Abstract
Resident macrophages are distributed in the network of interstitial cells of Cajal (ICC) and the myenteric nerve within the myenteric plexus. We evaluated changes in chemoattractant protein mRNA expression in macrophages and neutrophils, the ICC, nerve and macrophages in the myenteric plexus of model rats with TNBS-induced colitis. Chemoattractant proteins, MCP-1, GRO, MIP-2 and CINC-2α were upregulated in the colonic muscle layer after inflammation. Leukocyte infiltration and MPO activity were increased in the muscle layer. Electron microscopy indicated an irregular contour of the myenteric ganglia into which numerous macrophages had penetrated. Macrophages were also distributed near the ICC in the inflamed myenteric plexus. Immunohistochemistry showed that the ICC network and myenteric nerve system had disappeared from the inflamed region, whereas the number of resident macrophages was increased. TTX-insensitive, possibly ICC-mediated, rhythmic contractions of circular smooth muscle strips and enteric neuron-mediated TTX-sensitive peristalsis in the whole proximal colon tissue were significantly inhibited in the inflamed colon, indicating that the ICC-myenteric nerve system was dysfunctional in the inflamed muscle layer. Their accumulation around the myenteric nerve plexus and the ICC network suggests that macrophages play an important role in inducing intestinal dysmotility in gut inflammation.
Similar content being viewed by others
References
Allison MC, Poulter LW (1991) Changes in phenotypically distinct mucosal macrophage populations may be a prerequisite for the development of inflammatory bowel disease. Clin Exp Immunol 85:504–509
Annese V, Bassotti G, Napolitano G, Usai P, Andriulli A, Vantrappen G (1997) Gastrointestinal motility disorders in patients with inactive Crohn’s disease. Scand J Gastroenterol 32:1107–1117
Bossone C, Hosseini JM, Pineiro-Carrero V, Shea-Donohue T (2001) Alterations in spontaneous contractions in vitro after repeated inflammation of rat distal colon. Am J Physiol Gastrointest Liver Physiol 280:G949–G957
Bradley PP, Priebat DA, Christensen RD, Rothstein G (1982) Measurement of cutaneous inflammation: estimation of neutrophil content with an enzyme marker. J Investig Dermatol 78:206–209
Brewer DB, Thompson H, Haynes IG, Alexander-Williams J (1990) Axonal damage in Crohn’s disease is frequent, but non-specific. J Pathol 161:301–311
Burns AJ, Lomax AE, Torihashi S, Sanders KM, Ward SM (1996) Interstitial cells of Cajal mediate inhibitory neurotransmission in the stomach. Proc Nat Acad Sci USA 93:12008–12013
Daniel EE, Posey-Daniel V (1984) Neuromuscular structures in opossum esophagus: role of interstitial cells of Cajal. Am J Physiol 246:G305–G315
Dembowski C, Hofmann P, Koch T, Kamrowski-Kruck H, Riedesel H, Krammer HJ, Kaup FJ, Ehrenreich H (2000) Phenotype, intestinal morphology, and survival of homozygous and heterozygous endothelin B receptor–deficient (spotting lethal) rats. J Pediatr Surg 35:480–488
Depoortere I, Thijs T, van Assche G, Keith JC Jr, Peeters TL (2000) Dose-dependent effects of recombinant human interleukin-11 on contractile properties in rabbit 2,4,6-trinitrobenzene sulfonic acid colitis. J Pharmacol Exp Ther 294:983–990
Der T, Bercik P, Donnelly G, Jackson T, Berezin I, Collins SM, Huizinga JD (2000) Interstitial cells of Cajal and inflammation-induced motor dysfunction in the mouse small intestine. Gastroenterology 119:1590–1599
Dvorak AM, Onderdonk AB, McLeod RS, Monahan-Earley RA, Cullen J, Antonioli DA, Blair JE, Morgan ES, Cisneros RL, Estrella P (1993) Axonal necrosis of enteric autonomic nerves in continent ileal pouches. Possible implications for pathogenesis of Crohn’s disease. Ann Surg 217:260–271
