鸡成肌细胞50%、100%增殖期和1、2、3、4、5 d分化期的mRNA表达水平；随后，利用m6A甲基化试剂盒检测金茅花鸡E12和1日龄腿肌和胸肌组织中m6A甲基化修饰水平，与m6A甲基化相关基因表达水平进行相关性分析。结果显示，m6A去甲基化基因ALKBH5和FTO mRNA表达水平在骨骼肌发育过程中显著上调（P<0.05），即在E12、E14低表达，E16、E18逐渐上调，1日龄达到最高。m6A甲基化写入基因METTL14、METTL3和WTAP mRNA表达水平在E12、E14、E16逐渐上升，E18下降，随后至1日龄表达量回升。在细胞增殖过程中，ALKBH5、FTO、METTL14、METTL3和WTAP基因表达均上调；在细胞分化过程中ALKBH5和FTO基因表达水平显著上调（P<0.05），在分化第5天达到最高。METTL14、METTL3和WTAP基因mRNA表达水平在细胞诱导分化的1、2、3、4 d表达量呈下降趋势，而在诱导分化的第5天有所回升。甲基化水平检测结果显示，腿肌和胸肌m6A甲基化水平变化趋势一致，均在胚胎发育过程中显著下降（P<0.05），至1日龄达到最低。相关性分析结果显示，鸡骨骼肌RNA m6A甲基化水平与m6A去甲基化修饰基因ALKBH5、FTO mRNA表达水平呈显著负相关（P<0.05）。综合以上试验结果，推测m6A甲基化修饰与鸡骨骼肌发育相关，而去甲基化基因ALKBH5、FTO可能通过调控RNA m6A甲基化水平，影响鸡骨骼肌发育。本研究结果为进一步研究m6A甲基化修饰调控鸡骨骼肌生长发育的功能和分子机制提供理论依据。     

猪lncRNA TCONS_00791383对骨骼肌卫星细胞增殖分化的影响

为了探究lncRNA TCONS_00791383对猪骨骼肌卫星细胞增殖和分化的影响。本研究利用qRT-PCR技术检测出生7 d内大白仔猪6种组织（心、脾、肺、肾、背肌和腿肌）以及猪骨骼肌卫星细胞增殖分化前后TCONS_00791383的表达水平；通过设计反义核苷酸（antisense oligonucleotides，ASO）片段在猪骨骼肌卫星细胞中对TCONS_00791383进行敲低，检测敲低TCONS_00791383之后增殖分化标志基因的表达量变化；通过trans （co-expression）对TCONS_00791383进行靶基因预测，使用DAVID对其进行GO富集和KEGG通路分析。结果显示，TCONS_00791383在猪心脏中表达量最高，在脾和肾组织中不表达。在骨骼肌卫星细胞从增殖到分化的过程中，TCONS_00791383的表达量逐渐上升，且在分化后30 h表达量达到最高。在使用ASO片段敲低TCONS_00791383之后，与对照组相比，在分化24 h，增殖标志基因Pax3、Pax7表达量显著或极显著降低（P<0.05，P<0.01），分化标志基因MyoG表达量极显著降低（P<0.01），在分化48 h，增殖标志基因Pax3表达量极显著降低（P<0.01），Pax7表达量显著降低（P<0.05），分化标志基因MyHC表达量显著降低（P<0.05）。预测得到的相关靶基因富集到AMPK、ATP等多个与骨骼肌卫星细胞增殖和分化过程相关的重要信号通路。本研究表明，lncRNA TCONS_00791383可能促进猪骨骼肌卫星细胞的增殖和分化。     

表观遗传修饰在骨骼肌细胞增殖分化过程中的研究进展

骨骼肌是肌肉的主要构成部分，骨骼肌细胞发生增殖和分化的过程都是肌肉发育的基础，直接影响着家养动物的产肉性能。研究发现表观遗传修饰作用对骨骼肌细胞增殖分化具有重要的调控作用，表明该遗传修饰作用对家养动物肌肉发育具有重大的意义。作者从DNA甲基化对骨骼肌细胞增殖分化影响、组蛋白乙酰化所含因子调控基因选择表达作用、非编码RNA调控和染色体重塑作用所起的影响等方面分别介绍了表观遗传在骨骼肌细胞增殖分化过程中的研究进展，简述了不同修饰方式和不同作用因子对骨骼肌增殖和分化两个过程的影响。同时也回顾了前人在研究骨骼肌增殖分化过程所用到的方法和手段，进而分析了表观调控作用因子在骨骼肌生长过程中所起到的作用。旨在进一步阐述表观遗传修饰在骨骼肌增殖和分化过程中所起到的重要作用，增强对骨骼肌增殖分化调控过程的了解，为和动物生产实际相结合提供参考途径，同时也为骨骼肌生长发育等分子调控提供更多参考素材。     

