Document Detail


When NADPH diaphorase (NADPHd) works in the presence of formaldehyde, the enzyme appears to visualize selectively cells with constitutive nitric oxide synthase (NOS).
MedLine Citation:
PMID:  7531934     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The NADPH diaphorase (NADPHd) reaction for nitric oxide synthase (NOS) visualization suffers from the circumstance that the diaphorase activity of NOS represents only part of the total diaphorase activity, and so far all efforts to make the reaction more specific for routine studies failed. The present investigation describes a simple procedure for mouse tissue, which allows the selective staining primarily of neurons, vascular endothelial cells and macula densa cells, those cells where constitutive NOS has been described reliably. In this method unfixed cryosections and 0.5-1% phosphate-buffered formaldehyde containing 0.5 mg NADPH/1 ml and 1 mg nitro BT/1 ml are used. Compared to strong prefixation with formaldehyde after which many additional cells are still positive for NADPHd, presence of formaldehyde in the incubation medium obviously allows the selective reaction of NOS-positive cells. In conclusion, compared with the original technique a more specific method for the visualization of the NADPHd activity of NOS appears to be available now and can be used for NOS studies in all kinds of mammalian species.
Authors:
G Nakos; R Gossrau
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Acta histochemica     Volume:  96     ISSN:  0065-1281     ISO Abbreviation:  Acta Histochem.     Publication Date:  1994 Sep 
Date Detail:
Created Date:  1995-03-16     Completed Date:  1995-03-16     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0370320     Medline TA:  Acta Histochem     Country:  GERMANY    
Other Details:
Languages:  eng     Pagination:  335-43     Citation Subset:  IM    
Affiliation:
Department of Anatomy, Free University of Berlin, Germany.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Oxidoreductases / metabolism*
Animals
Epithelium / enzymology
Formaldehyde*
Histocytochemistry
Male
Mice
NADPH Dehydrogenase / metabolism*
Nitric Oxide Synthase
Staining and Labeling
Tissue Fixation
Chemical
Reg. No./Substance:
50-00-0/Formaldehyde; EC 1.14.13.39/Nitric Oxide Synthase; EC 1.4.-/Amino Acid Oxidoreductases; EC 1.6.99.1/NADPH Dehydrogenase

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