Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Nitrite production does not always reflect nitric oxide synthesis by isolated glomeruli


The goal of this paper was the measurement of nitric oxide (NO) production in isolated rat glomeruli using two different techniques. NO production was detected directly by a NO-specific electrode and the results were compared with data measured by the Griess reaction, an indirect index to evaluate NO production. The NO production, determined by both techniques, was dependent on the number of glomeruli. Pretreatment with Nw-nitro-L-arginine methyl ester, an inhibitor of the NO synthesis, reduced the NO concentration detected by the NO-sensor, but increased the NO2-concentration (when both results where compared with glomeruli without treatment). Preincubation with 1 mg/ml of Escherichia coli lipopolysaccharide significantly enhanced both NO and NO2— concentrations. Therefore, the present study provides direct evidence of NO generated in isolated glomeruli under physiological conditions and demonstrates that the measurement of NO2- by the Griess reaction, is not always an adequate technique to evaluate the actual NO production.

This is a preview of subscription content, log in to check access.


  1. 1.

    Raij L (1993) Nitric oxide and the kidney. Circulation 87 [Suppl 5]:V26–V29

  2. 2.

    Hill-Kapturczak N, Kapturczak MH, Malinski T, Gross P (1995) Nitric oxide and nitric oxide synthase in the kidney: potential roles in normal renal function and in renal dysfunction. Endothelium 3:253–929

  3. 3.

    Cattell V, Cook HT (1993) Nitric oxide: role in the physiology and pathology of the glomerulus. Exp Nephrol 1:265–280

  4. 4.

    Raij L, Baylis C (1995) Glomerular actions of nitric oxide. Kidney Int 48:20–32

  5. 5.

    Wilcox CS, Welch WJ, Murad F; Gross SS, Taylor G, Levi R, Schmidt HHHW (1992) Nitric oxide synthase in macula densa regulates glomerular capillary pressure. Proc Natl Acad Sci USA 89:1993–1997

  6. 6.

    Ito S, Ren Y-L (1993) Evidence for the role of nitric oxide in macula densa control of glomerular hemodynamics. J Clin Invest 92:1093–1098

  7. 7.

    Braam B, Koomans HA (1995) Reabsorption of nitro-L-arginine infused into the late proximal tubule participates in modulation of TGF responsiveness. Kidney Int 47:1252–1257

  8. 8.

    Pfeilschifter J (1995) Does nitric oxide, an inflammatory mediator of glomerular mesangial cells, have a role in diabetic nephropathy? Kidney Int 48 [Suppl 51]:S50–S60

  9. 9.

    Nakashima A, Matsuoka H, Yasukawa H, Kohno K, Nishida H, Nomura G, Imaizumi T, Morimatsu M (1996) Renal denervation prevents intraglomerular platelet aggregation and glomerular injury induced by chronic inhibition of nitric oxide synthesis. Nephron 73:34–40

  10. 10.

    Moncada S, Higgs EA (1995) Molecular mechanisms and therapeutic strategies related to nitric oxide. FASEB J 9:1319–1330

  11. 11.

    Michel T, Feron O (1997) Nitric oxide synthases: which, where, how and why? J Clin Invest 100:2146–2152

  12. 12.

    Moncada S, Palmer RMJ, Higgs EA (1991) Nitric oxide: Physiology, pathophysiology and pharmacology. Pharmacol Rev 43:109–142

  13. 13.

    Ujie K, Yuen J, Hogart L, Danzinger R, Star RA (1994) Localization and regulation of endothelial NO synthase mRNA expression in rat kidney. Am J Physiol 267:F296–F302

  14. 14.

    Morrisey JJ, McCraken R, Kaneto H, Vehaskari M, Montani D, Klahr S (1994) Location of an inducible nitric oxide synthase mRNA in the normal kidney. Kidney Int 45:998–1005

  15. 15.

    Archer S (1993) Measurement of nitric oxide in biological models. FASEB J 7:349–360

  16. 16.

    Leone AM, Rhodes P, Furst V, Moncada S (1995) Techniques for the measurement of nitric oxide. Methods Mol Biol 41:285–299

  17. 17.

    Tsikas D, Gutzki FM, Rossa S, Bauer H, Neumann C, Dockendorff K, Sandmann J, Frolich JC (1997) Measurement of nitrite and nitrate in biological fluids by gas chromatography, mass spectrometry and by the Griess assay: problems with the spectrometry. Ann Biochem 44:208–220

  18. 18.

    Rivas-Cabañero L, Valdivieso JM, López-Novoa JM (1996) NG-Nitro-L-arginine meth-yl ester causes glomerular nitrite production in long-term incubations. Nephron 73: 97–98

  19. 19.

