Allendoerfer K L, Magnani J L, Patterson P H
Division of Biology, California Institute of Technology, Pasadena 91125, USA.
Mol Cell Neurosci. 1995 Aug;6(4):381-95. doi: 10.1006/mcne.1995.1029.
A key problem in nervous system development is how distinct subpopulations of progenitor cells give rise to different adult brain structures. The labeling pattern of the FORSE-1 antibody subdivides the neuroepithelium of the embryonic forebrain into domains resembling those of certain transcription factors, suggesting that the FORSE-1 epitope may be involved in the specification of development compartments. Therefore, it is important to determine the identity of the antigen(s) recognized by FORSE-1. On immunoblots, FORSE-1 recognizes a single, high-molecular-weight species, which we have identified as phosphacan, a brain-specific chondroitin sulfate proteoglycan that binds neural cell adhesion molecules. This identification is based on cross-immunoprecipitations and immunoblotting using an anti-phosphacan antibody and FORSE-1. FORSE-1 also recognizes two major neutral glycolipids in embryonic brain. The FORSE-1 epitope is sensitive to endo-beta-galactosidase, suggesting that the epitope corresponds to a carbohydrate moiety. Moreover, immunoprecipitates of the proteoglycan bearing the FORSE-1 epitope bind antibodies that recognize the Le* carbohydrate, and immunostaining patterns of embryonic brain sections by FORSE-1 and a known anti-Le* antibody are identical. Finally, purified FORSE-1 specifically recognizes Le*-containing glycoconjugates in ELISAs. The pattern of FORSE-1 labeling, the identification of its epitope as Le*, which has implicated in cell adhesion, and the presence of Le* on phosphacan suggest that this carbohydrate epitope may play a role in adhesive interactions important for proliferation, cell migration, or axon guidance.
神经系统发育中的一个关键问题是祖细胞的不同亚群如何产生不同的成体脑结构。FORSE-1抗体的标记模式将胚胎前脑的神经上皮细分为类似于某些转录因子的结构域,这表明FORSE-1表位可能参与发育区室的特化。因此,确定FORSE-1识别的抗原身份很重要。在免疫印迹上,FORSE-1识别单一的高分子量物质,我们已将其鉴定为phosphacan,一种与神经细胞粘附分子结合的脑特异性硫酸软骨素蛋白聚糖。这一鉴定基于使用抗phosphacan抗体和FORSE-1的交叉免疫沉淀和免疫印迹。FORSE-1还识别胚胎脑中的两种主要中性糖脂。FORSE-1表位对内切β-半乳糖苷酶敏感,这表明该表位对应于一个碳水化合物部分。此外,带有FORSE-1表位的蛋白聚糖免疫沉淀物能结合识别Le碳水化合物的抗体,并且FORSE-1和已知的抗Le抗体对胚胎脑切片的免疫染色模式相同。最后,纯化的FORSE-1在酶联免疫吸附测定中特异性识别含Le的糖缀合物。FORSE-1的标记模式、其表位鉴定为Le(这与细胞粘附有关)以及phosphacan上存在Le*表明,这种碳水化合物表位可能在对增殖、细胞迁移或轴突导向很重要的粘附相互作用中发挥作用。