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


RESULTS
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 +).


CONCLUSIONS
This method may be useful to determine cells participating in glomerular damage induced by reactive oxygen radicals.
Romero
M
M
Instituto de Investigaciones Clínicas, Universidad del Zulia, Venezuela.
Mosquera
J
J
Rodríguez-Iturbe
B
B
D000818:Animals; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005609:Free Radicals / Q000378:metabolism; D006801:Humans; D007674:Kidney Diseases / Q000139:chemically induced / Q000378:metabolism / Q000473:pathology; D007678:Kidney Glomerulus / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D009393:Nephritis / Q000378:metabolism / Q000473:pathology; D009580:Nitroblue Tetrazolium / Q000378:metabolism*; D010088:Oxidoreductases / Q000037:antagonists & inhibitors; D010894:Piroxicam / Q000494:pharmacology; D011692:Puromycin Aminonucleoside / Q000633:toxicity; D051381:Rats; D011917:Rats, Inbred Lew; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013482:Superoxide Dismutase / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014517:Ureteral Obstruction / Q000378:metabolism / Q000473:pathology
D004791:Enzyme Inhibitors; D005609:Free Radicals; D017382:Reactive Oxygen Species; D010894:Piroxicam; D009580:Nitroblue Tetrazolium; D011692:Puromycin Aminonucleoside; D010088:Oxidoreductases; D013482:Superoxide Dismutase; D013755:Tetradecanoylphorbol Acetate