Reynolds R, Carey E M, Herschkowitz N
Department of Biochemistry, Imperial College, London, U.K.
Neuroscience. 1989;28(1):181-8. doi: 10.1016/0306-4522(89)90242-x.
Oligodendrocytes, the cells responsible for myelin sheath formation in the central nervous system, were isolated from primary dissociated mixed glial cultures prepared from newborn mouse forebrain, and further cultured in a serum-free defined culture medium. Single and double indirect immunofluorescence using antibodies against the myelin glycolipids, galactocerebroside and sulfatide, and the myelin proteins, myelin basic protein and 2',3'-cyclic nucleotide 3'-phosphohydrolase, was used to investigate the composition of the flat membrane extensions produced by some oligodendrocytes in culture. Galactocerebroside and sulfatide were both expressed on the external surface of the plasma membrane of oligodendrocyte cell bodies and processes and also the membrane expansions. Neither myelin basic protein nor 2',3'-cyclic nucleotide 3'-phosphohydrolase were expressed on the external surface of oligodendrocytes. Myelin basic protein could be localized to the cell body and the membrane expansions but not the major and fine processes. The localization of these myelin components suggests that the expansions have characteristics of the mature myelin membrane. 2',3'-Cyclic nucleotide 3'-phosphohydrolase was found to be localized in the cell body, and in total contrast to myelin basic protein, in the major processes and the fine interconnecting processes, but not the membrane expansions. In some of the cells 2',3'-cyclic nucleotide 3'-phosphohydrolase was present at the outer extremities of the flat membrane sheets, giving the appearance of an extending growth region. Our results thus clearly show that 2',3'-cyclic nucleotide 3'-phosphohydrolase is localized within oligodendrocytes in discrete regions of plasma membranes and suggest that this protein has a possible role in the early stages of myelin formation.
少突胶质细胞是负责中枢神经系统髓鞘形成的细胞，从新生小鼠前脑制备的原代解离混合神经胶质培养物中分离出来，并在无血清限定培养基中进一步培养。使用针对髓鞘糖脂、半乳糖脑苷脂和硫苷脂以及髓鞘蛋白、髓鞘碱性蛋白和2',3'-环核苷酸3'-磷酸水解酶的抗体进行单重和双重间接免疫荧光，以研究培养的一些少突胶质细胞产生的扁平膜延伸物的组成。半乳糖脑苷脂和硫苷脂均表达于少突胶质细胞胞体、突起以及膜扩张物的质膜外表面。髓鞘碱性蛋白和2',3'-环核苷酸3'-磷酸水解酶均未在少突胶质细胞的外表面表达。髓鞘碱性蛋白可定位于胞体和膜扩张物，但不在主要突起和细小突起中。这些髓鞘成分的定位表明，这些扩张物具有成熟髓鞘膜的特征。发现2',3'-环核苷酸3'-磷酸水解酶定位于胞体，与髓鞘碱性蛋白完全相反，它存在于主要突起和细小连接突起中，但不存在于膜扩张物中。在一些细胞中，2',3'-环核苷酸3'-磷酸水解酶存在于扁平膜片的外端，呈现出延伸生长区域的外观。因此，我们的结果清楚地表明，2',3'-环核苷酸3'-磷酸水解酶定位于少突胶质细胞内质膜的离散区域，并表明该蛋白在髓鞘形成的早期阶段可能具有作用。
