Igf1r+/CD34+ immature ICC are putative adult progenitor cells, identified ultrastructurally as fibroblast-like ICC in Ws/Ws rat colon
X.Y. Wang
Farncombe Family Digestive Health Research Institute, Department of Medicine, McMaster University, Hamilton, Ontario, Canada
Search for more papers by this authorE. Albertí
Department of Cell Biology, Physiology and Immunology, Universitat Autònoma de Barcelona, Barcelona, Spain
Centro de Investigación Biomédica en Red de enfermedades hepáticas y Digestivas (CIBERehd)
Search for more papers by this authorE.J. White
Farncombe Family Digestive Health Research Institute, Department of Medicine, McMaster University, Hamilton, Ontario, Canada
Search for more papers by this authorH.B. Mikkelsen
Department of Cellular and Molecular Medicine, University of Copenhagen, The Panum Institute, Copenhagen, Denmark
Search for more papers by this authorJ.O. Larsen
Department of Neuroscience and Pharmacology, University of Copenhagen, The Panum Institute, Copenhagen, Denmark
Search for more papers by this authorM. Jiménez
Department of Cell Biology, Physiology and Immunology, Universitat Autònoma de Barcelona, Barcelona, Spain
Search for more papers by this authorCorresponding Author
J.D. Huizinga
Farncombe Family Digestive Health Research Institute, Department of Medicine, McMaster University, Hamilton, Ontario, Canada
Correspondence to: Dr J.D. HUIZINGA, Department of Medicine, Farncombe Family Digestive Health Research Institute, McMaster University, Hamilton, L8N3Z5, Ontario, Canada.Tel.: 905-525-9140 ext. 22590Fax: 905 522 3454E-mail: [email protected]Search for more papers by this authorX.Y. Wang
Farncombe Family Digestive Health Research Institute, Department of Medicine, McMaster University, Hamilton, Ontario, Canada
Search for more papers by this authorE. Albertí
Department of Cell Biology, Physiology and Immunology, Universitat Autònoma de Barcelona, Barcelona, Spain
Centro de Investigación Biomédica en Red de enfermedades hepáticas y Digestivas (CIBERehd)
Search for more papers by this authorE.J. White
Farncombe Family Digestive Health Research Institute, Department of Medicine, McMaster University, Hamilton, Ontario, Canada
Search for more papers by this authorH.B. Mikkelsen
Department of Cellular and Molecular Medicine, University of Copenhagen, The Panum Institute, Copenhagen, Denmark
Search for more papers by this authorJ.O. Larsen
Department of Neuroscience and Pharmacology, University of Copenhagen, The Panum Institute, Copenhagen, Denmark
Search for more papers by this authorM. Jiménez
Department of Cell Biology, Physiology and Immunology, Universitat Autònoma de Barcelona, Barcelona, Spain
Search for more papers by this authorCorresponding Author
J.D. Huizinga
Farncombe Family Digestive Health Research Institute, Department of Medicine, McMaster University, Hamilton, Ontario, Canada
Correspondence to: Dr J.D. HUIZINGA, Department of Medicine, Farncombe Family Digestive Health Research Institute, McMaster University, Hamilton, L8N3Z5, Ontario, Canada.Tel.: 905-525-9140 ext. 22590Fax: 905 522 3454E-mail: [email protected]Search for more papers by this authorAbstract
The colon of Ws/Ws mutant rats shows impairment of pacemaker activity and altered inhibitory neurotransmission. The present study set out to find structural correlates to these findings to resolve mechanisms. In the colon of Ws/Ws rats, interstitial cells of Cajal associated with Auerbach’s plexus (ICC-AP) were significantly decreased and ICC located at the submuscular plexus and intramuscular ICC were rarely observed based on immunohistochemistry and electron microscopy. Ultrastructural investigations revealed that there was no overall loss of all types of interstitial cells combined. Where loss of ICC was observed, a marked increase in fibroblast-like ICC (FL-ICC) was found at the level of AP. Immunoelectron microscopy proved FL-ICC to be c-Kit– but gap junction coupled to each other and to c-Kit+ ICC; they were associated with enteric nerves and occupied space normally occupied by ICC in the wild-type rat colon, suggesting them to be immature ICC. In addition, a marked increase in immunoreactivity for insulin-like growth factor 1 receptor (Igf1r) occurred, co-localized with CD34 but not with c-Kit. A significantly higher number of Igf1r+/CD34+ cells were found in Ws/Ws compared to wild-type rat colons. These CD34+/Igf1r+ cells in the Ws/Ws colon occupied the same space as FL-ICC. Hence we propose that a subset of immature ICC (FL-ICC) consists of adult progenitor cells. Immunohistochemistry revealed a reduction of neurons positive for neuronal nitric oxide synthase. The functional capabilities of the immature ICC and the regenerative capabilities of the adult progenitor cells need further study. The morphological features described here show that the loss of pacemaker activity is not associated with failure to develop a network of interstitial cells around AP but a failure to develop this network into fully functional pacemaker cells. The reduction in nitrergic innervation associated with the Ws mutation may be the result of a reduction in nitrergic neurons.
