Summary
The distribution of nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) in the central grey region (lamina X of Rexed) of the rat upper thoracic cord was examined by LM and EM. Numerous NADPH-d positive neuronal somata and fibres were present in the subependymal areas of the central grey region at levels T1–T3. Most of the neurons were located dorsal to the central canal in horizontal sections through this region. Many medially-directed NADPH-d positive fibres arising from neurons in n. intermediolateralis pars principalis, n. intercalatus spinalis and longitudinally-directed NADPH-d positive fibres arising from neurons in n. intercalatus pars paraependymalis formed a subependymal plexus. In horizontal sections through the central canal, some NADPH-d positive nerve fibres appeared to traverse the ependyma to enter and run along the central canal. By EM, NADPH-d reaction products were localized on the nuclear membrane, outer mitochondrial membrane and Golgi apparatus of both neurons and ependymal cells and in some axon terminals containing pleomorphic and round agranular synaptic vesicles. Present results suggest that besides the traditional monoamine-, amino acid- and peptidecontaining axon terminals, the central grey region also contains fibres in which nitric oxide is utilized as a neurotransmitter or neuromodulator. The finding of NADPH-d positive fibres in the central canal suggests that nitric oxide may be released into the cerebrospinal fluid. Since some of the ependymal cells were NADPH-d positive, it is suggested that they may be involved in the modulation of nitric oxide levels in the cerebrospinal fluid.
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References
Anderson, C. R. (1992) NADPH diaphorase-positive neurons in the rat spinal cord include a subpopulation of autonomic preganglionic neurons.Neuroscience Letters 139, 280–4.
Barber, R. P., Vaughn, J. E., Slemmon, J. R., Salvaterra, P. M., Roberts, E. &Leeman, S. E. (1979) The origin, distribution and synaptic relationships of substance P axons in rat spinal cord.Journal of Comparative Neurology 184, 331–52.
Barber, R. P., Vaughn, J. E. &Roberts, E. (1982) The cytoarchitecture of GABAergic neurons in rat spinal cord.Brain Research 238, 303–28.
Blottner, D. &Baumgarten, H. G. (1992) Nitric Oxide synthetase (NOS)-containing sympathoadrenal cholinergic neurons of the rat IML-cell column: evidence from histochemistry, immunohistochemistry, and retrograde labelling.Journal of Comparative Neurology 316, 45–55.
Bredt, D. S., Hwang, P. M., Glatt, C. E., Lowenstein, C., Reed, R. R. &Snyder, S. H. (1991) Cloned and expressed nitric oxide synthase structurally resembles cytochrome P-450 reductase.Nature 351, 714–18.
Dun, N. J., Dun, S. L., Förstermann, U. &Tseng, L. F. (1992) Nitric oxide synthase immunoreactivity in rat spinal cord.Neuroscience Letters 147, 217–20.
Dun, N. J., Dun, S. L., Wu, S. Y., Förstermann, U., Schmidt, H. H. H. W. &Tseng, L. F. (1993) Nitric oxide synthase-immunoreactivity in the rat, mouse, cat and squirrel monkey spinal cord.Neuroscience 54, 845–57.
Gibson, S. J., Polak, J. M., Bloom, S. R. &Wall, P. D. (1981) The distribution of nine peptides in rat spinal cord with special emphasis on the substantia gelatinosa and on the area around the central canal (lamina X).Journal of Comparative Neurology 201, 65–79.
Glazer, E. J. &Basbaum, A. I. (1981) Immunohistochemical localization of leucine-enkephalin in the spinal cord of the cat: enkaphalin-containing marginal neurons and pain modulation.Journal of Comparative Neurology 196, 377–89.
Hökfelt, T., Elde, R., Johansson, O., Terenius, L. &Stein, L. (1977) The distribution of enkephalin-immunoreactive cell bodies in the rat central nervous system.Neuroscience Letters 5, 25–31.
Honda, C. N. (1985) Visceral and somatic afferent convergence onto neurons near the central canal in the sacral spinal cord of the cat,Journal of Neurophysiology 53, 1059–78.
Honda, C. N. &Lee, C. L. (1985) Immunohistochemistry of synaptic input and functional characterization of neurons near the spinal central canal.Brain Research 343, 120–8.
Honda, C. &Perl, E. R. (1985) Functional and morphological features of neurons in the midline region of the caudal spinal cord of the cat.Brain Research 340, 285–95.
Kemp, M. C., Kuonen, D. R., Sutton, A. &Roberts, P. J. (1988) Rat brain NADPH-dependent diaphorase. A possible relationship to cytochrome P-450 reductase.Biochemical Pharmacology. 37, 3063–70.
Knigge, K. M., Hoffman, G. E., Joseph, S. A., Scott, D. E., Sladek, C. D. &Sladek, J. R. JR. (1980) Recent advances in structure and function of the endocrine hypothalamus. In:Handbook of the Hypothalamus. II. Physiology of the Hypothalamus (edited byMorgan, P. J. &Panksepp, J.) pp. 63–164. New York: Marcel Dekker.
Lamotte, C. (1987) Vasoactive intestinal polypeptide cerebrospinal fluid-contacting neurons of the monkey and cat spinal central canal.Journal of Comparative Neurology 258, 527–41.
