Webb M
J Neurochem. 1983 Mar;40(3):769-76. doi: 10.1111/j.1471-4159.1983.tb08045.x.
The sialoglycoproteins of cultures of relatively pure rat cerebellar interneurons were labelled by NaIO4 oxidation/NaB 3H4 reduction. The labelled molecules were analysed by polyacrylamide gel electrophoresis in sodium dodecyl sulphate followed by fluorography. Faint labelling could be detected in three components if cells were labelled without any oxidation. In young cultures, oxidation by galactose oxidase alone failed to reveal any additional bands. After oxidation by NaIO4 or galactose oxidase in the presence of neuraminidase, many more components were labelled. After NaIO4 oxidation, about 80% of the cell-associated radioactivity could be removed by treating the cells with neuraminidase, which left the cells more than 95% viable. The majority of the bands seen after neuraminidase treatment were substantially reduced when compared with untreated controls, supporting a surface localisation of these molecules. Reproducible developmental changes were seen in the profiles of bands labelled by NaIO4/NaB 3H4 in time course studies of cultures up to 8 days in vitro. Some bands became more prominent, and others disappeared. The gel profiles of the neuron cultures were quite distinct from those of cerebellar astrocyte cultures, which contain all the cell types likely to be contaminants of the neuron cultures.
相对纯的大鼠小脑中间神经元培养物中的唾液糖蛋白通过高碘酸钠氧化/硼氢化钠还原进行标记。标记的分子通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳分析，随后进行放射自显影。如果细胞未经任何氧化进行标记，在三个组分中可检测到微弱的标记。在年轻培养物中，仅用半乳糖氧化酶氧化未能显示任何额外的条带。在神经氨酸酶存在下用高碘酸钠或半乳糖氧化酶氧化后，有更多的组分被标记。高碘酸钠氧化后，用神经氨酸酶处理细胞可去除约80%的细胞相关放射性，处理后细胞活力超过95%。与未处理的对照相比，神经氨酸酶处理后看到的大多数条带明显减少，这支持了这些分子位于细胞表面。在体外培养长达8天的时间进程研究中，通过高碘酸钠/硼氢化钠标记的条带图谱中观察到了可重复的发育变化。一些条带变得更加明显，而其他条带消失。神经元培养物的凝胶图谱与小脑星形胶质细胞培养物的凝胶图谱截然不同，小脑星形胶质细胞培养物包含所有可能是神经元培养物污染物的细胞类型。
