Izant J G, McIntosh J R
Proc Natl Acad Sci U S A. 1980 Aug;77(8):4741-5. doi: 10.1073/pnas.77.8.4741.
Hybridomas that secret IgG reacting specifically with the brain microtubule-associated protein MAP2 have been prepared with speen cells from BALB/c mice hyperimmunized with high molecular weight neurotubule-associated proteins. Immunofluorecence microscopy using dual fluorochrome labeling of tubulin and MAP2 antigens revealed identical patterns of interphase fiber networks in cells from explants of newborn mouse brain. The anti-MAP2 antibody did not stain primary mouse kidney cells or CHO, 3T3, HeLa, or PtK1 cell lines. Immunoprecipitation and antibody gel staining techniques failed to demonstrate any crossreacting antigen in these cells. MAP2 antigen was not seen in association with the mitotic spindle in any of the cells examined. Radioimmunoassay showed species crossreactivity of the anti-MAP2 antibody with mammalian but not avian neural cell extracts. Glial cells and some neuroblastoma cell lines did not appear to contain MAP2. However, in the B104 rat neuroblastoma cell line the MAP2 antigen appeared to be associated with the cytoskeleton concomitant with differentiation induced by dibutyryl cyclic AMP. In disagreement with most previously published reports, our data suggest that MAP2 is found only in differentiated neuronal cells and raises the possibility that MAP2 is involved in neuronal differentiation or neuron-specific processes.
用高分子量神经微管相关蛋白对BALB/c小鼠进行超免疫后获取的脾细胞，制备出了能分泌与脑微管相关蛋白MAP2特异性反应的IgG的杂交瘤。使用微管蛋白和MAP2抗原的双荧光染料标记的免疫荧光显微镜检查显示，新生小鼠脑外植体细胞中相间纤维网络的模式相同。抗MAP2抗体未对原代小鼠肾细胞或CHO、3T3、HeLa或PtK1细胞系染色。免疫沉淀和抗体凝胶染色技术未能在这些细胞中显示出任何交叉反应抗原。在所检查的任何细胞中，均未观察到MAP2抗原与有丝分裂纺锤体相关。放射免疫测定表明，抗MAP2抗体与哺乳动物而非禽类神经细胞提取物具有种属交叉反应性。神经胶质细胞和一些神经母细胞瘤细胞系似乎不含MAP2。然而，在B104大鼠神经母细胞瘤细胞系中，MAP2抗原似乎与由二丁酰环磷腺苷诱导分化的细胞骨架相关。与大多数先前发表的报告不同，我们的数据表明MAP2仅存在于分化的神经元细胞中，并增加了MAP2参与神经元分化或神经元特异性过程的可能性。
