Nussbaum J L, Espinosa de los Monteros A, Pari F M, Doerr-Schott J, Roussel G, Neskovic N M
Centre de Neurochimie du CNRS and U44 de l'INSERM, Strasbourg, France.
Int J Dev Neurosci. 1988;6(4):395-408. doi: 10.1016/0736-5748(88)90022-6.
This study reports the production of myelin-like membranes in oligodendrocyte subcultures derived from 20-day-old primary glial cell cultures of newborn rat brain. These multi-layered structures show a variable number of membrane turns; up to 10 concentric lamellae are found in 3- to 4-week-old subcultures. When they are compacted, alternate dense and intraperiodic lines with a periodicity of 11.2 nm are noticeable. The most typical myelin proteins were detected straight on the multi-lammellar structures by a gold immunocytochemical method. Subcellular fractions containing these myelin-like structures were isolated by ultracentrifugation on a discontinuous sucrose gradient. They were analysed by sodium dodecylsulfate-polyacrylamide gel electrophoresis and immunoblotting; UDP-galactose: ceramide galactosyltransferase and 2',3'-cyclic nucleotide 3'-phosphohydrolase activities were also measured. The results indicate that the multi-layered membrane profiles have many characteristics of the myelin found in vivo; nevertheless some differences were still apparent. Our data support the concept of the cultured oligodendrocytes expressing the intrinsic myelinogenic properties and possessing a basic developmental program of myelination, apparently in the absence of stimuli coming from other brain cells.
本研究报告了从新生大鼠脑20日龄原代神经胶质细胞培养物中获得的少突胶质细胞亚培养物中髓鞘样膜的产生情况。这些多层结构显示出数量可变的膜层；在3至4周龄的亚培养物中发现多达10个同心薄片。当它们压实后，可观察到具有11.2nm周期的交替致密线和周期内线。通过金免疫细胞化学方法直接在多层结构上检测到了最典型的髓鞘蛋白。通过在不连续蔗糖梯度上进行超速离心分离出含有这些髓鞘样结构的亚细胞组分。对其进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和免疫印迹分析；还测定了UDP-半乳糖：神经酰胺半乳糖基转移酶和2',3'-环核苷酸3'-磷酸水解酶的活性。结果表明，多层膜结构具有体内发现的髓鞘的许多特征；然而，一些差异仍然明显。我们的数据支持培养的少突胶质细胞表达内在髓鞘形成特性并具有基本髓鞘形成发育程序的概念，显然这是在没有来自其他脑细胞的刺激的情况下发生的。
