肌生成抑制素基因研究进展

肌生成抑制素是 1 997年发现的骨骼肌生长发育负调控因子 ,肌生成抑制素不仅在骨骼肌中表达 ,还可在心肌和浦肯野纤维等多种组织中表达。研究表明 ,大鼠骨骼肌在体内的再生 ,对 MSTN m RNA的表达表现出明显的时间依赖性 ,并且 m RNA的表达不必由功能性神经支配。通过对不同品种双肌表型牛 MSTN基因的研究 ,发现其骨胳肌增大的共同特点是肌生成抑制素基因发生了突变 ,如布鲁牛 MSTN基因在编码区外显子 3发生 1 1 bp的缺失 ,导致MSTN此位点后的阅读框架改变 ,并在此点后的 1 4个密码子处终止了阅读框架 ,从而使MSTN被截短 ,MSTN蛋白活性区域消失 ,活性丧失 ,结果肌肉大量增加。目前 ,已分别对人、牛和猪的 MSTN基因进行了定位研究 ,猪肌生成抑制素基因定位研究表明 ,猪 MSTN基因位于1 5号染色体上。     

Growth factors controlling muscle development.

The enlarged muscles of certain breeds of cattle, such as the Belgian Blue, have been shown to result from a marked increase in the number of normal sized muscle fibers. Originally insulin-like growth factors (IGFs) were implicated in this myofiber hyperplasia, as IGFs have been shown to stimulate myoblast proliferation as well as maintain fiber differentiation. Recently it has been reported that mice lacking a myostatin gene, a member of the TGFbeta superfamily, have enhanced skeletal mass resulting from increased muscle fiber number and size. Mutations in this gene have been found in double-muscled cattle, indicating that myostatin is an inhibitor of muscle growth. Myostatin is expressed early in gestation and then maintained to adulthood in certain muscles. Myostatin expression in bovine muscle is highest during gestation when muscle fibers are forming and some of the myogenic regulatory factors have elevated expression over the same period as myostatin. Molecular expression of the IGF axis does not differ between Belgian Blue and normal muscled cattle, and IGF-II mRNA is increased throughout formation of secondary fibers in both breeds. However, myostatin and MyoD expression in muscle differ between normal and hypertrophied muscle cattle breeds. This evidence strongly suggests that lack of myostatin is associated with an increase in fiber number which then results in a marked increase in potential muscle mass in double-muscled cattle.     

肌肉抑制素与“双肌”表型关系的研究进展

肌肉抑制素是特异性表达于骨骼肌的一种细胞因子,属于TGF-β超家族成员,可以抑制骨骼肌的发育,并同多种疾病有关。肌肉抑制素在不同物种间高度保守,其抑制功能的丧失可以导致骨骼肌显著增生,即动物产生"双肌"表型。具有"双肌"表型的动物则表现出肌肉产量显著增加、饲料转化率高、肌肉嫩度增加、脂肪含量少、而脂肪酸组成中不饱和脂肪酸比例显著增加等优秀性状,这些性状是动物育种计划中的理想育种目标。本文从"双肌"表型的起源及特点、肌肉抑制素的发现及功能等方面对肌肉抑制素与"双肌"表型关系的研究进展进行了综述。     

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。     

动物肌肉生长发育调控的功能基因研究进展

成肌细胞的增殖和分化是受生肌决定因子 (MyoD)调控 ,生肌决定因子包括 4个基因 ,MyoDl(myf3)、Myogenin(MyoG)、Myf5、Myf 6 (herculin或MRF4 )。MyoD家族基因属于碱性螺旋 环 螺旋 (bHLH)转录因子 ,它能激活肌肉生成的特定基因。肌细胞生长抑制素 (MSTN ,Myostatin ,又称GDF 8) ,属于转化生长因子超家族 ,它通过负向控制肌细胞的生长发育 ,GDF 8基因缺失的小鼠表现出比正常小鼠具有“双肌现象”。     

Expression profile of myostatin mRNA during the embryonic organogenesis of domestic chicken (Gallus gallus domesticus)

