Brain vascular transcriptomes of mouse hyperglycemia mutants and controls

Purpose:To take a comprehensive effort in characterizing the brain vasculature gene expression upon hyperglycemia. Methods: We extracted mRNA from brain microvasculature fragments isolated from a genetic mouse model of hyperglycemia (Ins2-AKITA) and WT mice and analyzed their transcriptome with RNA sequencing The samples were sequenced on an Illumina HiSeq 2500 sequencer at the SNP&SEQ sequencing facility (Science for Life laboratory (SciLifeLab), Uppsala sequencing node). The reads were aligned to the Ensembl mouse gene assembly (NCBIM37) using Tophat2 software (version 2.0.4). The duplicated reads were removed using the picard tool (version 1.92). To identify the genes significantly enriched in the pericyte samples as compared with microvascular samples, statistical tests were performed using the Cufflinks tool (version 2.2.1) Results: Twenty-three genes were significantly regulated in mutant when compared to WT (False Discovery Rate < 0.05) The microvascular RNA from two male heterozygous Ins2-AKITA mice and three littermate wild-type controls were processed and sequenced on the Illumina HiSeq 2500 platform in the sequencing facility in Uppsala University.研究目的：全面解析高血糖状态下的脑血管基因表达特征。 研究方法：我们从高血糖遗传小鼠模型（Ins2-AKITA）与野生型（Wild Type, WT）小鼠分离的脑微血管片段中提取mRNA，并通过RNA测序对其转录组进行分析。样本于瑞典乌普萨拉大学科学与生命实验室（Science for Life Laboratory, SciLifeLab）的SNP&SEQ测序中心（乌普萨拉测序节点），使用Illumina HiSeq 2500测序仪完成测序。采用Tophat2软件（版本2.0.4）将测序读段（reads）比对至Ensembl数据库小鼠基因组装版本NCBIM37；利用Picard工具（版本1.92）去除重复测序读段。为鉴定相较于微血管样本显著富集的周细胞样本基因，本研究使用Cufflinks工具（版本2.2.1）开展统计学检验。 研究结果：与野生型小鼠相比，突变型小鼠体内共有23个基因发生显著表达调控（错误发现率False Discovery Rate < 0.05）。本研究共处理2只雄性杂合Ins2-AKITA小鼠与3只同窝野生型对照小鼠的微血管RNA，并于乌普萨拉大学测序中心使用Illumina HiSeq 2500平台完成测序。