Abstract
The intramural projections of nerve cells containing serotonin (5-HT), calcitonin gene-related peptide (CGRP), vasoactive intestinal peptide (VIP) and nitric oxide synthase or reduced nicotinamide adenine dinucleotide phosphate diaphorase (NOS/NADPHd) were studied in the ascending colon of 5- to 6-week-old pigs by means of immunocytochemistry and histochemistry in combination with myectomy experiments. In control tissue of untreated animals, positive nerve cells and fibres were common in the myenteric and outer submucous plexus and, except for 5-HT-positive perikarya, immunoreactive cell bodies and fibres were also observed in the inner submucous plexus. VIP- and NOS/NADPHd-positive nerve fibres occurred in the ciruclar muscle layer while VIP was also abundant in nerve fibres of the mucosal layer. 5-HT- and CGRP-positive nerve fibres were virtually absent from the aganglionic nerve networks. In the submucosal layer, numerous paravascular CGRP-immunoreactive (IR) nerve fibres were encountered. Myectomy studies revealed that 5-HT-, CGRP-, VIP- and NOS/NADPHd-positive myenteric neurons all displayed anal projections within the myenteric plexus. In addition, some of the serotonergic myenteric neurons projected anally to the outer submucous plexus, whereas a great number of the VIP-ergic and nitrergic myenteric neurons send their axons towards the circular muscle layer. The possible function of these nerve cells in descending nerve pathways in the porcine colon is discussed in relation to the distribution pattern of their perikarya and processes and some of their morphological characteristics.
Similar content being viewed by others
References
Barbiers M, Timmermans J-P, Scheuermann DW, Adriaensen D, Mayer B, De Groodt-Lasseel MHA (1993a) Distribution and morphological features of nitrergic neurons in the porcine large intestine. Histochemistry 100:27–34
Barbiers M, Timmermans J-P, Adriaensen D, De Groodt-Lasseel MHA, Scheuermann DW (1993b) Topographical distribution and immunocytochemical features of colonic neurons that project to the cranial mesenteric ganglion in the pig. J Auton Nerv Syst 44:119–127
Barbiers M, Timmermans J-P, Scheuermann DW, Adriaensen D, Mayer B, De Groodt-Lasseel MHA (1994) Nitric oxide synthase-containing neurons in the pig large intestine: topography, morphology and viscerofugal projections. Microse Res Techn 29:72–78
Boeckxstaens GE, Pelckmans PA, Bult H, De Man JG, Herman AG, Van Maercke YM (1991) Evidence for nitric oxide as mediator of non-adrenergic, non-cholinergic relaxations induced by ATP and GABA in the canine gut. Br J Pharmacol 102:434–438
Bornstein JC, Hendriks R, Furness JB, Trussell DC (1991a) Ramifications of the axons of AH-neurons injected with the intracellular marker biocytin in the myenteric plexus of the guinea pig small intestine. J Comp Neurol 314:437–451
Bornstein JC, Furness JB, Smith TK, Trussell DC (1991b) Synaptic responses evoked by mechanical stimulation of the mucosa in morphologically characterized myenteric neurons of the guinea-pig ileum. J Neurosci 11:505–518
Costa M, Furness JB (1983) The origins, pathways and terminations of neurons with VIP-like immunoreactivity in the guinea-pig small intestine. Neuroscience 8:665–676
Costa M, Furness JB, Cuello AC, Verhofstad AAJ, Steinbusch HWJ, Elde RP (1982) Neurons with 5-hydroxytryptamine-like immunoreactivity in the enteric nervous system: their visualization and reactions to drug treatment. Neuroscience 7:351–363
Costa M, Furness JB, Brookes SJH, Bredt DS, Snyder SH (1991) Presence and chemical coding of neurons with nitric oxide synthase immunoreactivity in the guinea pig small intestine. Proc Aust Physiol Pharmacol Soc 22:97P
