Abstract
The neuropeptide substance P (SP) is involved in the regulation of epithelial secretion and motility in the rat small intestine. The morphology, chemical profiles and proportion of SP-containing enteric neurons in this tissue have been examined by immunohistochemical analysis of whole-mount preparations obtained from colchicine-treated rats. In the submucosal plexus of the duodenum, jejunum and ileum, the proportion of SP-positive neurons is 53%, 51% and 49%, respectively. All SP-positive submucosal neurons are positive for neurofilament 200 (NF-200) and calretinin. Immunoreactivity for calcitonin gene-related peptide (CGRP) is detectable in 55% of the SP-positive submucosal neurons. Some SP-positive submucosal neurons have two or more long processes emerging from an oval or round cell body, a characteristic of the Dogiel type II neuron (type II neuron; a putative intrinsic primary afferent neuron). About one-third of the neurons in the myenteric plexus are positive for SP and a majority of them are NF-200/calretinin-positive type II neurons. Immunoreactivity for the SP receptor neurokinin-1 receptor (NK1R) has been detected mainly in the submucosal and myenteric NF-200-positive neurons, which are expected to contain SP. These neurons possibly stimulate each other via SP release. Most of the submucosal and myenteric neurons, including type II neurons, show immunoreactive for the prostaglandin E2 receptor EP3 receptor (EP3R). Thus, SP/NF-200/calretinin/NK1R/EP3R is the common chemical profile of type II neurons in the rat small intestine. The proportion of SP-immunopositive submucosal neurons (49%–53%) is higher in the rat small intestine than in the colon (≤11%) and around 50% are positive for CGRP.
Similar content being viewed by others
References
Brehmer A, Schrödl F, Neuhuber W (1999) Morphological classifications of enteric neurons—100 years after Dogiel. Anat Embryol (Berl) 200:125–135
Brehmer A, Croner R, Dimmler A, Papadopoulos T, Schrödl F, Neuhuber W (2004) Immunohistochemical characterization of putative primary afferent (sensory) myenteric neurons in human small intestine. Auton Neurosci 112:49–59
Brodin E, Sjölund K, Håkanson R, Sundler F (1983) Substance P-containing nerve fibers are numerous in human but not in feline intestinal mucosa. Gastroenterology 85:557–564
Cassuto J, Siewert A, Jodal M, Lundgren O (1983) The involvement of intramural nerves in cholera toxin induced intestinal secretion. Acta Physiol Scand 117:195–202
Cooke HJ, Sidhu M, Fox P, Wang YZ, Zimmermann EM (1997) Substance P as a mediator of colonic secretory reflexes. Am J Physiol 272:G238–G245
Costa M, Furness JB, Llewellyn-Smith IJ, Cuello AC (1981) Projections of substance P-containing neurons within the guinea-pig small intestine. Neuroscience 6:411–424
Costa M, Brookes SJ, Hennig GW (2000) Anatomy and physiology of the enteric nervous system. Gut 47(Suppl 4):iv15–iv19
Dekkers JA, Akkermans LM, Kroese AB (1997) Effects of the inflammatory mediator prostaglandin E2 on myenteric neurons in guinea pig ileum. Am J Physiol 272:G1451–G1456
Desaki J, Fujiwara T, Komuro T (1984) A cellular reticulum of fibroblast-like cells in the rat intestine: scanning and transmission electron microscopy. Arch Histol Jpn 47:179–186
Ekblad E, Winther C, Ekman R, Håkanson R, Sundler F (1987) Projections of peptide-containing neurons in rat small intestine. Neuroscience 20:169–188
Furness JB (2006) The enteric nervous system. Blackwell, Oxford
Furuya S, Furuya K (2007) Subepithelial fibroblasts in intestinal villi: roles in intercellular communication. Int Rev Cytol 264:165–223
Furuya K, Furuya S, Yamagishi S (1994) Intracellular calcium responses and shape conversions induced by endothelin in cultured subepithelial fibroblasts of rat duodenal villi. Pflugers Arch 428:97–104
Furuya K, Sokabe M, Furuya S (2005) Characteristics of subepithelial fibroblasts as a mechano-sensor in the intestine: cell-shape-dependent ATP release and P2Y1 signaling. J Cell Sci 118:3289–3304
Giuliani S, Tramontana M, Lecci A, Maggi CA (1996) Tachykinin receptors mediate atropine-resistant rat duodenal reflex contractions in vivo. Naunyn Schmiedebergs Arch Pharmacol 354:327–335
Holzer P (1998) Implications of tachykinins and calcitonin gene-related peptide in inflammatory bowel disease. Digestion 59:269–283
Johnson PJ, Bornstein JC, Burcher E (1998) Roles of neuronal NK1 and NK3 receptors in synaptic transmission during motility reflexes in the guinea-pig ileum. Br J Pharmacol 124:1375–1384
Komuro T, Hashimoto Y (1990) Three-dimensional structure of the rat intestinal wall (mucosa and submucosa). Arch Histol Cytol 53:1–21