Lamotte, C. C., Johns, D. R. &DE Lanerolle, N. C. (1982) Immunohistochemical evidence of indolamine neurons in monkey spinal cord.Journal of Comparative Neurology 206, 359–70.
Llewellyn-Smith, I. J., Song, Z. M., Costa, M., Bredt, D. S. &Snyder, S. H. (1992) Ultrastructural localization of nitric oxide synthase immunoreactivity in guinea-pig enteric neurons.Brain Research 577, 337–42.
Mclaughlin, B. J., Barber, R., Saito, K., Roberts, E. &Wu, J. Y. (1975) Immunocytochemical localization of glutamate decarboxylase in rat spinal cord.Journal of Comparative Neurology 164, 305–21.
Miller, K. E. &Seybold, V. S. (1987) Comparison of metenkephalin, dynorphin A, and neurotensin immunoreactive neurons in the cat and rat spinal cords. I. Lumbar cord.Journal of Comparative Neurology 225, 293–304.
Mizukawa, K., Vincent, S. R., Mcgeer, P. L. &Mcgeer, E. G. (1989) Distribution of reduced-nicotinamide-adenine-dinucleotide-phosphate diaphorase-positive cells and fibres in the cat central nervous system.Journal of Comparative Neurology 279, 281–311.
Nahin, R. L., Madsen, A. M. &Giesler, G. J. Jr (1983) Anatomical and physiological studies of the grey matter surrounding the spinal cord central canal.Journal of Comparative Neurology 220, 321–35.
Nahin, R. L., Madsen, A. M., Micevych, P. E., Haist, F., Terman, G. W. &Geisler, G. J. Jr (1984) Studies of the area around the central canal: efferents and afferents.Society for Neuroscience Abstracts 10, 487.
Newton, B. &Hamill, R. W. (1985) Immunohistochemical localization of serotonin neurons in autonomic regions of rat spinal cord.Society for Neuroscience Abstracts 11, 1018.
Onstott, D., Mayer, B. &Beitz, A. (1993) Nitric Oxide synthase immunoreactive neurons anatomically define a longitudinal dorsolateral column within the midbrain periaqueductal grey of the rat: analysis using laser confocal microscopy.Brain Research 610, 317–24.
Sasek, C. A. &Elde, R. P. (1985) Distribution of neuropeptide Y-like immunoreactivity and its relationship to FMRF-amide-like immunoreactivity in the sixth lumbar and first sacral spinal cord segments of the rat.Journal of Neuroscience 5, 1729–39.
Sasek, C. A., Seybold, V. S. &Elde, R. P. (1984) The immunohistochemical localization of nine peptides in the sacral parasympathetic nucleus and the dorsal grey commissure in rat spinal cord.Neuroscience 12, 855–73.
Tang, F. R., Tan, C. K. &Ling, E. A. (1995a) A lightmicroscopic study of the intermediolateral nucleus following injection of CB-HRP and fluorogold into the superior cervical ganglion of the rat.Journal of the Autonomic Nervous System 50, 33–8.
Tang, F. R., Tan, C. K. &Ling, E. A. (1995b) A comparative study by retrograde neuronal tracing and substance P immunohistochemistry of sympathetic preganglionic neurons in spontaneously hypertensive rats and Wistar-Kyoto rats.Journal of Anatomy 186, 197–207.
Valtschanoff, J. C., Weinberg, R. J. &Rustioni, A. (1992) NADPH diaphorase in the spinal cord of rats.Journal of Comparative Neurology 321, 209–22.
Vanderhaeghen, J. J., Deschepper, C., Lotstra, F., Vierendeels, G. &Schoenen, J. (1982) Immunohistochemical evidence for cholecystokinin-like peptdes in neuronal cell bodies of the rat spinal cord.Cell and Tissue Research 223, 463–7.
Vigh, B. &Vigh-Teichmann, I. (1992) Cytochemistry of CSF-contacting neurons and pinealocytes.Progress in Brain Research 91, 299–306.
Vigh, B., Vigh-Teichmann, I., Manzanoe Silva, M. J. &Van Den Poi, A. N. (1983) Cerebrospinal fluidcontacting neurons of the central canal and terminal ventricle in various vertebrates.Cell and Tissue Research 231, 615–21.
Vincent, S. R. &Kimura, H. (1992) Histochemical mapping of nitric oxide synthase in the rat.Neuroscience 46, 722–84.
Wang, X. Y., Wong, W. C. &Ling, E. A. (1995) Localization of NADPH-diaphorase activity in the submucous plexus of the guinea-pig intestine: light and electron microscopic studies.Journal of Neurocytology 24 (in press).
Wolf, G., Würdig, S. &Schünzel, G. (1992) Nitric oxide synthase in rat brain is predominantly located at neuronal endoplasmic reticulum: an electron microscopic demonstration of NADPH-diaphorase activity.Neuroscience Letters 147, 63–6.
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Tang, F.R., Tan, C.K. & Ling, E.A. The distribution of NADPH-d in the central grey region (lamina X) of rat upper thoracic spinal cord. J Neurocytol 24, 735–743 (1995). https://doi.org/10.1007/BF01191210
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DOI: https://doi.org/10.1007/BF01191210