Wacker I U, Rickard J E, De Mey J R, Kreis T E
European Molecular Biology Laboratory, Heidelberg, Germany.
J Cell Biol. 1992 May;117(4):813-24. doi: 10.1083/jcb.117.4.813.
The establishment of epithelial cell polarity correlates with the formation of specialized cell-cell junctions and striking changes in the organization of microtubules. A significant fraction of the microtubules in MDCK cells become stabilized, noncentrosomally organized, and arranged in longitudinal bundles in the apical-basal axis. This correlation suggests a functional link between cell-cell junction formation and control of microtubule organization. We have followed the distribution of pp170, a recently described microtubule-binding protein, during establishment of epithelial cell polarity. This protein shows the typical patchy distribution along microtubules in subconfluent fibroblasts and epithelial cells, often associated with the peripheral ends of a subpopulation of microtubules. In contrast to its localization in confluent fibroblasts (A72) and HeLa cells, however, pp170 accumulates in patches delineating the regions of cell-cell contacts in confluent polarizing epithelial cells (MDCK and Caco-2). Double immunolocalization with antibodies specific for cell-cell junction proteins, confocal microscopy, and immunoelectron microscopy on polarized MDCK cells suggest that pp170 accumulates at desmosomal plaques. Furthermore, microtubules and desmosomes are found in close contact. Maintenance of the desmosomal association of pp170 is dependent on intact microtubules in 3-d-old, but not in 1-d-old MDCK cell cultures. This suggests a regulated interaction between microtubules and desmosomes and a role for pp170 in the control of changes in the properties of microtubules induced by epithelial cell-cell junction formation.
上皮细胞极性的建立与特殊细胞间连接的形成以及微管组织的显著变化相关。MDCK细胞中的很大一部分微管变得稳定,呈非中心体组织排列,并沿顶-基轴排列成纵向束状。这种相关性表明细胞间连接形成与微管组织控制之间存在功能联系。我们追踪了一种最近描述的微管结合蛋白pp170在上皮细胞极性建立过程中的分布。这种蛋白在亚汇合的成纤维细胞和上皮细胞中沿微管呈典型的斑点状分布,常与微管亚群的外周末端相关。然而,与它在汇合的成纤维细胞(A72)和HeLa细胞中的定位不同,pp170在汇合的极化上皮细胞(MDCK和Caco-2)中积聚在描绘细胞间接触区域的斑点中。对极化的MDCK细胞进行细胞间连接蛋白特异性抗体的双重免疫定位、共聚焦显微镜检查和免疫电子显微镜检查表明,pp170积聚在桥粒斑处。此外,发现微管和桥粒紧密接触。在3日龄的MDCK细胞培养物中,pp170与桥粒的结合维持依赖于完整的微管,但在1日龄的培养物中则不然。这表明微管和桥粒之间存在调节性相互作用,并且pp170在上皮细胞间连接形成诱导的微管特性变化的控制中发挥作用。