Reference genes for normalising gene expression data in collagenase-induced rat intracerebral haemorrhage.
Reference genes for normalising gene expression data in collagenase-induced rat intracerebral haemorrhage.
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DOI：
10.1186/1471-2199-11-7
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发表时间：
2010-01-20
影响因子：
--
通讯作者：
Vink R
中科院分区：
文献类型：
--
作者：
Cook NL;Kleinig TJ;van den Heuvel C;Vink R
脑出血(ICH)后脑损伤的机制尚不完全清楚。应用实时定量RT-PCR研究脑出血后的基因表达增加了我们对这些机制的理解，然而观察到的不一致的结果可能与不适当的参考基因选择有关。参考基因应该在不同的实验条件下稳定表达，然而，共同的参考基因的转录水平已经被证明有很大的差异。因此，已经提出了参考基因小组来克服这种潜在的混杂因素。本研究评估了胶原酶诱导的啮齿类动物脑出血后5小时和24小时内纹状体和上皮层7个候选参考基因的稳定性。使用geNorm对候选参考基因的转录水平进行量化，并按稳定性排序。当我们的感兴趣的基因，瞬时受体潜力Melastatin 2(TRPM2)，分别与每个参考基因正常化时，TRPM2的mRNA水平是高度可变的。当归一化为准确归一化数据选择的四个最稳定的参考基因时，我们发现ICH和Vehicle大鼠之间没有显著差异。本研究中确定的一组参考基因将使实验性脑出血急性期的基因表达数据更准确地正常化。
The mechanisms of brain injury following intracerebral haemorrhage (ICH) are incompletely understood. Gene expression studies using quantitative real-time RT-PCR following ICH have increased our understanding of these mechanisms, however the inconsistent results observed may be related to inappropriate reference gene selection. Reference genes should be stably expressed across different experimental conditions, however, transcript levels of common reference genes have been shown to vary considerably. Reference gene panels have therefore been proposed to overcome this potential confounder. The present study evaluated the stability of seven candidate reference genes in the striatum and overlying cortex of collagenase-induced ICH in rodents at survival times of 5 and 24 hours. Transcript levels of the candidate reference genes were quantified and ranked in order of stability using geNorm. When our gene of interest, transient receptor potential melastatin 2 (TRPM2), was normalised against each reference gene individually, TRPM2 mRNA levels were highly variable. When normalised to the four most stable reference genes selected for accurate normalisation of data, we found no significant difference between ICH and vehicle rats. The panel of reference genes identified in the present study will enable more accurate normalisation of gene expression data in the acute phase of experimental ICH.
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评估合适的参考基因用于牛肌肉组织中的基因表达研究。
影响因子：
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作者：
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