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Journal of Histochemistry and Cytochemistry, Vol. 47, 197-208, February 1999, Copyright © 1999, The Histochemical Society, Inc.

ARTICLE

NOS- and Non-NOS NADPH Diaphorases in the Insular Cortex of the Syrian Golden Hamster

Richard G. Wehbya and Marion E. Franka,b
a Program in Neuroscience, Schools of Medicine and Dental Medicine, University of Connecticut Health Center, Farmington, Connecticut
b Department of BioStructure and Function, Schools of Medicine and Dental Medicine, University of Connecticut Health Center, Farmington, Connecticut

Correspondence to: Richard G. Wehby, Dept. of Brain and Cognitive Sciences, MIT, E25-236, Cambridge, MA 02139.

We had previously shown NADPH diaphorase activity in fixed tissue slices of the insular cortex of the Syrian golden hamster (Mesocricetus auratus). The objective of this work was to determine the chemical identity of agents responsible for the observed NADPH diaphorase activities. Three different enzymatic NADPH diaphorase activities were distinguished in the insular cortex. (a) The activity seen in endothelial cells was not characterized histochemically, but it co-localized with eNOS-like immunoreactivity. (b) The neuronal Type I activity showed little sensitivity to 10-5 M dicoumarol, could use either {alpha}- or ß-NADPH with almost equal facility, and co-localized with nNOS-like immunoreactivity. This activity was primarily attributable to nNOS. (c) The neuronal Type II activity was greatly attenuated by 10-5 M dicoumarol, had a strong preference for ß-NADPH (rather than {alpha}-NADPH), and did not co-localize with any NOS-like immunoreactivity. These characteristics also apply to the NADPH diaphorase activity observed in the diffuse blue band in Layers II and III of agranular and dysgranular insular cortex and in the meshwork of cortical fibers. This staining was due primarily to a dicoumarol-sensitive dehydrogenase(s), either an isozyme of DT diaphorase (EC 1.6.99.2), or NADPH dehydrogenase (quinone) (EC 1.6.99.6), or to a novel dicoumarol-sensitive NADPH dehydrogenase. (J Histochem Cytochem 47:197–207, 1999)

Key Words: NADPH diaphorase, nitric oxide synthase, dicoumarol, dicumarol, NADPH dehydrogenase


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