A modified method allows for correlation between NADPH-diaphorase histochemistry and immunohistochemistry for the demonstration of neuronal nitric oxide synthase (nNOS).
Results obtained with the conventional nitro blue tetrazolium salt method for the visualization of the NADPH-diaphorase (NADPH-d) activity of nitric oxide synthase (NOS) are not specific for this particular enzyme, since this activity represents only a fraction of the total cellular NADPH-d pool. Therefore, the standard NADPH-d procedure was modified by performing the incubation in the presence of formaldehyde. Parallel application of the modified NADPH-d staining technique and the indirect immunofluorescence using an antibody against the neuronal isoenzyme (nNOS) on rat, mouse and guinea-pig tissues showed a correlation between histochemical and immunocytochemical staining. It can thus be concluded that the modified NADPH-d procedure allows for a more specific detection of the histochemical nNOS activity than the conventional method.
Grozdanovic
Z
Z
Department of Anatomy, Free University of Berlin, Germany.
Nakos
G
G
Mayer
B
B
Gossrau
R
R
D000818:Animals; D005260:Female; D005455:Fluorescent Antibody Technique; D005557:Formaldehyde; D006168:Guinea Pigs; D006651:Histocytochemistry / Q000379:methods*; D016591:Histocytological Preparation Techniques; D007150:Immunohistochemistry / Q000379:methods; D008297:Male; D051379:Mice; D009252:NADPH Dehydrogenase / Q000378:metabolism*; D009474:Neurons / Q000201:enzymology; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000032:analysis*; D051381:Rats; D017208:Rats, Wistar; D013379:Substrate Specificity
D005557:Formaldehyde; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D009252:NADPH Dehydrogenase