Notterpek L M, Bullock P N, Malek-Hedayat S, Fisher R, Rome L H
Department of Biological Chemistry, UCLA School of Medicine 90024-1737.
J Neurosci Res. 1993 Dec 15;36(6):621-34. doi: 10.1002/jnr.490360603.
Myelin deposition and maintenance are critical to proper function of the mammalian nervous system. Previous investigations of myelination in the central nervous system (CNS) were hampered by the lack of an in vitro system that can faithfully reproduce in vivo events yet is amenable to biochemical investigation. We have developed a procedure, based on organotypic cultures, which permits efficient preparation of large numbers of cerebellar slice cultures that can be easily manipulated. Cultures have been examined to document myelination biochemically (by incorporation of [35S]sulfate into sulfolipids), immunohistochemically (by labeling the myelin components myelin basic protein and galactocerebroside), and morphologically (by both light and electron microscopy). We tested the effects of biologically active peptides and antibodies on myelination in the thin slices. The results indicate that the cultures provide an in vitro system that can be used to examine specific cellular events that occur during CNS myelination.
髓磷脂的沉积和维持对于哺乳动物神经系统的正常功能至关重要。先前对中枢神经系统(CNS)髓鞘形成的研究因缺乏一种体外系统而受阻,该系统既能忠实地重现体内事件,又便于进行生化研究。我们基于器官型培养开发了一种方法,可高效制备大量易于操作的小脑切片培养物。已对培养物进行了生化(通过将[35S]硫酸盐掺入硫脂中)、免疫组织化学(通过标记髓磷脂成分髓磷脂碱性蛋白和半乳糖脑苷脂)和形态学(通过光学显微镜和电子显微镜)检查,以记录髓鞘形成过程。我们测试了生物活性肽和抗体对薄片中髓鞘形成的影响。结果表明,这些培养物提供了一种体外系统,可用于研究中枢神经系统髓鞘形成过程中发生的特定细胞事件。