Stevenson B R, Anderson J M, Goodenough D A, Mooseker M S
Department of Biology, Yale University School of Medicine, New Haven, Connecticut.
J Cell Biol. 1988 Dec;107(6 Pt 1):2401-8. doi: 10.1083/jcb.107.6.2401.
The relationship of tight junction permeability to junction structure and composition was examined using two strains of Madin-Darby canine kidney (MDCK) cells (I and II) which differ greater than 30-fold in transepithelial resistance. This parameter is largely determined by paracellular, and hence junctional, permeability under most conditions. When these two strains of cells were grown on permeable filter supports, they formed monolayers with equivalent linear amounts of junction/area of monolayer. Ultrastructural analysis of these monolayers by thin section EM revealed no differences in overall cellular morphology or in tight junction organization. Morphometric analysis of freeze-fractured preparations indicated that the tight junctions of these two cell strains were similar in both number and density of junctional fibrils. Prediction of transepithelial resistance for the two strains from this freeze-fracture data and a published structure-function formulation (Claude, P. 1978, J. Memb. Biol. 39:219-232) yielded values (I = 26.5 omega/cm2, II = 35.7 omega/cm2) that were significantly lower than those observed (I = 2,500-5,000 omega/cm2, II = 50-70 omega/cm2). Consistent with these structural studies, a comparison of the distribution and cellular content of ZO-1, a polypeptide localized exclusively to the tight junction, revealed no significant differences in either the localization of ZO-1 or the amount of ZO-1 per micron of junction (I = 1,415 +/- 101 molecules/micron, II = 1,514 +/- 215 molecules/micron).
利用两株Madin-Darby犬肾(MDCK)细胞(I和II)研究紧密连接通透性与连接结构和组成的关系,这两株细胞的跨上皮电阻相差30倍以上。在大多数情况下,这个参数很大程度上由细胞旁通透性决定,因此也由连接通透性决定。当这两株细胞在可渗透滤膜支持物上生长时,它们形成的单层细胞中连接的线性量与单层面积相当。通过超薄切片电镜对这些单层细胞进行超微结构分析,结果显示整体细胞形态或紧密连接组织没有差异。对冷冻断裂制剂的形态计量分析表明,这两株细胞的紧密连接在连接纤维的数量和密度上相似。根据这个冷冻断裂数据和已发表的结构-功能公式(Claude, P. 1978, J. Memb. Biol. 39:219-232)预测这两株细胞的跨上皮电阻,得到的值(I = 26.5Ω/cm2,II = 35.7Ω/cm2)明显低于观察到的值(I = 2500 - 5000Ω/cm2,II = 50 - 70Ω/cm2)。与这些结构研究一致,对仅定位于紧密连接的多肽ZO-1的分布和细胞含量进行比较,结果显示ZO-1的定位或每微米连接中ZO-1的量均无显著差异(I = 1415 ± 101分子/微米,II = 1514 ± 215分子/微米)。