Abstract
Double-label immunocytochemistry was used to investigate the colocalisation of various neuropeptides and the enzymes nitric oxide synthase (NOS) and tyrosine hydroxylase (TH) in intramural ganglia of the human male urinary bladder neck and trigone. Postmortem specimens were obtained from 7 male infants and children ranging in age from 2 mo to 3 y who had died as a result of cot death or accidental trauma. On average 60% of the intramural neurons were non-TH-immunoreactive (-IR) (i.e. presumptive cholinergic) and 40% were TH- and DbβH-IR (i.e. noradrenergic). Within the non-TH-IR population, calcitonin gene-related peptide (CGRP) was found in 65% of cells, neuropeptide Y (NPY) in 90%, nitric oxide synthase (NOS) in 45%, somatostatin (SOM) in 90%, and vasoactive intestinal polypeptide (VIP) in 40%. The corresponding values for the TH-IR neurons were CGRP (54%), NPY (70%), NOS (58%), SOM (73%) and VIP (40%). All the observed bombesin (BOM)-immunoreactivity was colocalised with TH while 90% of VIP and almost all the CGRP was colocalised with NPY. Less than 5% of neurons were immunoreactive for substance P (SP) or met-enkephalin (m-ENK) and some of these also contained TH. Varicose nerve fibres were seen in close proximity to some of the intramural neurons, the majority of such varicosities showing immunoreactivity to CGRP, VIP or TH. Less common were pericellular varicosities immunoreactive to NPY, SOM or SP. These results demonstrate the neurochemical heterogeneity of intramural neurons in the human bladder neck and provide indirect evidence for the complexity of the peripheral innervation of the human urinary bladder.
Keywords: Urinary bladder, intramural ganglia, neuropeptides, nitric oxide synthase, tyrosine hydroxylase
Full Text
The Full Text of this article is available as a PDF (416.0 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Alm P. Cholinergic innervation of the human urethra and urinary bladder: a histochemical study and review of methodology. Acta Pharmacol Toxicol (Copenh) 1978;43 (Suppl 2):56–62. doi: 10.1111/j.1600-0773.1978.tb03220.x. [DOI] [PubMed] [Google Scholar]
- Andersson K. E., Garcia Pascual A., Persson K., Forman A., Tøttrup A. Electrically-induced, nerve-mediated relaxation of rabbit urethra involves nitric oxide. J Urol. 1992 Jan;147(1):253–259. doi: 10.1016/s0022-5347(17)37208-7. [DOI] [PubMed] [Google Scholar]
- Blessing W. W., Howe P. R., Joh T. H., Oliver J. R., Willoughby J. O. Distribution of tyrosine hydroxylase and neuropeptide Y-like immunoreactive neurons in rabbit medulla oblongata, with attention to colocalization studies, presumptive adrenaline-synthesizing perikarya, and vagal preganglionic cells. J Comp Neurol. 1986 Jun 8;248(2):285–300. doi: 10.1002/cne.902480211. [DOI] [PubMed] [Google Scholar]
- 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]
- Burnett A. L., Tillman S. L., Chang T. S., Epstein J. I., Lowenstein C. J., Bredt D. S., Snyder S. H., Walsh P. C. Immunohistochemical localization of nitric oxide synthase in the autonomic innervation of the human penis. J Urol. 1993 Jul;150(1):73–76. doi: 10.1016/s0022-5347(17)35401-0. [DOI] [PubMed] [Google Scholar]
- Crowe R., Burnstock G. A histochemical and immunohistochemical study of the autonomic innervation of the lower urinary tract of the female pig. Is the pig a good model for the human bladder and urethra? J Urol. 1989 Feb;141(2):414–422. doi: 10.1016/s0022-5347(17)40785-3. [DOI] [PubMed] [Google Scholar]
- Crowe R., Burnstock G., Light J. K. Intramural ganglia in the human urethra. J Urol. 1988 Jul;140(1):183–187. doi: 10.1016/s0022-5347(17)41523-0. [DOI] [PubMed] [Google Scholar]
- 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]
- De Groat W. C. Nervous control of the urinary bladder of the cat. Brain Res. 1975 Apr 11;87(2-3):201–211. doi: 10.1016/0006-8993(75)90417-5. [DOI] [PubMed] [Google Scholar]
- Dixon J. S., Gilpin S. A., Gilpin C. J., Gosling J. A. Intramural ganglia of the human urinary bladder. Br J Urol. 1983 Apr;55(2):195–198. doi: 10.1111/j.1464-410x.1983.tb06554.x. [DOI] [PubMed] [Google Scholar]
