DYNAMIC ASPECTS OF THE SUPRAMOLECULAR ORGANIZATION OF INTERMEDIATE FILAMENT NETWORKS IN CULTURED EPIDERMAL-CELLS
DYNAMIC ASPECTS OF THE SUPRAMOLECULAR ORGANIZATION OF INTERMEDIATE FILAMENT NETWORKS IN CULTURED EPIDERMAL-CELLS
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DOI：
10.1002/cm.970020302
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发表时间：
1982-01-01
影响因子：
--
通讯作者：
GOLDMAN, RD
中科院分区：
文献类型：
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作者：
JONES, JCR;GOLDMAN, AE;GOLDMAN, RD
用小鼠角蛋白K2和EM抗血清进行间接免疫荧光显微镜观察，发现转化的(PAM)和原代(PME)小鼠表皮细胞具有广泛的IF[中间丝束]网络。在胰酶消化和PAM细胞复制后，IF束被视为一个连续的网络，从核周区进入外围细胞质区域。在含有Triton X-100和DNAase I的高离子强度溶液中裂解的PAM细胞中，如果束似乎与核膜残留物密切相关，在某些情况下，似乎附着在核孔复合体上。在低钙条件下培养的PME细胞具有核周的IF束双折射排列。在将细胞切换到正常钙水平的2小时内，PME IF束网络向细胞表面移动，并随着桥粒的形成而与细胞表面建立联系。相差显微镜观察活细胞，免疫荧光显微镜观察固定细胞，可将桥粒区分为隔开相邻细胞的暗带。从PME细胞分离的IF束网络的十二烷基硫酸钠-聚丙烯酰胺凝胶图谱中的主要蛋白质在钙离子转换前后几乎没有变化。钙离子转换后相对难溶的膜相关蛋白的重组可能参与桥粒的形成。与PME IF网络相比，PAM细胞中分离的IF网络在蛋白质组成上存在差异。这可能与PAM细胞桥粒数目显著减少有关。表皮细胞中的IF束系统似乎参与了形状的形成、形状的维持、桥粒的建立、核的集中和细胞与细胞的接触。
Indirect immunofluorescence microscopy using an antiserum against the mouse keratin subunit K2 and EM showed that transformed (PAM) and primary (PME) mouse epidermal cells possess extensive networks of IF [intermediate filament] bundles. Following trypsinization and replating of PAM cells, IF bundles are seen to move as a continuous network from a perinuclear zone into the peripheral cytoplasmic regions. In PAM cells lysed in high-ionic-strength solutions containing Triton X-100 and DNAase I, IF bundles appear to be closely associated with nuclear envelope remnants and, in some cases, appear to be attached to nuclear pore complexes. PME cells cultivated in low Ca2+-containing medium possess perinuclear birefringent arrays of IF bundles. Within 2 h of switching the cells to normal Ca2+ levels, the PME IF bundle network moves towards and establishes contact with the cell surface as desmosomes form. Live cells observed by phase contrast and fixed cells observed by immunofluorescence microscopy demonstrate that desmosomes can be distinguished as dark bands separating neighboring cells. There is little difference between the major proteins seen in SDS[sodium dodecyl sulfate]-polyacrylamide gel profiles of ioslated IF bundle networks from PME cells before and after the Ca2+ switch. A reorganization of relatively insoluble membrane-associated protein following the Ca2+ switch may be involved in desmosome formation. The isolated IF networks from PAM cells differ in protein composition compared to the PME IF networks. This may be related to the greatly reduced number of desmosomes in PAM cells. The IF bundle system in epidermal cells appears to be involved in shape formation, shape maintenance, the establishment of desmosomes, nuclear centration, and cell-cell contact.