NADPH‐diaphorase: A selective histochemical marker for striatal neurons containing both somatostatin‐ and avian pancreatic polypeptide (APP)‐like immunoreactivities

Abstract

Certain neurons in the brain are specifically and intensely stained by a histochemical method which demonstrates nicotinamide adenine dinucleotide phosphate NADPH-diaphorase activity. The cell types containing this enzyme in certain areas of the rat forebrain were examined by combining NADPH-diaphorase histochemistry with the indirect immunofluorescence technique. Neurons containing somatostatin- or avian pancreatic polypeptide (APP)-like immunoreactivities were found throughout the forebrain including the striatum and neocortex. These two neuropeptides were also found to coexist in many telencephalic neurons. After photography, the sections processed for immunohistochemistry were stained for NADPH-diaphorase activity by a histochemical method. It was found that within the striatum all of the neurons that were selectively stained by this technique also contained both somatostatin- and APP-like immunoreactivities. Also in the neocortex NADPH-diaphorase was found only in those neurons displaying somatostatin- or APP-like immunoreactivity. In other brain regions such as the nucleus laterodorsalis tegmenti, NADPH-diaphorase-containing cells did not contain these neuropeptides. The results indicate that NADPH-diaphorase histochemistry provides a simple, reliable, histochemical method to demonstrate those striatal neurons in which somatostatin- and APP-like immunoreactivities coexist. The selective occurrence of this enzyme within these neurons may provide a useful target for pharmacological studies of these neuropeptide-containing cells.

NMDA inhibitors cause apoptosis of pyramidal neurons in mature piriform cortex: Evidence for a nitric oxide-mediated effect involving inhibitory interneurons

“…Brain blocks containing the forebrain were postfixed overnight, treated with 30% sucrose till they sunk and sectioned coronally (40 μm) on a freezing microtome. Sections were stained in series for: cresyl violet; NADPH diaphorase histochemistry using the method of Vincent with minor modifications (Vincent et al, 1983); neuronal nitric oxide synthase (nNOS) ICC; and active caspase-3 ICC. All ICC reactions were based on the avidin-biotin-peroxidase (ABC) labeling protocol using commercially available kits (Vector Labs, Burlingame, CA) and goat anti-rabbit IgG as a linker (Vector Labs).…”

Hydrogen peroxide-mediated neuronal cell death induced by an endogenous neurotoxin, 3-hydroxykynurenine.

“…Whatever the precise cellular mechanisms, this observation serves an interesting coincidence with pathological characteristics of brains of patients with neurodegenerative disorders such as HD (25,26). The specific aspiny neuronal subpopulation containing NADPH diaphorase, which is colocalized with somatostatin and neuropeptide Y (33), is selectively spared in the degenerated striatum of HD patients (25,26).…”

Nitric Oxide and Synaptic Function

“…These cells also stain positive for somatostatin and neuropeptide Y (Vincent et al 1983, Dawson et al 1991.…”