Eskandari MK, Kalff JC, Billiar TR, Lee KK, Bauer AJ (1999) LPS-induced muscularis macrophage nitric oxide suppresses rat jejunal circular muscle activity. Am J Physiol Gastrointest Liver Physiol 277:G478–G486
Faussone-Pellegrini MS, Thuneberg L (1999) Guide to the identification of interstitial cells of Cajal. Microsc Res Tech 47:248–266
Frevert CW, Huang S, Danaee H, Paulauskis JD, Kobzik L (1995) Functional characterization of the rat chemokine KC and its importance in neutrophil recruitment in a rat model of pulmonary inflammation. J Immunol 154:335–344
Galeazzi F, Haapala EM, van Rooijen N, Vallance BA, Collins SM (2000) Inflammation-induced impairment of enteric nerve function in nematode-infected mice is macrophage dependent. Am J Physiol Gastrointest Liver Physiol 278:G259–G265
Galeazzi F, Lovato P, Blennerhassett PA, Haapala EM, Vallance BA, Collins SM (2001) Neural change in Trichinella-infected mice is MHC II independent and involves M-CSF-derived macrophages. Am J Physiol Gastrointest Liver Physiol 281:G151–158
Holschneider AM, Meier-Ruge W, Ure BM (1994) Hirschsprung’s disease and allied disorders—a review. Eur J Pediatr Surg 4:260–266
Hori M, Kita M, Torihashi S, Miyamoto S, Won KJ, Sato K, Ozaki H, Karaki H (2001) Upregulation of iNOS by COX-2 in muscularis resident macrophage of rat intestine stimulated with LPS. Am J Physiol Gastrointest Liver Physiol 280:G930–938
Howard ER, Garrett JR, Kidd A (1984) Constipation and congenital disorders of the myenteric plexus. J R Soc Med 77(Suppl 3):13–19
Huang S, Paulauskis JD, Godleski JJ, Kobzik L (1992) Expression of macrophage inflammatory protein-2 and KC mRNA in pulmonary inflammation. Am J Pathol 141:981–988
Huizinga JD (1998) Neural injury, repair, and adaptation in the GI tract, IV pathophysiology of GI motility related to interstitial cells of Cajal. Am J Physiol 275:G381–G386
Huizinga JD, Thuneberg L, Kluppel M, Malysz J, Mikkelsen HB, Bernstein A (1995) W/kit gene required for interstitial cells of Cajal and for intestinal pacemaker activity. Nature 373:347–349
Kanai T, Watanabe M, Okazawa A, Sato T, Yamazaki M, Okamoto S, Ishii H, Totsuka T, Iiyama R, Okamoto R, Ikeda M, Kurimoto M, Takeda K, Akira S, Hibi T (2001) Macrophage-derived IL-18-mediated intestinal inflammation in the murine model of Crohn’s disease. Gastroenterology 121:875–888
Kinoshita K, Sato K, Hori M, Ozaki H, Karaki H (2003) Decrease in activity of smooth muslce L-type Ca2+-channels and its reversal by NF-κB inhibitors in Crohn’s colitis model. Am J Physiol Gastrointest Liver Physiol 285:G483–G493
Komuro T (1999) Comparative morphology of interstitial cells of Cajal: ultrastructural characterization. Microsc Res Tech 47:267–285
Krishnamurthy S, Schuffler MD, Rohrmann CA, Pope CE 2nd (1985) Severe idiopathic constipation is associated with a distinctive abnormality of the colonic myenteric plexus. Gastroenterology 88:26–34
Linden DR, Couvrette JM, Ciolino A, McQuoid C, Blaszyk H, Sharkey KA, Mawe GM (2005a) Indiscriminate loss of myenteric neurones in the TNBS-inflamed guinea-pig distal colon. Neurogastroenterol Motil 17:751–760
Linden DR, Foley KF, McQuoid C, Simpson J, Sharkey KA, Mawe GM (2005b) Serotonin transporter function and expression are reduced in mice with TNBS-induced colitis. Neurogastroenterol Motil 17:565–574
Lu G, Qian X, Berezin I, Telford GL, Huizinga JD, Sarna SK (1997) Inflammation modulates in vitro colonic myoelectric and contractile activity and interstitial cells of Cajal. Am J Physiol Gastrointest Liver Physiol 273:G1233–G1245
Martinolle JP, Garcia-Villar R, Fioramonti J, Bueno L (1997) Altered contractility of circular and longitudinal muscle in TNBS-inflamed guinea pig ileum. Am J Physiol Gastrointest Liver Physiol 272:G1258–G1267
