摘要
【目的】通过在m RNA表达水平检测TGF-β/smad信号通路基因在湖羊肌肉组织中时空表达,来探究各基因的表达规律及内在联系。【方法】利用荧光定量PCR技术对湖羊Smads基因在出生后不同生长阶段(2日龄,2月龄和6月龄)不同性别的两种不同骨骼肌(腓肠肌、趾长伸肌)的相对表达进行分析,以及对湖羊Smad家族(Smad2、Smad3、Smad4、Smad7)基因、YAP1基因的表达相关性分析。【结果】湖羊肌肉TGF-β/smad信号通路中Smads基因时空表达规律研究发现:Smads在不同肌肉组织的表达分析中,Smads在腓肠肌中的表达高于趾长伸肌,可能与这两种骨骼肌分属不同部位有关;Smads在不同月龄的表达分析中,Smad2、Smad3、Smad4在2日龄的表达量均高于其它月龄,而Smad7在2日龄的表达量低于6月龄,2月龄时表达量最低;在不同性别的表达分析中,Smads在2日龄公羊中的表达量高于母羊,Smad2、Smad4、Smad7在2月龄和6月龄则低于母羊;Smad3在不同生长阶段中公羊表达量高于母羊。湖羊肌肉TGF-β/smad信号通路中Smads、YAP1基因表达相关性研究发现:在2日龄腓肠肌中,Smad2、Smad3、Smad4、Smad7的表达与YAP1相关不显著(P>0.05);在2月龄腓肠肌中,Smad2的表达与YAP1存在显著正相关(P<0.05),Smad3、Smad4、Smad7的表达与YAP1相关不显著(P>0.05);在6月龄腓肠肌中,Smad2、Smad3、Smad4、Smad7的表达与YAP1相关不显著(P>0.05);在腓肠肌的不同生长阶段中,Smad3的表达YAP1存在极显著负相关(P<0.01),Smad2、Smad4、Smad7的表达与YAP1相关不显著(P>0.05)。在2日龄趾长伸肌中,Smad3的表达与YAP1存在极显著正相关(P<0.01),Smad2、Smad4、Smad7的表达与YAP1存在显著正相关(P<0.05);在2月龄趾长伸肌中,Smad2、Smad3、Smad4、Smad7的表达与YAP1相关不显著(P>0.05);在6月龄趾长伸肌中,Smad7与YAP1存在极显著正相关(P<0.01);Smad2、Smad3、Smad4的表达与YAP1相关不显著(P>0.05);在趾长伸肌的不同生长阶段中,Smad2、Smad3、Smad4、Smad7的表达与YAP1存在极显著正相关(P<0.01)。【结论】由此推断不同组织、生长阶段以及性别等因素均可影响Smads在肌肉中的表达;在湖羊这两种不同肌肉组织中,Hippo通道中YAP1基因可通过参与调控TGF-β/smad信号通路而参与肌肉的增殖和分化过程。
【Objective】 In order to explore the expression pattern and inner connection between the different genes, the m RNA expression level was used to detect the spatial and temporal expression of genes in TGF-β/Smad signaling pathway of sheep muscle tissue. 【Method】 In this study, q-PCR was used to detect relative expression of Smads genes and Yes-associated protein 1（YAP1） in 2 different skeletal muscles（i.e., gastrocnemius muscle, extensor digitorum longus） and 3 different growth stages（i.e., 2-day-old, 2 and 6-month-old） of Hu Sheep, and analyze the relative expression between Smads（Smad2, Smad3, Smad4 and Smad7） and YAP1. 【Result】The results of the spatial and temporal expression of Smads in TGF-β/Smad signaling pathway showed that the expression of Smads in extensor digitorum longus were lower than in gastrocnemius muscle, which may be related to two different parts of the skeletal muscle. The expression of Smad2, Smad3 and Smad4 genes in the 2-day-old were higher than other growth stages, and Smad7 gene expression in the 2-day-old was lower than 6-month-old, and the expression in 2-month-old was the lowest. The expressions of Smads gene of ram in the 2-day-old was higher than ewe, and the expressions of Smad2, Smad4, and Smad7 genes of ram in 2-month-old and 6-month-old were lower than the ewe in 2-day-old, the expression of Smad3 gene of ram in 3 growth stages were higher than ewe. The correlation expression of Smads gene and YAP1 gene in TGF-β/smad signaling of sheep muscle was found that there was no significant correlation in 2-day-old of gastrocnemius muscle between YAP1 and smads（P0.05）. The expression between YAP1 and Smad2 had a significant positive correlation（P0.05） and the correlation of the expression between YAP1 and other Smads was not significant in 2-month-old of gastrocnemius muscle（P0.05）. There was a significant negative correlation between YAP1 and Smad3 in different growth stages of gastrocnemius muscle（P0.01）. YAP1 expression exhibited a high positive correlation with Smad3 and the expression of YAP1 with the others was significantiy correlated in extensor digitorum longus of 2-month-old（P0.05）. There was no significant correlation between YAP1 and Smads in 2-month-old of extensor digitorum longus（P0.05）. The expression between YAP1 and Smad7 had a high significant positive correlation（P0.01） and the correlation of expression between YAP1 and other Smads was not significant in 6-month-old of extensor digitorum longus（P0.05）. There were highly significant negative correlations between YAP1 and Smads in different growth stages of extensor digitorum longus（P0.01）. 【Conclusion】 It was suggested that different tissues, growth stages and gender could affect the expression of Smads in muscle. YAP1 of Hippo pathway may participate in regulating the TGF-β/smad pathway and the process of muscle proliferation and differentiation in gastrocnemius muscle and extensor digitorum longus of Hu sheep.
出处
《中国农业科学》
CAS
CSCD
北大核心
2016年第11期2203-2213,共11页
Scientia Agricultura Sinica
基金
江苏省农业科技支撑计划(BE2012331)
江苏省工程技术研究中心项目(BM2012308)
科技部家养动物平台项目
江苏高校优势学科建设工程资助项目
国家星火计划重点项目(2012GA690003)
江苏省苏北科技计划(BN2014003,BN2015013,BN2015014)
江苏省农业科技自主创新项目(CX(14)2073)
江苏省研究生科研创新计划(KYLX15_1375)
