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. 1997 Aug;191(Pt 2):161–175. doi: 10.1046/j.1469-7580.1997.19120161.x

Comparative analysis of nitric oxide synthase immunoreactivity in the sacral spinal cord of the cat, macaque and human

A H PULLEN 1 , P HUMPHREYS 1 , R G BAXTER 2
PMCID: PMC1467670  PMID: 9306194

Abstract

Nitric oxide synthase immunoreactivity (bNOS-ir) was examined in the sacral spinal cord of the cat, macaque monkey and human using an antibody to the c-terminal region of neuronal NOS. In S2 of all 3 species NOS-ir was identified in both dorsal and ventral horns. In cat, monkey and human, bNOS-r occurred in sensory neurons of superficial laminae and the base of the dorsal horn, in autonomic neurons around the central canal and in the intermediolateral sacral spinal nucleus. In all 3 species, a large proportion of somatic motor nuclei in the ventromedial (VM), ventrolateral (VL) nuclei, and Onuf's nucleus(ON) showed high bNOS-ir, while others exhibited markedly lower immunoreactivity. Validatory experiments showed separate cellular localisation for bNOS, inducible NOS(iNOS), and endothelial NOS(eNOS) with only bNOS being localised to neuronal perikarya. Comparative morphometric analyses of the relative proportions and diameters of motor neurons in the VL, VM and ON exhibiting high and low levels of bNOS-ir revealed statistically significant differences in proportions in individual nuclei, and differences in size were generally not statistically significant. Finally, a comparison between cat sacral and thoracic spinal cord showed bNOS-ir in motor neurons of S2 was subject to less animal and rostrocaudal segment variation than in T10.

Keywords: Sacral motor neurons, Onuf's nucleus, nitric oxide, NOS, anal and urethral sphincters

