TRPM2 contributes to antigen-stimulated Ca2+ influx in mucosal mast cells

  • Satoshi Oda
  • Kunitoshi Uchida
  • Xiaoyu Wang
  • Jaemin Lee
  • Yutaka Shimada
  • Makoto Tominaga
  • Makoto KadowakiEmail author
Molecular and Cellular Mechanisms of Disease


Food allergy (FA) is a common allergic disease without any currently available effective drug therapies. Mucosal mast cells (MMCs) play a particularly important role in FA, and the increase in their cytosolic Ca2+ concentration ([Ca2+]cyt) is considered to be a principal component of the degranulation process. However, the mechanisms governing Ca2+ influx remain poorly understood in MMCs. Recent reports have highlighted the functions of the transient receptor potential melastatin 2 (TRPM2) channel in immunocytes, including its role in monocyte chemokine production and macrophage phagocytic activity. Although TRPM2 gene expression has been demonstrated in mast cells, the significance of such expression remains virtually unknown. In this study, we found that antigen-stimulated degranulation was significantly reduced in mucosal-type bone marrow-derived mast cells (mBMMCs) prepared from TRPM2-knockout (TRPM2-KO) mice (TRPM2-KO mBMMCs) and was suppressed following the administration of three TRPM2 inhibitors with different chemical structures, including econazole, flufenamic acid (FFA), and 2-aminoethoxydiphenyl borate. Furthermore, the antigen-stimulated increase in [Ca2+]cyt was significantly decreased in TRPM2-KO mBMMCs and was also suppressed by the TRPM2 inhibitors econazole and FFA. In addition, thapsigargin-induced increase in [Ca2+]cyt was significantly decreased in TRPM2-KO mBMMCs. These results suggest that TRPM2 may participate in antigen-induced extracellular Ca2+ influx and subsequent degranulation. In addition, TRPM2 inhibitors were shown to improve food allergic reactions in a mouse model. Together, these results suggest that TRPM2 inhibitors suppress MMC degranulation via regulation of the increase in [Ca2+]cyt. Thus, TRPM2 may play a key role in degranulation by modulating intracellular Ca2+ in MMCs.


Mucosal mast cell mBMMC TRPM2 Food allergy Degranulation 



We thank H. Mihara for help with cell preparation and operation of measuring instruments, M. Kashio for animal breeding and management, and K. Tsuneyama and M. Fujimoto for help with immunohistochemical staining. This research was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan to M. Kadowaki (nos. 21590760 and 24590879) and by the Knowledge Cluster Initiative Program of the Ministry of Education, Culture, Sports, Science and Technology of Japan to M. Kadowaki.

Conflict of interest

The authors have declared that no conflict of interest exists.


