Advertisement

Journal of Gastroenterology

, Volume 44, Issue 8, pp 834–841 | Cite as

Restoration of gut motility in Kit-deficient mice by bone marrow transplantation

  • Shuji IshiiEmail author
  • Shingo Tsuji
  • Masahiko Tsujii
  • Tsutomu Nishida
  • Kenji Watabe
  • Hideki Iijima
  • Tetsuo Takehara
  • Sunao Kawano
  • Norio Hayashi
Original Article—Alimentary Tract

Abstract

Purpose

Interstitial cells of Cajal (ICC) play important roles in autonomic gut motility as electrical pacemakers and mediators of neural regulation of smooth muscle functions. Insufficiency of ICC has been reported in a wide range of gut dysmotilities. Thus, restoration of ICC may be a therapeutic modality in these diseases. Here we provide evidence that transplanted bone marrow (BM) cells can restore gut dysmotility in part via transdifferentiation to ICC.

Methods

Bone marrow cells obtained from Kit insufficient W/W v mice or syngeneic GFP-transgenic mice with wild-type Kit were transferred to W/W v recipients. Whole gut transit time and gastric emptying were examined 5 and 6 weeks after BM transplantation, respectively, and ICCs were identified in whole mounts, frozen sections and transmission electron immunomicroscopy of the gut smooth muscle layers using specific antibodies.

Results

Transplantation of wild-type BM into W/W v mice significantly improved whole gut transit time and gastric emptying. Fluorescent immunohistochemistry revealed GFP+Kit+ cells in the myenteric plexus, deep muscular plexus, and submucosal plexus smooth muscle layers of the stomach, small intestine, and colon, respectively. In the whole mounts, GFP+Kit+ cells were bipolar and spindle shaped, and transmission electron immunomicroscopy showed GFP+ cells rich in mitochondria and endoplasmic reticulum between gut smooth muscle layers, suggesting the presence of GFP+ cells with morphological characteristics of ICC.

Conclusions

These results suggest that BM contains cells that may incorporate into ICC networks and improve dysmotility in W/W v mice. Thus, BM transplantation may become to a new therapeutic modality for gut dysmotilities due to ICC insufficiency.

Keywords

Bone marrow Green fluorescent protein Interstitial cells of Cajal 

Abbreviations

BM

Bone marrow

ICC

Interstitial cells of Cajal

GFP

Green fluorescent protein

FBS

Fetal bovine serum

PBS

Phosphate buffered saline

AIC

Anti-ICC antibody

DAPI

4′, 6-Diamino-2-phenylindole

Notes

Acknowledgments

The authors thank Professors Shigeko Torihashi and Masaru Okabe for providing the AIC antibody and the GFP-Tg mice, respectively. Shuji Ishii conducted most of the experiments. Shingo Tsuji was the primary investigator who planned the study. Masahiko Tsujii, Tetsuo Takehara, Sunao Kawano, and Norio Hayashi contributed to the design of the study and interpretation of the results. Tsutomu Nishida performed bone marrow transplantation. Hideki Iijima participated in the flow cytometry. Kenji Watabe participated in histopathological analyses.

The study was supported by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (JSPS; no. 18590681).

