Skip to main content
NCBI home page
Journal of Anatomy logoLink to Journal of Anatomy
. 1997 Jan;190(Pt 1):135–145. doi: 10.1046/j.1469-7580.1997.19010135.x

Distribution of NADPH-diaphorase and nitric oxide synthase-containing neurons in the intramural ganglia of guinea pig urinary bladder

Y ZHOU 1, C K TAN 1, E A LING 1,✉
PMCID: PMC1467591  PMID: 9034889

Abstract

The cell population and distribution of NADPH-diaphorase positive and NOS immunoreactive intramural ganglion cells were examined on stretched whole-mount preparations of the guinea pig urinary bladder which was divided into 3 regions: base, body and dome. The results showed that the highest frequency both of NADPH-d and NOS positive neurons was observed in the bladder base. Cell counts in the whole bladder showed that the number of NADPH-d positive neurons was much more than that of NOS immunoreactive neurons. Using neuron specific enolase (NSE) positive neurons as a reference (100%), NADPH-d positive neurons accounted for 84% while NOS immunoreactive neurons only made up 45% of the total neuronal population. These results, along with previous studies on the function of nitric oxide, suggest that nitric oxide may be involved in the relaxation activity in the bladder base during micturition. The significant difference in the number of NADPH-d positive and NOS immunoreactive neurons suggests that the localisation of one enzyme does not necessarily reflect the presence of the other.

Keywords: NADPH-diaphorase, nitric oxide synthase, intramural ganglia, urinary bladder, guinea pig

