title: In vivo degradation of heparan sulfates in the glomerular basement membrane does not result in proteinuria.
abstract: Heparan sulfates (HS) are long, unbranched, negatively charged polysaccharides that are bound to core proteins. HS in the glomerular basement membrane (GBM) is reported to be important for charge-selective permeability. Aberrant GBM HS expression has been observed in several glomerular diseases, such as diabetic nephropathy and membranous glomerulopathy, and a decrease in HS generally is associated with proteinuria. This study, with the use of a controlled in vivo approach, evaluated whether degradation of HS in rat GBM resulted in acute proteinuria. Rats received two intravenous injections of either heparinase III to digest HS or neuraminidase to remove neuraminic acids (positive control). Urine samples were taken at various time points, and at the end of the experiment, kidneys were removed and analyzed. Injection with heparinase III resulted in a complete loss of glomerular HS as demonstrated by immunofluorescence staining using anti-HS antibodies and by electron microscopy using cupromeronic blue in a critical electrolyte concentration mode. In the urine, a strong increase in HS was found within 2 h after the first injection. Staining for agrin, the major HS proteoglycan core protein in the GBM, was unaltered. No urinary albumin or other proteins were detected at any time point, and no changes in glomerular morphology were noticed. Injection of rats with neuraminidase, however, resulted in a major increase of urinary albumin and was associated with an increase in urinary free neuraminic acid. An increased glomerular staining with Peanut agglutinin lectin, indicative of removal of neuraminic acid, was noted. In conclusion, removal of HS from the GBM does not result in acute albuminuria, whereas removal of neuraminic acid does.
authors.affiliation: Department of Matrix Biochemistry, Radboud University Nijmegen Medical Centre, Nijmegen Centre for Molecular Life Sciences, Nijmegen, PO Box 9101, 6500 HB Nijmegen, The Netherlands.
authors.forename: Tessa J M
authors.initials: TJ
authors.lastname: Wijnhoven
authors.forename: Joost F M
authors.initials: JF
authors.lastname: Lensen
authors.forename: Ronnie G P
authors.initials: RG
authors.lastname: Wismans
authors.forename: Mohammed
authors.initials: M
authors.lastname: Lamrani
authors.forename: Leo A H
authors.initials: LA
authors.lastname: Monnens
authors.forename: Ron A
authors.initials: RA
authors.lastname: Wevers
authors.forename: Angelique L W M M
authors.initials: AL
authors.lastname: Rops
authors.forename: Johan
authors.initials: J
authors.lastname: van der Vlag
authors.forename: Jo H M
authors.initials: JH
authors.lastname: Berden
authors.forename: Lambert P W J
authors.initials: LP
authors.lastname: van den Heuvel
authors.forename: Toin H
authors.initials: TH
authors.lastname: van Kuppevelt
mesh_terms: D000419:Albuminuria / Q000209:etiology / Q000378:metabolism; D000818:Animals; D050533:Glomerular Basement Membrane / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D006025:Glycosaminoglycans / Q000378:metabolism*; D006497:Heparan Sulfate / Q000032:analysis / Q000378:metabolism*; D007668:Kidney / Q000378:metabolism / Q000648:ultrastructure; D008297:Male; D008854:Microscopy, Electron; D009438:Neuraminic Acids / Q000378:metabolism*; D009439:Neuraminidase / Q000494:pharmacology; D011133:Polysaccharide-Lyases / Q000494:pharmacology; D011507:Proteinuria / Q000209:etiology*; D051381:Rats; D017208:Rats, Wistar
chemical_list: D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D009438:Neuraminic Acids; D009439:Neuraminidase; D011133:Polysaccharide-Lyases; C021349:heparitinsulfate lyase