Leskawa K C, Hogan E L
J Neurosci Res. 1985;13(4):539-50. doi: 10.1002/jnr.490130409.
Individual ganglioside species (possessing the gangliotetrose oligosaccharide) were purified from bovine brain gray matter and applied in varying concentrations to the culture medium of mouse neuroblastoma cells (N2A) in vitro. After 48 hr of incubation, the cells were stained, and the neuritogenic response quantitated with a video analysis system, employing a program to measure three parameters of neuroblastoma differentiation: neurites per cell (sprouting), neurite length (extension), and degree of neurite branching (arborization). All the individual gangliosides tested promoted neurite extension in a dose-dependent fashion. Asialogangliosides ("neutral" glycosphingolipids) were without effect, which suggests that sialic acid (N-acetylneuraminic acid) is necessary to elicit this cellular response. With increasing concentrations of GM1 (5 to 500 micrograms/ml), the average cellular neurite length increased significantly, whereas the number of neurites per cell decreased. With the trisialoganglioside GT1b, neurite length did not increase to the extent seen with GM1, but an increase in the number of neurites per cell (sprouting) and branch points per neurite (arborization) was observed. These results suggest that the in vitro neuronal response to exogenous gangliosides may combine specific responses to individual species making up the total.
从牛脑灰质中纯化出具有神经节四糖寡糖的单个神经节苷脂种类，并将其以不同浓度应用于体外培养的小鼠神经母细胞瘤细胞（N2A）的培养基中。孵育48小时后，对细胞进行染色，并用视频分析系统对神经突生成反应进行定量，该系统使用一个程序来测量神经母细胞瘤分化的三个参数：每个细胞的神经突数量（发芽）、神经突长度（延伸）和神经突分支程度（分支）。所有测试的单个神经节苷脂均以剂量依赖性方式促进神经突延伸。去唾液酸神经节苷脂（“中性”糖鞘脂）没有作用，这表明唾液酸（N-乙酰神经氨酸）是引发这种细胞反应所必需的。随着GM1浓度的增加（5至500微克/毫升），平均细胞神经突长度显著增加，而每个细胞的神经突数量减少。对于三唾液酸神经节苷脂GT1b，神经突长度没有增加到GM1所观察到的程度，但观察到每个细胞的神经突数量（发芽）和每个神经突的分支点数量（分支）增加。这些结果表明，体外神经元对外源神经节苷脂的反应可能结合了对构成总体的单个种类的特定反应。
