title: Hsp27 regulates podocyte cytoskeletal changes in an in vitro model of podocyte process retraction.
abstract: Nephrotic syndrome (NS) is characterized by structural changes in the actin-rich foot processes of glomerular podocytes. We previously identified high concentrations of the small heat shock protein hsp27 within podocytes as well as increased glomerular accumulation and phosphorylation of hsp27 in puromycin aminonucleoside (PAN) -induced experimental NS. Here we analyzed murine podocytes stably transfected with hsp27 sense, antisense, and vector control constructs using a newly developed in vitro PAN model system. Cell morphology and the microfilament structure of untreated sense and antisense transfectants were altered compared with controls. Vector cell survival, polymerized actin content, cell area, and hsp27 content increased after 1.25 microg/ml PAN treatment and decreased after 5.0 microg/ml treatment. In contrast, sense cells were unaffected by 1.25 microg/ml PAN treatment whereas antisense cells showed decreases or no changes in all parameters. Treatment of sense cells with 5.0 microg/ml PAN resulted in increased cell survival and cell area whereas antisense cells underwent significant decreases in all parameters. Hsp27 provided dramatic protection against PAN-induced microfilament disruption in sense > vector > antisense cells. We conclude that hsp27 is able to regulate both the morphological and actin cytoskeletal response of podocytes in an in vitro model of podocyte injury.
authors.affiliation: Pediatric Nephrology Division, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: William E
authors.initials: WE
authors.lastname: Smoyer
authors.forename: Richard F
authors.initials: RF
authors.lastname: Ransom
mesh_terms: D008841:Actin Cytoskeleton / Q000187:drug effects / Q000648:ultrastructure*; D000199:Actins / Q000032:analysis; D000818:Animals; D002460:Cell Line; D002461:Cell Line, Transformed; D048429:Cell Size / Q000187:drug effects; D002470:Cell Survival; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D006360:Heat-Shock Proteins; D007678:Kidney Glomerulus / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure*; D051379:Mice; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast; D018832:Molecular Chaperones; D009363:Neoplasm Proteins / Q000235:genetics / Q000502:physiology*; D009404:Nephrotic Syndrome / Q000378:metabolism / Q000473:pathology; D010766:Phosphorylation; D011692:Puromycin Aminonucleoside / Q000494:pharmacology; D014162:Transfection
chemical_list: D000199:Actins; D006360:Heat-Shock Proteins; C486113:Hsbp1 protein, mouse; D018832:Molecular Chaperones; D009363:Neoplasm Proteins; D011692:Puromycin Aminonucleoside