Crecelius Anna, Götz Andrea, Arzberger Thomas, Fröhlich Thomas, Arnold Georg J, Ferrer Isidro, Kretzschmar Hans A
Laboratory of Molecular Neuropathology, Centre of Neuropathology and Prion Research, LMU, Munich, Germany.
Proteomics. 2008 Mar;8(6):1276-91. doi: 10.1002/pmic.200700728.
The number of proteomics studies concerning human brain samples has been increasing in recent years, in particular in the discovery of biomarkers for neurological diseases. The human brain samples are obtained from brain banks, which are interested in providing high quality human nervous tissue. In order to provide brain banks as well as scientists working in the proteomics field with measures for tissue quality, the critical factors after death, the effect of post-mortem interval (PMI) and storage temperature on the human brain proteome were investigated. This study was focused on the gray matter of the frontal cortex. The PMI was artificially prolonged from the time of autopsy (2 h after death) by storing samples at 4 degrees C or room temperature over 18, 24, and 48 h. The samples were analyzed by 2-D DIGE using a pH 4-7 gradient, revealing a time course of quantitative protein changes. The degradation of three proteins, peroxiredoxin-1, stathmin, and glial fibrillary acidic protein were further confirmed by Western-blot analysis. Proteins vulnerable to PMI were analyzed by the 2-D DIGE analysis of cortex samples from three donors, and were derived from a variety of functional groups, including metabolic, structural, stress response, antioxidants, synaptosomal, and neuronal proteins.
近年来，有关人类脑样本的蛋白质组学研究数量不断增加，尤其是在神经疾病生物标志物的发现方面。人类脑样本取自脑库，这些脑库致力于提供高质量的人类神经组织。为了给脑库以及蛋白质组学领域的科研人员提供组织质量的衡量标准，研究了死后的关键因素、死后间隔时间（PMI）和储存温度对人类脑蛋白质组的影响。本研究聚焦于额叶皮质的灰质。通过将样本在4℃或室温下储存18、24和48小时，从尸检时间（死后2小时）开始人为延长PMI。使用pH 4 - 7梯度的二维差异凝胶电泳（2-D DIGE）对样本进行分析，揭示了蛋白质定量变化的时间进程。通过蛋白质免疫印迹分析进一步证实了三种蛋白质——过氧化物还原酶-1、微管相关蛋白和胶质纤维酸性蛋白的降解。通过对来自三名捐赠者的皮质样本进行二维差异凝胶电泳分析，分析了易受PMI影响的蛋白质，这些蛋白质来自各种功能组，包括代谢、结构、应激反应、抗氧化剂、突触体和神经元蛋白。
