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小鼠脑中肌动蛋白样蛋白定位的免疫荧光研究。

Immunofluorescence studies on localization of actin-like protein in the mouse brain.

作者信息

Ishii T, Haga S, Tokutake S

出版信息

Acta Neuropathol. 1978 May 24;42(2):99-103. doi: 10.1007/BF00690974.

Abstract

Localization of actin-like protein in the mouse brain was studied by the immunofluorescence method using FITC-labeled antiactin antibody. Antibody against actin was prepared according to Lazarides (1974). The problem of poor antigenicity and the presence of contaminants in the antigen preparation was circumvented by way of denaturation of the protein with sodium dodecyl sulfate (SDS) and subsequent purification of the antigen by SDS polyacrylamide gel electrophoresis (Lazarides, 1974). Intense specific fluorescence against actin was noted in the cytoplasma of the epithelial cells of the choroid plexus, of the ependymal cells of the ventricles, and a little less one in arachnoidea and in leptomeninges, as well as in the muscle layers, endothelial linings, and adventitia of the arteries. Diffuse but weak fluorescence was noted also in the neuropil of the grey matter of the brain. Among the latter, fluorescence in the granular layer of Ammon's horn and of the cerebellum appeared clearer than the diffuse one in the rest of the grey matter. Functional significance of the presence of actin-like protein in the above tissues was discussed.

摘要

利用异硫氰酸荧光素(FITC)标记的抗肌动蛋白抗体,通过免疫荧光法研究了小鼠脑中肌动蛋白样蛋白的定位。抗肌动蛋白抗体是根据拉扎里德斯(1974年)的方法制备的。通过用十二烷基硫酸钠(SDS)使蛋白质变性,随后通过SDS聚丙烯酰胺凝胶电泳纯化抗原,解决了抗原制备中抗原性差和存在污染物的问题(拉扎里德斯,1974年)。在脉络丛上皮细胞、脑室室管膜细胞的细胞质中发现了针对肌动蛋白的强烈特异性荧光,蛛网膜和软脑膜中的荧光稍弱一些,动脉的肌层、内皮衬里和外膜中也有荧光。在脑灰质的神经纤维网中也发现了弥漫但较弱的荧光。其中,海马角颗粒层和小脑的荧光比其余灰质中的弥漫性荧光更清晰。讨论了上述组织中肌动蛋白样蛋白存在的功能意义。

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