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首页> 外文期刊>Neurochemistry International: The International Journal for the Rapid Publication of Critical Reviews, Preliminary and Original Research Communications in Neurochemistry >Rapid and gentle extraction, reconstitution and characterization of microfilament and glia filament from rat astrocytes.
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Rapid and gentle extraction, reconstitution and characterization of microfilament and glia filament from rat astrocytes.

机译：从大鼠星形胶质细胞快速轻柔地提取，重建和表征微丝和神经胶质丝。

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摘要

We developed gentle and rapid methods for depolymerization and extraction of both microfilament and glia filament separately from a crude cytoskeletal fraction of rat astrocytes. Electron microscopy revealed that the filament reconstituted from the microfilament extract closely resembled F-actin that was formed from G-actin of rabbit skeletal muscle. It was found by immunoblotting analysis that even the reconstituted microfilament-like filaments, which had been purified by affinity chromatography with heavy meromyosin subfragment 1 (S1)-conjugated Sepharose, contained vimentin and glia fibrillary acidic protein (GFAP) besides actin, inferring the interaction between microfilament and glia filament. The filaments (9-10 nm thick) reconstituted from the glia filament extract were composed of actin and other minor components in addition to vimentin and GFAP. Actin, GFAP, 101, 34, 32.5, 30.5, 29.5 and 28 kDa proteins found in the reconstituted glia filament-like filaments were suggested to be glia filament-associated proteins.
机译：我们开发了温和，快速的方法，分别从大鼠星形胶质细胞的粗细胞骨架部分解聚和提取微丝和神经胶质丝。电子显微镜显示，由微丝提取物重构的细丝与由兔骨骼肌的G-肌动蛋白形成的F-肌动蛋白极为相似。通过免疫印迹分析发现，即使是重组的微丝状细丝，也就是用重肌球蛋白亚片段1（S1）偶联的琼脂糖凝胶经亲和层析纯化的，除肌动蛋白外，还含有波形蛋白和胶质纤维酸性蛋白（GFAP），从而推测相互作用在微丝和神经胶质丝之间。由胶质细丝提取物重构而成的细丝（9-10纳米厚）除肌动蛋白和GFAP外还由肌动蛋白和其他次要成分组成。重组胶质丝状丝中发现的肌动蛋白，GFAP，101、34、32.5、30.5、29.5和28 kDa蛋白被认为是胶质丝相关蛋白。

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