基于RNA-seq筛选不同饲养模式下肉鸡肌肉发育代谢相关差异基因

Application of RNA-Seq Technology for Screening Differentially Expressed Genes Related to Muscle Development and Metabolism of Broilers in Different Rearing Modes

为发掘饲养模式影响肌肉发育代谢的相关关键基因,本研究分别在笼养和地面平养2种饲养模式中随机选取5只42日龄Ross 308肉鸡进行屠宰,提取腿肌RNA,通过RNA-seq技术筛选出与肌肉发育代谢相关的差异基因,并利用qPCR进行验证。随后分离、鉴定并构建了鸡骨骼肌卫星细胞增殖分化模型,对关键差异基因PHGDH在鸡骨骼肌细胞中的表达模式进行检测。结果显示:在2种饲养模式中共筛选出3个差异表达基因(ASNS、JPH2、PHGDH),qPCR验证结果与RNA-Seq测序结果一致,差异基因均在平养模式鸡腿肌肌肉中高表达;在鸡骨骼肌细胞增殖期间,PHGDH表达逐渐上调,但在分化第2天至第6天,表达水平逐渐下降;EdU细胞增殖检测结果显示,抑制PHGDH的表达可显著抑制鸡骨骼肌细胞的增殖。以上结果表明,PHGDH可能在鸡骨骼肌形成过程中发挥重要作用。

In order to explore the key genes related to muscle development and metabolism of broilers in different rearing modes, this study used Ross 308 broiler as the research object, five broilers aged 42 days were randomly selected for slaughter in two rearing modes: cage and floor rearing, respectively, then extracted leg muscle RNA. Differentially expressed genes related to muscle development and metabolism were screened by RNA-Seq and verified by qPCR. Subsequently, a proliferation and differentiation model of chicken skeletal muscle satellite cells was isolated, identified and constructed. The expression pattern of PHGDH was detected. The results showed that there were three differentially expressed genes(ASNS, JPH2, PHGDH) between these two rearing modes. The mRNA expression levels which were verified by qPCR were in line with that identified by RNA-Seq technology. All the differentially expressed genes were highly expressed in the floor rearing mode. In addition, during the proliferation of chicken skeletal muscle cells, the expression of PHGDH was gradually increased. However, during the 2 to 6 day of cell differentiation, PHGDH expression level gradually decreased. The results of EdU cell proliferation test showed that inhibiting the expression of PHGDH could significantly inhibit the proliferation of chicken skeletal muscle cells. These results suggest that PHGDH might be a key gene in the formation of chicken skeletal muscle.