Müller-Husmann G, Reinhardt S, Stähle M, Lang E, Maelicke A
Laboratory of Molecular Neurobiology, Johannes Gutenberg University Medical School, Mainz, Germany.
Brain Res Dev Brain Res. 1994 Dec 16;83(2):262-77. doi: 10.1016/0165-3806(94)00148-0.
Using membrane fragments of PCC7-Mz1 embryonal carcinoma cells, an established in vitro model of neural differentiation (Lang et al., J. Cell Biol., 109 (1989) 2481-2493), we have raised monoclonal antibodies (mAb) against developmental stage-specific cell surface antigens. As shown by double-immunofluorescence labeling studies, employing differentiating PCC7-Mz1 cells, primary cultures of mouse cerebellum cells and cryosections of mouse brain and other tissues, rat mAb anti-mouse EX-1 recognizes a membrane protein which is exclusively expressed by cells of the neuronal cell lineage. EX-1-expressing neuronal precursor cells were identified by double labeling with antibodies directed against stem cell markers or BrdU, EX-1-expressing postmitotic neurons by labeling with antibodies directed against phenotypic markers. In the developing mouse brain, the EX-1 antigen is expressed in the neuroepithelium already at prenatal day 8, i.e. clearly before the onset of mature neuron-specific marker expression. Increasing co-expression with the latter is observed from embryonic day E10 throughout neuronal maturation, but not with markers of other cell types tested. From these studies, the EX-1 antigen is the earliest marker for the mouse brain neuronal cell lineage so far discovered.
利用PCC7-Mz1胚胎癌细胞的膜片段(一种已建立的神经分化体外模型,Lang等人,《细胞生物学杂志》,109 (1989) 2481-2493),我们制备了针对发育阶段特异性细胞表面抗原的单克隆抗体(mAb)。通过双免疫荧光标记研究表明,使用分化的PCC7-Mz1细胞、小鼠小脑细胞原代培养物以及小鼠脑和其他组织的冰冻切片,大鼠抗小鼠EX-1单克隆抗体识别一种仅由神经元细胞谱系的细胞表达的膜蛋白。通过用针对干细胞标志物或BrdU的抗体进行双重标记来鉴定表达EX-1的神经元前体细胞,通过用针对表型标志物的抗体进行标记来鉴定表达EX-1的有丝分裂后神经元。在发育中的小鼠脑中,EX-1抗原在产前第8天就在神经上皮中表达,即在成熟神经元特异性标志物表达开始之前就明显表达。从胚胎第E10天到整个神经元成熟过程中,观察到与后者的共表达增加,但与测试的其他细胞类型的标志物没有共表达。从这些研究来看,EX-1抗原是迄今为止发现的小鼠脑神经元细胞谱系的最早标志物。