Assessing quantitative post-mortem changes in the gray matter of the human frontal cortex proteome by 2-D DIGE
Assessing quantitative post-mortem changes in the gray matter of the human frontal cortex proteome by 2-D DIGE
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DOI：
10.1002/pmic.200700728
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发表时间：
2008-03-01
期刊：
影响因子：
3.4
通讯作者：
Kretzschmar, Hans A.
中科院分区：
文献类型：
--
作者：
Crecelius, Anna;Goetz, Andrea;Kretzschmar, Hans A.
近年来，有关人脑样本的蛋白质组学研究数量不断增加，特别是在发现神经系统疾病的生物标志物方面。人脑样本来自脑库，这些脑库对提供高质量的人类神经组织感兴趣。为了给脑库和蛋白质组学领域的科学家提供组织质量的测量方法，研究了死亡后的关键因素、死后间隔时间和储存温度对人脑蛋白质组的影响。这项研究的重点是额叶皮层的灰质。通过在4摄氏度或室温下保存样品超过18、24和48小时，人为延长尸检时间（死亡后2小时）的PMI。样品使用pH 4-7梯度进行二维DIGE分析，揭示定量蛋白质变化的时间过程。Western-blot分析进一步证实了三种蛋白的降解，即过氧化物还原素-1、stathmin和胶质纤维酸性蛋白。通过对来自三个供体的皮质样品的二维DIGE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基团，包括代谢蛋白、结构蛋白、应激反应蛋白、抗氧化剂、突触体蛋白和神经元蛋白。
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 postmortem 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.