A simple method to identify NBT-positive cells in isolated glomeruli

• Published in: Nephrology, dialysis, transplantation Vol. 12; no. 1; pp. 174 - 179
• Main Authors: ROMERO, M, MOSQUERA, J, RODRIGUEZ-ITURBE, B
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 1997
• Subjects: Animals; Biological and medical sciences; Enzyme Inhibitors - pharmacology; Free Radicals - metabolism; Humans; Kidney Diseases - chemically induced; Kidney Diseases - metabolism; Kidney Diseases - pathology; Kidney Glomerulus - cytology; Kidney Glomerulus - drug effects; Kidney Glomerulus - metabolism; Kidneys; Male; Medical sciences; Nephritis - metabolism; Nephritis - pathology; Nephrology. Urinary tract diseases; Nitroblue Tetrazolium - metabolism; Oxidoreductases - antagonists & inhibitors; Piroxicam - pharmacology; Puromycin Aminonucleoside - toxicity; Rats; Rats, Inbred Lew; Reactive Oxygen Species - metabolism; Superoxide Dismutase - pharmacology; Tetradecanoylphorbol Acetate - pharmacology; Ureteral Obstruction - metabolism; Ureteral Obstruction - pathology; Urinary system involvement in other diseases. Miscellaneous; Kidney disease; Oxygen; Urinary system disease; Rat; Rodentia; Free radical; Isolated organ; Renal glomerulus; Vertebrata; Experimental disease; Mammalia; Nephropathy; Urinary system; Animal; Production; Nitroblue tetrazolium test; Diagnosis
• ISSN: 0931-0509; 1460-2385
• DOI: 10.1093/ndt/12.1.174

Reactive oxygen radicals are probably involved in the pathogenesis of human and experimental models of renal disease, yet current methods are inadequate to quantify and identify the cells producing reactive oxygen radicals. We used the nitroblue tetrazolium reaction to determine superoxide anion production in glomerular cells in phorbol myristate-stimulated glomerular suspensions and in isolated glomeruli from rats with nephrotoxic nephritis, ureteral obstruction, and puromycin aminonucleoside nephrosis. We were also able to identify these nitroblue tetrazolium + cells using specific appropriate antibodies. When the technique was tested in conditions known to increase reactive oxygen radicals, as phorbol myristate-stimulated glomeruli and glomeruli from animals with nephrotoxic nephritis and ureteral obstruction, increased number of nitroblue tetrazolium + cells were found. These cells were identified as glomerular intrinsic cells (Thy-1 +) or infiltrating leukocytes (leukocyte common antigen + or antineutrophil +). This method may be useful to determine cells participating in glomerular damage induced by reactive oxygen radicals.