title: Retinal pigment epithelial cells from dystrophic rats form normal tight junctions in vitro.
abstract: OBJECTIVE
  In the genetically defective Royal College of Surgeons (RCS) rat model for retinal degeneration, a breakdown occurs in the retinal pigment epithelial (RPE) cell tight junctions just as the photoreceptors begin to degenerate. These experiments sought to determine the impact of the RPE genetic defect on this alteration in the RPE cell tight junctions.
  
  
  METHODS
  Retinal pigment epithelial cell cultures prepared from RCS and control rats were treated with hormonally defined medium (HDM), base medium conditioned by RCS or control retinas, or unconditioned base medium. The tight junctions formed by these cultures were assayed functionally by measuring transepithelial electrical resistance and permeability. Junction structure was evaluated by immunolocalization of the tight junction protein zonula occludens 1 and of the junction-associated actin microfilaments.
  
  
  RESULTS
  Retinal pigment epithelial cultures from dystrophic rats formed structurally and functionally normal tight junctions when maintained in hormonally defined medium. The junctions remained stable when the medium bathing the apical surface was switched to base medium preconditioned by normal retinas. In contrast, cultures treated with medium preconditioned by degenerating dystrophic retinas or with unconditioned medium exhibited a breakdown in their tight junctions.
  
  
  CONCLUSIONS
  Retinal pigment epithelial cells isolated from dystrophic RCS rats can form tight junctions normally in vitro. Normal, but not dystrophic, retinas release factors that support RPE tight junctions. Therefore, the junctional abnormality seen in dystrophic rat RPE cells in vivo is probably caused by the loss of trophic factors normally provided by the healthy neural retina rather than by a direct effect of the genetic defect on the tight junctions.
authors.affiliation: Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta 30912-3400, USA.
authors.forename: C W
authors.initials: CW
authors.lastname: Chang
authors.forename: D M
authors.initials: DM
authors.lastname: Defoe
authors.forename: R B
authors.initials: RB
authors.lastname: Caldwell
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D001813:Blood-Retinal Barrier; D018929:Cell Culture Techniques; D002463:Cell Membrane Permeability; D003470:Culture Media; D017077:Culture Media, Conditioned; D019084:Fluorescent Antibody Technique, Indirect; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008564:Membrane Potentials; D008565:Membrane Proteins / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D010857:Pigment Epithelium of Eye / Q000378:metabolism / Q000473:pathology*; D051381:Rats; D011922:Rats, Mutant Strains; D012162:Retinal Degeneration / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D019108:Tight Junctions / Q000378:metabolism / Q000473:pathology*; D062826:Zonula Occludens-1 Protein
chemical_list: D000199:Actins; D003470:Culture Media; D017077:Culture Media, Conditioned; D008565:Membrane Proteins; D010750:Phosphoproteins; C569044:Tjp1 protein, rat; D062826:Zonula Occludens-1 Protein; D007770:L-Lactate Dehydrogenase