MOLECULAR BIOLOGY OF ZONULAE OCCLUDENTES

生物上皮，其分隔的生理室充当 离子跨上皮通道的渗透性障碍 分子。 细胞之间的空间，称为细胞旁通路， 由类似垫圈的细胞间连接（闭锁带）密封 （ZO）。 ZO 是一系列细胞间质的最顶端成员 连接点，统称为连接复合体。 新颖的制剂 已制备出从啮齿动物肝脏中分离出来的连接复合物， 通过形态学技术证明包含结构成分 ZO 的。 这些复合物的聚丙烯酰胺凝胶分析揭示了 与整个多肽相比，大大简化了多肽的概况 肝细胞质膜。 使用单克隆和多克隆抗体 方法，在本申请中建议生成特定的 这些连接复合物制剂的抗血清并鉴定 通过免疫复制技术确定这些抗血清的特异性。 使用 免疫电子显微镜，抗原的解剖位置 被这些抗血清识别的将被映射到连接复合物内， 目的是专门鉴定该物质的分子成分 佐。 一旦确定，将研究 ZO 的生物发生和组装 在任一整体中合成后使用抗原的免疫沉淀 肝脏或粗糙微粒体富含多聚 A mRNA 的部分。 希望， 这些抗血清也将在生物运输研究中证明有价值 在运输上皮细胞的文化中。

Biological epithelia which separate physiological compartments act as permeability barriers to the transepithelial passage of ions and molecules. The spaces between the cells, called the paracellular pathway, are sealed by a gasket-like intercellular junction, the zonula occludens (ZO). The ZO is the apical-most member of a series of intercellular junctions, collectively termed the junctional complex. A novel preparation of isolated junctional complexes from rodent liver has prepared, which is demonstrated by morphological techniques to contain structural components of the ZO. Polyacrylamide gel analysis of these complexes reveals a greatly simplified profile of polypeptides, compared to the whole hepatocyte plasma membrane. Using monoclonal and polyclonal antibody approaches, it is proposed in this application to generate specific antisera to these junctional complex preparations and to identify the specificity of these antisera by immunoreplica techniques. Using immunoelectron microscopy, the anatomical locations of the antigens recognized by these antisera will be mapped within the junctional complex, with the goal of specifically identifying the molecular components of the ZO. Once identified, the biogenesis and assembly of the ZO will be studied using immune precipitation of antigens following synthesis in either whole liver or rough microsome enriched fractions of polyA mRNA. Hopefully, these antisera will also prove valuable in studies of biological transport in cultures of transporting epithelia.