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死后诱导的小鼠脑蛋白质组变化评估。

Assessment of post-mortem-induced changes to the mouse brain proteome.

作者信息

Hunsucker Stephen W, Solomon Benjamin, Gawryluk Jeremy, Geiger Jonathan D, Vacano Guido N, Duncan Mark W, Patterson David

机构信息

Proteomics Group, Department of Pediatrics, University of Colorado Denver School of Medicine, Aurora, Colorado 80045, USA.

出版信息

J Neurochem. 2008 May;105(3):725-37. doi: 10.1111/j.1471-4159.2007.05183.x. Epub 2007 Dec 12.

Abstract

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.

摘要

本研究旨在评估高能头部聚焦微波辐射及死后间隔时间对小鼠脑蛋白质组测量结果的影响。采用差异凝胶电泳法,比较断头处死(死后将组织在25摄氏度下保存选定时间间隔)及高能头部聚焦微波辐射后立即切除脑组织的小鼠脑蛋白水平。采用微波介导处死是因为它能全面迅速地使酶失活,同时很大程度上保留脑的细胞结构。在所分析的10块凝胶中至少8块共有的912个蛋白点中,35个(3.8%)蛋白点的水平根据动物是通过微波辐射还是断头处死而存在显著差异(t检验;p<0.05)。当动物通过断头处死时,43个蛋白点(4.7%)在死后间隔时间内水平发生变化(方差分析;p<0.05)。二维凝胶上近1000种明显的蛋白质中的绝大多数在长达4小时内是稳定的。这些数据对于脑蛋白研究具有重要意义,无论是基于对来自动物模型还是人类的组织分析。

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