Abstract
Nitric oxide synthase (NOS)- and vasoactive intestinal peptide (VIP)-immunoreactive neurons projecting to the upper rectum or penis were examined using retrograde tracing combined with immunohistochemistry in the major pelvic ganglion of male rats. Five days after injection of Fluoro-Gold (FG) into the upper rectum or penis, the major pelvic ganglion was treated with colchicine. FG injected into the upper rectum labelled many ganglion neurons in the major pelvic ganglion. Immunohistochemistry showed that 37% of FG-labelled neurons were immunoreactive for NOS and 33% for VIP. After injection of FG into the penis, 41% of FG-labelled neurons were immunoreactive for NOS and 25% for VIP. Serial cryostat sections stained for NOS and VIP, respectively, showed the co-localization of NOS and VIP in the ganglion cells projecting to the rectum and penis. In the major pelvic ganglion of the colchicinetreated animals, about 17% of the ganglion cells were immunoreactive for NOS and 32% were immunoreactive for VIP. These neurons were small in diameter (less than 30 μm). A histogram showing cell sizes in cross-sectional areas of NOS-immunoreactive neurons coincided with that of VIP-immunoreactive neurons. Most of the NOS-and VIP-immunoreactive neurons were less than 600 μm2. These results indicate that small neurons containing both NOS and VIP in the major pelvic ganglion project to the rectum and penis. In the penile erectile tissues and enteric ganglia, NO and VIP may be released from the same axons and may act concomitantly on the target tissue.
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Boeckxstaens GE, Pelckmans PA, Bogers JJ, Bult H, Man JG de, Oosterbosch L, Herman AG, Maercke YM van (1991) Release of nitric oxide upon stimulation of nonadrenergic noncholinergic nerves in the rat gastric fundus. J Pharmacol Exp Ther 256:441–447
Bredt DS, Hwang PM, Snyder SH (1990) Localization of nitric oxide synthase indicating a neural role for nitric oxide. Nature 347:768–770
Coons AH, Leduc EH, Connolly JM (1955) Studies on antibody production. I. A method for the histochemical demonstration of specific antibody and its application to a study of the hyperimmune rabbit. J Exp Med 102:42–60
Costa M, Furness JB (1983) The origins, pathways, and terminations of neurons with VIP-like immunoreactivity in the guineapig small intestine. Neuroscience 8:665–676
Costa M, Furness JB, Pompolo S, Brookes SJH, bornstein JC, Bredt DS, Snyder SH (1992) Projections and chemical coding of neurons with immunoreactivity for nitric oxide synthase in the guinea-pig small intestine. Neurosci Lett 148:121–125
Dail WG, Moll MA, Weber K (1983) Localization of vasoactive intestinal polypeptide in penile crectile tissue and in the major pelvic ganglion of the rat. Neuroscience 10:1379–1386
Dalziel HH, Thornbury KD, Ward SM, Sanders KM (1991) Involvement of nitric oxide synthetic pathway in inhibitory junction potentials in canine proximal colon. Am J Physiol 260:G789-G792
Daniel EE, Furness JB, Costa M, Belbeck L (1987) The projections of chemically identified nerve fibres in canine ileum. Cell Tissue Res 247:377–384
Dawson TM, Bredt DS, Fotuhi M, Hwang PM, Snyder SH (1991) Nitric oxide synthase and neuronal NADPH diaphorase are identical in brain and peripheral tissues. Proc Natl Acad Sci USA 88:7797–7801
Domoto T, Oki M (1991) Distribution of extrinsic neuropeptide Y-containing nerves in the rat upper rectum (in Japanese with English abstract). Auton Nerv Syst (Jpn) 28:18–25
Domoto T, Gonda T, Oki M (1992) Distribution of extrinsic nerves containing vasoactive intestinal polypeptide-like immunoreactivity in the aganglionic bowel of the congenital aganglionosis rat. Biomed Res (India) 3:65–77
Ekblad E, Winther C, Ekman R, H»kanson R, Sundler F (1987) Projections of peptide-containing neurons in rat small intestine. Neuroscience 20:169–188
Furchgott RF (1990) Studies on endothelium-dependent vasodilation and the endothelium-derived relaxing factor. Acta Physiol Scand 139:257–270
Furness JB, Pompolo S, Shuttleworth CWR, Burleigh DE (1992) Light- and electron-microscopic immunochemical analysis of nerve fibre types innervating the taenia of the guinea-pig caecum. Cell Tissue Res 270:125–137
Garthwaite J (1991) Glutamate, nitric oxide and cell-cell signalling in the nervous system. Trends Neurosci 14:60–67
Goyal RK, Rattan S, Said SI (1980) VIP as a possible neurotransmitter of non-cholinergic non-adrenergic inhibitory neurons. Nature 288:378–380
Grider JR, Murthy KS, Jin J-G, Makhlouf GM (1992) Stimulation of nitric oxide from muscle cells by VIP: prejunctional enhancement of VIP release. Am J Physiol 262:G774-G778
