Murray B A, Jensen J J
Department of Developmental and Cell Biology, University of California, Irvine 92717.
J Cell Biol. 1992 Jun;117(6):1311-20. doi: 10.1083/jcb.117.6.1311.
The adhesion of embryonic chicken retinal cells and mouse N2A neuroblastoma cells to purified embryonic chicken retinal NCAM adsorbed on a solid substratum was examined using a quantitative centrifugal adhesion assay. Both cell types adhered to NCAM and the adhesion was specifically inhibited by monovalent anti-NCAM antibody fragments. N2A cell adhesion depended on the amount of NCAM applied to the substratum, was cation independent, and was insensitive to treatment with the cytoskeletal perturbing drugs colchicine and cytochalasin D. These results indicated that the tubulin and actin cytoskeletons were not critically required for adhesion to NCAM and make it unlikely that the cell surface ligand for NCAM is an integrin. Adhesion was however temperature dependent, strengthening greatly after a brief incubation at 37 degrees C. CHO cells transfected with NCAM cDNAs did not adhere specifically to substratum-bound NCAM and pretreatment of N2A cells and retinal cells with anti-NCAM antibodies did not inhibit adhesion to substratum-bound NCAM. These results suggest that a heterophilic interaction between substratum-adsorbed NCAM and a non-NCAM ligand on the surface of the probe cells affects adhesion in this system and support the possibility that heterophilic adhesion may be a function of NCAM in vivo.
使用定量离心粘附试验检测了胚胎鸡视网膜细胞和小鼠N2A神经母细胞瘤细胞对吸附在固体基质上的纯化胚胎鸡视网膜神经细胞粘附分子(NCAM)的粘附情况。两种细胞类型均能粘附于NCAM,且这种粘附被单价抗NCAM抗体片段特异性抑制。N2A细胞的粘附取决于施加于基质上的NCAM量,不依赖阳离子,并且对细胞骨架干扰药物秋水仙碱和细胞松弛素D的处理不敏感。这些结果表明,微管蛋白和肌动蛋白细胞骨架对于粘附于NCAM并非至关重要,并且使得NCAM的细胞表面配体不太可能是整合素。然而,粘附是温度依赖性的,在37℃短暂孵育后显著增强。用NCAM cDNA转染的中国仓鼠卵巢(CHO)细胞不会特异性粘附于基质结合的NCAM,并且用抗NCAM抗体预处理N2A细胞和视网膜细胞不会抑制对基质结合的NCAM的粘附。这些结果表明,基质吸附的NCAM与探针细胞表面的非NCAM配体之间的异嗜性相互作用影响了该系统中的粘附,并支持异嗜性粘附可能是NCAM在体内的一种功能的可能性。