CRTC2基因在不同品种和月龄肉牛的表达分析

Analysis of CRTC2 gene expression in cattle of different varieties and months of age

为了研究肉牛CREB转录激活因子2(CRTC2)基因在肌肉组织中不同品种和月龄的表达情况,探讨CRTC2基因与生长发育的关系,利用qRT-PCR技术检测CRTC2基因在不同组织和不同月龄的表达规律;运用Western blot和免疫组化方法对CRTC2蛋白进行定量定位分析,探讨CRTC2蛋白的表达规律。结果显示:布莱凯特黑牛胰脏中CRTC2基因mRNA的表达量极显著高于其他组织(P<0.01),睾丸中CRTC2基因mRNA的表达量最低;CRTC2基因在不同时期随着月龄的增加表达量逐渐降低,其中2和6月龄mRNA表达量极显著高于10和18月龄(P<0.01),18月龄表达量最低;4个月龄布莱凯特黑牛的CRTC2表达量均高于鲁西黄牛,且2和6月龄品种间差异极限著(P<0.01);CRTC2蛋白随着月龄的增加呈现逐渐降低的趋势,其中2月龄表达量极显著高于其他月龄(P<0.01),18月龄相对表达量最低;4个月龄布莱凯特黑牛蛋白表达量均极显著高于鲁西黄牛(P<0.01)。提示:CRTC2基因主要在肌肉组织的肌原纤维中表达,可能参与动物的早期分化和生长相关过程,对调控肌肉生长发育发挥重要作用。

The aim of this study was to investigate the expression level of the CRTC2 gene in cattle of different breeds and months of age, and the relationship between growth and development of cattle, so as to lay a foundation for research on muscle growth and development of the animals. qRT-PCR was used to detect the mRNA expression of CRTC2 in their different tissues and at their different months of age. Western blot and immunohistochemistry were used to analyze the CRTC2 protein expression and positioning and to explore the expression patterns of the protein. The results showed that the expression of the CRTC2 gene was the highest in pancreas(P<0.01), followed by that of the thyroid gland and the longissimus dorsi muscle, but the expression was the lowest in testis. The expression of CRTC2 decreased as the months of age of the cattle increased, the expression level at two months and six months of age were the highest(P<0.01), but the lowest at 10 months and 18 months of age(P<0.01). The expression of CRTC2 of the Black cattle was higher than that of the Luxi cattle at all the four month age points, the expression was very significantly different between two and four months of age(P<0.01), and it the lowest at 18 months of age(P>0.05) mainly in muscle fibril and muscle tissue. In conclusion, the CRTC2 gene might be involved in early differentiation and growth of cattle, and play an important role in regulating of growth and development of their muscle.