  1. 1.
    Bradding P, Okayama Y, Kambe N, Saito H (2003) Ion channel gene expression in human lung, skin, and cord blood-derived mast cells. J Leukoc Biol 73:614–620PubMedCrossRefGoogle Scholar
  2. 2.
    Brandt EB (2003) Mast cells are required for experimental oral allergen-induced diarrhea. J Clin Invest 112:1666–1677PubMedGoogle Scholar
  3. 3.
    Carter RN, Tolhurst G, Walmsley G, Vizuete-Forster M, Miller N, Mahaut-Smith MP (2006) Molecular and electrophysiological characterization of transient receptor potential ion channels in the primary murine megakaryocyte. J Physiol (Lond) 576:151–162CrossRefGoogle Scholar
  4. 4.
    Gilfillan AM, Tkaczyk C (2006) Integrated signalling pathways for mast-cell activation. Nat Rev Immunol 6:218–230PubMedCrossRefGoogle Scholar
  5. 5.
    Hayama K, Suzuki Y, Inoue T, Ochiai T, Terui T, Ra C (2011) Gold activates mast cells via calcium influx through multiple H2O2-sensitive pathways including L-type calcium channels. Free Radic Biol Med 50:1417–1428PubMedCrossRefGoogle Scholar
  6. 6.
    Heiner I, Eisfeld J, Lückhoff A (2003) Role and regulation of TRP channels in neutrophil granulocytes. Cell Calcium 33:533–540PubMedCrossRefGoogle Scholar
  7. 7.
    Hill K, Benham CD, McNulty S, Randall AD (2004) Flufenamic acid is a pH-dependent antagonist of TRPM2 channels. Neuropharmacology 47:450–460PubMedCrossRefGoogle Scholar
  8. 8.
    Hill K, McNulty S, Randall AD (2004) Inhibition of TRPM2 channels by the antifungal agents clotrimazole and econazole. Naunyn Schmiedebergs Arch Pharmacol 370:227–237PubMedCrossRefGoogle Scholar
  9. 9.
    Inada H, Iida T, Tominaga M (2006) Different expression patterns of TRP genes in murine B and T lymphocytes. Biochem Biophys Res Commun 350:762–767PubMedCrossRefGoogle Scholar
  10. 10.
    Inoue T, Suzuki Y, Yoshimaru T, Ra C (2008) Reactive oxygen species produced up- or downstream of calcium influx regulate proinflammatory mediator release from mast cells: role of NADPH oxidase and mitochondria. Acta Bioenerg 1783:789–802CrossRefGoogle Scholar
  11. 11.
    Ishii M, Shimizu S, Hara Y, Hagiwara T, Miyazaki A, Mori Y, Kiuchi Y (2006) Intracellular-produced hydroxyl radical mediates H2O2-induced Ca2+ influx and cell death in rat beta-cell line RIN-5F. Cell Calcium 39:487–494PubMedCrossRefGoogle Scholar
  12. 12.
    Kageyama-Yahara N, Suehiro Y, Maeda F, Kageyama S-I, Fukuoka J, Katagiri T, Yamamoto T, and Kadowaki M (2010) Pentagalloylglucose down-regulates mast cell surface FcεRI expression in vitro and in vivo. FEBS Letters 584:111–118Google Scholar
  13. 13.
    Kageyama-Yahara N, Suehiro Y, Yamamoto T, Kadowaki M (2008) IgE-induced degranulation of mucosal mast cells is negatively regulated via nicotinic acetylcholine receptors. Biochem Biophys Res Commun 377:321–325PubMedCrossRefGoogle Scholar
  14. 14.
    Kageyama-Yahara N, Wang X, Katagiri T, Wang P, Yamamoto T, Tominaga M, Kadowaki M (2011) Suppression of phospholipase Cγ1 phosphorylation by cinnamaldehyde inhibits antigen-induced extracellular calcium influx and degranulation in mucosal mast cells. Biochem Biophys Res Commun 416:283–288PubMedCrossRefGoogle Scholar
  15. 15.
    Kashio M, Sokabe T, Shintaku K, Uematsu T, Fukuta N, Kobayashi N, Mori Y, Tominaga M (2012) Redox signal-mediated sensitization of transient receptor potential melastatin 2 (TRPM2) to temperature affects macrophage functions. Proc Natl Acad Sci USA 109:6745–6750PubMedCrossRefGoogle Scholar
  16. 16.
    Lange I, Penner R, Fleig A, Beck A (2008) Synergistic regulation of endogenous TRPM2 channels by adenine dinucleotides in primary human neutrophils. Cell Calcium 44:604–615PubMedCrossRefGoogle Scholar
  17. 17.
    Lange I, Yamamoto S, Partida-Sanchez S, Mori Y, Fleig A, Penner R (2008) TRPM2 functions as a lysosomal Ca2+-release channel in beta cells. Sci Signal 2:ra23–ra23CrossRefGoogle Scholar
  18. 18.
    McClain S, Bannon G (2006) Animal models of food allergy: opportunities and barriers. Curr Allergy Asthma Rep 6:141–144PubMedCrossRefGoogle Scholar
  19. 19.
    Metcalfe DD, Baram D, Mekori YA (1997) Mast cells. Physiol Rev 77:1033–1079PubMedGoogle Scholar
  20. 20.
    Morita T, Tanimura A, Baba Y, Kurosaki T, Tojyo Y (2009) A Stim1-dependent, noncapacitative Ca2+-entry pathway is activated by B-cell-receptor stimulation and depletion of Ca2+. J Cell Sci 122:1220–1228PubMedCrossRefGoogle Scholar
  21. 21.
    Perraud AL, Fleig A, Dunn CA, Bagley LA, Launay P, Schmitz C, Stokes AJ et al (2001) ADP-ribose gating of the calcium-permeable LTRPC2 channel revealed by Nudix motif homology. Nature 411:595–599PubMedCrossRefGoogle Scholar
