Calvert R, Anderton B H
EMBO J. 1985 May;4(5):1171-6. doi: 10.1002/j.1460-2075.1985.tb03756.x.
Monoclonal antibody (MAb) G10 labels a single high mol. wt. (HMW) band on Western blots of microtubule preparations from 2 day old rat brain. The G10 antigen is thermolabile and co-migrates with microtubule-associated protein (MAP)1 from young rat brain on low percentage (5%) polyacrylamide-SDS gels. The G10 antigen decreases by about five times from birth to adulthood in the rat cerebellum. The same single band is labelled on Western blots of homogenates of whole neonatal rat brain but no labelling is found using neonatal or adult kidney, lung or liver. We have therefore identified a brain-specific MAP1, designated MAP1(x). Immunofluorescence microscopy using MAb G10 on parasagittal sections of rat cerebella shows labelling of the newly formed molecular layer in 6 day old rats. Only a narrow band close to the pial surface is labelled in 18 day old animals, which disappears in the adult. Labelling of the cerebellar white matter found in young rats also disappears. Neurones but not flat cells in cerebellar cultures label with MAb G10. All staining patterns are consistent with an axonal distribution of the antigen. MAP1(x) may be part of a developmentally regulated microtubule structure.
单克隆抗体（MAb）G10在2日龄大鼠脑微管制剂的蛋白质免疫印迹上标记出一条单一的高分子量（HMW）条带。G10抗原不耐热，在低百分比（5%）聚丙烯酰胺 - SDS凝胶上与幼鼠脑微管相关蛋白（MAP）1共同迁移。在大鼠小脑中，G10抗原从出生到成年减少了约五倍。在新生大鼠全脑匀浆的蛋白质免疫印迹上也标记出了同一条带，但在新生或成年的肾、肺或肝中未发现标记。因此，我们鉴定出了一种脑特异性的MAP1，命名为MAP1(x)。使用MAb G10对大鼠小脑矢状旁切片进行免疫荧光显微镜检查显示，6日龄大鼠新形成的分子层有标记。在18日龄动物中，仅靠近软脑膜表面的一条窄带被标记，在成年动物中该标记消失。在幼鼠中发现的小脑白质标记也消失。小脑培养物中的神经元而非扁平细胞用MAb G10标记。所有染色模式均与抗原的轴突分布一致。MAP1(x)可能是发育调控的微管结构的一部分。
