The subcellular organization of Madin-Darby canine kidney cells during the formation of a polarized epithelium.
The subcellular organization of Madin-Darby canine kidney cells during the formation of a polarized epithelium.
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DOI：
10.1083/jcb.109.6.2817
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发表时间：
1989-12
期刊：
影响因子：
--
通讯作者：
Simons K
中科院分区：
文献类型：
--
作者：
Bacallao R;Antony C;Dotti C;Karsenti E;Stelzer EH;Simons K
对小鼠胚胎发育中的滋养外胚层的研究表明，在上皮形成过程中发生了广泛的细胞重塑。在这项研究中，共聚焦免疫荧光显微镜用于检查细胞结构的三维变化，这些变化发生在终末分化上皮细胞系的极化过程中。将Madin-Darby犬肾细胞低密度镀于可渗透滤膜支架上。特异性识别紧密连接、粘附连接、微管、中心体和高尔基复合体成分的抗体被用来研究极化细胞层形成过程中细胞结构的空间重构。免疫荧光数据与通过上皮抗性和细胞表面的反向交换测量的功能性紧密连接的建立相关，并用荧光脂质类似物N-（7-[4-硝基苯并-2-氧-1,3-二唑]）氨基丙基鞘氨醇的代谢物标记。镀后1 d，单个细胞有微管，从一个广泛的区域辐射，其中包含着中心体和高尔基复合体。2 d后，细胞已生长到融合，并在靠近基质处形成功能性紧密连接。中心粒已经分裂，不再组织在细胞核上下运行的微管。高尔基复合体已经扩散到细胞核周围。到电镀后的第五天，最终的极化状态已经达到。连接复合物从其基底位置向上移动了10微米以上。中心粒聚集在根尖膜下方，高尔基复合体位于细胞核上方的根尖区，呈带状卷曲结构。微管在柱状细胞的顶基轴上呈纵向微管束和顶端网状排列。纵向微管的排列方式为负端分布在细胞的顶端，正端分布在细胞的基部。这些发现表明，在细胞-细胞接触形成后，上皮细胞结构发生了广泛的重塑。微管网络的重组导致细胞质的功能性极化。
Studies of the developing trophectoderm in the mouse embryo have shown that extensive cellular remodeling occurs during epithelial formation. In this investigation, confocal immunofluorescence microscopy is used to examine the three-dimensional changes in cellular architecture that take place during the polarization of a terminally differentiated epithelial cell line. Madin-Darby canine kidney cells were plated at a low density on permeable filter supports. Antibodies that specifically recognize components of the tight junction, adherens junction, microtubules, centrosomes, and the Golgi complex were used to study the spatial remodeling of the cytoarchitecture during the formation of the polarized cell layer. The immunofluorescence data were correlated with establishment of functional tight junctions as measured by transepithelial resistance and back-exchange of the cell surface, labeled with metabolites of the fluorescent lipid analogue N-(7-[4- nitrobenzo-2-oxa-1,3-diazole]) aminocaproyl sphingosine. 1 d after plating, single cells had microtubules, radiating from a broad region, that contained the centrosomes and the Golgi complex. 2 d after plating, the cells had grown to confluence and had formed functional tight junctions close to the substratum. The centrioles had split and no longer organized the microtubules which were running above and below the nucleus. The Golgi complex had spread around the nucleus. By the fifth day after plating, the final polarized state had been achieved. The junctional complex had moved greater than 10 microns upward from its basal location. The centrioles were together below the apical membrane, and the Golgi complex formed a ribbon-like convoluted structure located in the apical region above the nucleus. The microtubules were organized in an apical web and in longitudinal microtubule bundles in the apical-basal axis of the columnar cell. The longitudinal microtubules were arranged with their minus ends spread over the apical region of the cell and their plus ends toward the basal region. These findings show that there is an extensive remodeling of epithelial cytoarchitecture after formation of cell-cell contacts. Reorganization of the microtubule network results in functional polarization of the cytoplasm.
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影响因子：
2.7
作者：
BURNSIDE, B
通讯作者：
BURNSIDE, B
DOI：
10.1083/jcb.100.1.327
发表时间：
1985-01
期刊：
The Journal of cell biology
影响因子：
--
作者：
Boller K;Vestweber D;Kemler R
通讯作者：
Kemler R
影响因子：
3.2
作者：
BEROD, A;HARTMAN, BK;PUJOL, JF
通讯作者：
PUJOL, JF
DOI：
10.1083/jcb.107.4.1575
发表时间：
1988-10
期刊：
The Journal of cell biology
影响因子：
--
作者：
Gumbiner B;Stevenson B;Grimaldi A
通讯作者：
Grimaldi A
DOI：
10.1083/jcb.108.4.1407
发表时间：
1989-04
期刊：
The Journal of cell biology
影响因子：
--
作者：
Fleming TP;McConnell J;Johnson MH;Stevenson BR
通讯作者：
Stevenson BR