Nitric oxide synthase in the brain of the turtle Pseudemys scripta elegans

The distribution pattern of nitric oxide synthase (NOS) was investigated in the brain of the turtle by NADPH-diaphorase histochemistry. The specificity of the histochemical staining was tested by immunocytochemical colocalization with an antiserum specific for NOS. In the forebrain, neurons staining intensely for nitric oxide synthase were localized in the olfactory tubercle, the basal ganglia complex, the basal amygdaloid nucleus, suprapeduncular nucleus, and the posterior hypothalamic area. Many positive fibers course in a tract connecting the basal amygdaloid nucleus with the hypothalamus, corresponding to the stria terminalis. Bundles of nitroxergic fibers were seen to course at the ventromedial edge of the optic tract and to cross in the supraoptic decussation, apparently consisting of tectothalamic and thalamotectal fibers. In the midbrain, strongly NOS-positive neurons were present in the substantia nigra, the nucleus profundus mesencephali, the periventricular grey of the optic tectum, the laminar nucleus of the torus semicircularis, and the nucleus of the lateral lemniscus. The area of the locus coeruleus harbored an accumulation of intensely stained neurons, which, as in mammals, might represent a cholinergic cell group of the reptilian brainstem. In the cerebellum, strong staining was confined to bundles of afferent fibers running in the lower molecular and in the Purkinje cell layer. These axons appeared to include ascending projections from the dorsal funicular nucleus or the spinal cord. NOS-positive cells in the caudal brainstem were found in the cerebellar nuclei, in the superior vestibular nucleus, in the reticular nuclei, ventrolateral to the nucleus of the solitary tract, in the perihypoglossal, and in the dorsal funicular nucleus. Taken together, these results suggest that nitric oxide acts as a messenger molecule in different areas of the reptilian brain and spinal cord. In certain areas, the pattern of expression of NOS appears to have evolved before radiation of present mammalian, avian, and reptilian species.