Grundke-Iqbal I, Iqbal K, Tung Y C, Wisniewski H M
Acta Neuropathol. 1984;62(4):259-67. doi: 10.1007/BF00687607.
Antisera to isolated Alzheimer neurofibrillary tangles (ANT) of paired helical filaments ( PHF ) were raised in rabbits. These anti- PHF sera immunolabeled both ANT in sections of Alzheimer hippocampus and ANT which were isolated and extracted with sodium dodecyl sulfate (SDS). The immunostaining of ANT in tissue sections was removed by absorption of the anti- PHF serum with small amounts of PHF and also with 40-fold the amount of a fraction prepared identically from normal brain; neurofilament and brain microtubule preparations used at the same concentration as the normal brain control fraction did not eliminate the tangle staining. Furthermore, the tangle staining was also not removed with glial filaments or actin and myosin filaments. No labeling of the neurofilaments of axons and cerebellar basket fibers by anti- PHF sera was observed in tissue sections from non-neurologic brain. On paper blots of SDS-polyacrylamide gels anti- PHF serum reacted with neither polypeptides of the normal brain control fraction nor major microtubule and neurofilament polypeptides. However, the immunoblots of PHF preparations with the anti- PHF serum revealed staining of several polypeptide bands in the 45,000-70,000 molecular weight (MW) region, material on top of the gel and diffuse staining of the high MW region. The tangles staining in tissue sections by the anti- PHF serum was abolished by its absorption with PHF polypeptides extracted from high and low molecular weight areas of SDS polyacrylamide gels but not with identically prepared neurofilament polypeptides.(ABSTRACT TRUNCATED AT 250 WORDS)
在兔体内制备了针对成对螺旋丝(PHF)中分离出的阿尔茨海默病神经原纤维缠结(ANT)的抗血清。这些抗PHF血清在阿尔茨海默病海马体切片中的ANT以及用十二烷基硫酸钠(SDS)分离提取的ANT上均产生免疫标记。通过用少量PHF以及与正常脑相同方法制备的、量为其40倍的组分吸收抗PHF血清,可去除组织切片中ANT的免疫染色;与正常脑对照组分相同浓度使用的神经丝和脑微管制剂并不能消除缠结染色。此外,用胶质丝或肌动蛋白和肌球蛋白丝也不能去除缠结染色。在非神经病变脑的组织切片中,未观察到抗PHF血清对轴突神经丝和小脑篮状纤维的标记。在SDS-聚丙烯酰胺凝胶的纸上印迹中,抗PHF血清与正常脑对照组分的多肽以及主要的微管和神经丝多肽均无反应。然而,用抗PHF血清对PHF制剂进行免疫印迹时,在分子量(MW)45,000 - 70,000区域显示出几条多肽带的染色以及凝胶上方物质和高分子量区域的弥漫性染色。抗PHF血清在组织切片中的缠结染色可通过用从SDS聚丙烯酰胺凝胶的高分子量和低分子量区域提取的PHF多肽吸收而消除,但用相同制备的神经丝多肽则不能。(摘要截短至250字)