Characterization of cell-matrix adhesion requirements for the formation of fascin microspikes
Characterization of cell-matrix adhesion requirements for the formation of fascin microspikes
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DOI：
10.1091/mbc.8.11.2345
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发表时间：
1997-11-01
影响因子：
3.3
通讯作者：
Adams, JC
中科院分区：
文献类型：
--
作者：
Adams, JC
细胞对凝血酶反应蛋白-1(TSP-1)的黏附与细胞-基质接触结构的组装有关，该结构含有筋膜蛋白微刺。在这项分析中，详细研究了组装束蛋白微刺所需的细胞基质。在6种细胞系中，细胞在TSP-1底物上的铺展与Fascin蛋白的表达和Fascin微刺的形成相关。H9c2细胞与纤维连接蛋白、玻璃体连接蛋白、IV型胶原蛋白或血小板因子4的黏附不能形成微刺。但是，粘附层粘连蛋白-1的细胞能组装成束蛋白微刺和局部接触。使用含有不同比例的TSP-1和纤维连接蛋白的混合底物，发现H9c2和C2C12细胞在含有25%纤维连接蛋白的底物上减少了束蛋白微棘的形成，而在含有75%纤维连接蛋白的底物上则完全消除了。与TSP-1和纤维连接蛋白的中间混合物的黏附导致了筋膜蛋白微刺和局灶性接触的共同组装，筋膜蛋白与肌动蛋白应力纤维束的共存，以及pi整合素、皮质α-肌动蛋白和原肌球蛋白的分布改变。在黏附于50%TSP-1：50%纤维连接蛋白的细胞中，GRGDSP多肽处理减少了焦点接触组装，改变了细胞骨架组织，但不抑制微棘突组装。用硫酸软骨素A或对硝基苯酚β-D-吡喃糖苷治疗可减少微刺的形成和改善细胞骨架组织，但不能抑制局灶性接触的形成。在极化的迁移和有丝分裂后的C2C12细胞中，束蛋白微刺和褶皱位于前沿，TSP基质沉积也集中在该区域。肝素处理耗尽基质TSP与减少微棘波的形成和细胞运动有关。因此，在最初的细胞-基质附着过程中，连接在细胞表面的黏附受体的平衡有助于调节组装的基质黏附接触的类型和随后的细胞骨架组织。表明了束蛋白微刺在细胞运动行为中的作用。
Cell adhesion to thrombospondin-1 (TSP-1) correlates with assembly of cell-substratum contact structures that contain fascin microspikes. In this analysis, cell-matrix requirements for assembly of fascin microspikes were examined in detail. In six cell lines, cell spreading on a TSP-1 substratum correlated with expression of fascin protein and formation of fascin microspikes. Microspikes were not formed by H9c2 cells adherent on fibronectin, vitronectin, collagen IV, or platelet factor 4. However, both fascin microspikes and focal contacts were assembled by cells adherent on laminin-1. Using mixed substrata containing different proportions of TSP-1, and fibronectin, fascin microspike formation by H9c2 and C2C12 cells was found to be reduced on substrata containing 25% fibronectin and abolished on substrata containing 75% fibronectin. Adhesion to intermediate mixtures of TSP-1 and fibronectin resulted in coassembly of fascin microspikes and focal contacts, colocalization of fascin with actin stress fiber bundles and altered distributions of pi integrins, cortical alpha-actinin, and tropomyosin. In cells adherent on 50% TSP-1:50% fibronectin, GRGDSP peptide treatment decreased focal contact assembly and altered cytoskeletal organization but did not inhibit microspike assembly. Treatment with chondroitin sulfate A or p-nitrophenol beta-D-xylopyranoside decreased microspike formation and modified cytoskeletal organization but did not inhibit focal contact formation. In polarized migratory and postmitotic C2C12 cells, fascin microspikes and ruffles were localized at leading edges and TSP matrix deposition was also concentrated in this region. Depletion of matrix TSP by heparin treatment correlated with decreased microspike formation and cell motility. Thus, the balance of adhesive receptors ligated at the cell surface during initial cell-matrix attachment serves to regulate the type of substratum adhesion contact assembled and subsequent cytoskeletal organization. A role for fascin microspikes in cell motile behavior is indicated.