Reynolds R, Wilkin G P
Department of Biochemistry, Imperial College of Science, Technology and Medicine, Kensington, London, UK.
J Neurocytol. 1991 Mar;20(3):216-24. doi: 10.1007/BF01186994.
To identify the stage in the life cycle of oligodendroglia at which they are mitotic during normal in vivo development [3H]thymidine autoradiography has been combined with immunocytochemistry on frozen sections of rat cerebellum. A panel of antibodies have been used that recognize antigens expressed by oligodendroglia at different stages of differentiation from progenitor to mature cell. It has been demonstrated that of the cells of the oligodendroglial lineage only the progenitors, identified by their expression of the ganglioside GD3, were seen to incorporate [3H]thymidine at all the developmental stages tested. Only few dividing GD3-positive cells were observed in the subventricular zones of the fourth-ventricle. The greatest number of dividing GD3-positive progenitors in the rat cerebellum was observed in the folia at postnatal day 7. Silver grains were never observed over cells that could be distinguished as oligodendroglia by their expression of galactocerebroside, 2'3'-cyclic nucleotide 3'-phosphohydrolase, or myelin basic protein. Mitotic astroglia were observed at all stages and could be clearly distinguished by their expression of glial fibrillary acidic protein and glutamine synthetase. When animals were injected with [3H]thymidine at postnatal day 7 and killed at 1-day intervals radiolabel was first observed in galactocerebroside-positive and 2'3'-cyclic nucleotide 3'-phosphohydrolase-positive oligodendroglia at day 9 and in myelin basic protein-positive cells at day 10-11, 3 days after the last cell division. Thus, we have demonstrated for the first time using in situ immunocytochemical techniques, a mitotic glial progenitor cell that is known to give rise to oligodendroglia both in vivo and in vitro.
为了确定少突胶质细胞在正常体内发育过程中进行有丝分裂的生命周期阶段,[3H]胸腺嘧啶核苷放射自显影技术已与大鼠小脑冰冻切片的免疫细胞化学相结合。使用了一组抗体,这些抗体可识别少突胶质细胞从祖细胞到成熟细胞不同分化阶段表达的抗原。结果表明,在少突胶质细胞谱系的细胞中,只有通过神经节苷脂GD3的表达鉴定出的祖细胞,在所有测试的发育阶段都能摄取[3H]胸腺嘧啶核苷。在第四脑室的室下区仅观察到少数分裂的GD3阳性细胞。在出生后第7天的小叶中观察到大鼠小脑中分裂的GD3阳性祖细胞数量最多。在通过半乳糖脑苷脂、2'3'-环核苷酸3'-磷酸水解酶或髓鞘碱性蛋白的表达可区分的少突胶质细胞上从未观察到银颗粒。在所有阶段都观察到有丝分裂的星形胶质细胞,并且可以通过其胶质纤维酸性蛋白和谷氨酰胺合成酶的表达清楚地加以区分。当在出生后第7天给动物注射[3H]胸腺嘧啶核苷并每隔1天处死时,在第9天首次在半乳糖脑苷脂阳性和2'3'-环核苷酸3'-磷酸水解酶阳性的少突胶质细胞中观察到放射性标记,在最后一次细胞分裂3天后的第10 - 11天在髓鞘碱性蛋白阳性细胞中观察到放射性标记。因此,我们首次使用原位免疫细胞化学技术证明了一种有丝分裂的神经胶质祖细胞,已知其在体内和体外均可产生少突胶质细胞。