The use of real-time PCR analysis in a gene expression study of Alzheimer's disease post-mortem brains
The use of real-time PCR analysis in a gene expression study of Alzheimer's disease post-mortem brains
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DOI：
10.1016/j.jneumeth.2003.09.005
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发表时间：
2004-01-15
影响因子：
3
通讯作者：
Reddy, PH
中科院分区：
文献类型：
--
作者：
Gutala, RV;Reddy, PH
神经退行性疾病患者大脑中基因表达的测量是大脑研究的一个主要领域。本研究的目的是确定定量实时PCR是否可以测量死后间隔超过12 h的脑中信使RNA（mRNA）的表达。在本文中，我们研究了RNA的质量从脑标本的阿尔茨海默病（AD）患者（n = 13）和非痴呆正常对照组（n = 6）。为了确定AD中不受调节的内源性参考基因，我们测量了常用参考基因β-肌动蛋白、18 S rRNA和GAPDH的mRNA表达。此外，我们确定是否死后间隔，脑重量，或死亡时的年龄影响mRNA的表达。我们的实时PCR分析结果表明，在所有脑标本中均可检测到β-肌动蛋白、18 S rRNA、GAPDH和突触素（AD的一种已知标记物）的mRNA表达。此外，使用实时PCR分析，我们发现β-肌动蛋白和18 S rRNA在AD和对照受试者的脑标本中差异表达，而GAPDH在AD和对照脑标本中表达相似。这些结果表明，GAPDH可以作为一个内源性的参考基因在AD脑的研究。比较基因表达分析还表明，突触体蛋白是下调AD脑标本相比，控制脑标本。(C)2003 Elsevier B. V.保留所有权利。
The measurement of gene expressions in brains with neurodegenerative diseases is a major area of brain research. The objective of our research was to determine whether quantitative real-time PCR could measure messenger RNA (mRNA) expression in brains with post-mortem intervals beyond 12 h. In the present paper, we examined the quality of RNA from brain specimens of both Alzheimer's disease (AD) patients (n = 13) and non-demented normal control subjects (n = 6). To determine a unregulated endogenous reference gene in AD, we measured mRNA expressions of the commonly used reference genes beta-actin, 18S rRNA, and GAPDH. In addition, we determined whether post-mortem interval, brain weight, or age at death influences mRNA expression. Our real-time PCR analysis results indicate that mRNA expression can be detected in all brain specimens for beta-actin, 18S rRNA, GAPDH, and also synaptophysin, a known marker for AD. Further, using real-time PCR analysis, we found that beta-actin and 18S rRNA are differentially expressed in the brain specimens of both AD and control subjects, while GAPDH is similarly expressed in AD and control brain specimens. These findings suggest that GAPDH can be used as a endogenous reference gene in the study of AD brains. A comparative gene expression analysis also suggests that synaptophysin is down-regulated in AD brain specimens compared to control brain specimens. (C) 2003 Elsevier B.V. All rights reserved.