Mikkelsen HB (1995) Macrophages in the external muscle layers of mammalian intestines. Histol Histopathol 10:719–736
Mikkelsen HB, Garbarsch C, Tranum-Jensen J, Thuneberg L (2004) Macrophages in the small intestinal muscularis externa of embryos, newborn and adult germ-free mice. J Mol Histol 35:377–387
Mikkelsen HB, Thuneberg L, Rumessen JJ, Thorball N (1985) Macrophage-like cells in the muscularis externa of mouse small intestine. Anat Rec 213:77–86
Modi WS, Yoshimura T (1999) Isolation of novel GRO genes and a phylogenetic analysis of the CXC chemokine subfamily in mammals. Mol Biol Evol 16:180–193
Nakagawa H, Komorita N, Shibata F, Ikesue A, Konishi K, Fujioka M, Kato H (1994) Identification of cytokine-induced neutrophil chemoattractants (CINC), rat GRO/CINC-2 a and CINC-2 b, produced by granulation tissue in culture: purification, complete amino acid sequences and characterization. Biochem J 301(Pt 2):545–550
Ozaki H, Kawai T, Shuttleworth CW, Won KJ, Suzuki T, Sato K, Horiguchi H, Hori M, Karaki H, Torihashi S, Ward SM, Sanders KM (2004) Isolation and characterization of resident macrophages from the smooth muscle layers of murine small intestine. Neurogastroenterol Motil 16:39–51
Poli E, Lazzaretti M, Grandi D, Pozzoli C, Coruzzi G (2001) Morphological and functional alterations of the myenteric plexus in rats with TNBS-induced colitis. Neurochem Res 26:1085–1093
Porcher C, Baldo M, Henry M, Orsoni P, Jule Y, Ward SM (2002) Deficiency of interstitial cells of Cajal in the small intestine of patients with Crohn’s disease. Am J Gastroenterol 97:118–125
Publicover NG, Hammond EM, Sanders KM (1993) Amplification of nitric oxide signaling by interstitial cells isolated from canine colon. Proc Nat Acad Sci USA 90:2087–2091
Rao SS, Read NW (1990) Gastrointestinal motility in patients with ulcerative colitis. Scand J Gastroenterol 172:22–28
Reddy SN, Bazzocchi G, Chan S, Akashi K, Villanueva-Meyer J, Yanni G, Mena I, Snape WJ Jr (1991) Colonic motility and transit in health and ulcerative colitis. Gastroenterology 101:1289–1297
Rogers DC, Burnstock G (1966) The interstitial cell and its place in the concept of the autonomic ground plexus. J Comp Neurol 126:255–284
Rollins BJ (1997) Chemokines. Blood 90:909–928
Rumessen JJ (1996) Ultrastructure of interstitial cells of Cajal at the colonic submuscular border in patients with ulcerative colitis. Gastroenterology 111:1447–1455
Rumessen JJ (1994) Identification of interstitial cells of Cajal. Significance for studies of human small intestine and colon. Dan Med Bull 41:275–293
Sanders KM (1996) A case for interstitial cells of Cajal as pacemakers and mediators of neurotransmission in the gastrointestinal tract. Gastroenterology 111:492–515
Sato D, Lai ZF, Tokutomi N, Tokutomi Y, Maeda H, Nishikawa S, Ogawa M, Nishi K (1996) Impairment of Kit-dependent development of interstitial cells alters contractile responses of murine intestinal tract. Am J Physiol Gastrointest Liver Physiol 271:G762–G771
Schroeder RA, delaTorre A, Kuo PC (1997) CD14-dependent mechanism for endotoxin-mediated nitric oxide synthesis in murine macrophages. Am J Physiol Gastrointest Liver Physiol 273:C1030–C1039
Seldenrijk CA, Drexhage HA, Meuwissen SG, Pals ST, Meijer CJ (1989) Dendritic cells and scavenger macrophages in chronic inflammatory bowel disease. Gut 30:486–491
Steinhoff MM, Kodner IJ, DeSchryver-Kecskemeti K (1988) Axonal degeneration/necrosis: a possible ultrastructural marker for Crohn’s disease. Mod Pathol 1:182–187
