In vivo enzymatic removal of alpha 2-->6-linked sialic acid from the glomerular filtration barrier results in podocyte charge alteration and glomerular injury.
The epithelial polyanion (podocalyxin) on the foot processes (pedicels) of podocytes plays a pivotal role in maintaining slit pore integrity and excluding proteins from the glomerular filtrate. Chromatographically purified recombinant sialidase from Vibrio cholerae, a corresponding heat-inactivated enzyme, truncated enzyme (missing the last 17 amino acids from the carboxyl terminus), and the sialidase from Salmonella typhimurium strain LT2 were inoculated intraperitoneally into mice, and the resultant renal alterations were documented by a variety of functional, morphologic, and histochemical techniques. Proteinuria and renal failure developed in a dose-dependent manner after a single inoculation of sialidase from Vibrio cholerae, but not with the corresponding heat-inactivated enzyme, truncated enzyme, or the sialidase from Salmonella typhimurium strain LT2. Biotinylated lectins of known sialyl linkage specificity demonstrated that Vibrio cholerae sialidase primarily removed alpha 2-->6-linked sialic acids from the glomerulus. Furthermore, the use of a poly-L-lysine cationic gold ultrastructural probe confirmed a transient loss of charge from the endothelium and epithelium of the glomerular filtration barrier. Loss of the epithelial polyanion was accompanied by the effacement of pedicels and the apparent formation of tight junctions between adjacent podocytes. The anionic charge returned to endothelial and epithelial sites within 2 days of sialidase inoculation, but the foot process loss remained. This animal model, in addition to providing an opportunity to study basic mechanisms of renal physiology, seems to mimic minimal change disease in children, diabetic nephropathy, and the renal effects of some bacterial infections.
Gelberg
H
H
Department of Pathobiology, College of Veterinary Medicine, University of Illinois, Urbana 61801, USA.
Healy
L
L
Whiteley
H
H
Miller
L A
LA
Vimr
E
E
D000419:Albuminuria; D000818:Animals; D003404:Creatinine / Q000652:urine; D005920:Glomerular Mesangium / Q000737:chemistry / Q000473:pathology; D007274:Injections, Intraperitoneal; D007674:Kidney Diseases / Q000139:chemically induced / Q000378:metabolism* / Q000473:pathology*; D007678:Kidney Glomerulus / Q000737:chemistry / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure; D037102:Lectins; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D019158:N-Acetylneuraminic Acid; D009439:Neuraminidase / Q000008:administration & dosage / Q000378:metabolism*; D011994:Recombinant Proteins; D012794:Sialic Acids / Q000502:physiology*
D037102:Lectins; D011994:Recombinant Proteins; D012794:Sialic Acids; D003404:Creatinine; D009439:Neuraminidase; D019158:N-Acetylneuraminic Acid