References

  1. 1.
    Sanders KM, Koh SD, Ward SM. Interstitial cells of Cajal as pacemakers in the gastrointestinal tract. Annu Rev Physiol. 2006;68:307–43.PubMedCrossRefGoogle Scholar
  2. 2.
    McHale N, Hollywood M, Sergeant G, Thornbury K. Origin of spontaneous rhythmicity in smooth muscle. J Physiol. 2006;570:23–8.PubMedCrossRefGoogle Scholar
  3. 3.
    Isozaki K, Hirota S, Miyagawa J, Taniguchi M, Shinomura Y, Matsuzawa Y. Deficiency of c-kit+ cells in patients with a myopathic form of chronic idiopathic intestinal pseudo-obstruction. Am J Gastroenterol. 1997;92:332–4.PubMedGoogle Scholar
  4. 4.
    Vanderwinden JM, Rumessen JJ. Interstitial cells of Cajal in human gut and gastrointestinal disease. Microsc Res Tech. 1999;47:344–60.PubMedCrossRefGoogle Scholar
  5. 5.
    Chen H, Isozaki K, Nakahara M, Kinoshita K, Shinomura Y, Matsuzawa Y, et al. Possible relationship between deficiency of interstitial cells of Cajal and colonic inertia in a patient with colon malfixation. J Gastroenterol. 2002;37:483–4.PubMedCrossRefGoogle Scholar
  6. 6.
    Kiyohara T, Shinomura Y, Isozaki K, Nakahara M, Tsutsui S, Nishibayashi H, et al. A decreased number of c-kit-expressing cells in a patient with afferent loop syndrome. J Gastroenterol. 2003;38:390–4.PubMedCrossRefGoogle Scholar
  7. 7.
    Piotrowska AP, Solari V, Puri P. Distribution of interstitial cells of Cajal in the internal anal sphincter of patients with internal anal sphincter achalasia and Hirschsprung disease. Arch Pathol Lab Med. 2003;127:1192–5.PubMedGoogle Scholar
  8. 8.
    Ordog T, Takayama I, Cheung WK, Ward SM, Sanders KM. Remodeling of networks of interstitial cells of Cajal in a murine model of diabetic gastroparesis. Diabetes. 2000;49:1731–9.PubMedCrossRefGoogle Scholar
  9. 9.
    Nakahara M, Isozaki K, Hirota S, Vanderwinden JM, Takakura R, Kinoshita K, et al. Deficiency of KIT-positive cells in the colon of patients with diabetes mellitus. J Gastroenterol Hepatol. 2002;17:666–70.PubMedCrossRefGoogle Scholar
  10. 10.
    Forster J, Damjanov I, Lin Z, Sarosiek I, Wetzel P, McCallum RW. Absence of the interstitial cells of Cajal in patients with gastroparesis and correlation with clinical findings. J Gastrointest Surg. 2005;9:102–8.PubMedCrossRefGoogle Scholar
  11. 11.
    Chang IY, Glasgow NJ, Takayama I, Horiguchi K, Sanders KM, Ward SM. Loss of interstitial cells of Cajal and development of electrical dysfunction in murine small bowel obstruction. J Physiol. 2001;536:555–68.PubMedCrossRefGoogle Scholar
  12. 12.
    Yanagida H, Yanase H, Sanders KM, Ward SM. Intestinal surgical resection disrupts electrical rhythmicity, neural responses, and interstitial cell networks. Gastroenterology. 2004;127:1748–59.PubMedCrossRefGoogle Scholar
  13. 13.
    Faussone-Pellegrini MS, Gay J, Vannucchi MG, Corsani L, Fioramonti J. Alterations of neurokinin receptors and interstitial cells of Cajal during and after jejunal inflammation induced by Nippostrongylus brasiliensis in the rat. Neurogastroenterol Motil. 2002;14:83–95.PubMedCrossRefGoogle Scholar
  14. 14.
    Young HM, Ciampoli D, Southwell BR, Newgreen DF. Origin of interstitial cells of Cajal in the mouse intestine. Dev Biol. 1996;180:97–107.PubMedCrossRefGoogle Scholar
  15. 15.
    Lorincz A, Redelman D, Horvath VJ, Bardsley MR, Chen H, Ordog T. Progenitors of interstitial cells of cajal in the postnatal murine stomach. Gastroenterology. 2008;134:1083–93.PubMedCrossRefGoogle Scholar
  16. 16.
    Petersen BE, Bowen WC, Patrene KD, Mars WM, Sullivan AK, Murase N, et al. Bone marrow as a potential source of hepatic oval cells. Science. 1999;284:1168–70.PubMedCrossRefGoogle Scholar
