A culture model of development reveals multiple properties of RPE tight junctions.

PURPOSE A culture model was used to examine the development of tight junctions in the retinal pigment epithelium (RPE). METHODS Chick RPE was isolated on embryonic day 7 (E7), E10 or E14 and cultured on laminin-coated filters. Barrier properties were stimulated with E14 retinal conditioned medium. Morphology was characterized by confocal microscopy. Permeability was determined by measuring the flux of horseradish peroxidase (HRP), radiolabeled inulin and mannitol, and the transepithelial electrical resistance (TER). Changes in the expression of ZO-1 and a related protein, ZO-1LP, were determined by immunoblotting. RESULTS RPE from each age formed epithelial monolayers of similar height, but the density of the cultures varied in parallel with density changes in vivo. The cultures appeared to regulate the permeability to ions and nonionic solutes independently. With embryonic age, there was a progressive decrease in permeability that first affected larger and then smaller tracers. Despite a small decrease in the permeability to mannitol, there was a large decrease in the permeability to ions. This suggests that in E14 cultures tight junctions discriminated by charge, as well as size. Although E14 retinal conditioned medium reduced the permeability to all solutes, it appeared to regulate size discrimination more than charge discrimination. Despite large effects on permeability, conditioned medium had no effect on the expression of ZO-1 or ZO-1LP. CONCLUSIONS The ability of tight junctions to discriminate on the basis of charge and size is regulated independently during development. The permeability of tight junctions cannot be predicted by the level of ZO-1 expression.

[1]  L. Rizzolo,et al.  Remodeling of junctional complexes during the development of the outer blood‐retinal barrier , 1997, The Anatomical record.

[2]  L. Rizzolo.  Polarity and the development of the outer blood-retinal barrier. , 1997, Histology and histopathology.

[3]  R. Caldwell,et al.  Serum Opens Tight Junctions and Reduces ZO‐1 Protein in Retinal Epithelial Cells , 1997, Journal of neurochemistry.

[4]  L. Rubin,et al.  Occludin as a possible determinant of tight junction permeability in endothelial cells. , 1997, Journal of cell science.

[5]  M. Itoh,et al.  Involvement of ZO-1 in Cadherin-based Cell Adhesion through Its Direct Binding to α Catenin and Actin Filaments , 1997, The Journal of cell biology.

[6]  C P Ponting,et al.  PDZ Domains: Targeting signalling molecules to sub‐membranous sites , 1997, BioEssays : news and reviews in molecular, cellular and developmental biology.

[7]  L. Ye,et al.  Serum inhibits tight junction formation in cultured pigment epithelial cells. , 1997, Investigative ophthalmology & visual science.

[8]  B. Gumbiner,et al.  A Synthetic Peptide Corresponding to the Extracellular Domain of Occludin Perturbs the Tight Junction Permeability Barrier , 1997, The Journal of cell biology.

[9]  J. Zieske,et al.  ZO1 in corneal epithelium: association to the zonula occludens and adherens junctions. , 1997, Experimental eye research.

[10]  R. D. Lynch,et al.  Occludin is a functional component of the tight junction. , 1996, Journal of cell science.

[11]  M. Balda,et al.  Functional dissociation of paracellular permeability and transepithelial electrical resistance and disruption of the apical- basolateral intramembrane diffusion barrier by expression of a mutant tight junction membrane protein , 1996, The Journal of cell biology.

[12]  A. Rajasekaran,et al.  Catenins and zonula occludens-1 form a complex during early stages in the assembly of tight junctions , 1996, The Journal of cell biology.

[13]  G. Firestone,et al.  Glucocorticoid-induced Functional Polarity of Growth Factor Responsiveness Regulates Tight Junction Dynamics in Transformed Mammary Epithelial Tumor Cells (*) , 1995, The Journal of Biological Chemistry.

[14]  C. V. Van Itallie,et al.  Tight junctions and the molecular basis for regulation of paracellular permeability. , 1995, The American journal of physiology.

