Halfter W, Liverani D, Vigny M, Monard D
Friedrich Miescher-Institut, Basel, Switzerland.
Cell Tissue Res. 1990 Dec;262(3):467-81. doi: 10.1007/BF00305243.
Fibroblasts from rat, mouse and chick embryos cultured on poly-lysine/fibronectin- or poly-lysine/laminin-coated dishes were stained with antibodies directed to extracellular matrix molecules. The staining showed that cells had migrated during culture and deposited extracellular matrix components along their migration trails. Depending on the antigen, the staining of the matrix revealed fibrils, spots or a diffuse smear along the migration pathways. The major matrix components were fibronectin and heparan sulfate proteoglycan; however, laminin nidogen, tenascin, glia-derived nexin (GDN) and chondroitin-4-sulfate proteoglycan were also found. The migration trails were also detectable by scanning electron microscopy. Here, the fibrils were the prominent structures. The deposition of matrix was independent from the substratum: fibronectin was deposited on laminin, plain poly-lysine, basal lamina and even on fibronectin. Functional assays using anti-fibronectin or an antiserum to embryonic pigment epithelium basement membrane disturbed the formation of matrix fibrils, but did not inhibit cell attachment and translocation. Likewise, heparin in the culture medium only partially inhibited cell migration, despite the fact that it disturbed the formation of proper matrix fibrils. Our results suggest that the deposition of extracellular matrix by cells may not be mandatory for attachment and translocation. However, the deposition of matrix along defined trails might be important for the pathfinding of cells or nerve fibers that appear later in development.
在聚赖氨酸/纤连蛋白或聚赖氨酸/层粘连蛋白包被的培养皿上培养的大鼠、小鼠和鸡胚胎成纤维细胞,用针对细胞外基质分子的抗体进行染色。染色结果显示,细胞在培养过程中发生了迁移,并沿着其迁移轨迹沉积了细胞外基质成分。根据抗原的不同,基质的染色显示出沿迁移路径的纤维、斑点或弥漫性涂片。主要的基质成分是纤连蛋白和硫酸乙酰肝素蛋白聚糖;不过,也发现了层粘连蛋白、巢蛋白、腱生蛋白、胶质衍生的神经连接蛋白(GDN)和硫酸软骨素-4-硫酸盐蛋白聚糖。迁移轨迹也可通过扫描电子显微镜检测到。在此,纤维是突出的结构。基质的沉积与底物无关:纤连蛋白沉积在层粘连蛋白、普通聚赖氨酸、基膜甚至纤连蛋白上。使用抗纤连蛋白或针对胚胎色素上皮基底膜的抗血清进行的功能分析扰乱了基质纤维的形成,但并未抑制细胞附着和移位。同样,尽管培养基中的肝素扰乱了正常基质纤维的形成,但它仅部分抑制了细胞迁移。我们的结果表明,细胞外基质的沉积对于细胞附着和移位可能不是必需的。然而,沿特定轨迹沉积基质对于发育后期出现的细胞或神经纤维的路径寻找可能很重要。