骨骼肌生长发育过程及调控研究现状

骨骼肌是哺乳动物最大的组织,其功能或再生特性的丧失会导致生长发育不良及肌肉骨骼疾病。骨骼肌有600多块单独的肌肉,起支撑和运动的作用。骨骼肌可以被机体自主控制,这也是其不同于平滑肌和心肌的一个特点。农业动物的骨骼肌是肉产品的主要来源,为人类提供优质的动物蛋白和营养物质。影响生长发育的因素众多,肌肉的生长和发育决定着肉的产量和品质,是农业动物生产中极其重要的经济性状。骨骼肌的生长、发育调控涉及众多基因及其相关通路的激活或沉默,是一个极其复杂的多层次调控网络。作者综述了骨骼肌的结构、组成和生长发育过程,介绍了包括生长因子和细胞因子等在内的多种调节因子对肌肉发育的调控作用,并结合现有研究结果预测到未来发展方向将是重要候选基因的体外和体内试验验证,最终转化到实际生产应用中,为骨骼肌生长发育的分子调控机制研究和肌肉遗传疾病的治疗提供理论基础和参考依据。     

Role of serum myostatin during the lactation period

Myostatin, also known as GDF-8 (Growth/Differentiation Factor-8), is a member of the TGF-beta superfamily that negatively regulates skeletal muscle mass in mammals. Mutation of the myostatin gene in mice, cattle, and humans causes a massively developed skeletal muscle, characterized by muscle hypertrophy and hyperplasia. Although myostatin is predominantly expressed in skeletal muscle tissue, several recent studies have shown the presence of myostatin protein in blood and suggested a possible role for circulating myostatin in the regulation of skeletal muscle mass. In the present study, we examined changes in the levels of active form myostatin (13 kDa) in serum after birth by Western blot analysis to predict the role of serum myostatin in early postnatal muscle growth in the rat. Interestingly, the amount of active form myostatin in serum increased after birth and then decreased along with ageing after weaning. To clarify the role of increased serum myostatin during the postnatal period, we administrated follistatin, an inhibitor of myostatin activity, into postnatal rats intraperitoneally just after birth. Follistatin-administration during the postnatal period caused selective hypertrophy of type II muscle fibers in the soleus muscle. These results demonstrate that myostatin in serum acts on skeletal muscle and negatively regulates early postnatal muscle growth.     

Feeding old cats--an update on new nutritional therapies

Aging is associated with a wide variety of physiological changes that compromise the ability of the body to respond to stress or change, and ultimately contribute to morbidity and mortality. Much research has been done on the aging process in humans and experimental animals, and the impact of diet, but there is relatively little data from companion animal studies. However, although many of the aging changes are likely to be similar between different mammals, it is known that there are some differences with cats. In contrast to dogs and humans, elderly cats experience an increase in their maintenance energy requirements rather than a continued decline, which may be mediated partly through a better-maintained basal metabolic rate but also through progressive compromise in fat and protein digestion. Old age is also associated with a progressive loss of lean body mass that may be mitigated in part by feeding higher protein levels. Diets designed for healthy elderly cats should therefore concentrate on being energy dense, highly digestible and with an increase in the proportion of protein calories, and being fed with the aim of supporting optimal body weight. There is also strong evidence emerging that dietary manipulation in cats may be able to modify aspects of the aging process, with a long-term longitudinal study of cats demonstrating both survival and health benefits from feeding a diet supplemented with antioxidants, fatty acids, and a prebiotic source.     

Foetal development of skeletal muscle in bovines as a function of maternal nutrition,foetal sex and gestational age

To determine the effects of maternal nutrition on modifications of foetal development of the skeletal muscle and possible increase in the potential of skeletal muscle growth in cattle, gestating cows were either fed 190% NRC recommendations (overnourished; ON) or 100% NRC recommendation (control; CO). Interaction between maternal nutrition (MN) and the foetal sex (FS) was also investigated. Foetuses were necropsied at four different time points throughout gestation (139, 199, 241 and 268 days of gestation) to assess the mRNA expression of myogenic, adipogenic and fibrogenic markers in skeletal muscle. Phenotypic indicators of the development of skeletal muscle fibres, intramuscular lipogenesis and collagen development were also evaluated. Modifications in mRNA expression of skeletal muscle of foetuses were observed in function of MN and FS despite the lack of effect of MN and FS on foetal weight at necropsy. Maternal ON increased the mRNA expression of the myogenic marker Cadherin‐associated protein, beta 1 (CTNNB1) and adipogenic markers Peroxissome proliferator‐activated receptor gamma (PPARG) and Zinc finger protein 423 (ZNF423) at midgestation. However, no differences on foetal skeletal muscle development were observed between treatments at late gestation indicating that a compensatory development may have occurred on CO foetuses making the effect of MN on skeletal muscle development not significant at late gestation. Moreover, our data have shown an evidence of sexual dimorphism during foetal stage with a greater skeletal muscle development in male than in female foetuses. In conclusion, providing a higher nutritional level to pregnant cows changes the trajectory of the development of skeletal muscle during midgestation, but apparently does not change the potential of post‐natal growth of muscle mass of the offspring, as no differences in skeletal muscle development were observed in late gestation.     

