Endothelial UCP2 regulates the neurogenic-to-astrogenic fate switch during brain development

Purpose: To gain futher insight into how endothelial UCP2 regulates the neurogenic-to-astrogenic fate switch ,RNA-seq was used to analyze the genome-wide changes resulting from isolated endothelial cells of E14.5 and E17.5 UCP2 endothelial conditional knock out mice and littermate wild-type. Methods: mRNA from E15 and E18 isolated endothelial cells of wild-type(WT) and UCP2f/f;Tie2-cre mice was extracted. Specifically, Agilent 2100 Bioanalyze was used to quality controlled and quantified. then, mRNA was converted to cDNA and bound the library. RNA-sequencing analysis was used by the Illumina HiSeq 2500 platform in Annoroad Genomics Results: Approximately one thousand transcripts showed differential expression between the wild-type(WT) and UCP2ECKO mice brain cortex, with a fold change >= 2 and p value <0.05.Geneontology analysis of the up-regulated genes showed obvious enrichment of biological processes related to development process and regulation of cell communication.The down-regulated genes exhibited enrichment of biological processes related to cell fate commitment. These results reflected endothelial UCP2 plays roles in cortex development. Conclusions: Endothelial UCP2 RNA-seq would provide a overall understanding how endothelial UCP2 regulates the neurogenic-to-astrogenic fate switch during brain development mRNA profiles of E15 and E18 wild type (WT) and UCP2ECKO mice were generated by deep sequencing, in triplicate, using Illumina HiSeq 2500.研究目的：为深入探究内皮UCP2（endothelial UCP2）如何调控神经发生向星形胶质发生的命运转换，本研究采用RNA测序（RNA-seq）技术，分析E14.5及E17.5胎龄的UCP2内皮条件性敲除小鼠与同窝野生型小鼠的分离内皮细胞所引发的全基因组表达变化。 实验方法：提取野生型（WT）及UCP2f/f;Tie2-cre小鼠E15、E18胎龄分离内皮细胞的mRNA。具体而言，采用安捷伦2100生物分析仪（Agilent 2100 Bioanalyzer）对RNA进行质量控制与定量；随后将mRNA反转录为cDNA并构建测序文库，依托Illumina HiSeq 2500平台于安诺优达基因科技（Annoroad Genomics）完成RNA测序分析。 实验结果：野生型（WT）与UCP2ECKO小鼠大脑皮层样本中，约有1000个转录本呈现差异表达，差异倍数≥2且P值<0.05。对上调基因的基因本体（Gene Ontology, GO）富集分析显示，其显著富集于发育过程及细胞通信调控相关的生物学过程；下调基因则显著富集于细胞命运决定相关的生物学过程。上述结果表明内皮UCP2在大脑皮层发育中发挥重要调控作用。 研究结论：本研究的内皮UCP2 RNA-seq数据可全面解析大脑发育过程中内皮UCP2如何调控神经发生向星形胶质发生的命运转换；本研究通过Illumina HiSeq 2500平台完成了三次生物学重复的深度测序，获取了E15、E18胎龄野生型（WT）及UCP2ECKO小鼠的内皮细胞转录组图谱。