PMID: 9539206Apr 16, 1998Paper

Histochemical differentiation between nitric oxide synthase-related and -unrelated diaphorase activity in the rat olfactory bulb

The Histochemical Journal
R Spessert, M Claassen

Abstract

The widely used NADPH-diaphorase reaction for demonstrating neuronal nitric oxide synthase is not as specific as previously thought, as it visualizes both a nitric oxide synthase-related activity and a nitric oxide synthase-unrelated diaphorase. In the present study, we used the rat olfactory bulb as a model to characterize the NADPH-diaphorase activity of neuronal nitric oxide synthase histochemically in comparison with neuronal nitric oxide-unrelated diaphorase activity. The NADPH-diaphorase activity of nitric oxide synthase peaked at pH 8 and at Triton X-100 concentrations of 1-2.5%. It was stable in an acidic environment but was reduced in the presence of Triton X-100 and was inactivated by the flavoprotein inhibitor, diphenyleneiodonium. It preferred beta-NADPH as the co-substrate to alpha-NADPH and alpha-NADH. In contrast, nitric oxide synthase-unrelated diaphorase peaked at pH 10, displayed a Triton X-100 optimum at a concentration of 1%, was unstable in an acidic environment and used beta-NADPH, alpha-NADPH and alpha-NADH to similar extents. Differences in the characteristics between neuronal nitric oxide synthase-related and nitric oxide synthase-unrelated NADPH-diaphorase can be used to increase the specificity of the...Continue Reading

Citations

Mar 19, 2003·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Swidbert R Ott, Maurice R Elphick
Mar 30, 2005·Journal of Neuroscience Research·Enrica BiagiottiPaolino Ninfali
Jun 22, 2002·Annals of the New York Academy of Sciences·Seng-Kee LeongZhi Zhang
Jan 21, 2012·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Jae-Youn ChoiJung-Ho Cha
Apr 7, 2005·Journal of Inorganic Biochemistry·Leonid L MorozPeter Lovell
Oct 31, 2006·European Journal of Protistology·Andrea AmaroliMaria Umberta Delmonte Corrado

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