- Dixon J. S., Jen P. Y. Development of nerves containing nitric oxide synthase in the human male urogenital organs. Br J Urol. 1995 Dec;76(6):719–725. doi: 10.1111/j.1464-410x.1995.tb00763.x. [DOI] [PubMed] [Google Scholar]
- Edyvane K. A., Smet P. J., Trussell D. C., Jonavicius J., Marshall V. R. Patterns of neuronal colocalisation of tyrosine hydroxylase, neuropeptide Y, vasoactive intestinal polypeptide, calcitonin gene-related peptide and substance P in human ureter. J Auton Nerv Syst. 1994 Aug;48(3):241–255. doi: 10.1016/0165-1838(94)90053-1. [DOI] [PubMed] [Google Scholar]
- Edyvane K. A., Trussell D. C., Jonavicius J., Henwood A., Marshall V. R. Presence and regional variation in peptide-containing nerves in the human ureter. J Auton Nerv Syst. 1992 Jun 15;39(2):127–137. doi: 10.1016/0165-1838(92)90053-j. [DOI] [PubMed] [Google Scholar]
- Ehrén I., Adolfsson J., Wiklund N. P. Nitric oxide synthase activity in the human urogenital tract. Urol Res. 1994;22(5):287–290. doi: 10.1007/BF00297196. [DOI] [PubMed] [Google Scholar]
- Ek A., Alm P., Andersson K. E., Persson C. G. Adrenergic and Cholinergic Nerves of the Human Urethra and Urinary Bladder. A histochemical study. Acta Physiol Scand. 1977 Mar;99(3):345–352. doi: 10.1111/j.1748-1716.1977.tb10387.x. [DOI] [PubMed] [Google Scholar]
- Fehér E., Csányi K., Vajda J. Ultrastructure of the nerve cells and fibres in the urinary bladder wall of the cat. Acta Anat (Basel) 1979;103(1):109–118. [PubMed] [Google Scholar]
- 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]
- Gabella G., Uvelius B. Urinary bladder of rat: fine structure of normal and hypertrophic musculature. Cell Tissue Res. 1990 Oct;262(1):67–79. doi: 10.1007/BF00327747. [DOI] [PubMed] [Google Scholar]
- Garcia-Pascual A., Costa G., Garcia-Sacristan A., Andersson K. E. Relaxation of sheep urethral muscle induced by electrical stimulation of nerves: involvement of nitric oxide. Acta Physiol Scand. 1991 Apr;141(4):531–539. doi: 10.1111/j.1748-1716.1991.tb09114.x. [DOI] [PubMed] [Google Scholar]
- 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]
- Gosling J. A., Dixon J. S. The structure and innervation of smooth muscle in the wall of the bladder neck and proximal urethra. Br J Urol. 1975 Oct;47(5):549–558. doi: 10.1111/j.1464-410x.1975.tb06260.x. [DOI] [PubMed] [Google Scholar]
- 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]
- Gu J., Blank M. A., Huang W. M., Islam K. N., McGregor G. P., Christofides N., Allen J. M., Bloom S. R., Polak J. M. Peptide-containing nerves in human urinary bladder. Urology. 1984 Oct;24(4):353–357. doi: 10.1016/0090-4295(84)90209-7. [DOI] [PubMed] [Google Scholar]
- HAMBERGER B., NORBERG K. A. ADRENERGIC SYNAPTIC TERMINALS AND NERVE CELLS IN BLADDER GANGLIA OF THE CAT. Int J Neuropharmacol. 1965 Feb;4:41–45. doi: 10.1016/0028-3908(65)90045-6. [DOI] [PubMed] [Google Scholar]
- Heym C., Common B., Yin S., Klimaschewski L., Couraud J. Y., Bachmann S. Neurochemistry, connectivity and plasticity of small intensely fluorescent (SIF) cells in the rat superior cervical ganglion. Ann Anat. 1993 Aug;175(4):309–319. doi: 10.1016/s0940-9602(11)80026-9. [DOI] [PubMed] [Google Scholar]
- James M. J., Birmingham A. T., Hill S. J. Partial mediation by nitric oxide of the relaxation of human isolated detrusor strips in response to electrical field stimulation. Br J Clin Pharmacol. 1993 Apr;35(4):366–372. doi: 10.1111/j.1365-2125.1993.tb04152.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- James S., Burnstock G. Colocalization of peptides and a catecholamine-synthesizing enzyme in intramural neurones of the newborn guinea-pig urinary bladder in culture. Regul Pept. 1990 Apr 24;28(2):177–188. doi: 10.1016/0167-0115(90)90016-p. [DOI] [PubMed] [Google Scholar]
- James S., Burnstock G. Neuropeptide Y-like immunoreactivity in intramural ganglia of the newborn guinea pig urinary bladder. Regul Pept. 1988 Nov;23(2):237–245. doi: 10.1016/0167-0115(88)90031-6. [DOI] [PubMed] [Google Scholar]
