Stainier D Y, Bilder D H, Gilbert W
Department of Cellular and Developmental Biology, Harvard University, Cambridge, Massachusetts 02138.
Dev Biol. 1991 Mar;144(1):177-88. doi: 10.1016/0012-1606(91)90489-p.
We have previously reported the isolation of a monoclonal antibody, mAb B30, that recognizes two minor gangliosides specifically expressed in a small subset of neurons in the developing mouse central nervous system (Stainier and Gilbert, 1989). B30 labels mesencephalic trigeminal neurons shortly after differentiation until about 2 weeks after birth. Postnatally, it also labels two specific monolayers of cerebellar neurons. In this study, we have characterized the B30 immunoreactivity in the developing peripheral nervous system of the mouse. We report that B30 is a marker for neural crest-derived neurons and have used it to follow the neuronal differentiation of neural crest cells in a serum-free chemically defined culture system. Within hours after plating, neural crest cells migrate away from the neural tube explant on a fibronectin or laminin substrate and by 24 hr, up to 15% of them have differentiated into morphologically identifable neurons. In vitro as in vivo, undifferentiated mouse neural crest cells express the GD3 ganglioside which is recognized by mAb B33, and neural crest-derived neurons can be labeled by mAbs B33, B30, and also E1.9, a specific neuronal cytoskeletal marker. We also show the unique biochemical specificity of mAb B30 and provide experimental evidence for the role of the B30 ganglioside in the cellular adhesion process.
我们之前报道过一种单克隆抗体mAb B30的分离,它能识别在发育中的小鼠中枢神经系统一小部分神经元中特异性表达的两种次要神经节苷脂(Stainier和Gilbert,1989)。B30在分化后不久便标记中脑三叉神经元,直至出生后约2周。出生后,它还标记小脑神经元的两个特定单层。在本研究中,我们对小鼠发育中的外周神经系统中的B30免疫反应性进行了表征。我们报告B30是神经嵴衍生神经元的标志物,并已在无血清化学限定培养系统中用它来追踪神经嵴细胞的神经元分化。接种后数小时内,神经嵴细胞在纤连蛋白或层粘连蛋白底物上从神经管外植体迁移离开,到24小时时,高达15%的细胞已分化为形态可识别的神经元。在体外和体内一样,未分化的小鼠神经嵴细胞表达被mAb B33识别的GD3神经节苷脂,神经嵴衍生的神经元可被mAb B33、B30以及E1.9(一种特异性神经元细胞骨架标志物)标记。我们还展示了mAb B30独特的生化特异性,并为B30神经节苷脂在细胞黏附过程中的作用提供了实验证据。