Full Text

The Full Text of this article is available as a PDF (549.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alm P., Elmér M. Adrenergic and cholinergic innervation of the rag urinary bladder. Acta Physiol Scand. 1975 May;94(1):36–45. doi: 10.1111/j.1748-1716.1975.tb05859.x. [DOI] [PubMed] [Google Scholar]
  2. Altschuler R. A., Reeks K. A., Marangos P. J., Fex J. Neuron-specific enolase-like immunoreactivity in inner hair cells but not outer hair cells in the guinea pig organ of Corti. Brain Res. 1985 Feb 18;327(1-2):379–384. doi: 10.1016/0006-8993(85)91541-0. [DOI] [PubMed] [Google Scholar]
  3. Belai A., Schmidt H. H., Hoyle C. H., Hassall C. J., Saffrey M. J., Moss J., Förstermann U., Murad F., Burnstock G. Colocalization of nitric oxide synthase and NADPH-diaphorase in the myenteric plexus of the rat gut. Neurosci Lett. 1992 Aug 31;143(1-2):60–64. doi: 10.1016/0304-3940(92)90233-w. [DOI] [PubMed] [Google Scholar]
  4. Bennett B. C., Kruse M. N., Roppolo J. R., Flood H. D., Fraser M., de Groat W. C. Neural control of urethral outlet activity in vivo: role of nitric oxide. J Urol. 1995 Jun;153(6):2004–2009. [PubMed] [Google Scholar]
  5. Bredt D. S., Hwang P. M., Snyder S. H. Localization of nitric oxide synthase indicating a neural role for nitric oxide. Nature. 1990 Oct 25;347(6295):768–770. doi: 10.1038/347768a0. [DOI] [PubMed] [Google Scholar]
  6. Bredt D. S., Snyder S. H. Nitric oxide, a novel neuronal messenger. Neuron. 1992 Jan;8(1):3–11. doi: 10.1016/0896-6273(92)90104-l. [DOI] [PubMed] [Google Scholar]
  7. Bult H., Boeckxstaens G. E., Pelckmans P. A., Jordaens F. H., Van Maercke Y. M., Herman A. G. Nitric oxide as an inhibitory non-adrenergic non-cholinergic neurotransmitter. Nature. 1990 May 24;345(6273):346–347. doi: 10.1038/345346a0. [DOI] [PubMed] [Google Scholar]
  8. Burnett A. L., Lowenstein C. J., Bredt D. S., Chang T. S., Snyder S. H. Nitric oxide: a physiologic mediator of penile erection. Science. 1992 Jul 17;257(5068):401–403. doi: 10.1126/science.1378650. [DOI] [PubMed] [Google Scholar]
  9. Burnstock G., Cocks T., Crowe R., Kasakov L. Purinergic innervation of the guinea-pig urinary bladder. Br J Pharmacol. 1978 May;63(1):125–138. doi: 10.1111/j.1476-5381.1978.tb07782.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Crowe R., Haven A. J., Burnstock G. Intramural neurons of the guinea-pig urinary bladder: histochemical localization of putative neurotransmitters in cultures and newborn animals. J Auton Nerv Syst. 1986 Apr;15(4):319–339. doi: 10.1016/0165-1838(86)90018-4. [DOI] [PubMed] [Google Scholar]
  11. Dawson T. M., Bredt D. S., Fotuhi M., Hwang P. M., Snyder S. H. Nitric oxide synthase and neuronal NADPH diaphorase are identical in brain and peripheral tissues. Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7797–7801. doi: 10.1073/pnas.88.17.7797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dokita S., Morgan W. R., Wheeler M. A., Yoshida M., Latifpour J., Weiss R. M. NG-nitro-L-arginine inhibits non-adrenergic, non-cholinergic relaxation in rabbit urethral smooth muscle. Life Sci. 1991;48(25):2429–2436. doi: 10.1016/0024-3205(91)90377-n. [DOI] [PubMed] [Google Scholar]
  13. Ehrén I., Iversen H., Jansson O., Adolfsson J., Wiklund N. P. Localization of nitric oxide synthase activity in the human lower urinary tract and its correlation with neuroeffector responses. Urology. 1994 Nov;44(5):683–687. doi: 10.1016/s0090-4295(94)80206-8. [DOI] [PubMed] [Google Scholar]
  14. Fletcher T. F., Bradley W. E. Neuroanatomy of the bladder-urethra. J Urol. 1978 Feb;119(2):153–160. doi: 10.1016/s0022-5347(17)57419-4. [DOI] [PubMed] [Google Scholar]
  15. Gabella G. Intramural neurons in the urinary bladder of the guinea-pig. Cell Tissue Res. 1990 Aug;261(2):231–237. doi: 10.1007/BF00318664. [DOI] [PubMed] [Google Scholar]
