Chen A, Haines S, Maxson K, Akeson R A
Division of Basic Research, Children's Hospital Research Foundation, Cincinnati, Ohio 45229-2899.
J Neurosci Res. 1994 Aug 1;38(5):483-92. doi: 10.1002/jnr.490380502.
The neural cell adhesion molecule (NCAM) is found on cells as several related polypeptides formed by alternative splicing of the single NCAM gene. The alternatively spliced 30-bp VASE exon in the fourth immunoglobulin-like domain is the structural variation nearest those portions of the polypeptide proposed to mediate cell-cell adhesion. To test the ability of distinct forms of the NCAM molecules to mediate cell adhesion, L cells were transfected with expression vectors encoding rat 140 kD NCAM +/- the VASE exon. L cell lines which expressed these polypeptides were isolated and tested for self-aggregation in a low shear, rapid aggregation assay. Increased cellular aggregation of the transfectants was observed to be a function of the NCAM molecule expressed. These transfected cells showed segregation in a long term co-aggregation assay: cells expressing NCAM--VASE formed aggregates which tended to exclude cells expressing NCAM+VASE and vice versa. These results provide direct evidence that this small difference in NCAM structure is sufficient to allow segregation of cells.
神经细胞黏附分子(NCAM)以几种相关多肽的形式存在于细胞上,这些多肽由单一的NCAM基因通过可变剪接形成。在第四个免疫球蛋白样结构域中可变剪接的30bp VASE外显子是最接近多肽中被认为介导细胞间黏附部分的结构变异。为了测试不同形式的NCAM分子介导细胞黏附的能力,用编码大鼠140kD NCAM ± VASE外显子的表达载体转染L细胞。分离出表达这些多肽的L细胞系,并在低剪切、快速聚集试验中测试其自我聚集能力。观察到转染细胞的细胞聚集增加是所表达的NCAM分子的函数。在长期共聚集试验中,这些转染细胞表现出分离现象:表达NCAM-VASE的细胞形成聚集物,倾向于排斥表达NCAM+VASE的细胞,反之亦然。这些结果提供了直接证据,表明NCAM结构上的这种微小差异足以使细胞分离。