Suzuki T, Won KJ, Horiguchi K, Kinoshita K, Hori M, Torihashi S, Momotani E, Itoh K, Hirayama K, Ward SM, Sanders KM, Ozaki H (2004) Muscularis inflammation and the loss of interstitial cells of Cajal in the endothelin ETB receptor null rat. Am J Physiol Gastrointest Liver Physiol 287:G638–G646
Szabo V, Feher E (1991) Ultrastructural changes in the nerve elements in Crohn’s disease. Acta Chir Hung 32:25–32
Thuneberg L (1982) Interstitial cells of Cajal: intestinal pacemaker cells? Adv Anat Embryol Cell Biol 71:1–130
Torihashi S, Ozaki H, Hori M, Kita M, Ohota S, Karaki H (2000) Resident macrophages activated by lipopolysaccharide suppress muscle tension and initiate inflammatory response in the gastrointestinal muscle layer. Histochem Cell Biol 113:73–80
Turler A, Schwarz NT, Turler E, Kalff JC, Bauer AJ (2002) MCP-1 causes leukocyte recruitment and subsequently endotoxemic ileus in rat. Am J Physiol Gastrointest Liver Physiol 282:G145–G155
Vanderwinden JM (1999) Role of interstitial cells of Cajal and their relationship with the enteric nervous system. Eur J Morphol 37:250–256
Vermillion DL, Huizinga JD, Riddell RH, Collins SM (1993) Altered small intestinal smooth muscle function in Crohn’s disease. Gastroenterology 104:1692–1699
von Boyen GB, Krammer HJ, Suss A, Dembowski C, Ehrenreich H, Wedel T (2002) Abnormalities of the enteric nervous system in heterozygous endothelin B receptor deficient (spotting lethal) rats resembling intestinal neuronal dysplasia. Gut 51:414–419
Walgenbach KJ, Kalff JC, Sonmez-Alpan E, Hirner A, Bauer AJ (1998) Upregulation of basic fibroblast growth factor during chronic rejection of rat intestinal allografts and its downregulation by FK 506. Transplant Proc 30:2587
Wang XY, Berezin I, Mikkelsen HB, Der T, Bercik P, Collins SM, Huizinga JD, Huizina JD (2002) Pathology of interstitial cells of Cajal in relation to inflammation revealed by ultrastructure but not immunohistochemistry. Am J Pathol 160:1529–1540
Wang XY, Sanders KM, Ward SM (1999) Intimate relationship between interstitial cells of Cajal and enteric nerves in the guinea-pig small intestine. Cell Tissue Res 295:247–256
Ward SM, Beckett EA, Wang X, Baker F, Khoyi M, Sanders KM (2000) Interstitial cells of Cajal mediate cholinergic neurotransmission from enteric motor neurons. J Neurosci 20:1393–1403
Ward SM, Burns AJ, Torihashi S, Sanders KM (1994) Mutation of the proto-oncogene c-kit blocks development of interstitial cells and electrical rhythmicity in murine intestine. J Physiol 480:91–97
Won KJ, Suzuki T, Hori M, Ozaki H (2006) Motility disorder in experimentally obstructed intestine: relationship between muscularis inflammation and disruption of the ICC network. Neurogastroenterol Motil 18:53–61
Won KJ, Torihashi S, Mitsui-Saito M, Hori M, Sato K, Suzuki T, Ozaki H, Karaki H (2002) Increased smooth muscle contractility of intestine in the genetic null of the endothelin ETB receptor: a rat model for long segment Hirschsprung’s disease. Gut 50:355–360
Zagorski J, DeLarco JE (1993) Rat CINC (cytokine-induced neutrophil chemoattractant) is the homolog of the human GRO proteins but is encoded by a single gene. Biochem Biophys Res Commun 190:104–110
Acknowledgments
This work was supported in part by a Grant-in Aid for scientific research from the Japanese Ministry of Education, the Program for the Promotion of Basic Research Activities for Innovative Biosciences (BRAIN), and Morinaga Hoshi-kai.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kinoshita, K., Horiguchi, K., Fujisawa, M. et al. Possible involvement of muscularis resident macrophages in impairment of interstitial cells of Cajal and myenteric nerve systems in rat models of TNBS-induced colitis. Histochem Cell Biol 127, 41–53 (2007). https://doi.org/10.1007/s00418-006-0223-0
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00418-006-0223-0