Cell adhesion to fibronectin and tenascin: quantitative measurements of initial binding and subsequent strengthening response.
Cell adhesion to fibronectin and tenascin: quantitative measurements of initial binding and subsequent strengthening response.
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DOI：
10.1083/jcb.109.4.1795
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发表时间：
1989-10
期刊：
影响因子：
--
通讯作者：
McClay DR
中科院分区：
文献类型：
--
作者：
Lotz MM;Burdsal CA;Erickson HP;McClay DR
细胞-基质粘附强度已经使用成纤维细胞和胶质瘤细胞与两种细胞外基质蛋白质纤连蛋白和腱生蛋白结合来定量。基于离心力的粘附测定用于粘附强度测量，并且通过干涉反射显微镜观察粘附的相应形态。在4 ℃下测量的初始粘附为10（-5）达因/细胞的数量级，并且不涉及细胞骨架。在37 ℃下15分钟后，粘附到纤连蛋白的强度超过一个数量级;强化反应需要细胞骨架参与。与FN粘附力的显著增强相反，在37 ℃下15分钟后，TN粘附力不变或减弱。所实现的粘附的绝对强度根据蛋白质和细胞类型而变化。当测试纤连蛋白和腱生蛋白的混合基质时，发现腱生蛋白的存在降低了通常在37 ℃下在单独的纤连蛋白基质上观察到的细胞粘附的增强水平。相应的干涉反射显微照片的平行分析表明，在10-15 nm的基底内的细胞表面的面积的差异密切相关的粘附观察到的每个变化：在37 ° C的纤连蛋白孵育15分钟后，神经胶质瘤细胞增加其表面积内密切接触的基板的积分到125倍。生腱蛋白上的细胞没有增加它们的接触表面积。细胞松弛素B的增加的接触表面积和抑制活性表明，在最初的结合后的15分钟内的粘合剂“加强”带来了额外的粘附分子进入粘合位点，并耦合肌动蛋白细胞骨架的粘附复合物。
Cell-substratum adhesion strengths have been quantified using fibroblasts and glioma cells binding to two extracellular matrix proteins, fibronectin and tenascin. A centrifugal force-based adhesion assay was used for the adhesive strength measurements, and the corresponding morphology of the adhesions was visualized by interference reflection microscopy. The initial adhesions as measured at 4 degrees C were on the order of 10(-5)dynes/cell and did not involve the cytoskeleton. Adhesion to fibronectin after 15 min at 37 degrees C were more than an order of magnitude stronger; the strengthening response required cytoskeletal involvement. By contrast to the marked strengthening of adhesion to FN, adhesion to TN was unchanged or weakened after 15 min at 37 degrees C. The absolute strength of adhesion achieved varied according to protein and cell type. When a mixed substratum of fibronectin and tenascin was tested, the presence of tenascin was found to reduce the level of the strengthening of cell adhesion normally observed at 37 degrees C on a substratum of fibronectin alone. Parallel analysis of corresponding interference reflection micrographs showed that differences in the area of cell surface within 10-15 nm of the substratum correlated closely with each of the changes in adhesion observed: after incubation for 15 min on fibronectin at 37 degrees C, glioma cells increased their surface area within close contact to the substrate by integral to 125- fold. Cells on tenascin did not increase their surface area of contact. The increased surface area of contact and the inhibitory activity of cytochalasin b suggest that the adhesive "strengthening" in the 15 min after initial binding brings additional adhesion molecules into the adhesive site and couples the actin cytoskeleton to the adhesion complex.