title: Apical and basal regulation of the permeability of the retinal pigment epithelium.
abstract: OBJECTIVE
  The functional characteristics of tight junctions in the outer blood-retinal barrier change during embryonic development and in the presence of disease. A culture model of developing retinal pigment epithelium (RPE) was used to examine the regulation of the tight junctions.
  
  
  METHODS
  RPE from chick embryos was cultured on filters that separated the apical and basal medium compartments. Cultures were maintained in various combinations of serum-free medium, serum-free medium that was conditioned by neural retinas, or serum-free medium that was supplemented with bovine pituitary extract, serum, or various hormones. Function was monitored by the transepithelial electrical resistance (TER) or the permeation of small organic tracers. Structure was monitored by immunofluorescence and freeze-fracture electron microscopy.
  
  
  RESULTS
  Functional analysis indicated differences in permeability among RPE of different embryonic age and culture conditions. In serum-free medium, the tight junctions were leaky or failed to form. Barrier properties increased if pituitary extract was added to the basal medium chamber or retina-conditioned medium was added to the apical chamber. Retina-conditioned medium was more effective at organizing tight junctional strands into a continuous network, but bovine pituitary extract appeared to modulate the permeability of that network. In combination, they synergistically elevated the TER to physiological levels. Although the thyroid hormone T3 had no effect, serum in the apical medium chamber inhibited the ability of RPE cells to respond to retina-conditioned medium.
  
  
  CONCLUSIONS
  Diffusible factors secreted by the neural retina acted synergistically with basolateral stimulation to regulate the structure and function of RPE tight junctions. Serum on the apical side of the RPE monolayer inhibited the ability of retinal factors to upregulate the tight junction barrier.
authors.affiliation: Department of Surgery, Yale University School of Medicine, New Haven, Connecticut 06520, USA.
authors.forename: Shaomin
authors.initials: S
authors.lastname: Peng
authors.forename: Christoph
authors.initials: C
authors.lastname: Rahner
authors.forename: Lawrence J
authors.initials: LJ
authors.lastname: Rizzolo
mesh_terms: D000818:Animals; D001769:Blood; D002452:Cell Count; D002463:Cell Membrane Permeability / Q000502:physiology*; D002478:Cells, Cultured; D002642:Chick Embryo; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D004553:Electric Conductivity; D005614:Freeze Fracturing; D008565:Membrane Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D062793:Occludin; D010857:Pigment Epithelium of Eye / Q000166:cytology* / Q000378:metabolism; D019108:Tight Junctions / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D008565:Membrane Proteins; D062793:Occludin