Characterization of a rat glomerular visceral epithelial cell line.
Cultures of glomerular epithelial cells (GEC) are currently used to identify important cellular and molecular mechanisms involved in the pathogenesis of renal diseases. However, there is still controversy in the literature as to the visceral or parietal origin of cultured GEC. Our aim was to firmly establish the nature of a GEC cell line. The reactivity of cultured GEC was investigated with a large panel of mono- and polyclonal antibodies by using immunofluorescent techniques and compared with literature data on the in vivo expression of these antigens on podocytes. In addition, the podocyte specific 5A (podocalyxin), 13A and 27A (9-O-acetylated GD3) antigen expression was investigated in immuno-overlay experiments with isolated gangliosides and in immunoprecipitations with metabolically labelled cells. In general, immunoreactivities between cultured GEC and literature data on GEC in vivo expressions were similar. Important podocyte epitopes in vivo were expressed by cultured GEC such as podocalyxin, gp330 and the 13A antigen. Cultured GEC however differed from their in vivo counterparts in their expression of keratin-18, their lack of expression of pp44 and no detectable immunohistological expression of the ganglioside 9-O-acetylated GD3. Interestingly, the podocyte-specific epitope 9-O-acetylated GD3 was detected by the 27A antibodies in immuno-overlays of isolated GEC gangliosides. Moreover, by using the 27A antibody, we were able to precipitate the podocyte-specific 103-kD protein from 35S-methionine metabolically labelled GEC. From our immunohistological data together with the detectability of the 27A antigen we conclude that the cell line we use very probably originates from glomerular visceral epithelial cells.
Coers
W
W
Department of Pathology, University of Groningen, The Netherlands.
Reivinen
J
J
Miettinen
A
A
Huitema
S
S
Vos
J T
JT
Salant
D J
DJ
Weening
J J
JJ
D000818:Animals; D002460:Cell Line; D003598:Cytoskeletal Proteins / Q000032:analysis; D004847:Epithelial Cells; D004848:Epithelium / Q000737:chemistry / Q000276:immunology; D016326:Extracellular Matrix Proteins / Q000032:analysis; D005455:Fluorescent Antibody Technique; D005732:Gangliosides / Q000032:analysis; D006801:Humans; D007118:Immunoassay; D007163:Immunosorbent Techniques; D007678:Kidney Glomerulus / Q000737:chemistry / Q000166:cytology* / Q000276:immunology; D008565:Membrane Proteins / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D014781:Viscera
D003598:Cytoskeletal Proteins; D016326:Extracellular Matrix Proteins; D005732:Gangliosides; D008565:Membrane Proteins; C049132:9-O-acetyl-GD3 ganglioside