Dahl D, Selkoe D J, Pero R T, Bignami A
J Neurosci. 1982 Jan;2(1):113-9. doi: 10.1523/JNEUROSCI.02-01-00113.1982.
Using anti-chicken brain neurofilament antisera, Alzheimer's neurofibrillary tangles from two patients with senile dementia were stained by immunofluorescence and by the peroxidase-antiperoxidase procedure in cryostat sections of hippocampus and frontal cortex. In sections of cerebellum obtained from the same patients, the distribution of immunostaining was the same as that observed in experimental animals: Purkinje cell baskets and nerve fibers in the inner half of the molecular layer were demonstrated selectively. The immunostaining of the tangles was abolished when the antisera were absorbed by their own antigen, by bovine brain filament preparations, or by the fraction of bovine brain filament preparations nonabsorbed on anti-glial fibrillary acidic (GFA) protein immunoaffinity columns. Absorption with a bovine microtubule preparation isolated by two cycles of the assembly-disassembly procedure did not abolish the staining. Immunostaining experiments conducted on bovine brain filament preparations resolved on sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the antisera staining the tangles reacted with the 200,000-, 150,000-, and 70,000-dalton neurofilament polypeptides. Antisera raised to the 150,000- dalton bovine neurofilament polypeptide isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis stained the tangle much less intensely, although Purkinje cell baskets in the cerebellum appeared well stained. No staining of neurofibrillary tangles was observed with antisera to other classes of 10-nm filament proteins (GFA protein, vimentin, and desmin).
使用抗鸡脑神经丝抗血清,对两名老年痴呆症患者的阿尔茨海默病神经原纤维缠结进行免疫荧光染色,并在海马体和额叶皮质的低温切片中通过过氧化物酶-抗过氧化物酶法进行染色。在从同一患者获取的小脑切片中,免疫染色的分布与在实验动物中观察到的相同:选择性地显示了浦肯野细胞篮和分子层内半部分的神经纤维。当抗血清被其自身抗原、牛脑丝制剂或未被抗胶质纤维酸性(GFA)蛋白免疫亲和柱吸附的牛脑丝制剂部分吸收时,缠结的免疫染色被消除。用通过两轮组装-拆卸程序分离的牛微管制剂进行吸收并没有消除染色。对在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳上分离的牛脑丝制剂进行的免疫染色实验表明,对缠结进行染色的抗血清与200,000、150,000和70,000道尔顿的神经丝多肽发生反应。针对通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离的150,000道尔顿牛神经丝多肽产生的抗血清对缠结的染色强度要低得多,尽管小脑中的浦肯野细胞篮染色良好。用针对其他类别的10纳米丝蛋白(GFA蛋白、波形蛋白和结蛋白)的抗血清未观察到神经原纤维缠结的染色。