  17. 17.
    Ferrari G, Cusella-De Angelis G, Coletta M, Paolucci E, Stornaiuolo A, Cossu G, et al. Muscle regeneration by bone marrow-derived myogenic progenitors. Science. 1998;279:1528–30.PubMedCrossRefGoogle Scholar
  18. 18.
    Zhang S, Jia Z, Ge J, Gong L, Ma Y, Li T, et al. Purified human bone marrow multipotent mesenchymal stem cells regenerate infarcted myocardium in experimental rats. Cell Transplant. 2005;14:787–98.PubMedCrossRefGoogle Scholar
  19. 19.
    Houghton J, Stoicov C, Nomura S, Rogers AB, Carlson J, Li H, et al. Gastric cancer originating from bone marrow-derived cells. Science. 2004;306:1568–71.PubMedCrossRefGoogle Scholar
  20. 20.
    Komori M, Tsuji S, Tsujii M, Murata H, Iijima H, Yasumaru M, et al. Involvement of bone marrow-derived cells in healing of experimental colitis in rats. Wound Repair Regen. 2005;13:109–18.PubMedCrossRefGoogle Scholar
  21. 21.
    Komori M, Tsuji S, Tsujii M, Murata H, Iijima H, Yasumaru M, et al. Efficiency of bone marrow-derived cells in regeneration of the stomach after induction of ethanol-induced ulcers in rats. J Gastroenterol. 2005;40:591–9.PubMedCrossRefGoogle Scholar
  22. 22.
    Torihashi S, Nishi K, Tokutomi Y, Nishi T, Ward S, Sanders KM. Blockade of kit signaling induces transdifferentiation of interstitial cells of cajal to a smooth muscle phenotype. Gastroenterology. 1999;117:140–8.PubMedCrossRefGoogle Scholar
  23. 23.
    Isozaki K, Hirota S, Nakama A, Miyagawa J, Shinomura Y, Xu Z, et al. Disturbed intestinal movement, bile reflux to the stomach, and deficiency of c-kit-expressing cells in Ws/Ws mutant rats. Gastroenterology. 1995;109:456–64.PubMedCrossRefGoogle Scholar
  24. 24.
    Nakama A, Hirota S, Okazaki T, Nagano K, Kawano S, Hori M, et al. Disturbed pyloric motility in Ws/Ws mutant rats due to deficiency of c-kit-expressing interstitial cells of Cajal. Pathol Int. 1998;48:843–9.PubMedCrossRefGoogle Scholar
  25. 25.
    Sanders KM, Ordog T, Ward SM. Physiology and pathophysiology of the interstitial cells of Cajal: from bench to bedside. IV. Genetic and animal models of GI motility disorders caused by loss of interstitial cells of Cajal. Am J Physiol Gastrointest Liver Physiol. 2002;282:G747–56.PubMedGoogle Scholar
  26. 26.
    Der-Silaphet T, Malysz J, Hagel S, Larry Arsenault A, Huizinga JD. Interstitial cells of cajal direct normal propulsive contractile activity in the mouse small intestine. Gastroenterology. 1998;114:724–36.PubMedCrossRefGoogle Scholar
  27. 27.
    Shimada M, Kitamura Y, Yokoyama M, Miyano Y, Maeyama K, Yamatodani A, et al. Spontaneous stomach ulcer in genetically mast-cell depleted W/Wv mice. Nature. 1980;283:662–4.PubMedCrossRefGoogle Scholar
  28. 28.
    Kitamura Y, Yokoyama M, Matsuda H, Shimada M. Coincidental development of forestomach papilloma and prepyloric ulcer in nontreated mutant mice of W/Wv and SI/SId genotypes. Cancer Res. 1980;40:3392–7.PubMedGoogle Scholar
  29. 29.
    Yokoyama M, Tatsuta M, Baba M, Kitamura Y. Bile reflux: a possible cause of stomach ulcer in nontreated mutant mice of W/WV genotype. Gastroenterology. 1982;82:857–63.PubMedGoogle Scholar
  30. 30.
    Yokoyama M, Kitamura Y, Kohrogi T, Miyoshi I. Necessity of bile for and lack of inhibitory effect of retinoid on development of forestomach papillomas in nontreated mutant mice of the W/Wv genotype. Cancer Res. 1982;42:3806–9.PubMedGoogle Scholar
  31. 31.
    Yokoyama M, Tomoi M, Taguchi T, Nakano T, Asai H, Ono T, et al. Fatal antral ulcer in conventionally fed W/Wv mutant mice given indomethacin by injection. Am J Pathol. 1985;119:367–75.PubMedGoogle Scholar
