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首页> 外文期刊>Journal of pharmacological sciences. >Screening for control genes in mouse hippocampus after transient forebrain ischemia using high-density oligonucleotide array.
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Screening for control genes in mouse hippocampus after transient forebrain ischemia using high-density oligonucleotide array.

机译：使用高密度寡核苷酸阵列筛选短暂性前脑缺血后小鼠海马中的对照基因。

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摘要

In conventional relative gene expression analysis (Northern blotting, RT-PCR, and in situ hybridization), housekeeping genes such as the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and beta-actin genes, whose expression levels are considered stable, have been used as control genes for normalization of RNA quantitation. However, it has been reported that the expression levels of these two control genes are affected by ischemia. Therefore, we have been searching for novel control genes whose expression levels are stable in a mouse model of transient forebrain ischemia. Using the GeneChip Mu6500 array set, we monitored the expression levels of approximately 6000 murine genes in the mouse hippocampus during 24 h of ischemia-reperfusion. To select stable genes, we applied the restricted criterion of a 1.5-fold change in expression level as the threshold. By adding statistical analysis with this criterion, we identified 10 genes as candidates for control genes from the GeneChip data. In this criterion, GAPDH and beta-actin genes were not included in the 10 genes as candidates for control genes. The present findings might be relevant to the use of control genes in quantitation of RNA, particularly in the study of mouse transient forebrain ischemia.
机译：在常规的相对基因表达分析中（Northern印迹，RT-PCR和原位杂交），使用了看守基因，例如3-磷酸甘油醛脱氢酶（GAPDH）和β-肌动蛋白基因，它们的表达水平被认为是稳定的。作为用于RNA定量标准化的对照基因。但是，据报道这两个对照基因的表达水平受局部缺血的影响。因此，我们一直在寻找其表达水平在短暂性前脑缺血的小鼠模型中稳定的新型控制基因。使用GeneChip Mu6500阵列组，我们监测了缺血再灌注24小时内小鼠海马中约6000个鼠类基因的表达水平。为了选择稳定的基因，我们将表达水平的1.5倍变化的限制性标准用作阈值。通过添加具有此标准的统计分析，我们从GeneChip数据中确定了10个基因作为对照基因的候选者。在此标准中，GAPDH和β-肌动蛋白基因不包含在10个基因中作为对照基因的候选基因。目前的发现可能与在RNA定量中使用控制基因有关，特别是在小鼠短暂性前脑缺血的研究中。

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