Groat AC de, Steers WD (1988) Neuroanatomy and neurophysiology of penile erection. In: Tanagho EA, Lue TF, McClure RD (eds) Contemporary management of impotence and fertility. Wiliams & Wilkins, Baltimore, pp 3–27
Hope BT, Michael GJ, Knigge KM, Vincent SR (1991) Neuronal NADPH diaphorase is a nitric oxide synthase. Proc Natl Acad Sci USA 88:2811–2814
Hsu S-M, Raine L, Fanger H (1981) Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabelled antibody (PAP) procedures. J Histochem Cytochem 29:577–580
Ignarro LJ, Bush PA, Buga GM, Wood KS, Fukuto JM, Rajfer J (1990) Nitric oxide and cyclic GMP formation upon electrical field stimulation cause relaxation of corpus cavernosum smooth muscle. Biochem Biophys Res Commun 170:843–850
Keast JR (1992) A possible neural source of nitric oxide in the rat penis. Neurosci Lett 143:69–73
Keast JR, Groat WC de (1989) Immunohistochemical characterization of pelvio neurons which project to the bladder, colon, or penis in rats. J Comp Neurol 288:387–400
Keast JR, Booth AM, Groat WC de (1989) Distribution of neurons in the major pelvic ganglion of the rat which supply the bladder, colon or penis. Cell Tissue Res 256:105–112
Knispel HH, Goessl C, Beckmann R (1991) Basal and acetylcholine-stimulated nitric oxide formation mediates relaxation of rabbit cavernous smooth muscle. J Urol 146:1429–1433
Li CG, Rand MJ (1990) Nitric oxide and vasoactive intestinal polypeptide mediate non-adrenergic, non-cholinergic inhibitory transmission to smooth muscle of the rat gastric fundus. Eur J Pharmacol 191:303–309
Li CG, Rand MJ (1991) Evidence that part of the NANC relaxant response of guinea-pig trachea to electrical field stimulation is mediated by nitric oxide. Br J Pharmacol 102:91–94
Maggi CA, Barbanti G, Turini D, Giuliani S (1991) Effect of NG-monomethyl L-arginine (L-NMMA) and NG-nitro L-arginine (L-NOARG) on non-adrenergic non-cholinergic relaxation in the circular muscle of the human ileum. Br J Pharmacol 103:1970–1972
McConalogue K, Furness JB (1993) Projections of nitric oxide synthesizing neurons in the guinea-pig colon. Cell Tissue Res 271:545–553
Morgan KG, Schmalz PF, Szurszewski HJ (1978) The inhibitory effects of vasoactive intestinal polypeptide on the mechanical and electrical activity of canine antral smooth muscle. J Physiol (Lond) 282:437–450
Palmer RMJ, Moncada S (1989) A novel citrulline-forming enzyme implicated in the formation of nitric oxide by vascular endothelial cells. Biochem Biophys Res Commun 158:348–352
Palmer RMJ, Ferrige AG, Moncada S (1987) Nitrie oxide release accounts for the biological activity of endothelium-derived relaxing factor. Nature 327:527–526
Sanders KM, Ward SM (1992) Nitric oxide as a mediator of nonad-renergic noncholinergic neurotransmission. Am J Physiol 262:G379-G392
Santer RM, Symons D (1993) Distribution of NADPH-diaphorase activity in rat paravertebral, prevertebral and pelvic sympathetic ganglia. Cell Tissue Res 271:115–121
Springall DR, Riveros-Moreno V, Buttery L, Suburo A, Bishop AE, Merrett M, Moncada S, Polak JM (1992) Immonological detection of nitric oxide synthase(s) in human tissues using heterologous antibodies suggesting different isoforms. Histochemistry 98:259–266
Stark ME, Szurszewski JH (1992) Role of nitric oxide in gastrointestinal and hepatic function and disease. Gastroenterology 103:1928–1949
Stefanini M, De Martino C, Zamboni L (1967) Fixation of ejaculated spermatozoa for electron microscopy. Nature 216:173–174
Vincent SR, Hope BT (1992) Neurons that say NO. Trends Neurosci 15:108–113
Ward SM, Xue C, Shuttleworth W, Bredt DS, Snyder SH, Sanders KM (1992) NADPH diaphorase and nitric oxide synthase colocalization in enteric neurons of canine proximal colon. Am J Physiol 263:G277-G284
Williams JT, North RA (1979) Vasoactive intestinal polypeptide excites neurones of the myenteric plexus. Brain Res 175:174–177
Willis E, Ottesen B, Wagner G, Sundler F, Fahrenkrug J (1981) Vasoactive intestinal polypeptide (VIP) as a possible neurotransmitter involved in penile erection. Acta Physiol Scand 113:545–547
Young HM, Furness JB, Shuttleworth CWR, Bredt DS, Snyder SH (1992) Co-localization of nitric oxide synthase immunoreactivity and AADPH diaphorase staining in neurons of the guinea-pig intestine. Histochemistry 97:375–378
Zafirov DH, Palmer JM, Nemeth PR, Wood JD (1985) Bombesin, gastrin releasing peptide and vasoactive intestinal peptide excite myenteric neurons. Eur J Pharmacol 115:103–107
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Domoto, T., Tsumori, T. Co-localization of nitric oxide synthase and vasoactive intestinal peptide immunoreactivity in neurons of the major pelvic ganglion projecting to the rat rectum and penis. Cell Tissue Res. 278, 273–278 (1994). https://doi.org/10.1007/BF00414170
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DOI: https://doi.org/10.1007/BF00414170