Choi A H, Siu C H
J Cell Biol. 1987 May;104(5):1375-87. doi: 10.1083/jcb.104.5.1375.
During the early phase of Dictyostelium discoideum development, cells undergo chemotactic migration to form tight aggregates. A developmentally regulated surface glycoprotein of Mr 80,000 (gp80) has been implicated in mediating the EDTA-resistant type of cell cohesion at this stage. We have used a monoclonal antibody directed against gp80 to study the topographical distribution of gp80 on the cell surface. Indirect immunofluorescence studies showed that gp80 was primarily localized on the cell surface, with a higher concentration at contact areas. Immunoelectron microscopy was carried out by indirect labeling using protein A-gold, and a nonrandom distribution of gp80 was revealed. In addition to contact regions, gold particles were found preferentially localized on filopodia. Quantitative analysis using transmission electron microscopy (TEM) showed that approximately 60% more gold particles were localized in contact regions in comparison with the noncontact regions, and the filopodial surfaces had a twofold higher gold density. Both TEM and scanning electron microscopy showed that contact areas were enriched in filopodial structures. Filopodia often appeared to adhere to either smooth surfaces or similar filopodial structures of an adjacent cell. These observations suggest that the formation of stable cell-cell contacts involves at least four sequential steps in which filopodia and gp80 probably play an important role in the initial stages of recognition and cohesion among cells.
在盘基网柄菌发育的早期阶段,细胞进行趋化性迁移以形成紧密的聚集体。一种分子量为80,000的发育调控表面糖蛋白(gp80)被认为在此阶段介导了对EDTA抗性的细胞黏附类型。我们使用了一种针对gp80的单克隆抗体来研究gp80在细胞表面的拓扑分布。间接免疫荧光研究表明,gp80主要定位于细胞表面,在接触区域浓度更高。通过使用蛋白A-金的间接标记进行免疫电子显微镜观察,发现gp80呈非随机分布。除了接触区域外,金颗粒还优先定位于丝状伪足上。使用透射电子显微镜(TEM)进行的定量分析表明,与非接触区域相比,接触区域的金颗粒数量大约多60%,丝状伪足表面的金密度高出两倍。TEM和扫描电子显微镜均显示接触区域富含丝状伪足结构。丝状伪足常常似乎粘附于光滑表面或相邻细胞的类似丝状伪足结构。这些观察结果表明,稳定的细胞间接触的形成至少涉及四个连续步骤,其中丝状伪足和gp80可能在细胞间识别和黏附的初始阶段发挥重要作用。