畜禽骨骼肌肌纤维特性及其发育机制研究进展

骨骼肌是生物机体的重要组成部分，约占产肉动物机体的40%，与畜禽的运动、发育及产肉能力等重要性状密切相关。骨骼肌是由平行排列的肌原纤维组成的多核肌纤维，肌纤维的数量、类型以及转变方式直接反映了个体肌肉发育情况以及机体的生理状况，因而在选育工作中作为重要的经济性状而被关注。对肌纤维的深入研究将有助于加快今后的选育进程，促进对近年频发的畜禽肌源性疾病的溯源与研究，从而缓解肌源性疾病对畜禽养殖业造成的经济损失，满足消费者对优质肉产品的需求。近年来，对肌纤维的认识与研究均取得重大进展，包括依照肌纤维的结构与功能特性对骨骼肌的生物学功能进行预测与验证、肌纤维的类型与肉品质的关联分析以及模式动物的应用，挖掘出肌纤维发生发育以及再生修复过程中的重要调控分子，并对它们的功能进行注释。作者参照现有研究成果对肌纤维的结构与功能进行系统的描述，并从分子层面对肌纤维胚胎期的发生发育以及出生后的再生修复进行综述，以期为今后提高肉用动物的产肉性能及培育高品质产肉动物新品种或新品系的选育工作提供理论参考。     

非编码RNA对猪骨骼肌发育的影响

骨骼肌发育与猪肉产量和品质密切相关，且受到各种因素的影响。近年来，非编码RNA （non-coding RNA，ncRNA）对骨骼肌发育的影响已成为新的研究热点之一。ncRNA主要包括微小RNA （microRNA，miRNA）、长链非编码RNA （long non-coding RNA，lncRNA）和环状RNA （circular RNA，circRNA），是一类不具有编码蛋白功能的RNA，最初被认为只是在转录或转录后水平调控基因的表达，但随着研究的深入，越来越多的ncRNA被证实参与骨骼肌细胞增殖、分化与凋亡等生物过程，其中miRNA可通过与靶基因互补序列结合发挥功能；lncRNA与circRNA主要作为分子海绵竞争性结合miRNA，解除其对靶基因的抑制作用。作者主要从ncRNA介绍及miRNA、lncRNA和circRNA对猪骨骼肌发育的影响等进行综述，并就ncRNA对猪骨骼肌生长发育的研究进行了展望。     

Expression and location of IGF binding proteins-2, -4, and -5 in developing fetal tissues.

Insulin-like growth factors are associated with myogenesis in vivo, and their actions are mediated by IGF binding proteins (IGFBP). Sites of IGFBP production and their location during early development are not clear. The objective of this research was to examine the developmental expression and location of IGFBP-2, -4, and -5 mRNA and peptides in developing porcine skeletal muscle and liver. Pregnant pigs were euthanatized at various times postconception (pc). Developmental expression of IGFBP was evaluated using total RNA extracted from skeletal muscle and liver of 30-, 44-, 59-, 68-, 75-, 89-, and 109-d pc fetuses and from adult and neonatal pigs. Localization of IGFBP-2, -4, and -5 mRNA and peptides was examined by in situ hybridization and immunocytochemistry of muscle samples from contralateral pelvic limbs of each pig. Overall muscle IGFBP gene expression decreased (P < .05) with increasing age. Moreover, expression of liver IGFBP-2 and -5, but not of IGFBP-4, was greater (P < .05) during prenatal than during postnatal periods. The majority of immunoreactive IGFBP was located in developing muscle cells, with little localized to connective tissue, except at later stages of development. These data show that IGFBP-2, -4, and -5 expression is time- and tissue-dependent in fetal liver and muscle.     

