Masco D, Flott B, Seifert W
Department of Neurobiology, Max-Planck Institut für Biophysikalische Chemie, Göttingen, Federal Republic of Germany.
Glia. 1989;2(4):231-40. doi: 10.1002/glia.440020404.
Ganglioside GM1 3H-labelled at the terminal galactose was added to astrocyte cell cultures. GM1 incorporation was studied in the two typical forms of astrocytes in cell culture of flat and stellate morphology. There was a strong time- and concentration-dependent increase in GM1 incorporation for both cell types of astrocytes. The incorporation of GM1 into the stellate form increased continuously up to 48 h (maximum time studied), while the incorporation into the flat form reached a plateau at the same time. After 2 h of GM1 incubation additional gangliosides appeared; the latter resulted from the metabolism of the GM1 incorporated, indicating that astrocytes in cell culture can biosynthesize more complex gangliosides. To confirm that GM1 was indeed incorporated into astrocytes, two other different approaches were used. Astrocyte cells treated with 3H-GM1 were visualized using autoradiography. The specific marker for GM1, rhodamine-labelled choleratoxin, was used to detect the incorporated GM1 using fluorescence microsocpy. In both cases GM1 treated cells were intensely labelled. These observations indicate that exogenous GM1 ganglioside can also be integrated into the astrocyte membranes as occurs in other types of cells and membranes.
将末端半乳糖用³H标记的神经节苷脂GM1添加到星形胶质细胞培养物中。在扁平形态和星状形态的细胞培养中的两种典型星形胶质细胞类型中研究了GM1的掺入情况。两种星形胶质细胞类型的GM1掺入均呈现出强烈的时间和浓度依赖性增加。GM1掺入星状形态细胞的量持续增加直至48小时(研究的最长时间),而掺入扁平形态细胞的量在同一时间达到平台期。GM1孵育2小时后出现了其他神经节苷脂;后者是由掺入的GM1代谢产生的,表明细胞培养中的星形胶质细胞可以生物合成更复杂的神经节苷脂。为了证实GM1确实掺入了星形胶质细胞,使用了另外两种不同的方法。用³H-GM1处理的星形胶质细胞通过放射自显影进行可视化。GM1的特异性标记物,罗丹明标记的霍乱毒素,用于通过荧光显微镜检测掺入的GM1。在这两种情况下,GM1处理的细胞都被强烈标记。这些观察结果表明,外源性神经节苷脂GM1也可以像在其他类型的细胞和膜中一样整合到星形胶质细胞膜中。