Gratecos Danielle, Krejci Eric, Sémériva Michel
Laboratoire de Génétique et Biologie cellulaires, Centre Universitaire de Luminy, CNRS, Case 907, 13288, Marseille cedex 9, France.
Rouxs Arch Dev Biol. 1990 May;198(7):411-419. doi: 10.1007/BF00376160.
By using an in vitro functional assay, we have shown that Drosophila embryonic cells possess Ca-dependent adhesive sites, which resemble in many respects those described for vertebrate cells and tissues. The cells, obtained by mechanical disruption of gastrulastage embryos, form aggregates within 30 min when maintained under constant rolling. The aggregation is completely dependent on the presence of Ca in the medium. In its absence, the cells remain dispersed but the process is reversible by readdition of Ca. In addition the aggregation is temperature-dependent. No aggregation occurs at 4° C but it can be restored by raising the temperature to 25° C. These properties are characteristic of these cells: established cell lines do not aggregate under the same conditions and mixing of cell lines and embryonic cells does not result in chimeric aggregates, thus pointing towards cell-type selectivity with respect to aggregability. Observations in electron microscopy have shown that the embryonic cells in the aggregates tightly adhere to one another and form, as early as after 30 min, maculae adherens junctions. Drosophila embryonic cells have adhesion sites that are protected from trypsin proteolysis in the presence of Ca and sensitive in its absence. The cells' aggregation can be inhibited by a mouse antiserum directed against cell-surface components and a good correlation exists between neutralization of the inhibitory activity of the antiserum and the presence of trypsin-sensitive sites on the cells. These data are in favour of cell-cell adhesion mediated by specific adhesion proteins.
通过使用体外功能分析,我们已经表明果蝇胚胎细胞拥有钙依赖性粘附位点,这些位点在许多方面类似于脊椎动物细胞和组织中所描述的那些。通过机械破坏原肠胚阶段胚胎获得的细胞,在持续滚动培养下30分钟内形成聚集体。聚集完全依赖于培养基中钙的存在。在没有钙的情况下,细胞保持分散状态,但通过重新添加钙,这个过程是可逆的。此外,聚集还依赖于温度。在4℃时不发生聚集,但将温度升至25℃可恢复聚集。这些特性是这些细胞的特征:已建立的细胞系在相同条件下不会聚集,细胞系与胚胎细胞混合不会形成嵌合聚集体,因此表明在聚集能力方面具有细胞类型选择性。电子显微镜观察表明,聚集体中的胚胎细胞彼此紧密粘附,早在30分钟后就形成了粘着斑连接。果蝇胚胎细胞具有在有钙时免受胰蛋白酶蛋白水解作用保护、在无钙时敏感的粘附位点。细胞的聚集可被针对细胞表面成分的小鼠抗血清抑制,并且抗血清抑制活性的中和与细胞上胰蛋白酶敏感位点的存在之间存在良好的相关性。这些数据支持由特定粘附蛋白介导的细胞间粘附。