  32. 32.
    Nocka K, Tan JC, Chiu E, Chu TY, Ray P, Traktman P, et al. Molecular bases of dominant negative and loss of function mutations at the murine c-kit/white spotting locus: W37, Wv, W41 and W. Embo J. 1990;9:1805–13.PubMedGoogle Scholar
  33. 33.
    Okabe M, Ikawa M, Kominami K, Nakanishi T, Nishimune Y. ‘Green mice’ as a source of ubiquitous green cells. FEBS Lett. 1997;407:313–9.PubMedCrossRefGoogle Scholar
  34. 34.
    Nagakura Y, Naitoh Y, Kamato T, Yamano M, Miyata K. Compounds possessing 5-HT3 receptor antagonistic activity inhibit intestinal propulsion in mice. Eur J Pharmacol. 1996;311:67–72.PubMedCrossRefGoogle Scholar
  35. 35.
    Mashimo H, Kjellin A, Goyal RK. Gastric stasis in neuronal nitric oxide synthase-deficient knockout mice. Gastroenterology. 2000;119:766–73.PubMedCrossRefGoogle Scholar
  36. 36.
    Ordog T, Redelman D, Horvath VJ, Miller LJ, Horowitz B, Sanders KM. Quantitative analysis by flow cytometry of interstitial cells of Cajal, pacemakers, and mediators of neurotransmission in the gastrointestinal tract. Cytometry A. 2004;62:139–49.PubMedCrossRefGoogle Scholar
  37. 37.
    Torihashi S, Yokoi K, Nagaya H, Aoki K, Fujimoto T. New monoclonal antibody (AIC) identifies interstitial cells of Cajal in the musculature of the mouse gastrointestinal tract. Auton Neurosci. 2004;113:16–23.PubMedCrossRefGoogle Scholar
  38. 38.
    Vassilopoulos G, Wang PR, Russell DW. Transplanted bone marrow regenerates liver by cell fusion. Nature. 2003;422:901–4.PubMedCrossRefGoogle Scholar
  39. 39.
    Nakagawa T, Misawa H, Nakajima Y, Takaki M. Absence of peristalsis in the ileum of W/W(V) mutant mice that are selectively deficient in myenteric interstitial cells of Cajal. J Smooth Muscle Res. 2005;41:141–51.PubMedCrossRefGoogle Scholar
  40. 40.
    Maeda H, Yamagata A, Nishikawa S, Yoshinaga K, Kobayashi S, Nishi K. Requirement of c-kit for development of intestinal pacemaker system. Development. 1992;116:369–75.PubMedGoogle Scholar
  41. 41.
    Beckett EA, Ro S, Bayguinov Y, Sanders KM, Ward SM. Kit signaling is essential for development and maintenance of interstitial cells of Cajal and electrical rhythmicity in the embryonic gastrointestinal tract. Dev Dyn. 2007;236:60–72.PubMedCrossRefGoogle Scholar
  42. 42.
    Takaki M, Misawa H, Shimizu J, Kuniyasu H, Horiguchi K. Inhibition of gut pacemaker cell formation from mouse ES cells by the c-kit inhibitor. Biochem Biophys Res Commun. 2007;359:354–9.PubMedCrossRefGoogle Scholar
  43. 43.
    Sanders KM. Interstitial cells of Cajal at the clinical and scientific interface. J Physiol. 2006;576:683–7.PubMedCrossRefGoogle Scholar
  44. 44.
    Kluppel M, Huizinga JD, Malysz J, Bernstein A. Developmental origin and Kit-dependent development of the interstitial cells of cajal in the mammalian small intestine. Dev Dyn. 1998;211:60–71.PubMedCrossRefGoogle Scholar

Copyright information

© Springer 2009

Authors and Affiliations

  • Shuji Ishii
    • 1
    Email author
  • Shingo Tsuji
    • 1
  • Masahiko Tsujii
    • 1
  • Tsutomu Nishida
    • 1
  • Kenji Watabe
    • 1
  • Hideki Iijima
    • 1
  • Tetsuo Takehara
    • 1
  • Sunao Kawano
    • 2
    • 3
  • Norio Hayashi
    • 1
  1. 1.Department of Gastroenterology and Hepatology (K1)Osaka University Graduate School of MedicineSuitaJapan
  2. 2.Department of Clinical Laboratory ScienceOsaka University Graduate School of MedicineSuitaJapan
  3. 3.Rinku General Medical CenterIzumisano HospitalIzumisanoJapan

Personalised recommendations