Zonulae occludentes in junctional complex-enriched fractions from mouse liver: preliminary morphological and biochemical characterization.
Zonulae occludentes in junctional complex-enriched fractions from mouse liver: preliminary morphological and biochemical characterization.
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DOI：
10.1083/jcb.98.4.1209
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发表时间：
1984-04
期刊：
影响因子：
--
通讯作者：
Goodenough DA
中科院分区：
文献类型：
--
作者：
Stevenson BR;Goodenough DA
使用亚细胞分级分离技术从小鼠肝脏中获得了含有连接复合物的胆小管衍生制剂。连接复合体包括结构完整的闭塞小带 (ZO)。用阴离子去污剂脱氧胆酸钠（DOC）提取该制剂，留下连接带，即连接复合物的去污剂不溶性小带残余物。当在负染色电子显微镜下观察时，这些带中的每一个都包含原纤维的分支和吻合网络，其看起来与冷冻断裂的整个肝脏中的 ZO 相似。冷冻断裂和负染色原纤维直径和网络密度的比较测量支持了这种关系。 SDS 聚丙烯酰胺凝胶分析显示，DOC 不溶性连接带的特征在于 37,000 和 48,000 处的主要多肽，以及 34,000、41,000、71,000、86,000、92,000 和 102,000 处的次要条带。在其他浓度的 EGTA 存在下，在其他人证明会破坏整个活体上皮细胞中正常 ZO 形态和生理学的条件下，已分离出含 ZO 的膜部分。通过负染色在这些级分中可视化的原纤维网络在结构上对 DOC 处理具有抗性，但会被 N-月桂酰肌氨酸溶解或破坏。
A bile canaliculus-derived preparation containing junctional complexes has been obtained from mouse livers using subcellular fractionation techniques. The junctional complexes include structurally intact zonulae occludentes (ZOs). Extraction of this preparation with the anionic detergent sodium deoxycholate (DOC) left junctional ribbons, the detergent-insoluble zonular remnants of the junctional complexes. When visualized in negative stain electron microscopy, each of these ribbons contained a branching and anastomosing network of fibrils which appears similar to that of ZOs in freeze-fractured whole liver. Comparative measurements of freeze-fracture and negative stain fibril diameters and network densities support this relationship. SDS polyacrylamide gel analysis shows the DOC-insoluble junctional ribbons to be characterized by major polypeptides at 37,000 and at 48,000, with minor bands at 34,000, 41,000, 71,000, 86,000, 92,000, and 102,000. The ZO-containing membrane fractions have been isolated in the presence of EGTA in concentrations and under conditions shown by others to disrupt normal ZO morphology and physiology in whole living epithelia. The network of fibrils visualized in these fractions by negative staining is structurally resistant to treatment with DOC, but is either solubilized or disrupted by N-lauroylsarcosine.