Transient receptor potential M3 channels are ionotropic steroid receptors in pancreatic β cells

Abstract

Transient receptor potential (TRP) cation channels are renowned for their ability to sense diverse chemical stimuli. Still, for many members of this large and heterogeneous protein family it is unclear how their activity is regulated and whether they are influenced by endogenous substances. On the other hand, steroidal compounds are increasingly recognized to have rapid effects on membrane surface receptors that often have not been identified at the molecular level. We show here that TRPM3, a divalent-permeable cation channel, is rapidly and reversibly activated by extracellular pregnenolone sulphate, a neuroactive steroid. We show that pregnenolone sulphate activates endogenous TRPM3 channels in insulin-producing β cells. Application of pregnenolone sulphate led to a rapid calcium influx and enhanced insulin secretion from pancreatic islets. Our results establish that TRPM3 is an essential component of an ionotropic steroid receptor enabling unanticipated crosstalk between steroidal and insulin-signalling endocrine systems.

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Figure 1: TRPM3 channels are activated by PS and nifedipine.
Figure 2: Properties and specificity of agonist-evoked currents through TRPM3 channels.
Figure 3: PS is active only from the extracellular side and monovalent cations inhibit PS-induced currents through TRPM3 channels.
Figure 4: PS activates Ca2+-permeable, outwardly rectifying channels in Ins1 cells and mouse pancreatic islet cells.
Figure 5: Analysis of TRPM3 expression in Ins1 cells and pancreatic islets.
Figure 6: Downregulation of TRPM3 induced by shRNA reduces steroid-induced Ca2+ signals.
Figure 7: PS-induced Ca2+ signals in pancreatic islet cells.
Figure 8: PS enhances insulin release from pancreatic islets.

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Acknowledgements

We thank S. Plant, M. Portz, H. Löhr, K. Kraushaar, U. Soltek and M. Simon-Thomas for technical support; U. Wissenbach for TRPV6 and TRPM8 expressing cells; V. Chubanov for the TRPM7 expression vector; A. Lückhoff and F.J.P. Kühn for the TRPM2 expression vector; M. Menger and P. Weißgerber for initial help with isolating pancreatic islets. We also thank A. Beck, D. Beech, B. Fakler, M. Flick, M. Freichel, B. Niemeyer and F. Zufall for discussions and for reading an earlier version of the manuscript. Financial support was provided by DFG (Emmy Noether program, J.O., and SFB530, S.E.P. and V.F.) and HOMFOR.

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Correspondence to Johannes Oberwinkler.

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Wagner, T., Loch, S., Lambert, S. et al. Transient receptor potential M3 channels are ionotropic steroid receptors in pancreatic β cells. Nat Cell Biol 10, 1421–1430 (2008). https://doi.org/10.1038/ncb1801

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