- Jen P. Y., Dixon J. S., Gearhart J. P., Gosling J. A. Nitric oxide synthase and tyrosine hydroxylase are colocalized in nerves supplying the postnatal human male genitourinary organs. J Urol. 1996 Mar;155(3):1117–1121. [PubMed] [Google Scholar]
- Jen P. Y., Dixon J. S., Gosling J. A. Co-localisation of tyrosine hydroxylase, nitric oxide synthase and neuropeptides in neurons of the human postnatal male pelvic ganglia. J Auton Nerv Syst. 1996 Jun 10;59(1-2):41–50. doi: 10.1016/0165-1838(96)00004-5. [DOI] [PubMed] [Google Scholar]
- Jen P. Y., Dixon J. S., Gosling J. A. Immunohistochemical localization of neuromarkers and neuropeptides in human fetal and neonatal urinary bladder. Br J Urol. 1995 Feb;75(2):230–235. doi: 10.1111/j.1464-410x.1995.tb07317.x. [DOI] [PubMed] [Google Scholar]
- Kawatani M., Rutigliano M., De Groat W. C. Selective facilitatory effect of vasoactive intestinal polypeptide (VIP) on muscarinic firing in vesical ganglia of the cat. Brain Res. 1985 Jun 17;336(2):223–234. doi: 10.1016/0006-8993(85)90649-3. [DOI] [PubMed] [Google Scholar]
- Lundberg J. M., Hökfelt T. Multiple co-existence of peptides and classical transmitters in peripheral autonomic and sensory neurons--functional and pharmacological implications. Prog Brain Res. 1986;68:241–262. doi: 10.1016/s0079-6123(08)60242-3. [DOI] [PubMed] [Google Scholar]
- Lundberg J. M., Terenius L., Hökfelt T., Goldstein M. High levels of neuropeptide Y in peripheral noradrenergic neurons in various mammals including man. Neurosci Lett. 1983 Dec 2;42(2):167–172. doi: 10.1016/0304-3940(83)90401-9. [DOI] [PubMed] [Google Scholar]
- Maggi C. A., Meli A. The role of neuropeptides in the regulation of the micturition reflex. J Auton Pharmacol. 1986 Jun;6(2):133–162. doi: 10.1111/j.1474-8673.1986.tb00640.x. [DOI] [PubMed] [Google Scholar]
- Maggi C. A. The role of peptides in the regulation of the micturition reflex: an update. Gen Pharmacol. 1991;22(1):1–24. doi: 10.1016/0306-3623(91)90304-o. [DOI] [PubMed] [Google Scholar]
- Papka R. E., McNeill D. L. Light- and electron-microscopic study of synaptic connections in the paracervical ganglion of the female rat: special reference to calcitonin gene-related peptide-, galanin- and tachykinin (substance P and neurokinin A)-immunoreactive nerve fibers and terminals. Cell Tissue Res. 1993 Mar;271(3):417–428. doi: 10.1007/BF02913724. [DOI] [PubMed] [Google Scholar]
- Polak J. M., Bloom S. R. Localisation and measurement of VIP in the genitourinary system of man and animals. Peptides. 1984 Mar-Apr;5(2):225–230. doi: 10.1016/0196-9781(84)90211-0. [DOI] [PubMed] [Google Scholar]
- Prieto D., Benedito S., Rodrigo J., Martínez-Murillo R., García-Sacristán A. Distribution and density of neuropeptide Y-immunoreactive nerve fibres and cells in the horse urinary bladder. J Auton Nerv Syst. 1989 Jul;27(2):173–180. doi: 10.1016/0165-1838(89)90099-4. [DOI] [PubMed] [Google Scholar]
- Rand M. J. Nitrergic transmission: nitric oxide as a mediator of non-adrenergic, non-cholinergic neuro-effector transmission. Clin Exp Pharmacol Physiol. 1992 Mar;19(3):147–169. doi: 10.1111/j.1440-1681.1992.tb00433.x. [DOI] [PubMed] [Google Scholar]
- 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]
- Schulman C. C., Duarte-Escalante O., Boyarsky S. The ureterovesical innervation. A new concept based on a histochemical study. Br J Urol. 1972 Dec;44(6):698–712. doi: 10.1111/j.1464-410x.1972.tb10144.x. [DOI] [PubMed] [Google Scholar]
- Scultéty S. Neuropeptides in urology. Int Urol Nephrol. 1989;21(1):39–45. doi: 10.1007/BF02549900. [DOI] [PubMed] [Google Scholar]
- Smet P. J., Edyvane K. A., Jonavicius J., Marshall V. R. Colocalization of nitric oxide synthase with vasoactive intestinal peptide, neuropeptide Y, and tyrosine hydroxylase in nerves supplying the human ureter. J Urol. 1994 Oct;152(4):1292–1296. doi: 10.1016/s0022-5347(17)32570-3. [DOI] [PubMed] [Google Scholar]