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

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.).

Author information

Authors and Affiliations

Authors

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.

Corresponding authors

Correspondence to Rudi Vennekens or Marc Freichel.

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The authors declare no competing financial interests.

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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