Cell Surface Sialoglycoproteins of Cultured Rat Cerebellar Interneurons

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Abstract
The sialoglycoproteins of cultures of relatively pure rat cerebellar interneurons were labeled by NaIO4 oxidation/NaB 3H4 reduction. The labeled molecules were analyzed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate followed by fluorography. Faint labeling could be detected in 3 components if cells were labeled 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 labeled. After NaIO4 oxidation, .apprx. 80% of the cell-associated radioactivity could be removed by treating the cells with neuraminidase, which left the cells > 95% viable. The majority of the bands seen after neuraminidase treatment were substantially reduced when compared with untreated controls, supporting a surface localization of these molecules. Reproducible developmental changes were seen in the profiles of bands labeled 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.