Sox6基因调控鸡骨骼肌分化和肌纤维类型的研究

Study on the Regulations of Sox6 Gene on Skeletal Muscle Differentiation and Muscle Fiber Types in Chicken

本实验室前期研究发现一个拷贝数变异区域与鸡的重要经济性状显著相关,并且该区域与Sox6编码区域重叠,提示Sox6基因对骨骼肌生长发育具有调控作用。研究首先采用RTqPCR分析Sox6基因在不同胚龄隐性白母鸡的胸肌和腿肌中的表达情况和表达差异。接着在原代成肌细胞中过表达Sox6基因,通过RT-qPCR、免疫荧光和HE染色观察Sox6对原代成肌细胞的肌纤维类型调控情况和分化的影响。结果表明,Sox6 mRNA表达量在不同胚龄的胸肌和腿肌上的表达趋势相近,但在胸肌上的表达量高于腿肌,并且在E14、E17和7W时,胸肌表达量显著高于腿肌(P<0.05),提示Sox6可能对快肌起到重要作用。原代成肌细胞上面过表达Sox6后定量快慢肌特异基因发现Sox6能够促进MyH1B,抑制sMyHC1的表达;免疫荧光和HE染色观察发现Sox6能够促进成肌细胞的分化和肌管的形成,从而证明了Sox6对骨骼肌的分化和肌纤维类型调控具有一定的作用。

In our previous research,we found a copy number variation area which overlapped with the coding sequence of Sox6 were significantly associated with important economic traits.So Sox6 might play a role in regulation of the development of skeletal muscle.In this study,the expression profiles and differences of Sox6 between pectoral and leg muscles in recessive white hens of different embryonic ages were analysed by RT-qPCR.Then Sox6 in primary myoblasts was overexpressed and the functions of Sox6 on myoblast muscle fiber types and differentiation were detected by RT-qPCR,immunofluorescence and HE stain.The results indicated that the expression trends of Sox6 between pectoral and leg muscle in different embryonic ages were similar,but the expression level in pectoral muscle was higher than that in leg muscle,and the differences were significant in E14、E17 and 7 weeks of age(P<0.05),which suggested that Sox6 might have an important role in fast-type muscle.Overexpression of Sox6 in primary myoblasts demonstrated that Sox6 could promote the expression of MyH1 B and inhibit the expressions MyHC1,while the promotions of Sox6 on myoblasts differentiation and myotubes formation were proved by immunofluoresence and HE stain.Taken together,these data proved the regulations of Sox6 on differentiation and muscle fiber types in skeletal muscle.