title: Vascular endothelial growth factor mRNA expression in minimal change, membranous, and diabetic nephropathy demonstrated by non-isotopic in situ hybridisation.
abstract: OBJECTIVE
  To investigate vascular endothelial growth factor (VEGF) mRNA expression in glomerular disease in the context of heavy proteinuria.
  
  
  METHODS
  Non-radioisotopic in situ hybridisation was performed using a cocktail of 12 deoxyoligonucleotides complementary to VEGF mRNA labelled during solid phase synthesis with 2,4-dinitrophenyl. Archival renal biopsies were studied from cases of minimal change nephropathy, membranous nephropathy, diabetic nephropathy, and controls, matched for age, sex, race, and storage time. Hybrid detection used NBT/BCIP colorimetric development.
  
  
  RESULTS
  More VEGF mRNA positive glomerular cells per unit cross sectional diameter were seen in minimal change nephropathy (mean (SEM), 19.35 (1.5)) compared with controls (12.6 (1.73)), p < 0.01. In contrast, fewer were seen in diabetic nephropathy (5.93 (0.97)) compared with controls (9.97 (1.25)), p < 0.03. Analysis of membranous nephropathy (10 (1.62)) showed no difference from controls (10.98 (1.51)), NS. In addition, in minimal change nephropathy there was a significant correlation between 24 hour protein excretion at the time of biopsy and the number of VEGF mRNA cells per glomerulus (r = 0.08, p = 0.01).
  
  
  CONCLUSIONS
  Using non-radioisotopic in situ hybridisation, VEGF mRNA is almost exclusively expressed by visceral glomerular epithelial cells. Abnormal numbers of cells are seen in both minimal change and diabetic nephropathy. As VEGF exists in a number of functionally distinct isoforms, further study of qualitative VEGF isoform expression in diagnostic groups is indicated.
authors.affiliation: Department of Nephrology, Leicester General Hospital, UK.
authors.forename: E
authors.initials: E
authors.lastname: Bailey
authors.forename: M J
authors.initials: MJ
authors.lastname: Bottomley
authors.forename: S
authors.initials: S
authors.lastname: Westwell
authors.forename: J H
authors.initials: JH
authors.lastname: Pringle
authors.forename: P N
authors.initials: PN
authors.lastname: Furness
authors.forename: J
authors.initials: J
authors.lastname: Feehally
authors.forename: P E
authors.initials: PE
authors.lastname: Brenchley
authors.forename: S J
authors.initials: SJ
authors.lastname: Harper
mesh_terms: D003928:Diabetic Nephropathies / Q000378:metabolism; D016228:Endothelial Growth Factors / Q000032:analysis* / Q000235:genetics; D004848:Epithelium / Q000378:metabolism; D015433:Glomerulonephritis, Membranous / Q000378:metabolism; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods; D007674:Kidney Diseases / Q000378:metabolism*; D007678:Kidney Glomerulus / Q000378:metabolism*; D016014:Linear Models; D008222:Lymphokines / Q000032:analysis* / Q000235:genetics; D009402:Nephrosis, Lipoid / Q000378:metabolism; D011507:Proteinuria / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis*; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
chemical_list: D016228:Endothelial Growth Factors; D008222:Lymphokines; D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors