QUALITATIVE AND QUANTITATIVE STUDIES ON HUMAN MYELIN BASIC PROTEIN IN SITU WITH RESPECT TO TIME INTERVAL BETWEEN DEATH AND AUTOPSY
QUALITATIVE AND QUANTITATIVE STUDIES ON HUMAN MYELIN BASIC PROTEIN IN SITU WITH RESPECT TO TIME INTERVAL BETWEEN DEATH AND AUTOPSY
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DOI：
10.1097/00005072-197603000-00005
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发表时间：
1976-03
影响因子：
3.2
通讯作者：
K. Ansari;A. Rand;H. Hendrickson;M. Bentley
中科院分区：
文献类型：
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作者：
K. Ansari;A. Rand;H. Hendrickson;M. Bentley
取自尸体解剖的正常人额叶白质被用于确定髓鞘碱性蛋白（BP）原位死后降解的程度。研究了以下两个因素的影响：1）死亡与尸检之间的时间间隔，2）组织的冷冻和解冻。从尸检材料中对BP的定量提取显示，可归因于死亡与尸检之间时间间隔（最长48小时）的BP损失极小。这些样本中纯化的BP在pH 4.3的丙烯酰胺凝胶上进行电泳，发现其电泳图谱与零小时的牛BP样本相当。从尸检样本中获得的BP在豚鼠中被发现具有致脑炎性。当用兔抗人BP血清进行检测时，纯化的BP制剂在免疫沉淀试验中产生单一弧。从冷冻一次且在解冻前处理的组织中提取和纯化的BP，其产量、致脑炎活性和丙烯酰胺圆盘凝胶电泳图谱与从未冷冻的组织中的BP相似。然而，解冻后在处理前于室温下孵育8小时的冷冻组织仅产生可提取总蛋白的13 - 25%。这种BP也具有致脑炎性，并且显示出与从未冷冻的组织中的BP相似的丙烯酰胺条带模式。通过电子显微镜检查白质样本。未冷冻的尸检材料显示髓鞘片层有一些分离。冷冻和解冻的白质切片中的髓鞘显示片层分离以及破坏。冷冻、解冻然后在室温下孵育8小时的样本除了片层分离外，还显示出致密线物质的零星缺失。
Normal human frontal lobe white matter obtained at autopsy was used to determine the extent of in situ post-mortem degradation of myelin basic protein (BP). Effects of the following two factors were studied: 1) time interval between death and autopsy, and 2) freezing and thawing the tissue. Quantita-tive extraction of BP from the autopsy material showed only minimal loss of BP that could be attributed to the time interval between death and autopsy (up to 48 h). The purified BP from these samples was electrophoresed on acrylamide gels at pH 4.3 and it was found that the electrophoretic patterns were comparable to zero hour bovine BP samples. The BP obtained from the autopsy samples was found to be encephalitogenic in guinea pigs. When tested against rabbit anti-human-BP serum, the purified BP preparations gave a single are in immunoprecipitin test. BP extracted and purified from tissue that was frozen once and processed before it could thaw showed yields, encephalitogenic activity and acrylamide disc gel electrophoretic patterns that were similar to those of BP from tissue that was never frozen. However, frozen tissue that was thawed and then incubated for 8 h at room temperature before processing yielded only 13–25% of the total extractable protein. This BP also was encephalitogenic and showed acrylamide banding pattern that was similar to BP from tissue that was never frozen. Samples of white matter were examined by electron microscope. Unfrozen autopsy material showed some separation of myelin lamellae. Myelin in sections of frozen and thawed white matter showed separation as well as disruption of the lamellae. Samples that were frozen, thawed and then incubated at room temperature for 8 h showed sporadic loss of dense line material in addition to lamellar separation.