Daniel EE, Furness JB, Costa M, Belbeck L (1987) The projections of chemically identified nerve fibers in canine ileum. Cell Tissue Res 247:377–384
Domoto T, Bishop AE, Oki M, Polak JM (1990) An in vitro study of the projections of enteric vascoactive intestinal polypeptide-immunoreactive neurons in the human colon. Gastroenterology 98:819–827
Ekblad E, Winther C, Ekman R, Håkanson R, Sundler F (1987) Projections of peptide-containing neurons in the rat small intestine. Neuroscience 20:169–188
Ekblad E, Ekman R, Håkanson R, Sundler F (1988) Projections of peptide-containing neurons in rat colon. Neuroscience 27:655–674
Ekblad E, Håkanson R, Sundler F (1991) Microanatomy and chemical coding of peptide-containing neurons in the digestive tract. In: Daniel EE (ed) Neuropeptide function in the gastrointestinal tract. CRC Press, Boston, pp 131–179
Furness JB, Costa M (1979) Projections of intestinal neurons showing immunoreactivity for vasoactive intestinal polypeptide are consistent with these neurons being the enteric inhibitory neurons. Neurosci Lett 15:199–204
Furness JB, Costa M (1982) Neurons with 5-hydroxytryptamine-like immunoreactivity in the enteric nervous system: their projections in the guinea-pig small intestine. Neuroscience 7:341–349
Furness JB, Costa M (1987) The enteric nervous system. Churchill Livingstone, London
Furness JB, Costa M, Gibbins IL, Llewellyn-Smith IJ, Oliver JR (1985) Neurochemically similar myenteric and submucous neurons directly traced to the mucosa of the small intestine. Cell Tissue Res 241:155–163
Furness JB, Costa M, Rökaeus A, McDonald TJ, Brooks B (1987) Galanin-immunoreactive neurons in the guinea-pig small intestine: their projections and relationships to other neurons. Cell Tissue Res 250:607–615
Furness JB, Lloyd KCK, Sternini C, Walsch JH (1990a) Projections of substance P, vasoactive intestinal peptide and tyrosine hydroxylase immunoreactive nerve fibres in the canine intestine, with special reference to the innervation of the circular muscle. Arch Histol Cytol 53:129–140
Furness JB, Trussell DC, Pompolo S, Bornstein JC, Smith TK (1990b) Calbinding neurons of the guinea-pig small intestine: quantitative analysis of their numbers and projections. Cell Tissue Res 260:261–272
Furness JB, Kuramoto H, Messenger JP (1990c) Morphological and chemical identification of neurons that project from the colon to the inferior mesenteric ganglia in the guinea pig. J Auton Nerv Syst 31:203–210
Furness JB, Costa M, Pompolo S, Bornstein JC, Bredt, DS, Snyder SH (1991) Projections of neurons with nitric oxide synthase immunoreactivity in the guinea-pig small intestine. Proc Aust Physiol Pharmacol Soc 22:99P
Grider JR (1990) Identification of calcitonin gene related peptide (CGRP) as a sensory neurotansmitter of stretch-induced peristaltic reflex. Gastroenterology 98:A355
Grider JR, Makhlouf GM (1986) Colonic peristaltic reflex: identification of vasoactive intestinal peptide as mediator of descending relaxation. Am J Physiol 251:G40-G45
Kirchgessner AL, Gershon MD (1989) Identification of vagal efferent fibers and putative target neurons in the enteric nervous system of the rat. J Comp Neurol 285:38–53
Klatt P, Heinzel B, John M, Kastner M, Böhme E, Mayer B (1992) Ca2+/calmodulin-dependent cytochrome c reductase activity of brain nitric oxide synthase. J Biol Chem 267:11374–11378
Llewellyn-Smith IJ, Costa M, Furness JB (1985) Light and electron microscopic immunocytochemistry of the same nerves from whole mount preparations. J Histochem Cytochem 33:857–866
McConalogue K, Furness JB (1993) Projections of nitric oxide synthesizing neurons in the guinea-pig colon. Cell Tissue Res 271:545–553
Messenger JP (1993) Immunohistochemical analysis of neurons and their projections in the proximal colon of the guinea-pig. Arch Histol Cytol 56:459–473