  22. 22.
    Prasad P, Yanagihara AA, Small-Howard AL, Turner H, Stokes AJ (2008) Secretogranin III directs secretory vesicle biogenesis in mast cells in a manner dependent upon interaction with chromogranin A. J Immunol 181:5024–5034PubMedGoogle Scholar
  23. 23.
    Rivera J, Gilfillan AM (2006) Molecular regulation of mast cell activation. J Allergy Clin Immun 117:1214–1226PubMedCrossRefGoogle Scholar
  24. 24.
    Sicherer SH, Sampson HA (2010) Food allergy. J Allergy Clin Immun 125:S116–S116PubMedCrossRefGoogle Scholar
  25. 25.
    Starkus J, Beck A, Fleig A, Penner R (2007) Regulation of TRPM2 by extra- and intracellular calcium. J Gen Physiol 130:427–440PubMedCrossRefGoogle Scholar
  26. 26.
    Sumoza-Toledo A, Penner R (2011) TRPM2: a multifunctional ion channel for calcium signalling. J Physiol (Lond) 589:1515–1525CrossRefGoogle Scholar
  27. 27.
    Swindle EJ, Metcalfe DD (2007) The role of reactive oxygen species and nitric oxide in mast cell-dependent inflammatory processes. Immunol Rev 217:186–205PubMedCrossRefGoogle Scholar
  28. 28.
    Takahashi N, Kozai D, Kobayashi R, Ebert M, Mori Y (2011) Roles of TRPM2 in oxidative stress. Cell Calcium 50:279–287PubMedCrossRefGoogle Scholar
  29. 29.
    Tegoshi T, Nishida M, Arizono N (2005) Expression and role of E-cadherin and CD103β7 (αEβ7 integrin) on cultured mucosal-type mast cells. APMIS 113:91–98Google Scholar
  30. 30.
    Togashi K, Inada H, Tominaga M (2009) Inhibition of the transient receptor potential cation channel TRPM2 by 2-aminoethoxydiphenyl borate (2-APB). Br J Pharmacol 153:1324–1330CrossRefGoogle Scholar
  31. 31.
    Togashi K, Hara Y, Tominaga T, Higashi T, Konishi Y, Mori Y, Tominaga M (2006) TRPM2 activation by cyclic ADP-ribose at body temperature is involved in insulin secretion. EMBO J 25:1804–1815PubMedCrossRefGoogle Scholar
  32. 32.
    Uchida K, Dezaki K, Damdindorj B, Inada H, Shiuchi T, Mori Y, Yada T, Minokoshi Y, Tominaga M (2011) Lack of TRPM2 impaired insulin secretion and glucose metabolisms in mice. Diabetes 60:119–126PubMedCrossRefGoogle Scholar
  33. 33.
    Vennekens R, Olausson J, Meissner M, Bloch W, Mathar I, Philipp SE, Schmitz F et al (2007) Increased IgE-dependent mast cell activation and anaphylactic responses in mice lacking the calcium-activated nonselective cation channel TRPM4. Nat Immunol 8:312–320PubMedCrossRefGoogle Scholar
  34. 34.
    Wang X, Kageyama-Yahara N, Hayashi S, Yamamoto T, Kadowaki M (2012) Sphingosine kinase-1-dependent and -independent inhibitory effects of zanthoxyli fructus to attenuate the activation of mucosal mast cells and ameliorate food allergies in mice. Evid Based Complement Alternat Med 2012:862743PubMedGoogle Scholar
  35. 35.
    Westerberg CM, Ullerås E, Nilsson G (2012) Differentiation of mast cell subpopulations from mouse embryonic stem cells. J Immunol Methods 382:160–166PubMedCrossRefGoogle Scholar
  36. 36.
    Wissenbach U, Philipp SE, Gross SA, Cavalié A, Flockerzi V (2006) Primary structure, chromosomal localization and expression in immune cells of the murine ORAI and STIM genes. Cell Calcium 42:439–446CrossRefGoogle Scholar
  37. 37.
    Yamamoto S, Shimizu S, Kiyonaka S, Takahashi N, Wajima T, Hara Y, Negoro T et al (2008) TRPM2-mediated Ca2+ influx induces chemokine production in monocytes that aggravates inflammatory neutrophil infiltration. Nat Med 14:738–747PubMedCrossRefGoogle Scholar
  38. 38.
    Yamamoto T, Fujiwara K, Yoshida M, Kageyama-Yahara N, Kuramoto H, Shibahara N, Kadowaki M (2009) Therapeutic effect of Kakkonto in a mouse model of food allergy with gastrointestinal symptoms. Int Arch Allergy Immunol 148:175–185PubMedCrossRefGoogle Scholar
  39. 39.
    Zhang D, Spielmann A, Wang L, Ding G, Huang F, Gu Q, Schwarz W (2012) Mast-cell degranulation induced by physical stimuli involves the activation of transient-receptor-potential channel TRPV2. Physiol Res 61:113–124PubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Satoshi Oda
    • 1
    • 2
  • Kunitoshi Uchida
    • 3
  • Xiaoyu Wang
    • 1
  • Jaemin Lee
    • 1
  • Yutaka Shimada
    • 2
  • Makoto Tominaga
    • 3
  • Makoto Kadowaki
    • 1
    Email author
  1. 1.Division of Gastrointestinal Pathophysiology, Institute of Natural MedicineUniversity of ToyamaToyamaJapan
  2. 2.Department of Japanese Oriental Medicine, Faculty of MedicineUniversity of ToyamaToyamaJapan
  3. 3.Division of Cell Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological Sciences)National Institutes of Natural SciencesOkazakiJapan

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