Salamanders completely regenerate their limbs after amputation. Thus, these animals are unique models to investigate the mechanisms modulating regeneration in vertebrates. To investigate the influence of microRNAs (miRNAs) on newt limb regeneration, the miRNAs and mRNAs were simultaneously profiled using Illumina HiSeq 2500 System during limb regeneration of Cynops orientalis at 3, 7, 14, 30 and 42 days postamputation. A total of 203 miRNAs and 4230 mRNAs were identified to be differentially expressed. Together with the proteomic data obtained from our previous study, integrative analysis of multiple profiling data sets was performed to construct an interaction network of differentially expressed miRNAs, mRNAs and proteins. Results of GO and KEGG analyses showed that the differentially expressed miRNA targets were mainly directed to cytoskeletal remodeling and carbohydrate metabolism. The stage-specific regulation of miRNAs on their targets was analyzed by hierarchical clustering analysis and validated by qRT-PCR. The negative regulation of miR-223 and miR-133a on their targets was tested by performing dual luciferase reporter assay. The integration analysis will provide a powerful tool to identify the regulatory mechanisms of miRNAs and their targets. The results may have implications in understanding the complex mechanisms underlying newt limb regeneration.

中文翻译：

---

东方夜蛾肢体再生过程中MicroRNAome，转录组和蛋白质组的综合分析。

put截肢后，completely完全再生。因此，这些动物是研究脊椎动物中再生机制的独特模型。为了研究microRNA（miRNA）对new肢再生的影响，在截肢后第3、7、14、30和42天，使用Illumina HiSeq 2500系统同时分析了东方侧柏的肢体再生过程中的miRNA和mRNA。总共鉴定出203个miRNA和4230个mRNA被差异表达。结合从我们先前的研究中获得的蛋白质组学数据，对多个分析数据集进行了综合分析，以构建差异表达的miRNA，mRNA和蛋白质的相互作用网络。GO和KEGG分析的结果表明，差异表达的miRNA靶标主要针对细胞骨架重塑和碳​​水化合物代谢。通过分级聚类分析来分析miRNA在其靶标上的阶段特异性调控，并通过qRT-PCR进行验证。miR-223和miR-133a在其靶标上的负调控通过双重荧光素酶报告基因检测法进行检测。整合分析将提供强大的工具来识别miRNA及其靶标的调控机制。该结果可能对理解limb肢再生的复杂机制有影响。miR-223和miR-133a在其靶标上的负调控通过双重荧光素酶报告基因检测法进行检测。整合分析将提供强大的工具来识别miRNA及其靶标的调控机制。该结果可能对理解limb肢再生的复杂机制有影响。miR-223和miR-133a在其靶标上的负调控通过双重荧光素酶报告基因检测法进行检测。整合分析将提供强大的工具来识别miRNA及其靶标的调控机制。该结果可能对理解limb肢再生的复杂机制有影响。