title: Podocyte depletion and glomerulosclerosis have a direct relationship in the PAN-treated rat.
abstract: BACKGROUND
  Podocytes are highly differentiated glomerular epithelial cells with limited potential to divide. They are responsible for maintaining and supporting the glomerular basement membrane so as to facilitate efficient filtration. The hypothesis tested was whether the development of glomerulosclerosis in the puromycin aminonucleoside (PAN)-treated rat could be attributed to podocyte depletion.
  
  
  METHODS
  PAN was injected in Sprague-Dawley rats once, twice, or three times at 30-day intervals. Podocytes were counted in glomeruli using immunoperoxidase histochemistry and antibodies to both GLEPP1 (PTPRO) and WT-1. Podocytes were assayed in urine using reverse transcription-quantitative polymerase chain reaction (RT-QPCR). Glomerular areas were measured by computerized morphometry.
  
  
  RESULTS
  In a preliminary experiment, a single injection of PAN caused a reduction in the glomerular podocyte count by 25%. Additional independent confirmation that podocytes were lost from glomeruli after PAN injection was obtained identifying detached podocytes in Bowman's space, measurement of nephrin and GLEPP1 mRNAs in urine, ultrastructural analysis of glomeruli, and identification of TUNEL-positive apoptotic podocytes in glomeruli. In a second experiment, sequential podocyte depletion by 15, 31, and 53% was achieved by the administration of one, two, or three injections of PAN at 30-day intervals. The region of the glomerulus devoid of podocytes developed glomerulosclerosis, and this area progressively increased as podocytes were progressively depleted. The correlation coefficient (r(2)) value for the relationship between percent podocyte depletion and glomerulosclerotic area was 0.99. The Y intercept of this plot showed that glomerulosclerosis was initiated when only 10 to 20% of podocytes were lost.
  
  
  CONCLUSIONS
  This report supports the growing body of data linking glomerulosclerosis directly to a reduction in relative podocyte number [increased glomerular area per podocyte (GAPP)]. It raises important questions related to the mechanisms of podocyte loss, strategies for prevention of podocyte depletion, and the prevention of progression of glomerular diseases.
authors.affiliation: Division of Nephrology, Department of Internal Medicine, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Y H
authors.initials: YH
authors.lastname: Kim
authors.forename: M
authors.initials: M
authors.lastname: Goyal
authors.forename: D
authors.initials: D
authors.lastname: Kurnit
authors.forename: B
authors.initials: B
authors.lastname: Wharram
authors.forename: J
authors.initials: J
authors.lastname: Wiggins
authors.forename: L
authors.initials: L
authors.lastname: Holzman
authors.forename: D
authors.initials: D
authors.lastname: Kershaw
authors.forename: R
authors.initials: R
authors.lastname: Wiggins
mesh_terms: D000818:Animals; D002452:Cell Count; D004847:Epithelial Cells / Q000187:drug effects; D005923:Glomerulosclerosis, Focal Segmental / Q000139:chemically induced / Q000503:physiopathology*; D007678:Kidney Glomerulus / Q000503:physiopathology*; D008297:Male; D011692:Puromycin Aminonucleoside; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D011692:Puromycin Aminonucleoside