An ultrastructural study of NADPH-diaphorase and nitric oxide synthase in the perivascular nerves and vascular endothelium of the rat basilar artery

Abstract

This is the first report on the ultrastructural distribution of nicotinamide adenine dinucleotide phosphate-diaphorase activity and neuronal isoform (Type I) of nitric oxide synthase immunoreactivity in perivascular nerves (axons) and vascular endothelial cells. In the Sprague-Dawley rat cerebral basilar artery, positive labelling for nicotinamide adenine dinucleotide phosphate-diaphorase and nitric oxide synthase was localized in axons and the endothelium. Over half (approximately 53%) of the axon profiles examined were positive for nicotinamide adenine dinucleotide phosphate-diaphorase. Labelling of nicotinamide adenine dinucleotide phosphate-diaphorase activity in the axons and endothelial cells was mostly distributed in patches within the cytoplasm. In endothelial cells, a relation between the nicotinamide adenine dinucleotide phosphate-diaphorase-labelling and cytoplasmic vesicle-like structures was seen. In both axons and the endothelium, nitric oxide synthase immunoreactivity was seen throughout the cell cytoplasm and in association with the membranes of mitochondria, endoplasmic reticulum and cytoplasmic/synaptic vesicles (the lumen/content of the vesicles was negative for nitric oxide synthase). Also, microtubules were labelled in nitric oxide synthase positive axon profiles. The nitric oxide synthase-positive axon varicosities were characterized by the presence of spherical agranular vesicles with a diameter of 40-50 nm. Approximately 30% of the axon profiles examined were positive for nitric oxide synthase. The nicotinamide adenine dinucleotide phosphate-diaphorase-positive endothelial cells (approximately 20% of all observed endothelial cell profiles) were more frequently seen than those positive for nitric oxide synthase (approximately 7%). It is suggested that nitric oxide released from both perivascular nerves and endothelial cells may be involved in vasomotor control of cerebral circulation.