Ghandour M S, Skoff R P
Department of Anatomy and Cell Biology, Wayne State University, School of Medicine, Detroit, MI 48201.
J Neurocytol. 1988 Aug;17(4):485-98. doi: 10.1007/BF01189804.
Intense and specific immunostaining of oligodendrocytes in vivo has been obtained for the first time using antibodies to galactocerebroside. We have examined the differentiation of oligodendrocytes in normal mice and then compared their differentiation to the myelin-deficient mouse jimpy, using immunoperoxidase, immunogold and immunofluorescence labelling techniques. We also compared staining for galactocerebroside with staining obtained using antibodies to myelin basic protein, carbonic anhydrase II, 2', 3'-cyclic nucleotide 3'-phosphohydrolase and proteolipid protein. The results of this comparative study confirm previous tissue culture studies and show that galactocerebroside is specific for oligodendrocytes in situ. As in tissue culture, galactocerebroside is one of the earliest oligodendrocyte markers to be expressed, making it an important marker for studying the differentiation of this cell type. The shape of oligodendrocytes in situ changes distinctly with time, shifting from an early stellate form with numerous spidery processes to a cell with a few processes radiating from the perikaryon. These morphological changes are observed for both normal and jimpy mice and they parallel those described in vitro. Oligodendrocytes in jimpy mice express most myelin markers, but the staining within the cells is generally less intense than in normal oligodendrocytes and the antigens are restricted to the cell body and processes without being incorporated into myelin sheaths. Quantification of the number of oligodendrocytes stained for galactocerebroside in normal and jimpy mice show that their number is not reduced in the corpus callosum and cerebellum during the first 2 weeks postnatal. This finding shows that many cells in jimpy mice which were considered to be unclassifiable by the application of morphological criteria have, in fact, differentiated to the stage where they are galactocerebroside-positive.
首次使用抗半乳糖脑苷脂抗体在体内获得了少突胶质细胞强烈而特异的免疫染色。我们研究了正常小鼠少突胶质细胞的分化情况，然后使用免疫过氧化物酶、免疫金和免疫荧光标记技术，将它们的分化与髓鞘缺陷型小鼠jimpy进行了比较。我们还将半乳糖脑苷脂染色与使用抗髓鞘碱性蛋白、碳酸酐酶II、2',3'-环核苷酸3'-磷酸水解酶和蛋白脂蛋白抗体获得的染色进行了比较。这项比较研究的结果证实了先前的组织培养研究，并表明半乳糖脑苷脂在原位对少突胶质细胞具有特异性。与组织培养一样，半乳糖脑苷脂是最早表达的少突胶质细胞标志物之一，使其成为研究这种细胞类型分化的重要标志物。原位少突胶质细胞的形状随时间明显变化，从早期具有许多蜘蛛状突起的星状形态转变为具有从核周放射出的少数突起的细胞。在正常小鼠和jimpy小鼠中都观察到了这些形态变化，并且它们与体外描述的变化相似。jimpy小鼠中的少突胶质细胞表达大多数髓鞘标志物，但细胞内的染色通常不如正常少突胶质细胞强烈，并且抗原仅限于细胞体和突起，而不整合到髓鞘中。对正常小鼠和jimpy小鼠中用半乳糖脑苷脂染色的少突胶质细胞数量进行定量分析表明，在出生后的前2周内，它们在胼胝体和小脑中的数量没有减少。这一发现表明，jimpy小鼠中许多被认为无法通过形态学标准分类的细胞实际上已经分化到了半乳糖脑苷脂阳性的阶段。