Myostatin is a potent growth and differentiation factor involved in skeletal muscle tissue formation in vertebrates. However, recent studies in chicken embryo suggested that the myostatin was expressed even before the establishment of myogenic lineage. No studies have thus far been reported in birds to define the role of myostatin during the embryonic organogenesis. The present experiment was designed for studying the expression profiles of myostatin mRNA in the chicken liver, heart, brain, and intestine during their morphogenesis, using real-time PCR. The myostatin mRNA expression was significantly upregulated in liver during E15-E18. Similar results were observed during the development of chicken heart. In brain, the expression of myostatin was upregulated from E4 onwards. In intestine, the expression of myostatin was significantly increased many folds on E9-E18. Therefore, the increase in myostatin expression might be related to the growth of liver and heart on days E12-E18; morphogenesis and growth of brain during E15-E18; and morphogenesis and differentiation of intestine during E9-E18. In the present study, the tissue-specific expression of myostatin gene in chicken is similar to fishes, but different from that in mammals. Further, the inspection of chicken genome also suggested that there is no differentiation of GDF-8 and -11. A recent finding suggests that the chicken myostatin gene is closely related to mammals than fishes. Therefore, we propose that the chicken myostatin gene might have diverged in its function between teleosts and mammals. Indeed it is possible that its function might have only become fully differentiated to serve as a control of muscle mass in mammals.     

MSTN基因的研究进展

肌生成抑素(MSTN)是1997年发现的一种新的生长因子,其基因产物对骨骼肌的生长具有负调控作用,该基因缺失会导致骨骼肌增生.本文综述了MSTN基因的结构、在畜禽中的表达情况、作用机理以及研究意义.     

Effects of birth weight and postnatal nutrition on neonatal sheep: II. Skeletal muscle growth and development

This study investigated effects of birth weight and postnatal nutrition on growth and development of skeletal muscles in neonatal lambs. Low (L; mean +/- SD 2.289 +/- .341 kg, n = 28) and high (H; 4.840 +/- .446 kg, n = 20) birth weight male Suffolk x (Finnsheep x Dorset) lambs were individually reared on a liquid diet to grow rapidly (ad libitum fed, ADG 337 g, n = 20) or slowly (ADG 150 g, n = 20) from birth to live weights (LW) up to approximately 20 kg. At birth, weight of semitendinosus (ST) muscle in L lambs was 43% that in H lambs; aggregate weights of ST and seven other dissected muscles were similarly reduced. In ST muscle of L lambs, mass of DNA, RNA, and protein were also significantly reduced to levels 67, 60, and 34%, respectively, of those in H lambs. However, myofiber numbers of ST, tibialis caudalis, or soleus muscles did not differ between the L and H birth weight lambs and did not change during postnatal growth. During postnatal rearing, daily accretion rate of dissected muscle was lower in L than in H lambs. Accretion of muscle per kilogram of gain in empty body weight (EBW) was reduced in the slowly grown L lambs compared with their H counterparts, although the difference was less pronounced between the rapidly grown L and H lambs. Throughout the postnatal growth period, ST muscle of L lambs contained less DNA with a higher protein:DNA ratio at any given muscle weight than that of H lambs. Slowly grown lambs had heavier muscles at any given EBW than rapidly grown lambs. Content of DNA and protein:DNA ratio in ST muscle were unaffected by postnatal nutrition, but RNA content and RNA:DNA were greater and protein:RNA was lower at any given muscle weight in rapidly grown lambs. Results suggest that myofiber number in fetal sheep muscles is established before the presumed, negative effects of inadequate fetal nutrient supply on skeletal muscle growth and development become apparent. However, proliferation of myonuclei may be influenced by fetal nutrition in late pregnancy. Reduced myonuclei number in severely growth-retarded newborn lambs may limit the capacity for postnatal growth of skeletal muscles.     