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Selected References

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  1. Armand J., Olivier E., Edgley S. A., Lemon R. N. Postnatal development of corticospinal projections from motor cortex to the cervical enlargement in the macaque monkey. J Neurosci. 1997 Jan 1;17(1):251–266. doi: 10.1523/JNEUROSCI.17-01-00251.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blottner D., Baumgarten H. G. Nitric oxide synthetase (NOS)-containing sympathoadrenal cholinergic neurons of the rat IML-cell column: evidence from histochemistry, immunohistochemistry, and retrograde labeling. J Comp Neurol. 1992 Feb 1;316(1):45–55. doi: 10.1002/cne.903160105. [DOI] [PubMed] [Google Scholar]
  3. Bredt D. S., Hwang P. M., Glatt C. E., Lowenstein C., Reed R. R., Snyder S. H. Cloned and expressed nitric oxide synthase structurally resembles cytochrome P-450 reductase. Nature. 1991 Jun 27;351(6329):714–718. doi: 10.1038/351714a0. [DOI] [PubMed] [Google Scholar]
  4. Burleigh D. E. Non-cholinergic, non-adrenergic inhibitory neurons in human internal anal sphincter muscle. J Pharm Pharmacol. 1983 Apr;35(4):258–260. doi: 10.1111/j.2042-7158.1983.tb02927.x. [DOI] [PubMed] [Google Scholar]
  5. Dun N. J., Dun S. L., Wu S. Y., Förstermann U., Schmidt H. H., Tseng L. F. Nitric oxide synthase immunoreactivity in the rat, mouse, cat and squirrel monkey spinal cord. Neuroscience. 1993 Jun;54(4):845–857. doi: 10.1016/0306-4522(93)90579-5. [DOI] [PubMed] [Google Scholar]
  6. Gibson S. J., Polak J. M., Katagiri T., Su H., Weller R. O., Brownell D. B., Holland S., Hughes J. T., Kikuyama S., Ball J. A comparison of the distributions of eight peptides in spinal cord from normal controls and cases of motor neurone disease with special reference to Onuf's nucleus. Brain Res. 1988 Dec 6;474(2):255–278. doi: 10.1016/0006-8993(88)90440-4. [DOI] [PubMed] [Google Scholar]
  7. Holstege G., Tan J. Supraspinal control of motoneurons innervating the striated muscles of the pelvic floor including urethral and anal sphincters in the cat. Brain. 1987 Oct;110(Pt 5):1323–1344. doi: 10.1093/brain/110.5.1323. [DOI] [PubMed] [Google Scholar]
  8. Hope B. T., Michael G. J., Knigge K. M., Vincent S. R. Neuronal NADPH diaphorase is a nitric oxide synthase. Proc Natl Acad Sci U S A. 1991 Apr 1;88(7):2811–2814. doi: 10.1073/pnas.88.7.2811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Johnson I. P. A quantitative ultrastructural comparison of alpha and gamma motoneurons in the thoracic region of the spinal cord of the adult cat. J Anat. 1986 Aug;147:55–72. [PMC free article] [PubMed] [Google Scholar]
  10. Katagiri T., Gibson S. J., Su H. C., Polak J. M. Composition and central projections of the pudendal nerve in the rat investigated by combined peptide immunocytochemistry and retrograde fluorescent labelling. Brain Res. 1986 May 7;372(2):313–322. doi: 10.1016/0006-8993(86)91139-x. [DOI] [PubMed] [Google Scholar]
  11. Kawatani M., Nagel J., de Groat W. C. Identification of neuropeptides in pelvic and pudendal nerve afferent pathways to the sacral spinal cord of the cat. J Comp Neurol. 1986 Jul 1;249(1):117–132. doi: 10.1002/cne.902490109. [DOI] [PubMed] [Google Scholar]
  12. Koh J. Y., Peters S., Choi D. W. Neurons containing NADPH-diaphorase are selectively resistant to quinolinate toxicity. Science. 1986 Oct 3;234(4772):73–76. doi: 10.1126/science.2875522. [DOI] [PubMed] [Google Scholar]
  13. Kojima H., Furuta Y., Fujita M., Fujioka Y., Nagashima K. Onuf's motoneuron is resistant to poliovirus. J Neurol Sci. 1989 Oct;93(1):85–92. doi: 10.1016/0022-510x(89)90163-9. [DOI] [PubMed] [Google Scholar]
  14. Mannen T., Iwata M., Toyokura Y., Nagashima K. Preservation of a certain motoneurone group of the sacral cord in amyotrophic lateral sclerosis: its clinical significance. J Neurol Neurosurg Psychiatry. 1977 May;40(5):464–469. doi: 10.1136/jnnp.40.5.464. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Matsumoto T., Nakane M., Pollock J. S., Kuk J. E., Förstermann U. A correlation between soluble brain nitric oxide synthase and NADPH-diaphorase activity is only seen after exposure of the tissue to fixative. Neurosci Lett. 1993 May 28;155(1):61–64. doi: 10.1016/0304-3940(93)90673-9. [DOI] [PubMed] [Google Scholar]
  16. Mawe G. M., Bresnahan J. C., Beattie M. S. A light and electron microscopic analysis of the sacral parasympathetic nucleus after labelling primary afferent and efferent elements with HRP. J Comp Neurol. 1986 Aug 1;250(1):33–57. doi: 10.1002/cne.902500104. [DOI] [PubMed] [Google Scholar]