肌抑素的研究进展

由于长期以来对优良肉品质的孜孜以求,人们发现了抑制肌原性细胞的增殖和分化,最终表现为肌肉组织和肌肉量减少的肌抑素。本文介绍了肌抑素基因的结构、不同动物肌抑素基因的差异、肌抑素基因表达、肌抑素在体内的分布以及不同时间的表达和肌抑素基因的生物学功能,并提出了其应用前景和可能研究的方向。     

猪骨骼肌卫星细胞体外分离培养研究进展

猪骨骼肌是动物机体重要的运动组织及人类主要的肉食来源,也是研究肌肉生长发育和疾病的良好模型。猪出生后,骨骼肌的生长发育、损伤修复都需要肌卫星细胞的参与,体外分离培养猪骨骼肌卫星细胞是深入研究骨骼肌生长发育及疾病发生机理的基础,是在细胞水平进行分子功能验证的前提。随着肌肉发育和病理分子机制研究的不断深入,猪骨骼肌卫星细胞的体外分离培养技术也迅速发展起来。背最长肌、后腿肌和半腱肌常用于分离骨骼肌卫星细胞,1日龄猪背最长肌的分离效果最好。常用于分离骨骼肌卫星细胞的酶包括链酶蛋白酶、胶原酶、胰蛋白酶、胶原蛋白酶等,各酶及酶联合消化的时间不同,最优的过滤方式是200目+400目联合过滤,3次离心法可获得纯度较高的细胞。常使用的培养基为DMEM/F12+10%胎牛血清(FBS)+1%青-链霉素(P/S)。骨骼肌肌卫星细胞常见标记物有配对盒基因3(PAX3)、PAX7、生肌决定因子5(Myf5)、Myf4、肌分化因子(MyoD)、肌细胞生成素(MyoG)等。作者通过对猪骨骼肌肌卫星细胞的分离、培养及鉴定等方面进行综述,梳理出各步骤中最佳参数,为建立规范猪骨骼肌卫星细胞分离程序提供参考,以期为肌肉发育和疾病研究提供理论及技术支持。     

microRNA-21和TGFBI基因在猪骨骼肌生长发育中的表达分析

试验旨在研究microRNA-21(miR-21)和转化生长因子β诱导(TGFBI)基因在长白猪骨骼肌中的表达相关性。采用实时荧光定量PCR(qPCR)方法分析了miR-21和TGFBI基因在长白成年猪中的组织表达谱,同时比较分析了其在长白猪不同发育阶段(出生前和出生后共28个发育点)背最长肌中的表达相关性。结果表明,miR-21在长白成年猪的各个组织中均表达,且分布相对平衡,在不同发育阶段的背最长肌中呈现波浪式的表达趋势;TGFBI基因在长白猪胚胎期的背最长肌中高表达,在整个背最长肌发育过程中呈现出不同的表达丰度。相关性分析表明,在胚胎期骨骼肌发育过程中,miR-21和TGFBI表达之间为显著的负相关,TGFBI可能是miR-21的调控靶标基因。     

长链非编码RNA(H19)在小鼠骨骼肌生长发育过程中的表达

本试验旨在研究长链非编码RNA(H19)在骨骼肌发育和损伤修复过程中表达量的变化,为研究H19在小鼠骨骼肌发育中的作用机制奠定基础。以C2C12细胞系和ICR小鼠为试验素材,通过生物信息学方法分析H19的非编码特性和在物种间的低保守性,通过实时荧光定量PCR(qRT-PCR)方法检测H19在C2C12细胞分化,ICR小鼠骨骼肌发育不同时期和成体小鼠骨骼肌损伤修复过程中表达量的动态变化。结果表明,H19在小鼠骨骼肌中的表达量随出生后日龄的增长逐渐下降;在诱导C2C12细胞分化过程中,H19的表达量先是逐渐升高,进而在较高水平维持不变;在肌肉损伤模型中修复的第4~6天,H19的表达量维持在较高水平。鉴于上述H19在C2C12细胞分化和肌损伤修复过程中的表达特点,推断H19可能具有促进肌纤维形成和成体骨骼肌损伤后修复的功能。     

