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培养的少突胶质细胞中的UDP半乳糖:神经酰胺半乳糖基转移酶:一项酶学和免疫学研究。

UDPgalactose:ceramide galactosyltransferase in cultured oligodendrocytes: an enzymological and immunological study.

作者信息

Neskovic N M, Espinosa de los Monteros A, Roussel G, Nussbaum J L

机构信息

Centre de Neurochimie du CNRS, Strasbourg, France.

出版信息

Int J Dev Neurosci. 1988;6(6):535-46. doi: 10.1016/0736-5748(88)90061-5.

Abstract

The developmental expression of UDPgalactose:ceramide galactosyltransferase (CGalT), an enzyme marker of one myelinogenic activity in nervous tissue, was studied in cultured oligodendrocytes. The activity of CGalT in cultures followed a characteristic pattern of developmental changes. In the primary cultures these changes could be represented by a biphasic curve with a maximum of enzymatic activity at about the 25th day in culture. After purifying the oligodendrocytes from the primary cultures and replating them in culture dishes, similar developmental changes of CGalT were observed. In the subcultures prepared from 20-day-old primary cultures the activity of CGalT per oligodendrocyte increased from 1.3 x 10(-6) nmol/hr on day 4 to 3.7 x -6 nmol/hr on day 21. Immunocytochemical studies with the antiserum against rat brain CGalT showed the presence of CGalT+ oligodendrocytes after 7 days in the primary culture (earliest time studied), later on the number of CGalT+ oligodendrocytes increased until 28 days (latest time examined). In the subcultures of purified oligodendrocytes the bulk of oligodendrocytes was stained by the anti-CGalT antibodies after 15 days. These results suggest that the initial expression of CGalT in oligodendroglial cultures involves an increase of the number of CGalT+ oligodendrocytes and of the amount of enzyme protein per cell.

摘要

在培养的少突胶质细胞中研究了UDP半乳糖:神经酰胺半乳糖基转移酶(CGalT)的发育表达,CGalT是神经组织中一种髓鞘生成活性的酶标志物。培养物中CGalT的活性遵循发育变化的特征模式。在原代培养物中,这些变化可用双相曲线表示,在培养约第25天时酶活性最高。从原代培养物中纯化少突胶质细胞并将其重新接种到培养皿中后,观察到CGalT有类似的发育变化。在由20日龄原代培养物制备的传代培养物中,每个少突胶质细胞的CGalT活性从第4天的1.3×10^(-6) nmol/小时增加到第21天的3.7×10^(-6) nmol/小时。用抗大鼠脑CGalT抗血清进行的免疫细胞化学研究表明,原代培养7天后(研究的最早时间)存在CGalT+少突胶质细胞,随后CGalT+少突胶质细胞的数量增加,直至28天(检查的最晚时间)。在纯化少突胶质细胞的传代培养物中,15天后大部分少突胶质细胞被抗CGalT抗体染色。这些结果表明,少突胶质细胞培养物中CGalT的初始表达涉及CGalT+少突胶质细胞数量的增加以及每个细胞中酶蛋白量的增加。

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