[15]  G. Firestone,et al.  Transforming Growth Factor-α Abrogates Glucocorticoid-stimulated Tight Junction Formation and Growth Suppression in Rat Mammary Epithelial Tumor Cells (*) , 1995, The Journal of Biological Chemistry.

[16]  R. Sormunen,et al.  The polarity of the membrane skeleton in retinal pigment epithelial cells of developing chicken embryos and in primary culture. , 1995, Differentiation; research in biological diversity.

[17]  Gardner Tw.  Histamine, ZO-1 and increased blood-retinal barrier permeability in diabetic retinopathy. , 1995 .

[18]  T. Gardner.  Histamine, ZO-1 and increased blood-retinal barrier permeability in diabetic retinopathy. , 1995, Transactions of the American Ophthalmological Society.

[19]  S. Vinores.  Assessment of blood-retinal barrier integrity. , 1995, Histology and histopathology.

[20]  Y. Konishi,et al.  Localization of the 7H6 antigen at tight junctions correlates with the paracellular barrier function of MDCK cells. , 1994, Experimental cell research.

[21]  M. Hirsch,et al.  Tight junctions of the human ciliary epithelium: regional morphology and implications on transepithelial resistance. , 1994, Experimental eye research.

[22]  L. Rizzolo,et al.  Diffusible, retinal factors stimulate the barrier properties of junctional complexes in the retinal pigment epithelium. , 1993, Journal of cell science.

[23]  C. Hamel,et al.  Molecular cloning and expression of RPE65, a novel retinal pigment epithelium-specific microsomal protein that is post-transcriptionally regulated in vitro. , 1993, The Journal of biological chemistry.

[24]  M. Itoh,et al.  The 220-kD protein colocalizing with cadherins in non-epithelial cells is identical to ZO-1, a tight junction-associated protein in epithelial cells: cDNA cloning and immunoelectron microscopy , 1993, The Journal of cell biology.

[25]  C. Barnstable,et al.  RET-PE10: a 61 kD polypeptide epitope expressed late during vertebrate RPE maturation. , 1993, Investigative ophthalmology & visual science.

[26]  H. Wolburg,et al.  Tight junction complexity in the retinal pigment epithelium of the chicken during development , 1993, Neuroscience Letters.

[27]  J. Anderson,et al.  Localization and differential expression of two isoforms of the tight junction protein ZO-1. , 1992, The American journal of physiology.

[28]  M. Hughes,et al.  Detection of the tight junction-associated protein ZO-1 in astrocytes and other nonepithelial cell types. , 1992, The American journal of physiology.

[29]  L. Rubin,et al.  A cell culture model of the blood-brain barrier , 1991, The Journal of cell biology.

[30]  R. Dermietzel,et al.  Molecular anatomy of the blood-brain barrier as defined by immunocytochemistry. , 1991, International review of cytology.

[31]  C. Barnstable,et al.  Expression of the cell surface antigens RET-PE2 and N-CAM by rat retinal pigment epithelial cells during development and in tissue culture. , 1990, Experimental eye research.

[32]  L. Rizzolo.  The distribution of Na+,K(+)-ATPase in the retinal pigmented epithelium from chicken embryo is polarized in vivo but not in primary cell culture. , 1990, Experimental eye research.

[33]  N J Abbott,et al.  Electrical resistance across the blood‐brain barrier in anaesthetized rats: a developmental study. , 1990, The Journal of physiology.

[34]  James M. Anderson,et al.  Tight junction structure and ZO-1 content are identical in two strains of Madin-Darby canine kidney cells which differ in transepithelial resistance , 1988, The Journal of cell biology.

[35]  James M. Anderson,et al.  Characterization of ZO-1, a protein component of the tight junction from mouse liver and Madin-Darby canine kidney cells , 1988, The Journal of cell biology.

[36]  D. Beebe,et al.  Developmental changes in the blood-ocular barriers in chicken embryos. , 1984, Experimental eye research.

[37]  A. Coulombre,et al.  Correlations of structural and biochemical changes in the developing retina of the chick. , 1955, The American journal of anatomy.