Ervin John F, Heinzen Erin L, Cronin Kenneth D, Goldstein David, Szymanski Mari H, Burke James R, Welsh-Bohmer Kathleen A, Hulette Christine M
Department Medicine (Neurology), Joseph and Kathleen Bryan Alzheimer's Disease Research Center, Duke University Medical Center, Durham, North Carolina 27710, USA.
J Neuropathol Exp Neurol. 2007 Dec;66(12):1093-9. doi: 10.1097/nen.0b013e31815c196a.
The Bryan Alzheimer Disease Research Center obtains postmortem human brain tissue from patients with Alzheimer disease (AD) and cognitively normal control subjects for molecular and genetic research programs. A growing body of research suggests that variations in gene transcript levels may contribute to the onset and progression of disease. Identifying how the regulation of gene expression may affect AD requires the use of high-quality mRNA from banked human brains. The present study was conducted to establish the quality and suitability of available banked brain tissue for future gene expression studies. We chose 32 AD cases with Braak stage IV, V, or VI. These AD cases were matched to 36 normal control cases by age and sex when possible. Multiple regions from each brain were sampled, including frontal cortex, temporal cortex, occipital cortex, and cerebellum. Hippocampus was also available for study from 14 control cases. A comparison of several antemortem and postmortem variables, such as postmortem interval, agonal state, ventricular cerebrospinal fluid pH, and cause of death were analyzed. RNA was isolated from at least 1 area from every brain and most brains yielded intact RNA from all regions tested. Analysis of the clinical variables did not reveal any features that correlated with the ability to recover intact mRNA. We conclude that undegraded mRNA may be isolated from most brain regions many hours postmortem and that neither the pH of ventricular fluid nor postmortem interval is predictive of mRNA integrity.
布莱恩阿尔茨海默病研究中心从阿尔茨海默病（AD）患者和认知正常的对照受试者处获取死后人类脑组织，用于分子和基因研究项目。越来越多的研究表明，基因转录水平的变化可能导致疾病的发生和发展。确定基因表达调控如何影响AD需要使用来自储存人脑的高质量mRNA。本研究旨在确定现有储存脑组织的质量和适用性，以供未来的基因表达研究使用。我们选择了32例Braak分期为IV、V或VI期的AD病例。这些AD病例尽可能按年龄和性别与36例正常对照病例匹配。从每个大脑的多个区域取样，包括额叶皮质、颞叶皮质、枕叶皮质和小脑。14例对照病例的海马体也可供研究。分析了几个生前和死后变量，如死后间隔、濒死状态、脑室脑脊液pH值和死因。从每个大脑至少1个区域分离RNA，大多数大脑在所有测试区域都产生了完整的RNA。对临床变量的分析未发现与完整mRNA恢复能力相关的任何特征。我们得出结论，死后数小时可从大多数脑区分离出未降解的mRNA，脑室液pH值和死后间隔均不能预测mRNA的完整性。