Larsson MH, Sapnara M, Thomas EA, Bornstein JC, Lindström E, Svensson DJ, Sjövall H (2008) Pharmacological analysis of components of the change in transmural potential difference evoked by distension of rat proximal small intestine in vivo. Am J Physiol Gastrointest Liver Physiol 294:G165–G173
Lin Z, Gao N, Hu HZ, Liu S, Gao C, Kim G, Ren J, Xia Y, Peck OC, Wood JD (2002) Immunoreactivity of Hu proteins facilitates identification of myenteric neurons in guinea-pig small intestine. Neurogastroenterol Motil 14:197–204
Lundgren O (2002) Enteric nerves and diarrhoea. Pharmacol Toxicol 90:109–120
Mann PT, Furness JB, Southwell BR (1999a) Choline acetyltransferase immunoreactivity of putative intrinsic primary afferent neurons in the rat ileum. Cell Tissue Res 297:241–248
Mann PT, Southwell BR, Furness JB (1999b) Internalisation of the neurokinin 1 receptor in rat myenteric neurons. Neuroscience 91:353–362
Manning BP, Sharkey KA, Mawe GM (2002) Effects of PGE2 in guinea pig colonic myenteric ganglia. Am J Physiol Gastrointest Liver Physiol 283:G1388–G1397
Mitsui R (2009) Characterisation of calcitonin gene-related peptide-immunoreactive neurons in the myenteric plexus of rat colon. Cell Tissue Res 337:37–43
Mitsui R (2010) Immunohistochemical characteristics of submucosal Dogiel type II neurons in rat colon. Cell Tissue Res 340:257–265
Mongardi Fantaguzzi C, Thacker M, Chiocchetti R, Furness JB (2009) Identification of neuron types in the submucosal ganglia of the mouse ileum. Cell Tissue Res 336:179–189
Pan H, Gershon MD (2000) Activation of intrinsic afferent pathways in submucosal ganglia of the guinea pig small intestine. J Neurosci 20:3295–3309
Pataky DM, Curtis SB, Buchan AM (1990) The co-localization of neuropeptides in the submucosa of the small intestine of normal Wistar and non-diabetic BB rats. Neuroscience 36:247–254
Patton D, O’Reilly M, Vanner S (2005) Sensory peptide neurotransmitters mediating mucosal and distension evoked neural vasodilator reflexes in guinea pig ileum. Am J Physiol Gastrointest Liver Physiol 289:G785–G790
Peterson JW, Whipp SC (1995) Comparison of the mechanisms of action of cholera toxin and the heat-stable enterotoxins of Escherichia coli. Infect Immun 63:1452–1461
Phillips RJ, Hargrave SL, Rhodes BS, Zopf DA, Powley TL (2004) Quantification of neurons in the myenteric plexus: an evaluation of putative pan-neuronal markers. J Neurosci Methods 133:99–107
Rivera LR, Thacker M, Furness JB (2009) High- and medium-molecular-weight neurofilament proteins define specific neuron types in the guinea-pig enteric nervous system. Cell Tissue Res 335:529–538
Roza C, Reeh PW (2001) Substance P, calcitonin gene related peptide and PGE2 co-released from the mouse colon: a new model to study nociceptive and inflammatory responses in viscera, in vitro. Pain 93:213–219
Schultzberg M, Hökfelt T, Nilsson G, Terenius L, Rehfeld JF, Brown M, Elde R, Goldstein M, Said S (1980) Distribution of peptide- and catecholamine-containing neurons in the gastro-intestinal tract of rat and guinea-pig: immunohistochemical studies with antisera to substance P, vasoactive intestinal polypeptide, enkephalins, somatostatin, gastrin/cholecystokinin, neurotensin and dopamine beta-hydroxylase. Neuroscience 5:689–744
Sternini C, Su D, Gamp PD, Bunnett NW (1995) Cellular sites of expression of the neurokinin-1 receptor in the rat gastrointestinal tract. J Comp Neurol 358:531–540
Timmermans JP, Scheuermann DW, Barbiers M, Adriaensen D, Stach W, Van Hee R, De Groodt-Lasseel MH (1992) Calcitonin gene-related peptide-like immunoreactivity in the human small intestine. Acta Anat (Basel) 143:48–53
Turvill JL, Connor P, Farthing MJ (2000) Neurokinin 1 and 2 receptors mediate cholera toxin secretion in rat jejunum. Gastroenterology 119:1037–1044
Vanner S, MacNaughton WK (2004) Submucosal secretomotor and vasodilator reflexes. Neurogastroenterol Motil 16(Suppl 1):39–43
Vannucchi MG, Faussone-Pellegrini MS (2000) NK1, NK2 and NK3 tachykinin receptor localization and tachykinin distribution in the ileum of the mouse. Anat Embryol (Berl) 202:247–255
Weber E, Neunlist M, Schemann M, Frieling T (2001) Neural components of distension-evoked secretory responses in the guinea-pig distal colon. J Physiol (Lond) 536:741–751
Acknowledgements
The author thanks Prof. Terumasa Komuro (Waseda University) for helpful comments on the manuscript.
Author information
Authors and Affiliations
Corresponding author
Additional information
This study was supported by a Waseda University Grant for Special Research Projects (2009B-285, 2010B-294).
Rights and permissions
About this article
Cite this article
Mitsui, R. Immunohistochemical analysis of substance P-containing neurons in rat small intestine. Cell Tissue Res 343, 331–341 (2011). https://doi.org/10.1007/s00441-010-1080-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00441-010-1080-7