Spatiotemporal Expression Profiles of miR-25 in Siniperca chuatsi and Prediction Analysis of Its Targeting Core Circadian Clock Genes
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摘要:
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。 Abstract: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. -
Key words:
- Siniperca chuatsi /
- miR-25 /
- Expression pattern /
- Target gene prediction
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图 1 miR-25在鳜不同组织中的相对表达量
n = 3；*P < 0.05。
Figure 1. Relative expression of miR-25 in different tissues of Siniperca chuatsi
图 2 miR-25在鳜不同发育时期胚胎的相对表达量
n = 3；*P < 0.05。
Figure 2. Relative expression level of miR-25 in different embryonic stages of Siniperca chuatsi
图 3 miR-25与Arntl2结合靶位点预测
Figure 3. The prediction of target site on target gene Arntl2 of miR-25
图 4 miR-25与Nr1d2b结合靶位点预测
Figure 4. The prediction of target site on target gene Nr1d2b of miR-25
表 1 实时荧光定量PCR引物
Table 1. The Primer Sequences for RT-qPCR
名称 引物序列（5’-3’） sch-miR-25 F CATTGCACTTGTCTCGGTCTG U6 R TTTGGCTCTCTTGGCACGGAT U6 F ACGGACAGCTGAAGAACAAGT 
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