猪背最长肌中肌肉生长和脂肪沉积相关基因的差异表达和主成分分析

骨骼肌细胞和脂肪细胞在分化生长速度上相对竞争的平衡点是猪肉质和胴体性状的决定因素.利用Oligo功能分类芯片检测了瘦肉型的长白猪和脂肪型的太湖猪在初生、1、2、3、4和5月龄间背最长肌中肌肉生长和脂肪沉积相关基因的动态表达变化.差异表达分析结果显示,在初生至5月龄的品种间分别有15、16、11、13、18和20个基因的表达差异倍数大于2倍.品种内的方差分析表明,长白猪分别有18和22个基因,太湖猪分别有3和7个基因在月龄间的表达差异达极显著(Ｐ<0.01)和显著水平(Ｐ<0.05).主成分分析结果显示,先降后升是两品种内最具代表性的基因表达模式,且长白猪和太湖猪分别有7和6个基因的表达模式明显偏离其他基因,提示其可能受到了重要的调控. 此外,5个差异表达基因的荧光定量RT-PCR验证结果均与芯片结果呈正相关趋势.以上结果筛选出了对于猪肉质和胴体性状可能具有重要影响,值得深入研究的一些候选基因,为深入研究生长发育过程中参与肌纤维生长和脂肪酸合成关键基因的表达变化规律和互作调控机制提供了基础数据.     

长白猪和太湖猪不同生长阶段脂肪中肉质和胴体性状相关基因的表达谱分析

动物个体的皮下脂肪厚度（subcutaneous fat thickness,SFT）和肌内脂肪（intramuscular fat,IMF）含量取决于活体内脂肪酸合成与氧化竞争的动态平衡点．利用包含140个与猪脂肪沉积、代谢和肌肉生长密切相关基因的Oligo功能分类基因芯片检测了脂肪型的太湖猪和瘦肉型的长白猪在1,2,3,4和5月龄间背部皮下脂肪中这些基因的动态表达变化,发现长白猪有25个基因在月龄间的表达差异达显著水平（P〈0．05）,且包含23个基因的“参与脂肪或类固醇代谢的酶和调节蛋白”基因分组在品种间具有极显著意义（P〈0．01）．提示两猪种脂肪沉积能力的巨大表型差异可能与这些基因的差异表达规律密切相关．STEM聚类分析显示长白猪和太湖猪各有两个表达模式分别具有极显著（P〈0．01）和显著性意义（P〈0．05）,但太湖猪基因表达谱比较分散,占主导优势的表达模式没有长白猪明显．提示太湖猪脂肪细胞内参与相关代谢活动的基因间的调控关系较长白猪复杂．基于动态贝叶斯模型构建的基因调控网络从一定程度上反映了两猪种在脂肪沉积代谢相关生理生化活动方面存在的明显差异,可从中挖掘相关性状潜在的关键特征基因．此外,对5个差异表达基因的荧光定量RT-PCR验证显示两种实验方法结果的相关系数平均高达0．874±0．071．以上结果筛选出了对于猪脂肪相关性状可能具有重要影响并值得深入研究的一些基因,初步揭示了生长发育过程中脂肪细胞基因表达调控的分子互作机制．     

放牧藏猪、舍饲藏猪与商品猪粪便真菌群落组成及其与饲粮纤维消化的相关性研究

[目的]研究饲养在西藏高原的放牧藏猪、舍饲藏猪和商品猪(杜长大猪,DLY猪)粪便中真菌群落组成的差异性,获取与饲粮粗纤维消化相关的真菌群落.[方法]以饲养在西藏高原的5月龄放牧藏猪、舍饲藏猪和DLY猪为研究对象,采用消化试验测定放牧藏猪、舍饲藏猪和DLY猪对饲粮粗纤维的表观消化率.采集粪便样品利用单分子实时测序技术(S...     

