摘要
[目的]验证鸡环状RNA(circMICAL2)的真实性,研究circMICAL2 在鸡肌肉生长发育中的潜在功能及调控机制.[方法]基于circMICAL2 环状序列设计特异性引物采用PCR扩增完成其真实性验证,并测定RNase R和放线菌素D处理检测circMICAL2 的内源稳定性.采集 16 日龄三黄鸡(雏鸡)和 180 日龄三黄鸡(成年鸡)的心脏、肝脏、肺、肾脏、肠、皮肤、胸肌和腿肌等8 个组织样本,采用实时荧光定量PCR(qRT-PCR)分析circMICAL2 在鸡不同发育时期的组织表达图谱;运用生物信息学对circMICAL2 靶向miRNA和mRNA进行预测,并开展GO功能和KEGG通路富集分析.[结果]鸡circMICAL2 真实存在,其环化稳定性强;circ-MICAL2 在成年鸡和雏鸡各组织中广泛表达,且circMICAL2 均在成年鸡和雏鸡的胸肌和腿肌肌肉中表达量最高,circMICAL2 在成年鸡腿肌和胸肌中的表达量均显著高于雏鸡(P<0.05),circMICAL2 与鸡肌肉生长发育调控密切相关.circMICAL2 可靶向gga-miR-103-3p和gga-miR-130b-3p,调控下游 225 个潜在靶基因.circMICAL2 的靶基因主要富集于TGF-β信号通路、MAPK信号通路、细胞周期等相关信号通路.[结论]鸡circMICAL2 真实存在,circMICAL2 的表达对鸡不同生长发育时期肌肉的生长具有重要调控作用.
Abstract
[Objective]The purpose of this study is to verify the authenticity of chicken circular RNA(circMICAL2)and explore the function and regulation mechanism of circMICAL2 in chicken muscle growth and development.[Methods]Specific primers based on circMICAL2 circular sequences were designed by PCR amplification,and RNase R and actinomycin D treatments were performed to detect the endogenous sta-bility of circMICAL2.Eight tissue samples from heart,liver,lung,kidney,intestine,skin,chest muscle and leg muscles of 16-day-old and 180-day-old chickens(adult chickens)were collected,and the tissue ex-pression profile of circMICAL2 in different development periods was completed by real-time PCR(qRT-PCR);Bioinformatics was used to predict circMICAL2-targeted miRNA and mRNA,and to carry out GO function and KEGG pathway enrichment analysis.[Results]Chicken circMICAL2 really existed,and its cycli-zation stability was strong;circMICAL2 was widely expressed in all tissues of adults and chicks,and circMI-CAL2 was most highly expressed in their chest and leg muscles of adults and chicks,and the expression of cir-cMICAL2 in both adult leg and thorax muscles was significantly higher than that in chicks(P<0.05),indica-ting that circMICAL2 was closely related to the regulation of chicken muscle growth and development.circMI-CAL2 targets gga-miR-103-3p and gga-miR-130b-3p regulated 225 downstream potential target genes.The target genes of circMICAL2 were mainly enriched in TGF-β signaling pathway,MAPK signaling pathway,cell cycle and other related signaling pathways.[Conclusion]Chicken circMICAL2 really exists,and the expression of circMICAL2 plays an important role in regulating the growth of muscle during different growth and development periods.
基金项目
四川省科技计划-重点研发项目(2022YFN0039)
乐山师范学院科技计划项目(DGZZ202002)
乐山师范学院科技计划项目(22HX00051)
国家科技期刊平台
NSTL
万方数据