Gene expression analysis of Peyers patches after infection of C57BL/6 mice with Yersinia enterocolitica. Mus musculus

Orogastral infection of mice with Yersinia enterocolitical leads to HIF-1 alpha activation.To elucidate whether this HIF-1 alpha activation also results in a HIF-1 dependent gene programming, the transcriptomes from Peyers Patches of uninfected and Yersinia enterocolitica infected mice were analyzed by means of of microarray analyses using Affymetrix GeneChip probe arrays (MG-U74Av2). In total, 288 genes were differentially regulated three day after infection in PP compared with the expression of uninfected control mice. Of these 288 genes, 217 were found to be differentially upregulated and from these, 14 genes ( 6.5% of all upregulated genes) are well described to be regulated via HIF-1. These data indicate that orogatral infection with Y. enterocolitica results in HIF-1 dependent gene programmning Keywords: Time course Overall design: Per group five C57BL/6 mice were infected orogastrally with 500 Million Yersinia enterocolitica. 1 and 3 days after infection, Peyers Patches were removed and total RNA was prepared. In parallel, RNA was isolated from uninfected mice. The generation of fragmented cRNA was performed following the manufacturers instructions and used for hybridization onto GeneChip arrays MG-U74Avs2. Analysis of microarray data was performed using the Affymetrix Microarray Suite 5.0, Affymetrix Mining Tool 3.0. A median signal log2 ratio (SLR) grater than 1.5 or less than -1.5 was considered a significant change.

小鼠经口感染小肠结肠炎耶尔森菌（Yersinia enterocolitica）可诱导缺氧诱导因子-1α（HIF-1α）激活。为明确该HIF-1α激活是否可介导HIF-1依赖型基因表达程序，本研究采用Affymetrix GeneChip探针阵列（MG-U74Av2）开展微阵列分析，对未感染及经小肠结肠炎耶尔森菌感染小鼠的派尔集合淋巴结（Peyer's Patches）转录组进行检测。感染后3天，与未感染对照组小鼠相比，派尔集合淋巴结中共计288个基因的表达水平存在显著差异。其中217个基因表达上调，在上述上调基因中，有14个（占全部上调基因的6.5%）已被证实为HIF-1依赖型调控基因。以上结果表明，经口感染小肠结肠炎耶尔森菌可触发HIF-1依赖型基因表达程序。

关键词：时间进程

实验设计：每组选取5只C57BL/6小鼠，经口感染5×10^8株小肠结肠炎耶尔森菌；分别于感染后1天和3天摘取其派尔集合淋巴结并提取总RNA，同时设置未感染对照组小鼠并提取对应RNA。按照试剂生产商的说明书制备片段化互补RNA（cRNA），将其用于MG-U74Av2型GeneChip阵列杂交。采用Affymetrix Microarray Suite 5.0及Affymetrix Mining Tool 3.0软件进行微阵列数据分析，以中位数信号对数比值（signal log2 ratio, SLR）的绝对值大于1.5作为基因表达显著变化的判定标准。