References
- 1 Huizinga JD, Thuneberg L, Klüppel M, et al . The W/kit gene required for interstitial cells of Cajal and for intestinal pacemaker activity. Nature. 1995; 373: 347–9.
- 2 Ward SM, Burns AJ, Torihashi S, et al . Mutation of the proto-oncogene c-kit blocks development of interstitial cells and electrical rhythmicity in murine intestine. J Physiol. 1994; 480: 91–7.
- 3 Koh SD, Sanders KM, Ward SM. Spontaneous electrical rhythmicity in cultured interstitial cells of cajal from the murine small intestine. J Physiol. 1998; 513: 203–13.
- 4 Thomsen L, Robinson TL, Lee JCF, et al . Interstitial cells of Cajal generate a rhythmic pacemaker current. Nat Med. 1998; 4: 848–51.
- 5 Der-Silaphet T, Malysz J, Arsenault AL, et al . Interstitial cells of Cajal direct normal propulsive contractile activity in the small intestine. Gastroenterology. 1998; 114: 724–36.
- 6 Huizinga JD, Ambrous K, Der-Silaphet TD. Co-operation between neural and myogenic mechanisms in the control of distension-induced peristalsis in the mouse small intestine. J Physiol. 1998; 506: 843–56.
- 7 Boddy G, Willis A, Galante G, et al . Sodium-, chloride-, and mibefradil-sensitive calcium channels in intestinal pacing in wild-type and W/WV mice. Can J Physiol Pharmacol. 2006; 84: 589–99.
- 8 Liu LWC, Thuneberg L, Huizinga JD. Cyclopiazonic acid, inhibiting the endoplasmic reticulum calcium pump, reduces the canine colon pacemaker frequency. J Pharmacol Exp Ther. 1995; 275: 1058–68.
- 9 Liu LWC, Huizinga JD. Electrical coupling of circular muscle to longitudinal muscle and interstitial cells of Cajal in canine colon. J Physiol. 1993; 470: 445–61.
- 10 Langton P, Ward SM, Carl A, et al . Spontaneous electrical activity of interstitial cells of Cajal isolated from canine proximal colon. Proc Natl Acad Sci USA. 1989; 86: 7280–4.
- 11 Huizinga JD. Motor physiology of the human colon. In: M Kamm, J Lennard-Jones, editors. Gastrointestinal transit: pathophysiology and pharmacology. Wrightson : Biomedical Publishing Ltd.; 1991. pp. 65–75.
- 12 Alberti E, Mikkelsen HB, Wang XY, et al . Pacemaker activity and inhibitory neurotransmission in the colon of Ws/Ws mutant rats. Am J Physiol Gastrointest Liver Physiol. 2007; 292: G1499–510.
- 13 Pluja L, Alberti E, Fernandez E, et al . Evidence supporting presence of two pacemakers in rat colon. Am J Physiol Gastrointest Liver Physiol. 2001; 281: G255–66.
- 14 Burns AJ, Lomax AE, Torihashi S, et al . Interstitial cells of Cajal mediate inhibitory neurotransmission in the stomach. Proc Natl Acad Sci USA. 1996; 93: 12008–13.
- 15 Ward SM. Interstitial cells of Cajal in enteric neurotransmission. Gut. 2000; 47: iv40–3.
- 16 Suzuki H, Ward SM, Bayguinov YR, et al . Involvement of intramuscular interstitial cells in nitrergic inhibition in the mouse gastric antrum. J Physiol. 2003; 546: 751–63.
- 17 Ward SM, Sanders KM, Hirst GD. Role of interstitial cells of Cajal in neural control of gastrointestinal smooth muscles. Neurogastroenterol Motil. 2004; 16: 112–7.
