Levine J M, Beasley L, Stallcup W B
J Neurosci. 1984 Mar;4(3):820-31. doi: 10.1523/JNEUROSCI.04-03-00820.1984.
A monoclonal antibody against a cell surface ganglioside present on neuroepithelial cells was produced by immunizing mice with the B49 cell line, a clonal line with properties of both neurons and glial cells. The expression of this antigen, designated as D1.1, was analyzed in the developing rat cerebellum. The D1.1 antigen was localized by the immunofluorescent staining method to germinal cells of the external granule layer (EGL). Fluorescent labeling of cells comprising the EGL was apparent on embryonic day 18 when the EGL first forms, and the labeling was present throughout the period of postnatal cerebellar development. No cells within the adult cerebellum were labeled with the anti-D1.1 antibody. The D1.1-labeled cells of the EGL synthesize DNA, as demonstrated by [3H] thymidine autoradiography. However, within 48 hr after their final mitosis, nascent cerebellar cells that had migrated away from the external granule layer were no longer labeled with antibody. Some of the neurons and some of the astrocytes in cerebellar cell cultures were fluorescently labeled with the anti-D1.1 antibody. The number of anti-D1.1-labeled neurons in the cultures decreased over the first 10 days in vitro in agreement with the findings that in vivo the fluorescent labeling of the D1.1 antigen disappears from postmitotic cells. The antibody recognizes a ganglioside that in thin layer chromatographic experiments has a mobility between that of the GM1 and GM2 ganglioside. These data suggest that the D1.1 ganglioside antigen is a cell surface marker for germinal cells and that the acquisition and subsequent loss of this antigen is an aspect of the biochemical maturation of neurons and glial cells.
通过用B49细胞系免疫小鼠制备了一种针对神经上皮细胞表面神经节苷脂的单克隆抗体,B49细胞系是一种具有神经元和神经胶质细胞特性的克隆系。在发育中的大鼠小脑中分析了这种命名为D1.1的抗原的表达。通过免疫荧光染色法将D1.1抗原定位到外颗粒层(EGL)的生发细胞。当EGL首次形成时,即胚胎第18天,构成EGL的细胞的荧光标记就很明显,并且这种标记在出生后小脑发育期间一直存在。成年小脑中没有细胞被抗D1.1抗体标记。如[3H]胸苷放射自显影所示,EGL中被D1.1标记的细胞合成DNA。然而,在它们最后一次有丝分裂后的48小时内,从小脑外颗粒层迁移出来的新生小脑细胞不再被抗体标记。小脑细胞培养物中的一些神经元和一些星形胶质细胞被抗D1.1抗体荧光标记。培养物中抗D1.1标记的神经元数量在体外培养的前10天内减少,这与体内D1.1抗原的荧光标记从有丝分裂后细胞中消失的发现一致。该抗体识别一种神经节苷脂,在薄层色谱实验中其迁移率介于GM1和GM2神经节苷脂之间。这些数据表明,D1.1神经节苷脂抗原是生发细胞的细胞表面标志物,并且这种抗原的获得和随后的丧失是神经元和神经胶质细胞生化成熟的一个方面。