Tóvári József, Futosi Krisztina, Bartal Alexandra, Tátrai Enikő, Gacs Alexandra, Kenessey István, Paku Sándor
a Department of Experimental Pharmacology; National Institute of Oncology ; Budapest , Hungary.
Cell Adh Migr. 2014;8(5):509-16. doi: 10.4161/cam.28734. Epub 2014 Oct 31.
A 2D model was previously presented that describes the gliding motility of human fibrosarcoma cells. The model was based on the observation that adhesions are present only on the outer rim of the leading lamella of the semicircular cell. The present model describes the organization of adhesions and the cytoskeleton of migrating HT1080 fibrosarcoma and LX2 hepatic stellate cells in three dimensions. The migration assays were performed in a modified Boyden chamber using fibronectin, Matrigel, or collagen I as chemoattractants. The distribution of the adhesions was analyzed by confocal laser scanning microscope, and following decoration with heavy meromyosin, the organization of actin filaments was analyzed by electron microscopy. Double labeling was performed to study the relationship of the actin and vimentin filament network in the moving cells. Vinculin containing adhesions were observed only at the front of the cell in the form of a ring while passing through a filter pore of the Boyden chamber. Actin filaments were present only below the plasma membrane, except the very tip of the leading lamella. Vimentin intermediate filaments were localized around the cell nucleus behind the actin filament-rich lamella. This paper describes a model of the organization of adhesions and the cytoskeleton of migrating cells in the Boyden chamber. The model is based on the observation that adhesions are present only at the leading edge of the cell. The results extend the earlier 2D model of cell locomotion into 3D.
之前曾提出一个二维模型，用于描述人纤维肉瘤细胞的滑行运动。该模型基于这样的观察结果：黏附仅存在于半圆形细胞前缘薄片的外缘。当前模型描述了迁移的HT1080纤维肉瘤细胞和LX2肝星状细胞在三维空间中黏附及细胞骨架的组织情况。迁移实验在改良的博伊登小室中进行，使用纤连蛋白、基质胶或I型胶原作为趋化因子。通过共聚焦激光扫描显微镜分析黏附的分布，在用重酶解肌球蛋白进行标记后，通过电子显微镜分析肌动蛋白丝的组织情况。进行双重标记以研究运动细胞中肌动蛋白和波形蛋白丝网络的关系。当细胞穿过博伊登小室的滤孔时，含纽蛋白的黏附仅以环的形式出现在细胞前端。肌动蛋白丝仅存在于质膜下方，但前缘薄片的最顶端除外。波形蛋白中间丝位于富含肌动蛋白丝的薄片后方的细胞核周围。本文描述了博伊登小室中迁移细胞黏附及细胞骨架组织的模型。该模型基于黏附仅存在于细胞前缘的观察结果。这些结果将早期细胞运动的二维模型扩展到了三维。
