Cell and Tissue Research

, Volume 337, Issue 1, pp 37–43 | Cite as

Characterisation of calcitonin gene-related peptide-immunoreactive neurons in the myenteric plexus of rat colon

  • Retsu MitsuiEmail author
Regular Article


A mechanical or chemical stimulus applied to the intestinal mucosa induces motility reflexes in the rat colon. Enteric neurons containing calcitonin gene-related peptide (CGRP) have been suggested as intrinsic primary afferent neurons responsible for mediating such reflexes. In the present study, immunohistochemistry was performed on whole-mount stretch preparations to investigate chemical profiles, morphological characteristics and projections of CGRP-containing neurons in the myenteric plexus of the rat colon. CGRP-positive neuronal cell bodies were detected in preparations incubated with colchicine-containing medium, whereas CGRP-positive nerve fibres were found in colchicine-untreated preparations. These neurons had large oval or round cell bodies that were also immunoreactive for the calcium-binding protein calretinin and neurofilament 200. Myenteric neurons positive for both calretinin and neurofilament 200 had several long processes that emerged from the cell body, consistent with Dogiel type II morphology. Application of the neural tracer DiI to the intestinal mucosa revealed that DiI-labelled myenteric neurons each had an oval or round cell body immunoreactive for calretinin. Thus, CGRP-containing myenteric neurons are Dogiel type II neurons and are immunoreactive for calretinin and neurofilament 200 in the rat colon. These neurons probably project to the intestinal mucosa.


Enteric nervous system Intestine Neuropeptide Reflex Immunohistochemistry Rat (male, Wistar) 



The author thanks Prof. Terumasa Komuro (Waseda University) for useful discussions and helpful suggestions.


  1. Bertrand PP, Kunze WA, Bornstein JC, Furness JB (1998) Electrical mapping of the projections of intrinsic primary afferent neurones to the mucosa of the guinea-pig small intestine. Neurogastroenterol Motil 10:533–541PubMedCrossRefGoogle Scholar
  2. Brehmer A, Schrodl F, Neuhuber W (1999) Morphological classifications of enteric neurons—100 years after Dogiel. Anat Embryol (Berl) 200:125–135CrossRefGoogle Scholar
  3. 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 112:49–59PubMedCrossRefGoogle Scholar
  4. Browning KN, Lees GM (1996) Myenteric neurons of the rat descending colon: electrophysiological and correlated morphological properties. Neuroscience 73:1029–1047PubMedCrossRefGoogle Scholar
  5. Chiocchetti R, Grandis A, Bombardi C, Lucchi ML, Dal Lago DT, Bortolami R, Furness JB (2006) Extrinsic and intrinsic sources of calcitonin gene-related peptide immunoreactivity in the lamb ileum: a morphometric and neurochemical investigation. Cell Tissue Res 323:183–196PubMedCrossRefGoogle Scholar
  6. Clague JR, Sternini C, Brecha NC (1985) Localization of calcitonin gene-related peptide-like immunoreactivity in neurons of the rat gastrointestinal tract. Neurosci Lett 56:63–68PubMedCrossRefGoogle Scholar
  7. Dogiel AS (1899) Über den Bau der Ganglien in den Geflechten des Darmes und der Gallenblase des Menschen und der Säugetiere. Arch Anat Physiol Leipzig Anat Abt Jg 130–158Google Scholar
  8. Ekblad E, Ekman R, Håkanson R, Sundler F (1988) Projections of peptide-containing neurons in rat colon. Neuroscience 27:655–674PubMedCrossRefGoogle Scholar
  9. Furness JB, Costa M (1987) The enteric nervous system. Churchill Livingstone, EdinburghGoogle Scholar
  10. Furness JB, Robbins HL, Xiao J, Stebbing MJ, Nurgali K (2004) Projections and chemistry of Dogiel type II neurons in the mouse colon. Cell Tissue Res 317:1–12PubMedCrossRefGoogle Scholar
  11. Grider JR (1994) CGRP as a transmitter in the sensory pathway mediating peristaltic reflex. Am J Physiol 266:G1139–G1145PubMedGoogle Scholar
  12. Grider JR, Piland BE (2007) The peristaltic reflex induced by short-chain fatty acids is mediated by sequential release of 5-HT and neuronal CGRP but not BDNF. Am J Physiol Gastrointest Liver Physiol 292:G429–G437PubMedCrossRefGoogle Scholar
  13. Hens J, Schrödl F, Brehmer A, Adriaensen D, Neuhuber W, Scheuermann DW, Schemann M, Timmermans JP (2000) Mucosal projections of enteric neurons in the porcine small intestine. J Comp Neurol 421:429–436PubMedCrossRefGoogle Scholar
  14. Hukuhara T, Yamagami M, Nakayama S (1958) On the intestinal intrinsic reflexes. Jpn J Physiol 8:9–20PubMedGoogle Scholar
  15. Kunze WA, Bornstein JC, Furness JB (1995) Identification of sensory nerve cells in a peripheral organ (the intestine) of a mammal. Neuroscience 66:1–4PubMedCrossRefGoogle Scholar
  16. Mann PT, Southwell BR, Ding YQ, Shigemoto R, Mizuno N, Furness JB (1997) Localisation of neurokinin 3 (NK3) receptor immunoreactivity in the rat gastrointestinal tract. Cell Tissue Res 289:1–9PubMedCrossRefGoogle Scholar
  17. Mitsui R, Ono S, Karaki S, Kuwahara A (2005) Neural and non-neural mediation of propionate-induced contractile responses in the rat distal colon. Neurogastroenterol Motil 17:585–594PubMedCrossRefGoogle Scholar
  18. Mulderry PK, Ghatei MA, Spokes RA, Jones PM, Pierson AM, Hamid QA, Kanse S, Amara SG, Burrin JM, Legon S, Polak JM, Bloom SR (1988) Differential expression of alpha-CGRP and beta-CGRP by primary sensory neurons and enteric autonomic neurons of the rat. Neuroscience 25:195–205PubMedCrossRefGoogle Scholar
  19. 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 133:99–107PubMedCrossRefGoogle Scholar
  20. Scheuermann DW, Stach W, De Groodt-Lasseel MH, Timmermans JP (1987) Calcitonin gene-related peptide in morphologically well-defined type II neurons of the enteric nervous system in the porcine small intestine. Acta Anat (Basel) 129:325–328CrossRefGoogle Scholar
  21. Song ZM, Brookes SJ, Costa M (1991) Identification of myenteric neurons which project to the mucosa of the guinea-pig small intestine. Neurosci Lett 129:294–298PubMedCrossRefGoogle Scholar
  22. Sternini C, Reeve JR Jr, Brecha N (1987) Distribution and characterization of calcitonin gene-related peptide immunoreactivity in the digestive system of normal and capsaicin-treated rats. Gastroenterology 93:852–862PubMedGoogle Scholar
  23. Sundler F, Ekblad E, Håkanson R (1989) Projections of enteric peptide-containing neurons in the rat. Arch Histol Cytol 52(Suppl):181–189PubMedCrossRefGoogle Scholar
  24. Thomas JE, Baldwin MV (1971) The intestinal mucosal reflex in the unanesthetized dog. Am J Dig Dis 16:642–647PubMedCrossRefGoogle Scholar
  25. 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) 143:48–53CrossRefGoogle Scholar
  26. Yajima T (1985) Contractile effect of short-chain fatty acids on the isolated colon of the rat. J Physiol (Lond) 368:667–678Google Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  1. 1.Department of Health Science and Social Welfare, Faculty of Human SciencesWaseda UniversityTokorozawaJapan

Personalised recommendations