MicroRNA(miR)-27b在山羊生长过程中的组织表达谱分析

摘要

本研究旨在分析microRNA(miR)-27b在安徽白山羊(Capra hircus)和波尔山羊生长发育过程中的表达变化,以了解其在山羊肌肉发育中的重要作用.选用安徽白山羊和波尔山羊不同发育时期(初生,6月龄和12月龄3个时期)背最长肌为材料,利用Real-time quantitative PCR (qRT-PCR)方法检测miR-27b的表达变化.同时,分析了miR-27b在6月龄山羊各组织中(心脏、肝脏、脾脏、肺、肾脏、腿肌和胸肌)的表达.并用MEGA3.1分析miR-27b在不同物种中的保守性.结果显示,miR-27b在安徽白山羊和波尔山羊骨骼肌发育过程中差异表达,呈现不同的表达模式.安徽白山羊12月龄背最长肌miR-27b表达最高,初生时最低.在山羊不同组织中,骨骼肌和心脏中miR-27b表达量较高,肝脏和肾脏中miR-27表达处于中度水平,而在另外4种组织中表达量较低.实验结果表明,miR-27b可能参与山羊肌细胞增殖和分化,从而影响山羊骨骼肌生长发育,与山羊肌肉的生长类型有关.同时序列分析显示,miR-27b在10个物种中具有较高的保守性.山羊miR-27b的表达具有的组织和时序差异性实验结果为进一步研究miR-27b的功能提供基础资料.%The aim of this study is to detect the expression of microRNA(miR)-27b in skeletal muscle development in goats(Capra hircus),and explore the roles of miR-27b in muscle development.Real-time quantitative PCR(qRT-PCR) was performed to detect the miR-27b expression in the longissimus dorsi muscle from the skeletal muscle tissues of different stages (neonate,6 and 12 months) in Anhui White and Boer goats(C.hircus).Meanwhile,the expression of miR-27b in various tissues of 6 months goats was detected.And the conservation of miR-27b in different varieties was analysed by MEGA3.1.The miR-27b differential expression in skeletal muscle development exhibited different expression pattems between Anhui White and Boer goats.The expression level in longissimus dorsi muscle from 12 months goats was the highest,while,the expression level in neonate was the lowest.In 6 months goats,miR-27b was abundantly expressed in longissimus dorsi muscle and heart,moderately in liver and kidney,and weakly in other collected tissues.The results indicated that miR-27b may be involved in myofibril proliferation and differentiation and further contribute to skeletal muscle development and muscle phenotype with different genotypes,and the miR-27b was conservative in 10 different species.The expression of miR-27b has tissue and time specific,and the present result provides the theory base for study ofmiR-27b function.