    Rivas-Cabañero L, Montero A, López-Novoa JM (1994) Nitric oxide-dependent cyclic GMP synthesis by isolated rat glomeruli. Endothelium 1:259–261

  20. 20.

    Tsukahara H, Krivenko Y, Moore LC, Goligorsky MS (1994) Decrease in ambient [Cl] stimulates nitric oxide release from cultured rat mesangial cells. Am J Physiol 267:F190–F195

  21. 21.

    Ketteler M, Border WA, Noble NA (1994) Citokines and L-arginine in renal injury and repair. Am J Physiol 267:F197–F207

  22. 22.

    Shibuki K (1990) An electrochemical microprobe for detecting nitric oxide release in brain tissue. Neurosci Res 9:69–76

  23. 23.

    Ignarro LJ, Buga GM, Wood KS, Byrns RE (1987) Endothelium-derived relaxing factor produced and released from artery and vein is nitric oxide. Proc Natl Acad Sci USA 84:9265–9269

  24. 24.

    Fleming I, Gray GA, Schott C, Stoclet JC (1991) Inducible but not constitutive production of nitric oxide by vascular smooth muscle cells. Eur J Pharmacol 200:375–376

  25. 25.

    Heyman SM, Karmeli F, Rachmilewitz D, Haj-Yehia A, Brezio M (1997) Intrarenal nitric oxide monitoring with a Clark-type electrode: potential pitfalls. Kidney Int 51:1619–1623

  26. 26.

    Ignarro LJ, Buga GM, Woods KS, Byrns RE (1987) Endothelium-derived relaxing factor produced and released from artery and vein is nitric oxide. Proc Natl Acad Sci (USA) 84:9265–9269

  27. 27.

    Butler AR, Flitney FW, Williams DLH (1995) NO, nitrosonium ions, nitroxide ions, nitrosothiols and iron-nitrosyls in biology: a chemist’s perspective. Trends Pharmacol Sci 16:18–22

  28. 28.

    Cattell V, Cook T, Moncada S (1990) Glomeruli synthetize nitrite in experimental nephrotoxic nephritis. Kidney Int 38:1056–1060

  29. 29.

    Schott CA, Gray GA, Stoclet J-C (1993) Dependence of endotoxin-induced vascular hyporeactivity on extracellular L-arginine. Br J Pharmacol 108:38–43

  30. 30.

    Hom GJ, Grant SK, Wolfe G, Bach TJ, Macintyre DE, Hutchinson NI (1995) Lipopolysaccharide-induced hypotension and vascular hyporeactivity in the rat: tissue absence of dexamethasone, NG-monomethyl-L-arginine or indomethacine. J Pharmacol Exp Ther 272:452–459

  31. 31.

    Stoclet J-C, Fleming I, Gray G, Julou-Schaeffer G, Schneider F, Schott C, Schott C, Parratt JR (1993) Nitric oxide and endotoxemia. Circulation 87 [Suppl 5]:V–77–80

  32. 32.

    Schneider F, Buchner B, Schott C, Andre A, Julou-Schaeffer G, Stoclet J-C (1994) Effect of bacterial lipopolysaccharide on function of rat small femoral arteries. Am J Physiol 266:H191–198

  33. 33.

    Yaqoob M, Edelstein CL, Wieder ED, Alkhunaizi AM, Gengaro PE, Nemenoff RA, Schrier RW (1996) Nitric oxide kinetics during hypoxia in proximal tubules: effects of acidosis and glycine. Kidney Int 49:1314–1319

  34. 34.

    Bogle RG, Moncada S, Pearson JD, Mann GE (1992) Identification of inhibitors of nitric oxide synthase that do not interact with the endothelial cell L-arginine transporter. Br J Pharmacol 105:768–770

  35. 35.

    Bogle RG, Baydoum AR, Pearson JD, Mann GE (1996) Regulation of l-arginine transport and nitric oxide release in superfused porcine aortic endothelial cells. J Physiol 490:229–241

  36. 36.

    Ogawa T, Kimoto M, Sasaoka K (1989) Purification and properties of a new enzyme, NG-, NG-dimethylarginine dimethylamino-hydrolase, from rat kidney. J Biol Chem 264:10205–10219

Download references

Author information

Correspondence to J. M. López-Novoa.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Reverte, M., López-Novoa, J.M. Nitrite production does not always reflect nitric oxide synthesis by isolated glomeruli. Res. Exp. Med. 198, 55–62 (1998).

Download citation


  • NO-sensor
  • Rat glomeruli
  • Inducible nitric oxide synthase
  • Nitric oxide production
  • Griess reaction