静原鸡胸肌和腿肌肌苷酸特异性沉积相关circRNA的联合分析

Combined analysis of circRNA related to inosine monphosphate deposition in thoracic and leg muscle of Jingyuan chicken

旨为分析静原鸡胸肌和腿肌中circRNA的表达模式与肌苷酸(Inosine monphosphate,IMP)在肌肉组织特异性沉积的关系。利用RNA-seq技术对选取的具有相同遗传背景180日龄静原鸡胸肌和腿肌进行全转录组测序,对获得的测序数据进行拼接、比对、注释和差异分析等,并对差异表达的circRNA和mRNA进行GO功能和KEGG信号通路富集分析,使用qRT-PCR对转录组结果进行表达量验证。结果表明:共筛选到circRNA 3283个,组织特异性表达circRNA 242个。以|log2Fold change|≥1,P<0.05为筛选条件,共得到差异circRNA 446个,DEGs 1100个,差异miRNA 36个;对差异表达的circRNA和mRNA进行GO功能和KEGG信号通路富集分析,两者都显著富集于糖酵解/糖异生通路和心肌细胞的肾上腺素信号传导,并且差异circRNA显著富集于嘌呤代谢通路,DEGs显著富集于氨基酸的生物合成、磷酸戊糖途径、丙氨酸、天冬氨酸和谷氨酸代谢等与IMP合成高度相关的通路。对差异circRNA进行靶基因预测,构建circRNA-miRNA-mRNA可视化共表达网络,筛选到以novelcirc0013580、novelcirc0012931、novelcirc0011886、novelcirc0013588、novelcirc0007737和novelcirc0008274为核心的转录后调控因子和以PKM2、GART为核心的靶基因,靶向基因分析发现IMP合成关键调控因子可能来源于同一亲本基因的novelcirc0013580和novelcirc0013588,两者能够同时结合novel409作用于靶基因GART。对6个circRNA和6个miRNA qRT-PCR检测结果与全转录组测序结果趋势一致。综上,circRNA在静原鸡胸肌和腿肌中的差异性表达与IMP的特异性沉积具有相关性,为研究地方鸡种IMP在肌肉组织中特异性沉积的分子机制提供参考信息。

In order to explore the relationship between the expression pattern of circRNA in thoracic and leg muscle of Jingyuan chicken and the specific deposition of inosine monphosphate(IMP)in muscle tissue,the whole transcriptome sequencing of the thoracic and leg muscle of 180-day-old Jingyuan chicken with the same genetic background was carried out by RNA-seq.Then,the sequencing data were spliced,compared,annotated and differentially analyzed.The GO function and KEGG signal pathway enrichment analysis of differentially expressed circRNA and mRNA were carried out.qRT-PCR technology was used to verify the expression level of transcriptome results.The results showed that a total of 3283 circRNA were screened and 242 circRNA were tissue-specific.Using|log2 fold change|≥1,P<0.05 as screening conditions,446 differential circRNA,1100 DEGs and 36 differential miRNA were obtained.The GO function and KEGG signal pathway enrichment analysis of differentially expressed circRNA and mRNA were carried out.They were significantly enriched in glycolysis/gluconeogenesis pathway and adrenaline signal transduction in cardiomyocytes,and differential circRNA was significantly enriched in purine metabolic pathway.DEGs was significantly enriched in amino acid biosynthesis,pentose phosphate pathway,alanine,aspartic acid and glutamate metabolism and were highly related to IMP synthesis.The target genes of differential circRNA were predicted,the visual co-expression network of circRNA-miRNA-mRNA was constructed,and the post-transcriptional regulatory factors with novelcirc0013580,novelcirc0012931,novelcirc0011886,novelcirc0013588,novelcirc0007737 and novelcirc0008274 as the core and the target genes with PKM2 and GART as the core were screened.Targeted gene analysis showed that the key regulatory factors of IMP synthesis may be that both novelcirc0013580 and novelcirc0013588,which are derived from the same parent gene,can bind to novel409 and act on the target gene GART at the same time.The detection results of 6 circRNA and 6 miRNA qRT-PCR were consistent with the results of whole transcriptome sequencing.In conclusion,the differential expression of circRNA in thoracic and leg muscle of Jingyuan chicken is related to the specific deposition of IMP,which provides reference information for the study of the molecular mechanism of specific deposition of IMP in muscle tissue of local chicken breeds.