  16. Gilpin C. J., Dixon J. S., Gilpin S. A., Gosling J. A. The fine structure of autonomic neurons in the wall of the human urinary bladder. J Anat. 1983 Dec;137(Pt 4):705–713. [PMC free article] [PubMed] [Google Scholar]
  17. Grozdanovic Z., Baumgarten H. G., Brüning G. Histochemistry of NADPH-diaphorase, a marker for neuronal nitric oxide synthase, in the peripheral autonomic nervous system of the mouse. Neuroscience. 1992;48(1):225–235. doi: 10.1016/0306-4522(92)90351-2. [DOI] [PubMed] [Google Scholar]
  18. Grozdanovic Z., Brüning G., Baumgarten H. G. Nitric oxide--a novel autonomic neurotransmitter. Acta Anat (Basel) 1994;150(1):16–24. doi: 10.1159/000147598. [DOI] [PubMed] [Google Scholar]
  19. Hassall C. J., Saffrey M. J., Belai A., Hoyle C. H., Moules E. W., Moss J., Schmidt H. H., Murad F., Förstermann U., Burnstock G. Nitric oxide synthase immunoreactivity and NADPH-diaphorase activity in a subpopulation of intrinsic neurones of the guinea-pig heart. Neurosci Lett. 1992 Aug 31;143(1-2):65–68. doi: 10.1016/0304-3940(92)90234-x. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. James S., Burnstock G. Localization of muscarinic receptors on somatostatin-like immunoreactive neurones of the newborn guinea pig urinary bladder in culture. Neurosci Lett. 1989 Nov 20;106(1-2):13–18. doi: 10.1016/0304-3940(89)90194-8. [DOI] [PubMed] [Google Scholar]
  22. Jänig W., McLachlan E. M. Organization of lumbar spinal outflow to distal colon and pelvic organs. Physiol Rev. 1987 Oct;67(4):1332–1404. doi: 10.1152/physrev.1987.67.4.1332. [DOI] [PubMed] [Google Scholar]
  23. Lee J. H., Price R. H., Williams F. G., Mayer B., Beitz A. J. Nitric oxide synthase is found in some spinothalamic neurons and in neuronal processes that appose spinal neurons that express Fos induced by noxious stimulation. Brain Res. 1993 Apr 16;608(2):324–333. doi: 10.1016/0006-8993(93)91474-7. [DOI] [PubMed] [Google Scholar]
  24. Lowenstein C. J., Dinerman J. L., Snyder S. H. Nitric oxide: a physiologic messenger. Ann Intern Med. 1994 Feb 1;120(3):227–237. doi: 10.7326/0003-4819-120-3-199402010-00009. [DOI] [PubMed] [Google Scholar]
  25. Marangos P. J., Parma A. M., Goodwin F. K. Functional properties of neuronal and glial isoenzymes of brain enolase. J Neurochem. 1978 Sep;31(3):727–732. doi: 10.1111/j.1471-4159.1978.tb07847.x. [DOI] [PubMed] [Google Scholar]
  26. Matsumoto N., Kawano F. Simplified technique of recording edentulous jaw relationship. J Prosthet Dent. 1993 Apr;69(4):448–450. doi: 10.1016/0022-3913(93)90202-y. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. McNeill D. L., Traugh N. E., Jr, Vaidya A. M., Hua H. T., Papka R. E. Origin and distribution of NADPH-diaphorase-positive neurons and fibers innervating the urinary bladder of the rat. Neurosci Lett. 1992 Nov 23;147(1):33–36. doi: 10.1016/0304-3940(92)90768-3. [DOI] [PubMed] [Google Scholar]
  29. Mevorach R. A., Bogaert G. A., Kogan B. A. Role of nitric oxide in fetal lower urinary tract function. J Urol. 1994 Aug;152(2 Pt 1):510–514. doi: 10.1016/s0022-5347(17)32784-2. [DOI] [PubMed] [Google Scholar]
  30. Palmer R. M., Ferrige A. G., Moncada S. Nitric oxide release accounts for the biological activity of endothelium-derived relaxing factor. Nature. 1987 Jun 11;327(6122):524–526. doi: 10.1038/327524a0. [DOI] [PubMed] [Google Scholar]
  31. Persson K., Andersson K. E. Nitric oxide and relaxation of pig lower urinary tract. Br J Pharmacol. 1992 Jun;106(2):416–422. doi: 10.1111/j.1476-5381.1992.tb14349.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Pickel V. M., Reis D. J., Marangos P. J., Zomzely-Neurath C. Immunocytochemical localization of nervous system specific protein (NSP-R) in rat brain. Brain Res. 1976 Mar 19;105(1):184–187. doi: 10.1016/0006-8993(76)90936-7. [DOI] [PubMed] [Google Scholar]