Expression levels of candidate genes for intramuscular fat deposition in two Banna mini-pig inbred lines divergently selected for fatness traits

Intramuscular fat (IMF) content plays an important role in meat quality. Many genes involved in lipid and energy metabolism were identified as candidate genes for IMF deposition, since genetic polymorphisms within these genes were associated with IMF content. However, there is less information on the expression levels of these genes in the muscle tissue. This study aimed at investigating the expression levels of sterol regulating element binding protein-1c (SREBP-1c), diacylglycerol acyltransferase (DGAT-1), heart-fatty acids binding protein (H-FABP), leptin receptor (LEPR) and melanocortin 4 receptor (MC4R) genes and proteins in two divergent Banna mini-pig inbred lines (BMIL). A similar growth performance was found in both the fat and the lean BMIL. The fat meat and IMF content in the fat BMIL were significantly higher than in the lean BMIL, but the lean meat content was lower. The serum triacylglycerol (TAG) and free fatty acid (FFA) contents were significantly higher in the fat than in the lean BMIL. The expression levels of SREBP-1c, DGAT-1 and H-FABP genes and proteins in fat BMIL were increased compared to the lean BMIL. However, the expression levels of LEPR and MC4R genes and proteins were lower.     

The porcine fat mass and obesity associated (FTO) gene is associated with fat deposition in Italian Duroc pigs

In humans, common variants in the fat mass and obesity associated ( FTO ) gene are associated with body mass index and obesity. Here we sequenced exon 4, parts of introns 3 and 4 and two portions of the 3'-untranslated region of the porcine FTO gene in a panel of nine pigs of different breeds and identified three SNPs. Allele frequencies of the g.276T>G ( AM931150 ) mutation were studied in seven pig breeds. This mutation was used to linkage-map FTO to SSC6. Association analyses between the g.276T>G polymorphism and several traits [pH of semimembranosus muscle and estimated breeding values (EBV) for average daily gain, back fat thickness, lean cuts, ham weight and feed:gain ratio] were carried out in 257 sib-tested Italian Large White pigs. Only feed:gain ratio showed P  <   0.05. A selective genotyping approach was applied, analysing two extreme and divergent groups of Italian Large White pigs selected on the basis of back fat thickness EBV (50 with most positive and 50 with most negative values). Fisher's exact test (two-tailed) was not significant when comparing the allele frequencies of these two groups. The same approach was used in the Italian Duroc breed for which two extreme and divergent groups of animals were selected according to visible intermuscular fat EBV. Differences of allele frequencies between these two groups were highly significant ( P  <   0.00001, P  <   0.001 and P  <   0.0001, considering all animals or only two- or three-generation unrelated animals respectively), indicating association between the analysed FTO marker and intermuscular fat deposition.     

藏猪小肠形态、消化酶及微生物多样性研究

[目的]肠道是动物的主要消化器官,同时也是机体抵抗外源病原菌的重要屏障,已有研究表明,动物的品种、饲养方式、生长阶段均会影响动物的肠道菌群结构,但对舍饲和放牧饲养条件下藏猪的肠道菌群结构,以及藏猪和长白、约克与杜洛克三元杂交猪(DLY猪)的肠道菌群结构是否有差异,尚未见报道.[方法]本研究选取6-7月龄的放牧藏猪、舍饲...     

二花脸猪与大约克猪生长期肌内脂肪合成与水解基因表达特征的比较研究

大约克公猪于15kg、40kg、60kg和90kg时随机屠宰,二花脸公猪于18kg、40kg、60kg、80kg和90kg时随机屠宰,每阶段每品种各屠宰4头。采集背最长肌,用乙醚抽提法分析肌内脂含量,同时取冈上肌和半膜肌提取组织的总RNA,用相对定量RT-PCR方法,以18S作内标,分析脂肪合成基因(LPL、ME、ACX)与水解基因(HSL)表达的发育性变化。结果表明：(1)两品种猪肌内脂生长规律存在差异：二花脸猪生长前期(40kg前)肌内脂含量为2％左右,但在60kg阶段肌内脂水平显著升高到4％以上,90kg阶段达到5％以上而大约克猪在整个生长期肌内脂含量稳定在2％左右;(2)冈上肌和半膜肌肌内脂肪组织脂肪合成与水解基因表达规律相似;(3)二花脸猪20-60kg时LPL和MEmRNA表达水平的变化与肌内脂沉积趋势相吻合;结果提示：二花脸猪与大约克猪肌内脂含量的差异主要是由二花脸猪在40—60kg阶段肌内脂快速沉积引起的,而且ME和LPL基因表达水平的增加可能在二花脸猪肌内脂快速沉积过程中发挥了作用。     