去乙酰化酶3对线粒体功能和猪生长发育影响的研究进展

去乙酰化酶3（Sirt3）是去乙酰化酶家族的一员,具有高度的去乙酰基酶活性,对动物体的生长发育、衰老、代谢等生理过程具有调控作用。目前的研究表明,Sirt3可以激活超氧化物歧化酶、调控三羧酸循环和促进电子传递链3个途径,消除细胞内的活性氧（ROS）,避免细胞的氧化损伤和线粒体膜电位的损伤,进而影响细胞凋亡与自噬;并可通过调控ROS、降低各种退行性疾病的发病率等途径减缓衰老。同时,Sirt3通过激活AMP依赖的蛋白激酶（AMPK）等途径影响肌肉的发育。对猪的研究表明,不同Sirt3突变型猪的肌肉色值和失水率等肉质指标存在差异,且Sirt3对脂肪酸氧化和酮体生成有一定的影响。在对与猪Sirt3蛋白有较高同源性的牛进行的研究中发现,不同Sirt3突变型的牛体长、体高、肌肉脂肪含量等指标存在显著差异。作者总结了Sirt3对线粒体的能量代谢和细胞氧化损伤的调控机制及其对细胞凋亡、细胞自噬和机体衰老的影响,阐述了猪Sirt3蛋白的部分理化性质及其对猪生长发育的作用机制,以期为了解和挖掘该基因在猪肌肉和脂肪发育中的生物学作用,以及家畜生产性状改良提供理论依据。     

The Expression of Long Noncoding RNA (H19) in Mouse Skeletal Muscle Development

This experiment was intend to study the changes of long non-coding RNA (H19) expression levels in skeletal muscle development and regeneration,and lay the foundation of its mechanism reach in skeletal muscle development.C2C12 cell line and ICR mice were used as experimental material,bioinformatics assay was used to exploit its non-coding character and low conservatism in different species,and the expressions of H19 in C2C12 cell differentiation,skeletal muscle development and the phase of muscle regeneration were detected by quantitative Real-time PCR (qRT-PCR).The results showed that,H19 expression levels in postnatal mouse skeletal muscle decreased with increasing age;during C2C12 cell differentiation,H19 mRNA increased gradually,and then maintained a high level;The expression of H19 was maintained at a high level through days 4 to 6 after injury.In consideration of its express character in C2C12 cell differentiation and skeletal muscle damage repair model,H19 may play an important role in promoting myogenesis and skeletal muscle regeneration.     

Effects of eccentric exercise on branched‐chain amino acid profiles in rat serum and skeletal muscle

Supplementation of branched‐chain amino acid (BCAA) is often used to attenuate exercise‐induced skeletal muscle damage and promote adaptation, but no definitive conclusion on the benefits of BCAA on muscle recovery after injurious exercise can be drawn. Exploration of the systematic BCAA alteration in muscular injury‐repair stage per se without any BCAA supplement should provide some useful information in favour of BCAA application in muscle regeneration after injury. One bout of 90‐min downhill‐running exercise was performed to cause rat skeletal muscle injury. After exercise, myofibrillar BCAA concentrations showed minor changes compared with exercise before, while serum concentrations of BCAA were lower after exercise. Especially, serum leucine, isoleucine and total BCAA concentrations 2 weeks post‐run were significantly lower than normal values of exercise before (p = 0.008, p = 0.041, p = 0.015). The data demonstrate that a single eccentric exercise can significantly decrease the serum BCAA concentrations, which mean high utilization of BCAA for myogenesis after injurious exercise.     

Spontaneous extraskeletal osteosarcoma in the neck skeletal muscle of a Crlj:CD1 (ICR) mouse

Extraskeletal osteosarcoma is extremely rare in humans and animals, especially in rodents. This is the first case report on spontaneous extraskeletal osteosarcoma in the neck skeletal muscle of a Crlj:CD1 (ICR) mouse (36 weeks, dead). Necropsy revealed a solid white mass located in the neck skeletal muscle (scalenus muscle). Histological examination showed that the tumor consisted of atypical polygonal cells, a small osteoid clump, and bone tissue. Mitotic figures were observed. Serial sections showed that neoplastic cells lacked clear invasive proliferation to adjacent normal skeletal muscle and continuity with normal bone tissue. Immunohistochemical analysis showed that the neoplastic cells were positive for osteocalcin, osterix, vimentin, and S-100. Based on these results, the tumor was diagnosed as extraskeletal osteosarcoma in the neck skeletal muscle.     

Identification of immunohistochemical localization of irisin in the dwarf hamster (Phodopus roborovskii) tissues

Irisin is mainly secreted by heart and skeletal muscle cells. It is an exercise‐induced protein that converts white adipose tissue to brown. Increased irisin expression was lead to weight loss and improved glucose tolerance. We investigated irisin immunoreactivity in various tissues of the dwarf hamsters (Phodopus roborovskii). Tissues were processed, embedded in paraffin, sectioned at 5 μm and stained immunohistochemically for irisin. In the retina, irisin was found almost all layers, except outer nuclear layer. Also, irisin immunoreactivity was observed in the skin, cornea, striated muscle, parotid gland, tongue, oesophagus, stomach and small intestine. The findings from this study support the notion that skeletal muscle is not the primary source of irisin.