Hall D E, Neugebauer K M, Reichardt L F
J Cell Biol. 1987 Mar;104(3):623-34. doi: 10.1083/jcb.104.3.623.
Cell attachment and neurite outgrowth by embryonic neural retinal cells were measured in separate quantitative assays to define differences in substrate preference and to demonstrate developmentally regulated changes in cellular response to different extracellular matrix glycoproteins. Cells attached to laminin, fibronectin, and collagen IV in a concentration-dependent fashion, though fibronectin was less effective for attachment than the other two substrates. Neurite outgrowth was much more extensive on laminin than on fibronectin or collagen IV. These results suggest that different substrates have distinct effects on neuronal differentiation. Neural retinal cell attachment and neurite outgrowth were inhibited on all three substrates by two antibodies, cell substratum attachment antibody (CSAT) and JG22, which recognize a cell surface glycoprotein complex required for cell interactions with several extracellular matrix constituents. In addition, retinal cells grew neurites on substrates coated with the CSAT antibodies. These results suggest that cell surface molecules recognized by this antibody are directly involved in cell attachment and neurite extension. Neural retinal cells from embryos of different ages varied in their capacity to interact with extracellular matrix substrates. Cells of all ages, embryonic day 6 (E6) to E12, attached to collagen IV and CSAT antibody substrates. In contrast, cell attachment to laminin and fibronectin diminished with increasing embryonic age. Age-dependent differences were found in the profile of proteins precipitated by the CSAT antibody, raising the possibility that modifications of these proteins are responsible for the dramatic changes in substrate preference of retinal cells between E6 and E12.
通过单独的定量分析来测量胚胎神经视网膜细胞的细胞附着和神经突生长,以确定底物偏好的差异,并证明细胞对不同细胞外基质糖蛋白的反应在发育过程中受到调控的变化。细胞以浓度依赖的方式附着于层粘连蛋白、纤连蛋白和IV型胶原,尽管纤连蛋白在促进细胞附着方面不如其他两种底物有效。在层粘连蛋白上的神经突生长比在纤连蛋白或IV型胶原上广泛得多。这些结果表明不同的底物对神经元分化有不同的影响。两种抗体,细胞基质附着抗体(CSAT)和JG22,抑制了神经视网膜细胞在所有三种底物上的附着和神经突生长,这两种抗体识别细胞与几种细胞外基质成分相互作用所需的一种细胞表面糖蛋白复合物。此外,视网膜细胞在包被有CSAT抗体的底物上长出神经突。这些结果表明该抗体识别的细胞表面分子直接参与细胞附着和神经突延伸。来自不同年龄胚胎的神经视网膜细胞与细胞外基质底物相互作用的能力有所不同。所有年龄的细胞,从胚胎第6天(E6)到E12,都能附着于IV型胶原和CSAT抗体底物。相反,随着胚胎年龄的增加,细胞对层粘连蛋白和纤连蛋白的附着减少。在CSAT抗体沉淀的蛋白质谱中发现了年龄依赖性差异,这增加了这些蛋白质的修饰是导致E6和E12之间视网膜细胞底物偏好发生显著变化的原因的可能性。