The Blonde d’Aquitaine T3811>G3811 mutation in the myostatin gene: association with growth,carcass, and muscle phenotypes in veal calves

The mutation T3811 → G3811 (TG3811) discovered in the myostatin gene of the Blonde d’Aquitaine breed is suspected of contributing to the outstanding muscularity of this breed. An experiment was designed to estimate the effect of this mutation in an F2 and back-cross Blonde d’Aquitaine × Holstein population. By genotyping all known mutations in the myostatin gene, it was ensured that the TG3811 mutation was indeed the only known mutation segregating in this population. Fifty-six calves (43 F2, 13 back-cross) were intensively fattened and slaughtered at 24.0 ± 1.4 wk of age. The effects of the mutation were estimated by comparing the calves with the [T/T] (n = 18), [T/G] (n = 30), and [G/G] (n = 8) genotypes. Highly significant substitution effects (P < 0.001), above + 1.2 phenotypic SD, were shown on carcass yield and muscularity scores. Birth weight (P < 0.001) was positively affected by the mutation (+0.8 SD) but not growth rate (P = 0.97), while carcass length (P = 0.03), and fatness (P ≤ 0.03) were negatively affected (–0.5 to –0.7 SD). The characteristics of the Triceps brachii muscle were affected by the mutation (P < 0.001), with lower ICDH activity (oxidative) and a higher proportion of myosin type 2X muscle fibers (fast twitch). The effects of the TG3811 mutation were similar to those of other known myostatin mutations, although the Blonde d’Aquitaine animals, which are predominantly [G/G] homozygous, do not exhibit extreme double muscling.     

Evaluation of functional muscle anatomy scalability in the canine hind limb

In contrast to other mammals, the large variation in dog sizes is not accompanied by any significant genetic re-organization. In order to study the relationship between body mass, limb length and the functional anatomical muscle parameters of the canine hind limb, a large dataset comprising of muscle masses, optimal muscle fibre lengths and physiological cross-sectional area's (PCSA) were acquired for twenty-five muscles in ten dogs of sizes varying between 20 kg and 52 kg. The potential of body mass and limb length for reliably scaling individual muscle masses, optimal muscle fibre lengths and PCSA's were examined. For the majority of the muscles of the canine hind limb, neither body mass nor limb length were reliable scaling parameter for either muscle masses, PCSA's and optimal fibre length. These results indicate the need of a breed-specific approach to musculoskeletal modelling in future canine musculoskeletal research.     

过表达MyBPC1对牛骨骼肌卫星细胞增殖分化的影响

旨在探究肌球蛋白结合蛋白C1（myosin binding protein C1,MyBPC1）对牛骨骼肌卫星细胞增殖与成肌分化的影响,为进一步研究MyBPC1在细胞分化和肌肉发育过程中的调控作用提供依据。本研究利用西门塔尔胎牛原代牛骨骼肌卫星细胞体外诱导成肌分化模型模拟牛骨骼肌的生长发育过程。采用qRT-PCR和Western blot检测MyBPC1的细胞时序表达谱。试验分为两组。在RNA水平每组4个重复,每个重复20 μL;在蛋白水平每组3个重复,每个重复15 μg。采用qRT-PCR和Western blot检测牛骨骼肌卫星细胞转染MyBPC1的过表达效果,并进一步检测细胞增殖期标志因子Pax7、Ki67以及细胞分化期标志因子MyHC、MyOG的表达变化情况,观察牛骨骼肌卫星细胞肌管形成状态。结果,MyBPC1在牛骨骼肌卫星细胞分化前后表达水平存在极显著差异,牛骨骼肌卫星细胞诱导分化后MyBPC1的mRNA和蛋白表达量均极显著高于增殖期（P<0.01）。过表达MyBPC1后,细胞分化形成的肌管数量明显多于对照组,增殖标志因子Pax7的mRNA水平和蛋白表达水平无显著差异,分化标志因子MyHC的mRNA水平和蛋白表达水平极显著高于对照组（P<0.01）。过表达MyBPC1可以促进牛骨骼肌卫星细胞体外成肌分化,为进一步开展MyBPC1对牛骨骼肌卫星细胞的调控机制奠定基础。