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体外大鼠小脑细胞中岩藻糖基共轭物的分布:四角豆凝集素和欧洲荆豆凝集素的结合模式

Distribution of fucosyl-conjugates in rat cerebellar cells in vitro: binding patterns of Lotus tetragonolobus and Ulex europeus lectins.

作者信息

Minier L N, Erickson P F, Lasher R S

出版信息

Brain Res. 1982 May;256(1):13-21. doi: 10.1016/0165-3806(82)90092-x.

Abstract

Light microscope radioautographic procedures were employed to investigate the binding patterns of lectins derived from Lotus tetragonolobus (Lotus A) and Ulex europeus (UEA I) to the surfaces of rat cerebellar cells maintained in dispersed cell culture. Lotus A bound extensively to the surfaces of neuronal cell somata and neuronal processes at all time points investigated; the lectin also bound to nonneuronal cell surfaces, but to an extent much less than that for neurons. Lectin binding was completely inhibited by the presence of 0.1 M alpha-L-fucose in the reaction medium. Conversely, UEA I did not appear to bind to any significant extent to the surfaces of cerebellar cells in vitro at any stage of maintenance. The incorporation of [3H]fucose into fucosyl-conjugates by cerebellar neurons in culture was investigated using light microscope radioautography and polyacrylamide gel electrophoretic methods. Radioautographs indicated that the fucose came to be distributed throughout neuronal cell somata and processes and that, with increasing time, the grain density appeared to be highest over areas of neuropil and fascicles of cellular processes. The electrophoresis data demonstrated that, among others, the label was incorporated into prominent classes of polypeptides having nominal molecular weights of 400,000, 125,000, 91,000 and 46,000 daltons.

摘要

采用光学显微镜放射自显影技术,研究了来自四角豆(莲花凝集素A)和欧洲荆豆(荆豆凝集素I)的凝集素与分散细胞培养中大鼠小脑细胞表面的结合模式。在所有研究的时间点,莲花凝集素A广泛结合于神经元细胞体和神经元突起的表面;该凝集素也结合于非神经元细胞表面,但程度远低于神经元。反应介质中存在0.1 M α-L-岩藻糖时,凝集素结合被完全抑制。相反,在培养的任何阶段,荆豆凝集素I在体外似乎都没有明显结合于小脑细胞表面。利用光学显微镜放射自显影和聚丙烯酰胺凝胶电泳方法,研究了培养的小脑神经元将[3H]岩藻糖掺入岩藻糖缀合物的情况。放射自显影片表明,岩藻糖分布于整个神经元细胞体和突起,并且随着时间的增加,颗粒密度在神经毡区域和细胞突起束上似乎最高。电泳数据表明,除其他外,标记物掺入了标称分子量为400,000、125,000、91,000和46,000道尔顿的显著多肽类中。

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