Assessment of post-mortem-induced changes to the mouse brain proteome
Assessment of post-mortem-induced changes to the mouse brain proteome
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DOI：
10.1111/j.1471-4159.2007.05183.x
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发表时间：
2008-05-01
影响因子：
4.7
通讯作者：
Patterson, David
中科院分区：
文献类型：
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作者：
Hunsucker, Stephen W.;Solomon, Benjamin;Patterson, David
本研究旨在评估高能头部微波辐照和死后时间对小鼠脑蛋白质组测量的影响。差异凝胶电泳用于比较被斩首杀死的小鼠脑蛋白水平，其中组织在死后25摄氏度的选定时间间隔中保存，并通过高能头部聚焦微波照射，然后立即切除。微波介导的杀伤被使用，因为它全面地使酶快速失活，同时在很大程度上保留了脑细胞结构。在被分析的10种凝胶中，至少8种凝胶共有912个蛋白点，其中35个（3.8%）显示出显著的水平差异（t检验；p < 0.05），这取决于动物是被微波辐射杀死还是被斩首。当动物被斩首杀死时，43个蛋白斑点（4.7%）在死后出现水平变化（方差分析，p < 0.05）。2D凝胶上近1000种蛋白质中的绝大多数在长达4小时的时间内都是稳定的。这些数据对研究大脑中的蛋白质具有重要意义，无论是基于动物模型还是人类组织的分析。
This study was designed to assess the influence of high-energy head-focused microwave irradiation and the post-mortem interval on measurements of the mouse brain proteome. Difference gel electrophoresis was used to compare mouse brain protein levels in animals killed by decapitation, where the tissue was held at 25 degrees C for selected time intervals post-mortem, and by high-energy head-focused microwave irradiation followed by immediate resection. Microwave-mediated killing was used because it comprehensively snap-inactivates enzymes while largely retaining brain cytoarchitecture. Of the 912 protein spots common to at least eight of 10 gels analyzed, 35 (3.8%) showed significant differences in levels (t-test; p < 0.05) depending on whether animals were killed by microwave irradiation or decapitation. When animals were killed by decapitation, 43 protein spots (4.7%) showed changes in levels over the post-mortem interval (ANOVA; p < 0.05). The vast majority of the near 1000 proteins evident on a 2D gel were stable for up to 4 h. These data have important implications for studies of proteins in the brain, whether based on analysis of tissue derived from animal models or from humans.