title: Ultrastructural changes of extracellular matrices in diabetic nephropathy revealed by high resolution scanning and immunoelectron microscopy.
abstract: BACKGROUND
  Diabetic nephropathy is invariably associated with proteinuria.
  
  
  METHODS
  To delineate the mechanism(s) of proteinuria in diabetic nephropathy, ultrastructural changes of the glomerular basement membranes (GBMs) were studied by high resolution scanning and immunoelectron microscopy. Acellular glomeruli from diabetic and age-matched control human subjects were prepared by detergent method and subjected to conductive staining, the technique in which the tissues are impregnated with metals rather than surface-coated with metallic alloys for visualization by electron microscopy. Subsequent to conductive staining, the tissues were examined by in-lens field emission scanning electron microscopy.
  
  
  RESULTS
  Thirty glomeruli, each from the control and diabetic groups, were examined by scanning microscopy. In diabetic GBMs, a loose meshwork structure consisting of numerous pores of approximately 8 nm in diameter and distinct strands was observed. In contrast, meshwork structure was not readily discernible in controls and a few pores were observed. Five glomerular capillary loops, each from control and diabetic groups, were examined by immunoelectron microscopy. In controls, heparan sulfate-proteoglycan was localized in the lamina rara interna and externa, and type-IV collagen was distributed throughout the whole width of the GBM. In diabetic GBMs, a relative loss of staining of heparan sulfate-proteoglycan, both in the lamina rara interna and externa of the GBM, was observed. Type IV collagen was distributed in all layers of the thickened GBM, and the absolute number of the immunogold particles was increased. However, immunogold particle density of type IV collagen per unit area was decreased as compared with the control.
  
  
  CONCLUSIONS
  These ultrastructural and immunoelectron microscopic changes in the GBM may explain the loss of charge as well as size selectivities of the glomerulus, as observed in diabetic nephropathy associated with proteinuria.
authors.affiliation: Third Department of Internal Medicine, Okayama University Medical School, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Makino
authors.forename: Y
authors.initials: Y
authors.lastname: Yamasaki
authors.forename: T
authors.initials: T
authors.lastname: Haramoto
authors.forename: K
authors.initials: K
authors.lastname: Shikata
authors.forename: K
authors.initials: K
authors.lastname: Hironaka
authors.forename: Z
authors.initials: Z
authors.lastname: Ota
authors.forename: Y S
authors.initials: YS
authors.lastname: Kanwar
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D001485:Basement Membrane / Q000378:metabolism / Q000648:ultrastructure; D003094:Collagen / Q000145:classification / Q000378:metabolism; D003928:Diabetic Nephropathies / Q000473:pathology*; D005109:Extracellular Matrix / Q000648:ultrastructure*; D005260:Female; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000378:metabolism; D006801:Humans; D007678:Kidney Glomerulus / Q000378:metabolism / Q000648:ultrastructure; D008297:Male; D008855:Microscopy, Electron, Scanning; D016253:Microscopy, Immunoelectron; D011509:Proteoglycans / Q000378:metabolism; D012016:Reference Values
chemical_list: D003094:Collagen; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D011509:Proteoglycans