title: Distribution of alpha-chains of type IV collagen in glomerular basement membranes with ultrastructural alterations suggestive of Alport syndrome.
abstract: BACKGROUND
  In Alport syndrome (AS) impaired production and/or assembly of col IV alpha-chain isoforms results in abnormal structure of glomerular basement membrane (GBM), haematuria and, frequently, progressive renal disease. We investigated the relationship between col IV alpha-chains expression and morphology of GBM, as a possible key to the better understanding of the pathogenesis of renal disease in AS.
  
  
  METHODS
  GBM distribution of col IV alpha1-, alpha3-, and alpha5-chain was investigated by immunohistochemistry in 32 patients (21 males and 11 females, mean age at biopsy of 11.5 years) with ultrastructural findings suggestive of AS. Ten patients had a proven COL4A5 mutation. Based on the severity of ultrastructural findings, the biopsies were grouped in three (I-III) electron microscopy (EM) classes. Significant EM changes of GBM (thinning, thickening, splitting, basket weaving of the lamina densa) were singularly evaluated using a semiquantitative scale (0-3).
  
  
  RESULTS
  Col IV alpha1-chain was demonstrated in GBM of all patients. Three patterns of staining for col IValpha3- and alpha5-chains were observed: positive, negative, and alpha3(IV)-positive/alpha5(IV)-negative. By chi(2)-test, EM class III lesions and complete loss of alpha3(IV)- and alpha5(IV)-antigen were significantly more frequent (P<0.05 and P<0.01) in male patients, but no significant relation was observed between EM classes and immunohistochemical patterns. GBM alterations did not correlate with staining for alpha5(IV)-chain. Intensity of alpha3(IV)-chain staining, however, had a negative correlation (P<0.05) with the severity of GBM basket weaving.
  
  
  CONCLUSIONS
  Our results suggest that the alpha3(IV)-chain-containing col IV-network plays a fundamental role in structural and, possibly, functional organization of GBM. Absence of alpha3(IV)-chain in GBM could indicate a more severe renal disease in AS.
authors.affiliation: Dipartimento di Medicina Sperimentale e Patologia, Università La Sapienza, Viale Regina Elena, 324 (Policlinico Umberto I), I-00161 Rome, Italy.
authors.forename: P
authors.initials: P
authors.lastname: Barsotti
authors.forename: A O
authors.initials: AO
authors.lastname: Muda
authors.forename: G
authors.initials: G
authors.lastname: Mazzucco
authors.forename: L
authors.initials: L
authors.lastname: Massella
authors.forename: B
authors.initials: B
authors.lastname: Basolo
authors.forename: M
authors.initials: M
authors.lastname: De Marchi
authors.forename: G
authors.initials: G
authors.lastname: Rizzoni
authors.forename: G
authors.initials: G
authors.lastname: Monga
authors.forename: T
authors.initials: T
authors.lastname: Faraggiana
mesh_terms: D000293:Adolescent; D000328:Adult; D001485:Basement Membrane / Q000378:metabolism* / Q000648:ultrastructure; D002648:Child; D002675:Child, Preschool; D003094:Collagen / Q000235:genetics / Q000378:metabolism*; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D007678:Kidney Glomerulus / Q000378:metabolism* / Q000648:ultrastructure*; D008297:Male; D008854:Microscopy, Electron; D008875:Middle Aged; D009394:Nephritis, Hereditary / Q000378:metabolism* / Q000473:pathology*; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D014018:Tissue Distribution
chemical_list: D020033:Protein Isoforms; D003094:Collagen