Analysis of myostatin and its related factors in various porcine tissues

Myostatin is expressed in skeletal muscle tissue where it functions to suppress myoblast proliferation and myofiber hypertrophy. Recently, myostatin was detected in the tendon, mammary gland, and adipose tissue of mice. We sought to determine whether myostatin is expressed in the liver, spleen, lung, and kidney of pigs. Real-time PCR and Western blots demonstrated that myostatin, follistatin, decorin, and activin receptor IIB (ActRIIB) mRNA and proteins were expressed in skeletal muscle, heart muscle, and adipose tissue, and also in liver, spleen, lung, kidney, and cultured fibroblasts. The relative abundance of myostatin was closely related to follistatin and decorin in porcine tissues. Immunohistochemical analysis further demonstrated the presence of myostatin, follistatin, and decorin in the skeletal muscle, adipose tissue, heart muscle, liver, spleen, lung, and kidney of pigs. These results suggest that myostatin could be associated with certain functions of the internal organs, such as energy metabolism or fibrosis. We conclude that myostatin is a factor broadly expressed in the internal organs and muscle tissues of pigs.     

Temporal expression of transforming growth factor-beta2 and myostatin mRNA during embryonic myogenesis in Indian broilers

TGF-beta2 and myostatin, the members of TGF family, act through both autocrine and paracrine mechanisms to regulate the growth and differentiation at various developmental stages in chicken. The kinetics and expression profile of these two growth factors were investigated by semi-quantitative RT-PCR, during the myogenesis of Indian broiler chickens. Total RNA was isolated from whole embryos on each of embryonic days (E) 0-6 (n=3 per day) and from the biceps femoris muscle at E7-E18 (n=3 per day). The expression of TGF-beta2 was noticed on E2 that remained at the same level until E6. In biceps femoris muscle, higher level of TGF-beta2 expression was observed during E7-E12, which decreased gradually thereafter. These findings suggested that TGF-beta2 might be a regulatory factor participating in the myogenesis of chicken embryos. Initial myostatin expression was noticed on E1, even before the myogenic lineage is established in embryo. This finding suggested an additional role of myostatin in early chicken embryo development, other than myogenesis. Furthermore, myostatin expression was significantly higher on E3 as compared to earlier studies, where initial higher level was observed at E2, suggesting the differential expression of myostatin among breeds. Higher and almost static myostatin expression was noticed in biceps femoris muscle during the entire period of myogenesis (E7-E18). In the present study, the ontogeny of myostatin expression coincided with myogenesis of chicken. Therefore, it may be hypothesized that myostatin is not only a major determinant of muscle mass, but also involved in early embryogenesis in chickens.     

Laminin binds to myostatin and attenuates its signaling

Myostatin is a growth and differentiation factor and acts as a negative regulator of skeletal muscle mass. Although the mechanism whereby myostatin controls muscle cell growth is mostly clarified, the regulation of myostatin activity after its secretion into the extracellular matrix (ECM) is still unclear. In the present study, we investigated the interaction between laminin and myostatin and the effect of laminin on myostatin signaling in vitro. The surface plasmon resonance assay showed that laminin bound to mature myostatin and activin receptor type IIB (ActRIIB), but did not bind to latency‐associated protein, which remains non‐covalently linked to mature myostatin. Furthermore, kinetic analysis demonstrated that the affinity of mature myostatin for laminin was similar to that for ActRIIB. Next, we examined the action of laminin on the myostatin signaling pathway using a conventional reporter assay. The luciferase activity of myostatin‐treated cells was repressed significantly (P < 0.05) by coincubation of laminin. These results suggest that laminin has a potential to regulate myostatin activity through binding to mature myostatin and/or its receptor ActRIIB.     

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.     

Myostatin expression and possible functions in animal muscle growth

Myostatin (also known as growth/differentiation factor-8) is a recently identified member of the transforming growth factor-β family of secreted regulatory factors. Mice having targeted disruption of the myostatin gene displayed a marked increase in muscle mass, up to three times normal size. Additionally, a myostatin mutation has been linked to double muscled cattle breeds characterized by a visible, generalized increase in muscle mass. Therefore, it is suggested that myostatin in muscle may be one of the long sought inhibitors that specifically control the growth of individual tissues or organs. In the present paper, we review involvement of myostatin in muscle growth of different species.     

cDNA encoding sequences for myostatin and FGF6 in sea bass (Dicentrarchus labrax, L.) and the effect of fasting and refeeding on their abundance levels