  33. Saffrey M. J., Hassall C. J., Hoyle C. H., Belai A., Moss J., Schmidt H. H., Förstermann U., Murad F., Burnstock G. Colocalization of nitric oxide synthase and NADPH-diaphorase in cultured myenteric neurones. Neuroreport. 1992 Apr;3(4):333–336. doi: 10.1097/00001756-199204000-00011. [DOI] [PubMed] [Google Scholar]
  34. Saffrey M. J., Hassall C. J., Moules E. W., Burnstock G. NADPH diaphorase and nitric oxide synthase are expressed by the majority of intramural neurons in the neonatal guinea pig urinary bladder. J Anat. 1994 Dec;185(Pt 3):487–495. [PMC free article] [PubMed] [Google Scholar]
  35. Sanders K. M., Ward S. M. Nitric oxide as a mediator of nonadrenergic noncholinergic neurotransmission. Am J Physiol. 1992 Mar;262(3 Pt 1):G379–G392. doi: 10.1152/ajpgi.1992.262.3.G379. [DOI] [PubMed] [Google Scholar]
  36. Scheuermann D. W., Stach W., Timmermans J. P., Adriaensen D., De Groodt-Lasseel M. H. Neuron-specific enolase and S-100 protein immunohistochemistry for defining the structure and topographical relationship of the different enteric nerve plexuses in the small intestine of the pig. Cell Tissue Res. 1989 Apr;256(1):65–75. doi: 10.1007/BF00224719. [DOI] [PubMed] [Google Scholar]
  37. Schmechel D. E., Brightman M. W., Barker J. L. Localization of neuron-specific enolase in mouse spinal neurons grown in tissue culture. Brain Res. 1980 Jan 13;181(2):391–400. doi: 10.1016/0006-8993(80)90621-6. [DOI] [PubMed] [Google Scholar]
  38. Schmechel D., Marangos P. J., Zis A. P., Brightman M., Goodwin F. K. Brain endolases as specific markers of neuronal and glial cells. Science. 1978 Jan 20;199(4326):313–315. doi: 10.1126/science.339349. [DOI] [PubMed] [Google Scholar]
  39. Schuman E. M., Madison D. V. Nitric oxide and synaptic function. Annu Rev Neurosci. 1994;17:153–183. doi: 10.1146/annurev.ne.17.030194.001101. [DOI] [PubMed] [Google Scholar]
  40. Traub R. J., Solodkin A., Meller S. T., Gebhart G. F. Spinal cord NADPH-diaphorase histochemical staining but not nitric oxide synthase immunoreactivity increases following carrageenan-produced hindpaw inflammation in the rat. Brain Res. 1994 Dec 30;668(1-2):204–210. doi: 10.1016/0006-8993(94)90525-8. [DOI] [PubMed] [Google Scholar]
  41. Valtschanoff J. G., Weinberg R. J., Rustioni A. NADPH diaphorase in the spinal cord of rats. J Comp Neurol. 1992 Jul 8;321(2):209–222. doi: 10.1002/cne.903210204. [DOI] [PubMed] [Google Scholar]
  42. Vera P. L., Nadelhaft I. Afferent and sympathetic innervation of the dome and the base of the urinary bladder of the female rat. Brain Res Bull. 1992 Nov;29(5):651–658. doi: 10.1016/0361-9230(92)90134-j. [DOI] [PubMed] [Google Scholar]
  43. 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]
  44. Wang X. Y., Wong W. C., Ling E. A. Studies of the lymphatic vessel-associated neurons in the intestine of the guinea pig. J Anat. 1994 Aug;185(Pt 1):65–74. [PMC free article] [PubMed] [Google Scholar]
  45. Wells D. G., Talmage E. K., Mawe G. M. Immunohistochemical identification of neurons in ganglia of the guinea pig sphincter of Oddi. J Comp Neurol. 1995 Jan 30;352(1):106–116. doi: 10.1002/cne.903520108. [DOI] [PubMed] [Google Scholar]
  46. Young H. M., Furness J. B., Shuttleworth C. W., Bredt D. S., Snyder S. H. Co-localization of nitric oxide synthase immunoreactivity and NADPH diaphorase staining in neurons of the guinea-pig intestine. Histochemistry. 1992 May;97(4):375–378. doi: 10.1007/BF00270041. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Anatomy are provided here courtesy of Anatomical Society of Great Britain and Ireland

RESOURCES