Effects of two supplementation levels of linseed combined with CLA or tallow on meat quality traits and fatty acid profile of adipose and different muscle tissues in slaughter pigs

Dietary linseed supply efficiently elevates the linolenic acid concentration of pork. The main problem of increasing the n-3 fatty acid tissue levels arises from a higher susceptibility to lipid oxidation. Increasing the saturation level of tissue lipids by the dietary inclusion of conjugated linoleic acids (CLA) or tallow might prevent oxidation. Thus, the aim of the study was to evaluate the impact of dietary CLA or tallow supplementation combined with extruded linseed on the growth performance, carcass characteristics and fatty acid profile of muscles (longissimus, semimembranosus, biceps femoris) and subcutaneous fat (SF). The enzyme activity of the de novo lipogenesis and stearoyl-CoA desaturase in the SF was also assessed. From 18 to 104 kg BW, 32 Swiss Large White barrows were fed a diet supplemented with either: (1) 2% linseed (L2); (2) 3% linseed (L3); (3) 2% linseed + 1% CLA (L2-C) or (4) 2% linseed + 1% tallow (L2-T). The linolenic and eicosatrienoic acid concentrations were higher (P < 0.01) and the ∑n-6/∑n-3 ratio was lower (P < 0.01) in all tissues of L3 than L2 and L2-T barrows. Only in the SF the docosapentaenoic acid concentration was increased (P < 0.01) in L3 barrows. Compared with the other three diets, feeding the L2-C diets increased (P < 0.01) the amount of myristic, palmitic, stearic and palmitoleic acid at the expense of the oleic and eicosenoic acid content in the intramuscular and SF lipids. Except for the lower (P < 0.05) eicosadienoic acid concentration in the muscles, feeding the L2-C treatment resulted in similar polyunsaturated fatty acid concentrations and ∑n-6/∑n-3 ratio than feeding L2 or L2-T diets. Both the c9,t11- and t10,c12-CLA isomers found in the CLA-supplemented diet were also detected in the tissues, but the c9,t11-isomer was more abundant than the t10,c12-isomer. De novo lipogenesis was not (P > 0.05) affected by the dietary fats, whereas Δ9-desaturase activity was depressed (P < 0.05) by CLA inclusion (L2-C). Only when oxidation was challenged by cooking and subsequent storage for 4 days at 4°C values of thiobarbituric acid-reactive substances were lower (P < 0.05) in longissimus muscle chops of L2-C compared with L2, L3 and L2-T barrows. The present findings revealed that CLA, but not tallow, combined with extruded linseed enhanced the oxidative stability of pork probably by lowering the degree of unsaturation of the lipids without affecting the improved ∑n-6/∑n-3 ratio.     

The single nucleotide polymorphisms of the chicken myostatin gene are associated with skeletal muscle and adipose growth

Myostatin, a new member of the TGF-ß superfamily, is predominantly expressed in skeletal muscle cells and functions as a negative regulator of skeletal muscle growth in animals. Recently, we have reported three single nucleotide polymorphisms (SNPs) in the chicken myostatin gene. Herein, we investigate the association of those SNPs with the production traits in a F₂ chicken line derived from Broilers crossing to Silky with the least square analysis. The results show that the BB and AA genotypes are strongly associated with abdominal fat weight (AFW), abdominal fat percentage (AFP), and birth weight (BW) (P < 0.05). Breast muscle percentage (BMP) of the AA type is higher than that of the AB type. The breast muscle weight and breast muscle percentages of F₂ individuals have significant difference between CC and DD genotypes (P < 0.05). Breast muscle weight (BMW) of EF birds is higher than that of EE birds (P < 0.05). In this report, we present the first genetic evidence to show that chicken myostatin not only plays an important role in controlling skeletal muscle growth and differentiation, but also may be involved in regulation of adipose growth in chicken.     