- 18 De Lorijn F, De Jonge WJ, Wedel T, et al . Interstitial cells of Cajal are involved in the afferent limb of the rectoanal inhibitory reflex. Gut. 2005; 54: 1107–13.
- 19 Huizinga JD, Liu LW, Fitzpatrick A, et al . Deficiency of intramuscular ICC increases fundic muscle excitability but does not impede nitrergic innervation. Am J Physiol Gastrointest Liver Physiol. 2008; 294: G589–94.
- 20 Faussone-Pellegrini MS. Cytodifferentiation of the interstitial cells of Cajal related to the myenteric plexus of mouse intestinal muscle coat. An E.M. study from foetal to adult life. Anat Embryol. 1985; 171: 163–9.
- 21 Lorincz A, Redelman D, Horvath VJ, et al . Progenitors of interstitial cells of cajal in the postnatal murine stomach. Gastroenterology. 2008; 134: 1083–93.
- 22 Ishikawa K, Komuro T, Hirota S, et al . Ultrastructural identification of the c-kit-expressing interstitial cells in the rat stomach: a comparison of control and Ws/Ws mutant rats. Cell Tissue Res. 1997; 289: 137–43.
- 23 Alberti E, Mikkelsen HB, Larsen JO, et al . Motility patterns and distribution of interstitial cells of Cajal and nitrergic neurons in the proximal, mid- and distal-colon of the rat. Neurogastroenterol Motil. 2005; 17: 133–47.
- 24 Toth C, Brussee V, Martinez JA, et al . Rescue and regeneration of injured peripheral nerve axons by intrathecal insulin. Neuroscience. 2006; 139: 429–49.
- 25 Gundersen HJ. Stereology of arbitrary particles. A review of unbiased number and size estimators and the presentation of some new ones, in memory of William R. Thompson. J Microsc. 1986; 143: 3–45.
- 26 Larsen J.O. Qualitative and quantitative assessment of peripheral nerves with implanted nerve cuff electrodes. Faculty of Health Sciences, Denmark : University of Aarhus ; 1998.
- 27 Mayhew TM. An efficient sampling scheme for estimating fibre number from nerve cross sections: the fractionator. J Anat. 1988; 157: 127–34.
- 28 Horiguchi K, Komuro T. Ultrastructural characterization of interstitial cells of Cajal in the rat small intestine using control and Ws/Ws mutant rats. Cell Tissue Res. 1998; 293: 277–84.
- 29 Mitsui R, Komuro T. Distribution and ultrastructure of interstitial cells of Cajal in the gastric antrum of wild-type and Ws/Ws rats. Anat Embryol. 2003; 206: 453–60.
- 30 Horiguchi K, Komuro T. Ultrastructural observations of fibroblast-like cells forming gap junctions in the W/W(nu) mouse small intestine. J Auton Nerv Syst. 2000; 80: 142–7.
- 31 Fujita A, Takeuchi T, Jun H, et al . Localization of Ca2+-activated K+ channel, SK3, in fibroblast-like cells forming gap junctions with smooth muscle cells in the mouse small intestine. J Pharmacol Sci. 2003; 92: 35–42.
- 32 Vanderwinden JM, Rumessen JJ, de Kerchove dA Jr, et al . Kit-negative fibroblast-like cells expressing SK3, a Ca2+-activated K+ channel, in the gut musculature in health and disease. Cell Tissue Res. 2002; 310: 349–58.
- 33 Daniel EE, El-Yazbi A, Cho WJ. Caveolae and calcium handling, a review and a hypothesis. J Cell Mol Med. 2006; 10: 529–44.
- 34 Zarate N. Interstitial cells of Cajal: key players in gastrointestinal motor disorders. Barcelona : Autonomous University of Barcelona ; 2008.
- 35 Long JM, Mouton PR, Jucker M, et al . What counts in brain aging? Design-based stereological analysis of cell number. J Gerontol A Biol Sci Med Sci. 1999; 54: B407–17.
- 36 Young HM, Ciampoli D, Southwell BR, et al . Origin of interstitial cells of Cajal in the mouse intestine. Dev Biol. 1996; 96: 97–107.
- 37 Fox EA, Phillips RJ, Martinson FA, et al . C-Kit mutant mice have a selective loss of vagal intramuscular mechanoreceptors in the forestomach. Anat Embryol. 2001; 204: 11–26.