Fish have the ability to compensate for set-backs in growth as a result of fasting. When food levels are restored, growth in these fish can increase over and above normal rates. This phenomenon, known as “compensatory growth”, has been studied with respect to enhancing food conversion efficiency. However, the mechanisms by which food intake activates an increase in somatic growth, and especially in muscle growth, are not well understood. In this study, we report first on the isolation of two complete cDNAs sequences encoding sea bass (Dicentrarchus labrax) myostatin and fibroblast growth factor 6 (FGF6), which have been shown to be major genetic determinants of skeletal muscle growth. The open reading frames of myostatin (376 amino acids) and FGF6 (209 amino acids) showed 97–63% and 87–62% sequence identity with other vertebrate myostatins and FGF6s, respectively. We also report on the expression profile of myostatin and FGF6 in sea bass skeletal muscle in response to different feeding regimens, as quantified by real-time RT-PCR. Nutritional status significantly influenced the myostatin expression levels in muscle, inducing an up-regulation during fasting and a down-regulation during the recovery from fasting, whereas the muscular FGF6 mRNA levels were not significantly affected by the feeding status of the animals. These findings suggest that myostatin has an inhibitory role in muscle growth in response to different feeding regimens, whereas FGF6 is not involved in the muscle compensatory growth induced by refeeding.     

Decreased colostral immunoglobulin absorption in calves with postnatal respiratory acidosis

The effect of postnatal acid-base status on the absorption of colostral immunoglobulins by calves was examined in 2 field studies. In study 1, blood pH at 2 and 4 hours after birth was related to serum IgG1 concentration 12 hours after colostrum feeding (P less than 0.05). Decreased IgG1 absorption from colostrum was associated with respiratory, rather than metabolic, acidosis, because blood PCO2 at 2 and 4 hours after birth was negatively related to IgG1 absorption (P less than 0.05), whereas serum bicarbonate concentration was not significantly related to IgG1 absorption. Acidosis was frequently observed in the 30 calves of study 1. At birth, all calves had venous PCO2 value greater than or equal to 60 mm of Hg, 20 of the calves had blood pH less than 7.20, and 8 of the calves had blood bicarbonate concentration less than 24 mEq/L. Blood pH values were considerably improved by 4 hours after birth; only 7 calves had blood pH values less than 7.20. Calves lacking risk factors for acidosis were examined in study 2, and blood pH values at 4 hours after birth ranged from 7.25 to 7.39. Blood pH was unrelated to IgG1 absorption in the calves of study 2. However, blood PCO2 was again found to be negatively related to colostral IgG1 absorption (P less than 0.005). Results indicate that postnatal respiratory acidosis in calves can adversely affect colostral immunoglobulin absorption, despite adequate colostrum intake early in the absorptive period.     

Differential ammonia metabolism and toxicity between avian and mammalian species,and effect of ammonia on skeletal muscle: A comparative review

Comparative aspects of ammonia toxicity, specific to liver and skeletal muscle and skeletal muscle metabolism between avian and mammalian species are discussed in the context of models for liver disease and subsequent skeletal muscle wasting. The purpose of this review is to present species differences in ammonia metabolism and to specifically highlight observed differences in skeletal muscle response to excess ammonia in avian species. Ammonia, which is produced during protein catabolism and is an essential component of nucleic acid and protein biosynthesis, is detoxified mainly in the liver. While the liver is consistent as the main organ responsible for ammonia detoxification, there are evolutionary differences in ammonia metabolism and nitrogen excretory products between avian and mammalian species. In patients with liver disease and all mammalian models, inadequate ammonia detoxification and successive increased circulating ammonia concentration, termed hyperammonemia, leads to severe skeletal muscle atrophy, increased apoptosis and reduced protein synthesis, altogether having deleterious effects on muscle size and strength. Previously, an avian embryonic model, designed to determine the effects of increased circulating ammonia on muscle development, revealed that ammonia elicits a positive myogenic response. Specifically, induced hyperammonemia in avian embryos resulted in a reduction in myostatin, a well‐known inhibitor of muscle growth, expression, whereas myostatin expression is significantly increased in mammalian models of hyperammonemia. These interesting findings imply that species differences in ammonia metabolism allow avians to utilize ammonia for growth. Understanding the intrinsic physiological mechanisms that allow for ammonia to be utilized for growth has potential to reveal novel approaches to muscle growth in avian species and will provide new targets for preventing muscle degeneration in mammalian species.