title: Expression of E-cadherin by human retinal pigment epithelium: delayed expression in vitro.
abstract: OBJECTIVE
  To determine whether retinal pigment epithelial (RPE) cells, which reportedly express N-cadherin as their major cadherin cell adhesion protein, also express the more common epithelial cadherin, E-cadherin.
  
  
  METHODS
  Cadherins expressed by human RPE cells in situ were examined by western blot analysis of extracts prepared from the RPE of human adult eyes. Cadherins expressed in vitro were examined by analysis of confluent and postconfluent human RPE cultures, using the methods of reverse transcription-polymerase chain reaction (RT-PCR) and western blot analysis. Protein distribution was examined by conventional fluorescence microscopy, confocal imaging, or both. Proteins whose expression, distribution, or both correlated with E-cadherin expression in other epithelial cells were examined by similar methods in cultured RPE cells.
  
  
  RESULTS
  In addition to N-cadherin, E-cadherin (and P-cadherin) was found in adult human RPE in situ. In cultured human RPE cells, N-cadherin was ubiquitous, but E-cadherin was limited to patches of cells and was not expressed until several weeks after confluence, a time when several phenotypic variants become prominent. E-cadherin was absent from RPE cells of fusiform shape but was found in only a subset of epithelioid RPE cells. Unlike epithelial cell lines expressing E-cadherin, cultured RPE cells with E-cadherin did not show diminished coexpression of N-cadherin, increased expression of desmosomal proteins, or a preferential expression of the alphaE- (rather than alpha-N) isoform of the cadherin linker protein alpha-catenin. Na/K ATPase distributed to both apical and basolateral membranes in RPE cells with junctional E-cadherin and not preferentially to the basolateral domain as in most epithelial cells with E-cadherin.
  
  
  CONCLUSIONS
  RPE cells express E-cadherin, a cadherin found in most other epithelial cells, but which was believed to be absent from RPE. In RPE in vitro, E-cadherin expression is a late developmental event, occurring in late confluence in cells that already express N-cadherin. E-cadherin is an established epithelial morphoregulatory protein, but it does not induce the same properties in RPE cells as in other epithelial cells, suggesting tissue-specific differences in the potential of E-cadherin to determine an epithelial phenotype.
authors.affiliation: Department of Ophthalmology, Medical College of Wisconsin, Milwaukee, USA. jburke@mcw.edu
authors.forename: J M
authors.initials: JM
authors.lastname: Burke
authors.forename: F
authors.initials: F
authors.lastname: Cao
authors.forename: P E
authors.initials: PE
authors.lastname: Irving
authors.forename: C M
authors.initials: CM
authors.lastname: Skumatz
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015153:Blotting, Western; D015820:Cadherins / Q000096:biosynthesis* / Q000235:genetics; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D051180:Desmoplakins; D004847:Epithelial Cells / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007223:Infant; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D010857:Pigment Epithelium of Eye / Q000166:cytology / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism
chemical_list: D015820:Cadherins; D003598:Cytoskeletal Proteins; D017931:DNA Primers; D051180:Desmoplakins; D012333:RNA, Messenger; D000254:Sodium-Potassium-Exchanging ATPase