Messenger JP, Furness JB (1990) Projections of chemically-specified neurons in the guinea-pig colon. Arch Histol Cytol 53:467–495
Messenger JP, Furness JB (1993) Distribution of enteric nerve cells projecting to the superior and inferior mesenteric ganglia of the guinea pig. Cell Tissue Res 271:333–339
Parkman HP, Ma RC, Stapelfeldt WH, Szurszewski JH (1993) Direct and indirect mechanosensory pathways from the colon to the inferior mesenteric ganglion. Am J Physiol 265:G499-G505
Sanders KM, Smith TK (1986) Motoneurons of the submucous plexus regulate electrical activity of the circular muscle of canine proximal colon. J Physiol 280:293–310
Scherer-Singler U, Vincent SR, Kimura H, McGeer EG (1983) Demonstration of a unique population of neurons with NADPH-diaphorase histochemistry. J Neurosci Methods 9:229–234
Scheuermann DW, Stach W, De Groodt-Lasseel MHA, Timmermans J-P (1987) Calcitonin gene-related peptide in morphologically well-defined type II neurons of the enteric nervous system in the porcine small intestine. Acta Anat 129:325–328
Scheuermann DW, Stach W, Timmermans J-P (1990) Serotonin-immunoreactivity in the wall of the porcine small intestine. Verh Anat Ges 83 [Anat Anz Suppl 166]:551–552
Stach W (1989) A revised morphological classification of neurons in the enteric nervous system. In: Singer MV, Goebell H (eds) Nerves and the gastrointestinal tract. MTP Press, Dordrecht, pp 29–45
Timmermans J-P, Scheuermann DW, Stach W, Barbiers M, Adriaensen D, Krammer H-J, De Groodt-Lasseel MHA (1991) Projections of serotonin-containing neurons in the enteric nerve plexus, an experimental study in the porcine small intestine. Anat Anz Suppl 170:89–90
Timmermans J-P, Scheuermann DW, Stach W, Adriaensen D, De Groodt-Lasseel MHA (1992a) Functional morphology of the enteric nervous system with special references to large mammals. Eur J Morphol 30:113–122
Timmermans J-P, Scheuermann DW, Barbiers M, Adriaensen D, Stach W, Van Hee R, De Groodt-Lasseel MHA (1992b) Calcitonin gene-related peptide-like immunoreactivity in the human small intestine. Acta Anat 143:48–53
Timmermans J-P, Barbiers M, Scheuermann DW, Stach W, Adriaensen D, De Groodt-Lasseel MHA (1993) Occurrence, distribution and neurochemical features of small intestinal neurons projecting to the cranial mesenteric ganglion in the pig. Cell Tissue Res 272:49–58
Timmermans J-P, Barbiers M, Scheuermann DW, Stach W, Adriaensen D, Mayer B, De Groodt-Lasseel MHA (1994a) Distributional pattern, neurochemical features and projections of nitrergic neurons in the pig small intestine. Ann Anat, in press
Timmermans J-P, Barbiers M, Scheuermann DW, Bogers JJ, Adriaensen D, Fekete E, Mayer B, Van Marck EA, De Groodt-Lasseel MHA (1994b) Nitric oxide synthase immunoreactivity in the enteric nervous system of the developing human digestive tract. Cell Tissue Res 275:235–245
Vitadello M, Triban C, Fabris M, Gorio A, Schiaffino S (1986) Heterogeneity of rat neurofilament polypeptides revealed by a monoclonal antibody. J Neurochem 46:665–670
Wilson AJ, Llewellyn-Smith IJ, Furness JB, Costa M (1987) The source of the nerve fibres forming the deep muscular and circular muscle plexuses in the small intestine of the guinea-pig. Cell Tissue Res 247:497–504
Young HM, Furness JB, Shuttleworth CWR, Bredt DS, Snyder SH (1992) Co-localization of nitric oxide synthase immunoreactivity and NADPH diaphorase staining in neurons of the guinea-pig intestine. Histochemistry 97:375–378
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Barbiers, M., Timmermans, JP., Adriaensen, D. et al. Projections of neurochemically specified neurons in the porcine colon. Histochem Cell Biol 103, 115–126 (1995). https://doi.org/10.1007/BF01454008
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF01454008