The single nucleotide polymorphisms of the chicken myostatin gene are associated with skeletal muscle and adipose growth

Myostatin, a new member of the TGF-p superfamily, is predominantly expressed in skeletal muscle cells and functions as a negative regulator of skeletal muscle growth in animals. Recently, we have reported three single nucleotide polymorphisms (SNPs) in the chicken my-ostatin gene. Herein, we investigate the association of those SNPs with the production traits in a F2 chicken line derived from Broilers crossing to Silky with the least square analysis. The results show that the BB and AA genotypes are strongly associated with abdominal fat weight (AFW), abdominal fat percentage (AFP), and birth weight (BW) (P < 0.05). Breast muscle percentage (BMP) of the AA type is higher than that of the AB type. The breast muscle weight and breast muscle percentages of F2 individuals have significant difference between CC and DD genotypes (P< 0.05). Breast muscle weight (BMW) of EF birds is higher than that of EE birds (P< 0.05). In this report, we present the first genetic evidence to show that chicken myostatin not o     

Identification of extracellular matrix and cell adhesion molecule genes associated with muscle development in pigs

Extracellular matrix (ECM) and cell adhesion molecule (CAM) genes are involved in the regulation of skeletal muscle development; however, their roles in skeletal muscle development in pigs are still poorly understood. 65 days postcopulation (dpc) is a critical time point in pig development. Therefore, we analyzed expression of ECM and CAM genes in the longissimus dorsi muscles at 65?dpc from Landrace (lean-type: L65), Tongcheng (obese-type: T65), and Wuzhishan pigs (miniature-type: W65) using microarray technology. A total of 35 genes were differently expressed between the breeds, and of them, 18, 18, and 20 genes, were observed in the comparisons of L65 versus T65, L65 versus W65, and T65 versus W65 (L65/T65, L65/W65, and T65/W65), respectively. In L65/T65, differently expressed genes were widely distributed, whereas in L65/W65 and T65/W65, they mostly focused on the genes encoding CAMs and ECMs proteins. Moreover, the largest number of up-regulated genes involved in skeletal muscle development was detected in L65, a moderate number in W65, and the smallest number was in T65. Cluster analysis suggested that T65 showed a more similar expression pattern to L65 than W65. In addition, we validated that five genes from microarray data were more highly expressed in the prenatal as compared to postnatal periods in Landrace and Tongcheng pigs and showed a greater range of high-level expression during gestation in Landrace than Tongcheng pigs. Our data indicated that ECM and CAM genes are differently expressed among the three breeds, and more complicated molecular events involving CAMs and ECMs were observed in Wuzhishan pigs. This study advances our knowledge of the molecular basis of phenotypic variation and provides a helpful resource for the identification of candidate genes associated with meat production traits in pigs.     

Expression of proto-oncogenes and muscle specific genes during cardiac hypertrophy and development in rats and humans

A regulatory interdependence of expression of proto-oncogenes and muscle specific genes observed in smooth muscle was examined in cardiac muscle during normal development and hypertrophy both in rats and humans. During normal development in rats, myosin light chain 2 expression is very low at prenatal stages, while c-fos expression starts from the early stages of embryonic development. In aorta constricted rats c-fos induction occurs within 30 min whereas myosin light chain 2 expression is sufficiently high only after 3 or 4 days of post operative period. In the case of humans, the expression of myosin light chain 2 as well as c-fos occurs at high levels during embryonic development. Similar results were obtained with tissue samples obtained from patients with cardiac abnormalities. Induction of the c-fos gene in cultured myocytes by 12-O-tetradeeanoylphorbol 13-acetate has no influence on the expression of myosin light chain 2. These studies were extended with studies on c-myc and Β-myosin heavy chain gene expression which revealed a similar pattern of expression as that of c-fos and myosin light chain 2. These results have indicated that the expression of proto-oncogenes in cardiac muscle may be independently regulated from the expression of muscle specific genes.