Zhou L, Szigeti V, Miller R H
Department of Neurosciences, School of Medicine, Case Western Reserve University,Cleveland, OH 44106, USA.
J Neurosci Res. 1995 Nov 1;42(4):504-15. doi: 10.1002/jnr.490420409.
Cell-type-specific reagents have proven useful in the analysis of central nervous system (CNS) development and function. Most markers of oligodendrocytes are components of myelin sheath, which in the CNS is the specific product of oligodendrocytes. We have isolated a novel monoclonal antibody termed 2B10 which was raised against embryonic rat spinal cord tissue. In adult rat cerebrum, cerebellum, and spinal cord, 2B10 immunoreactivity is predominantly localized in white matter. 2B10 immunoreactivity is absent from peripheral nerve, suggesting that in the nervous system the 2B10 antigen is restricted to the CNS. Dissociated cell culture studies indicate that 2B10 labels a cell surface molecule, and its cellular distribution is coincident with O1 and myelin basic protein-positive oligodendrocytes. By contrast, 2B10 does not label GFAP-positive astrocytes. These data suggest that in the CNS the 2B10 antigen is expressed specifically on oligodendrocytes. Biochemical analysis indicates that 2B10 recognizes a protein with an apparent molecular weight of approximately 79,000 in reducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Developmentally, 2B10 immunoreactivity is detectable in rat spinal cord at as early as embryonic day 14. The relative abundance of this molecule decreases during myelination, but is maintained at a sustained level throughout adulthood. The 2B10 antigen appears not to be a myelin-associated protein since it is not detected in purified myelin fractions. 2B10 immunoreactivity is not detectable in extracts of rat sciatic nerve, heart, kidney, muscle, and skin, but is detectable in extracts of spleen and thymus. These data suggest that the 2B10 antigen is a novel cell surface molecule that is expressed on oligodendrocyte lineage cells throughout development.
细胞类型特异性试剂已被证明在中枢神经系统(CNS)发育和功能分析中很有用。少突胶质细胞的大多数标志物是髓鞘的组成部分,在中枢神经系统中,髓鞘是少突胶质细胞的特定产物。我们分离出一种名为2B10的新型单克隆抗体,它是针对胚胎大鼠脊髓组织产生的。在成年大鼠的大脑、小脑和脊髓中,2B10免疫反应性主要定位于白质。外周神经中不存在2B10免疫反应性,这表明在神经系统中,2B10抗原仅限于中枢神经系统。解离细胞培养研究表明,2B10标记一种细胞表面分子,其细胞分布与O1和髓鞘碱性蛋白阳性的少突胶质细胞一致。相比之下,2B10不标记GFAP阳性的星形胶质细胞。这些数据表明,在中枢神经系统中,2B10抗原在少突胶质细胞上特异性表达。生化分析表明,在还原型十二烷基硫酸钠-聚丙烯酰胺凝胶电泳中,2B10识别一种表观分子量约为79,000的蛋白质。在发育过程中,早在胚胎第14天就能在大鼠脊髓中检测到2B10免疫反应性。这种分子的相对丰度在髓鞘形成过程中降低,但在成年期一直维持在持续水平。2B10抗原似乎不是一种髓鞘相关蛋白,因为在纯化的髓鞘组分中未检测到它。在大鼠坐骨神经、心脏、肾脏、肌肉和皮肤的提取物中未检测到2B10免疫反应性,但在脾脏和胸腺的提取物中可检测到。这些数据表明,2B10抗原是一种新型细胞表面分子,在整个发育过程中在少突胶质细胞谱系细胞上表达。