Brain protein preservation largely depends on the postmortem storage temperature:: Implications for study of proteins in human neurologic disease and managemetn of brain banks:: A BrainNet Europe Study
Brain protein preservation largely depends on the postmortem storage temperature:: Implications for study of proteins in human neurologic disease and managemetn of brain banks:: A BrainNet Europe Study
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DOI：
10.1097/nen.0b013e31802c3e7d
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发表时间：
2007-01-01
影响因子：
3.2
通讯作者：
Kretzschmar, Hans
中科院分区：
文献类型：
--
作者：
Ferrer, Isidre;Santpere, Gabriel;Kretzschmar, Hans
本研究旨在揭示与死后延迟和储存温度相关的对照病例的蛋白质修饰，在三种范式中，相同的死后组织样本（额叶皮层）在死后短时间内冷冻或在1摄氏度，4摄氏度或室温下储存，然后在不同的时间间隔在-80摄氏度冷冻。单维凝胶电泳和Western blotting显示，在PC人工保存的样品中，在死亡后不同时间间隔冷冻50小时，未观察到蛋白质降解的证据。然而，几种蛋白质的水平在4℃下储存的样品中发生了变化，这种影响在室温下储存的样品中更为明显。二维凝胶电泳和质谱进一步证实了这些观察结果，并允许鉴定其他易受或耐死后延迟的蛋白质。最后，对阿尔茨海默病中萨科齐不溶性组分的凝胶电泳和免疫印迹分析显示，随着死后延迟，磷酸化tau特异性条带的强度降低，当大脑样本在室温下长时间保存时，这种影响更为显著。这些结果强调了在死亡后降低体温以减少蛋白质降解的必要性。
The present study was designed to reveal protein modifications in control cases related with postmortem delay and temperature of storage in 3 paradigms in which the same postmortem tissue sample (frontal cortex) was frozen a short time after death or stored at 1 degrees C, 4 degrees C, or room temperature and then frozen at -80 degrees C at different intervals. No evidence of protein degradation as revealed with monodimensional gel electrophoresis and Western blotting was observed in samples artificially stored at PC and then frozen at different intervals up to 50 hours after death. However, the levels of several proteins were modified in samples stored at 4 degrees C and this effect was more marked in samples stored at room temperature. Two-dimensional gel electrophoresis and mass spectrometry further corroborated these observations and permitted the identification of other proteins vulnerable or resistant to postmortem delay. Finally, gel electrophoresis and Western blotting of sarkosyl-insoluble fractions in Alzheimer disease showed reduced intensity of phospho-tau-specific bands with postmortem delay with the effects being more dramatic when the brain samples were stored at room temperature for long periods. These results emphasize the necessity of reducing the body temperature after death to minimize protein degradation.