- 38 Powley TL, Wang XY, Fox EA, et al . Ultrastructural evidence for communication between intramuscular vagal mechanoreceptors and interstitial cells of Cajal in the rat fundus. Neurogastroenterol Motil. 2008; 20: 69–79.
- 39 Maurer M, Theoharides T, Granstein RD, et al . What is the physiological function of mast cells Exp Dermatol. 2003; 12: 886–910.
- 40
Klüppel M,
Huizinga JD,
Malysz J,
et al
.
Developmental origin and Kit-dependent development of the interstitial cells of Cajal in the mammalian small intestine.
Dev Dyn.
1998; 211: 60–71.
10.1002/(SICI)1097-0177(199801)211:1<60::AID-AJA6>3.0.CO;2-5 CAS PubMed Web of Science® Google Scholar
- 41 Ward SM, Sanders KM. Physiology and pathophysiology of the interstitial cell of Cajal: from bench to bedside. I. Functional development and plasticity of interstitial cells of Cajal networks. Am J Physiol Gastrointest Liver Physiol. 2001; 281: G602–11.
- 42
Liu LWC,
Thuneberg L,
Huizinga JD.
Development of pacemaker activity and interstitial cells of Cajal in the neonatal mouse small intestine.
Dev Dyn. 1998; 213: 271–82.
10.1002/(SICI)1097-0177(199811)213:3<271::AID-AJA4>3.0.CO;2-R CAS PubMed Web of Science® Google Scholar
- 43 Torihashi S, Ward SM, Nishikawa S, et al . c-kit-dependent development of interstitial cells and electrical activity in the murine gastrointestinal tract. Cell Tissue Res. 1995; 280: 97–111.
- 44 Bernex F, De SP, Kress C, et al . Spatial and temporal patterns of c-kit-expressing cells in WlacZ/+ and WlacZ/WlacZ mouse embryos. Development. 1996; 122: 3023–33.
- 45
Thuneberg L,
Peters S.
Toward a concept of stretch-coupling in smooth muscle. I. Anatomy of intestinal segmentation and sleeve contractions.
Anat Rec.
2001; 262: 110–24.
10.1002/1097-0185(20010101)262:1<110::AID-AR1016>3.0.CO;2-0 CAS PubMed Web of Science® Google Scholar
- 46 Daniel EE, Willis A, Cho WJ, et al . Comparisons of neural and pacing activities in intestinal segments from W/W++ and W/W(V) mice. Neurogastroenterol Motil. 2005; 17: 355–65.
- 47 Horvath VJ, Vittal H, Lorincz A, et al . Reduced stem cell factor links smooth myopathy and loss of interstitial cells of Cajal in murine diabetic gastroparesis. Gastroenterology. 2006; 130: 759–70.
- 48 Vanderwinden JM, Rumessen JJ, De Laet MH, et al . CD34+ cells in human intestine are fibroblasts adjacent to, but distinct from, interstitial cells of Cajal. Lab Invest. 1999; 79: 59–65.
- 49 Ordog T. Interstitial cells of Cajal in diabetic gastroenteropathy. Neurogastroenterol Motil. 2008; 20: 8–18.
- 50 Huizinga JD, White EJ. Progenitor cells of interstitial cells of Cajal: on the road to tissue repair. Gastroenterology. 2008; 134: 1252–4.
- 51 Wang XY, Zarate N, Soderholm JD, et al . Ultrastructural injury to interstitial cells of Cajal and communication with mast cells in Crohn’s disease. Neurogastroenterol Motil. 2007; 19: 349–64.
- 52 Boeckxstaens GE, Rumessen JJ, de Wit L, et al . Abnormal distribution of the interstitial cells of cajal in an adult patient with pseudo-obstruction and megaduodenum. Am J Gastroenterol. 2002; 97: 2120–6.
- 53 Rumessen JJ, Vanderwinden JM. Interstitial cells in the musculature of the gastrointestinal tract: Cajal and beyond. Int Rev Cytol. 2003; 229: 115–208.
- 54 Der T, Bercik P, Donnelly G, et al . Interstitial cells of Cajal and inflammation-induced motor dysfunction in the mouse small intestine. Gastroenterology. 2000; 119: 1590–9.
- 55 Vanderwinden JM, Liu H, Menu R, et al . The pathology of infantile hypertrophic pyloric stenosis after healing. J Pediatr Surg. 1996; 31: 1530–4.
Citing Literature
