鳜miR-25的时空表达特征及其靶向核心生物钟基因的预测分析

Spatiotemporal Expression Profiles of miR-25 in Siniperca chuatsi and Prediction Analysis of Its Targeting Core Circadian Clock Genes

目的研究miR-25在鳜的时空表达特征,预测其靶向核心生物钟基因,对其可能存在的生物学意义进行探讨。方法采用实时荧光定量PCR分析miR-25在鳜各组织以及不同阶段胚胎中的表达特征,并利用TargetScan对其靶向核心生物钟基因进行预测分析。结果miR-25在鳜红肌和心肌中的表达量较高(P<0.05),而在鳜脾、肾、肠以及肝中的表达量较低,且这4个组织中的表达量,差异无统计学意义(P>0.05);miR-25在鳜胚胎的2细胞期表达量较高(P<0.05),随着胚胎发育,其表达水平逐渐降低。靶基因预测结果显示,节律基因Arntl2、Nr1d2b mRNA的3’UTR区域存在miR-25的靶位点。结论miR-25在胚胎发育过程中呈母源性表达,并且可能在鳜红肌和心肌中具有重要的调控意义,此外miR-25可能通过靶向调节Arntl2、Nr1d2b的表达来调控鳜生物节律,为后续研究miR-25的生理作用以及调控昼夜节律从而预防相关疾病提供参考依据。

Objective To study the spatiotemporal expression profiles of miR-25 in the Chinese perch(Siniperca chuatsi),to predict its targeting core Circadian clock genes,and to explore its possible biological significance.Methods Real-time fluorescence quantitative PCR was used to analyze the expression characteristics of miR-25 in various tissues and embryos at different stages of Siniperca chuatsi,and TargetScan was used to predict its targeted core clock genes.Results The expression level of miR-25 was higher in the red muscle and heart(P<0.05),while it was lower in the spleen,kidney,intestine,and liver of the Chinese perch.However,the expression level difference in these four tissues was not statistically significant(P>0.05).The expression level of miR-25 was higher in the phase 2 of cell cycle in embryos(P<0.05),and it gradually decreased as the embryos developed.Target gene prediction results showed that the 3’UTR region of the rhythmic genes Arntl2 and Nr1d2b mRNA contained target sites for miR-25.Conclusion miR-25 exhibited maternal expression during embryonic development and may have important regulatory significance in the red muscle and heart of Chinese perch.Additionally,miR-25 may regulate the circadian rhythm by targeting the expression of Arntl2 and Nr1d2b,providing a reference for further research on the physiological effects of miR-25 and the regulation of circadian rhythm to prevent related diseases.