  17. Nadelhaft I., de Groat W. C., Morgan C. The distribution and morphology of parasympathetic preganglionic neurons in the cat sacral spinal cord as revealed by horseradish peroxidase applied to the sacral ventral roots. J Comp Neurol. 1986 Jul 1;249(1):48–56. doi: 10.1002/cne.902490105. [DOI] [PubMed] [Google Scholar]
  18. Parlani M., Conte B., Manzini S. Nonadrenergic, noncholinergic inhibitory control of the rat external urethral sphincter: involvement of nitric oxide. J Pharmacol Exp Ther. 1993 May;265(2):713–719. [PubMed] [Google Scholar]
  19. Pullen A. H., Humphreys P. Diversity in localisation of nitric oxide synthase antigen and NADPH-diaphorase histochemical staining in sacral somatic motor neurones of the cat. Neurosci Lett. 1995 Aug 18;196(1-2):33–36. doi: 10.1016/0304-3940(95)11831-g. [DOI] [PubMed] [Google Scholar]
  20. Pullen A. H., Martin J. E., Swash M. Ultrastructure of pre-synaptic input to motor neurons in Onuf's nucleus: controls and motor neuron disease. Neuropathol Appl Neurobiol. 1992 Jun;18(3):213–231. doi: 10.1111/j.1365-2990.1992.tb00784.x. [DOI] [PubMed] [Google Scholar]
  21. Pullen A. H. Quantitative synaptology of feline motoneurones to external anal sphincter muscle. J Comp Neurol. 1988 Mar 15;269(3):414–424. doi: 10.1002/cne.902690308. [DOI] [PubMed] [Google Scholar]
  22. REXED B. A cytoarchitectonic atlas of the spinal cord in the cat. J Comp Neurol. 1954 Apr;100(2):297–379. doi: 10.1002/cne.901000205. [DOI] [PubMed] [Google Scholar]
  23. Roppolo J. R., Nadelhaft I., de Groat W. C. The organization of pudendal motoneurons and primary afferent projections in the spinal cord of the rhesus monkey revealed by horseradish peroxidase. J Comp Neurol. 1985 Apr 22;234(4):475–488. doi: 10.1002/cne.902340406. [DOI] [PubMed] [Google Scholar]
  24. Saito S., Kidd G. J., Trapp B. D., Dawson T. M., Bredt D. S., Wilson D. A., Traystman R. J., Snyder S. H., Hanley D. F. Rat spinal cord neurons contain nitric oxide synthase. Neuroscience. 1994 Mar;59(2):447–456. doi: 10.1016/0306-4522(94)90608-4. [DOI] [PubMed] [Google Scholar]
  25. Sato M., Mizuno N., Konishi A. Localization of motoneurons innervating perineal muscles: a HRP study in cat. Brain Res. 1978 Jan 20;140(1):149–154. doi: 10.1016/0006-8993(78)90244-5. [DOI] [PubMed] [Google Scholar]
  26. Schrøder H. D. Somatostatin in the caudal spinal cord: an immunohistochemical study of the spinal centers involved in the innervation of pelvic organs. J Comp Neurol. 1984 Mar 1;223(3):400–414. doi: 10.1002/cne.902230306. [DOI] [PubMed] [Google Scholar]
  27. Snyder S. H. Nitric oxide and neurons. Curr Opin Neurobiol. 1992 Jun;2(3):323–327. doi: 10.1016/0959-4388(92)90123-3. [DOI] [PubMed] [Google Scholar]
  28. Sung J. H., Mastri A. R., Segal E. Pathology of Shy-Drager syndrome. J Neuropathol Exp Neurol. 1979 Jul;38(4):353–368. doi: 10.1097/00005072-197907000-00001. [DOI] [PubMed] [Google Scholar]
  29. Tashiro T., Satoda T., Matsushima R., Mizuno N. Convergence of serotonin-, enkephalin- and substance P-like immunoreactive afferent fibers on single pudendal motoneurons in Onuf's nucleus of the cat: a light microscope study combining the triple immunocytochemical staining technique with the retrograde HRP-tracing method. Brain Res. 1989 Mar 6;481(2):392–398. doi: 10.1016/0006-8993(89)90821-4. [DOI] [PubMed] [Google Scholar]
  30. Terenghi G., Riveros-Moreno V., Hudson L. D., Ibrahim N. B., Polak J. M. Immunohistochemistry of nitric oxide synthase demonstrates immunoreactive neurons in spinal cord and dorsal root ganglia of man and rat. J Neurol Sci. 1993 Aug;118(1):34–37. doi: 10.1016/0022-510x(93)90242-q. [DOI] [PubMed] [Google Scholar]
  31. Thor K. B., Morgan C., Nadelhaft I., Houston M., De Groat W. C. Organization of afferent and efferent pathways in the pudendal nerve of the female cat. J Comp Neurol. 1989 Oct 8;288(2):263–279. doi: 10.1002/cne.902880206. [DOI] [PubMed] [Google Scholar]
  32. Vincent S. R., Hope B. T. Neurons that say NO. Trends Neurosci. 1992 Mar;15(3):108–113. doi: 10.1016/0166-2236(92)90021-y. [DOI] [PubMed] [Google Scholar]
  33. Vizzard M. A., Erdman S. L., Erickson V. L., Stewart R. J., Roppolo J. R., De Groat W. C. Localization of NADPH diaphorase in the lumbosacral spinal cord and dorsal root ganglia of the cat. J Comp Neurol. 1994 Jan 1;339(1):62–75. doi: 10.1002/cne.903390107. [DOI] [PubMed] [Google Scholar]
  34. Vizzard M. A., Erdman S. L., Förstermann U., de Groat W. C. Differential distribution of nitric oxide synthase in neural pathways to the urogenital organs (urethra, penis, urinary bladder) of the rat. Brain Res. 1994 May 23;646(2):279–291. doi: 10.1016/0006-8993(94)90090-6. [DOI] [PubMed] [Google Scholar]

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