Skrivanek J A, Ledeen R W, Margolis R U, Margolis R K
J Neurobiol. 1982 Mar;13(2):95-106. doi: 10.1002/neu.480130202.
To study ganglioside distribution within subcellular components and test the hypothesis that they are localized at the nerve ending, microsomes and synaptic plasma membranes were isolated from young adult rat brains and compared with respect to ganglioside composition. These were shown to be heterogeneous preparations by fractionation on a discontinuous sucrose gradient into subfractions which had differing ganglioside concentrations. The highest ganglioside concentrations occurred in membranes banding at the 0.8M/1.0M and 1.0M/1.3M interfaces for both microsomes and synaptic plasma membranes. These subfractions had closely similar ganglioside concentrations and pattern distributions. In addition, the kinetics of ganglioside labeling following administration of [3H]-glucosamine were similar for the two preparations. The fact that microsomal subfractions representing heterogeneous mixtures of brain cell membranes showed close similarity to synaptosomal plasma membranes argues against localization of gangliosides at the nerve ending. These results, together with other lines of evidence, support the concept that gangliosides are distributed over large portions of the neuron (and perhaps other brain cells). Data concerning the labeling of gangliosides in different microsomal subfractions indicated a movement of label over time from the more dense to the less dense membranes, as was also noted for the glycoproteins in the same subfractions. Specific radioactivity of the gangliosides increased relative to that of the glycoproteins with time.
为了研究神经节苷脂在亚细胞成分中的分布,并验证它们定位于神经末梢的假说,从成年幼鼠大脑中分离出微粒体和突触质膜,并比较它们的神经节苷脂组成。通过在不连续蔗糖梯度上分级分离成具有不同神经节苷脂浓度的亚组分,结果表明这些是异质制剂。对于微粒体和突触质膜,最高的神经节苷脂浓度出现在0.8M/1.0M和1.0M/1.3M界面处的膜中。这些亚组分具有非常相似的神经节苷脂浓度和模式分布。此外,给予[3H]-葡糖胺后神经节苷脂标记的动力学在两种制剂中相似。代表脑细胞膜异质混合物的微粒体亚组分与突触体质膜显示出密切相似性,这一事实反对神经节苷脂定位于神经末梢。这些结果与其他证据一起,支持了神经节苷脂分布在神经元的大部分区域(也许还有其他脑细胞)的概念。关于不同微粒体亚组分中神经节苷脂标记的数据表明,随着时间的推移,标记物从密度较高的膜向密度较低的膜移动,同一亚组分中的糖蛋白也有此现象。随着时间的推移,神经节苷脂的比放射性相对于糖蛋白增加。