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Increased IgE-dependent mast cell activation and anaphylactic responses in mice lacking the calcium-activated nonselective cation channel TRPM4

Nature Immunology volume 8, pages 312–320 (2007)Cite this article

Abstract

Mast cells are key effector cells in allergic reactions. Aggregation of the receptor FcεRI in mast cells triggers the influx of calcium (Ca2+) and the release of inflammatory mediators. Here we show that transient receptor potential TRPM4 proteins acted as calcium-activated nonselective cation channels and critically determined the driving force for Ca2+ influx in mast cells. Trpm4−/− bone marrow–derived mast cells had more Ca2+ entry than did TRPM4+/+ cells after FcεRI stimulation. Consequently, Trpm4−/− bone marrow–derived mast cells had augmented degranulation and released more histamine, leukotrienes and tumor necrosis factor. Trpm4−/− mice had a more severe IgE-mediated acute passive cutaneous anaphylactic response, whereas late-phase passive cutaneous anaphylaxis was not affected. Our results establish the physiological function of TRPM4 channels as critical regulators of Ca2+ entry in mast cells.

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Figure 1: BMMCs from Trpm4+/+ and Trpm4−/− mice.
Figure 2: A CAN channel in Trpm4+/+ but not Trpm4−/− BMMCs.
Figure 3: Ca2+ signaling in Trpm4+/+ and Trpm4−/− BMMCs.
Figure 4: Simultaneous measurement of membrane potential and [Ca2+]cyt in Trpm4+/+ and Trpm4−/− BMMCs.
Figure 5: Release of mediators from Trpm4+/+ and Trpm4−/− BMMCs.
Figure 6: PCA in Trpm4+/+ and Trpm4−/− mice.

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Acknowledgements

We thank S. Collins, R. Ramracheya and P. Rorsman for measuring insulin release from pancreatic islets; M. Thomas, M. Wymann, G. Opdenakker and T. Voets for discussions and for help with capacitance measurements (T.V.) and the PCA protocol (M.T. and M.W.); and J. Prenen, E. Martens, S. Buchholz, K. Fischer and C. Wesely for technical assistance. Supported by the Deutsche Forschungsgemeinschaft (M.F., S.P. and V.F.), Fonds der Chemischen Industrie and Sander-Stiftung (V.F.), Forschungsausschuss der Universität des Saarlandes (M.F., V.F.), the Human Frontiers Science Programme (RGP 32/2004 to R.V. and B.N.), the Belgian and Flemish Federal Government (GOA 2004/07, F.W.O., G.0136.00, F.W.O., G.0172.03 and IUAP Nr.3P4/23; Excellentiefinanciering EF/95/010 to R.V. and B.N.), the Flemish fund of scientific research (FWO-Vlaanderen; R.V.) and the Alexander von Humboldt-Stiftung (R.V.).

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Authors and Affiliations

  1. Division of Physiology, Department of Molecular Cell Biology, Laboratory of Ion Channel Research, Campus Gasthuisberg, O&N1, KU Leuven, Herestraat 49 bus 802, Leuven, B-3000, Belgium

    Rudi Vennekens & Bernd Nilius

  2. Experimentelle und Klinische Pharmakologie und Toxikologie, Universität des Saarlandes, Homburg, D-66421, Germany

    Rudi Vennekens, Jenny Olausson, Marcel Meissner, Ilka Mathar, Stephan E Philipp, Petra Weissgerber, Veit Flockerzi & Marc Freichel

  3. Institut für Kreislaufforschung und Sportmedizin, Molekulare und Zelluläre Sportmedizin, Deutsche Sporthochschule, Köln, 50927, Germany

    Wilhelm Bloch

  4. Anatomisches Institut, Universität des Saarlandes, Homburg, D-66421, Germany

    Frank Schmitz

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Contributions

R.V., M.F., B.N. and V.F. contributed to all aspects of the manuscript (conceptual design, experimentation, mouse work, writing); J.O. contributed to gene targeting; M.M. to contributed protein chemistry; F.S. and P.W. contributed to morphological characterization of BMMCs; W.B. contributed to histology, immunocytochemistry, immunohistochemistry and electron microscopy, I.M. contributed to anaphylaxis and glucose tolerance experiments; and S.E.P. performed cell-sorting and RT-PCR of B and T cells.

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Correspondence to Rudi Vennekens or Marc Freichel.

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Supplementary information

Supplementary Fig. 1

Targeted disruption of the Trpm4 gene. (PDF 95 kb)

Supplementary Fig. 2

Pancreatic β-cell function and expression analysis of Trpm4 and Trpm5 in Trpm4+/+ and Trpm4-/- mice. (PDF 168 kb)

Supplementary Fig. 3

Permeation properties of the endogenous Ca2+-activated cation current in BMMCs. (PDF 130 kb)

Supplementary Fig. 4

Ca2+ signaling and membrane potential in Trpm4+/+ and Trpm4-/- mast cells following DNP and combined adenosine+DNP stimulation in Ca2+-free and 156 mM K+ medium. (PDF 162 kb)

Supplementary Fig. 5

Ca2+ release–activated Ca2+ currents and FcεRI induced signaling in Trpm4+/+ and Trpm4-/- mast cells. (PDF 181 kb)

Supplementary Fig. 6

Membrane potential measurements in BMMCs after antigen stimulation. (PDF 143 kb)

Supplementary Fig. 7

Ca2+-activated Cl– and K+ currents in Trpm4+/+ and Trpm4-/- mast cells and a model for the role of TRPM4 in FcεRI-induced Ca2+ signaling. (PDF 185 kb)

Supplementary Fig. 8

GTP-γ-S-induced capacitance changes and LPS-induced activation of Trpm4+/+ and Trpm4-/- mast cells. (PDF 209 kb)

Supplementary Methods (PDF 165 kb)

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Vennekens, R., Olausson, J., Meissner, M. et al. Increased IgE-dependent mast cell activation and anaphylactic responses in mice lacking the calcium-activated nonselective cation channel TRPM4. Nat Immunol 8, 312–320 (2007). https://doi.org/10.1038/ni1441

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