Anandamide and arachidonic acid use epoxyeicosatrienoic acids to activate TRPV4 channels

Abstract

TRPV4 is a widely expressed cation channel of the ‘transient receptor potential’ (TRP) family1 that is related to the vanilloid receptor VR1 (TRPV1). It functions as a Ca2+ entry channel2 and displays remarkable gating promiscuity by responding to both physical stimuli (cell swelling, innoxious heat2,3,4,5,6,7) and the synthetic ligand 4αPDD8. An endogenous ligand for this channel has not yet been identified. Here we show that the endocannabinoid anandamide and its metabolite arachidonic acid activate TRPV4 in an indirect way involving the cytochrome P450 epoxygenase-dependent formation of epoxyeicosatrienoic acids. Application of 5′,6′-epoxyeicosatrienoic acid at submicromolar concentrations activates TRPV4 in a membrane-delimited manner and causes Ca2+ influx through TRPV4-like channels in vascular endothelial cells. Activation of TRPV4 in vascular endothelial cells might therefore contribute to the relaxant effects of endocannabinoids and their P450 epoxygenase-dependent metabolites on vascular tone9,10,11,12.

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Figure 1: Effect of AA on [Ca2+]i and currents in TRPV4-expressing HEK-293 cells.
Figure 2: AEA and 2-AG increase [Ca2+]i and activate currents in TRPV4-transfected cells.
Figure 3: Unravelling the activation cascade of TRPV4.
Figure 4: Activation of TRPV4 by EETs.
Figure 5: 5′,6′-EET-induced increase of [Ca2+]i and activation of a TRPV4-like current in native endothelial cells from mouse aorta.

References

  1. 1

    Clapham, D. E., Runnels, L. W. & Strübing, C. The TRP ion channel family. Nature Rev. Neurosci. 2, 387–396 (2001)

    CAS  Article  Google Scholar 

  2. 2

    Strotmann, R., Harteneck, C., Nunnenmacher, K., Schultz, G. & Plant, T. D. OTRPC4, a nonselective cation channel that confers sensitivity to extracellular osmolarity. Nature Cell Biol. 2, 695–702 (2000)

    CAS  Article  PubMed Central  Google Scholar 

  3. 3

    Güler, A. et al. Heat-evoked activation of the ion channel, TRPV4. J. Neurosci. 22, 6408–6414 (2002)

    Article  PubMed Central  Google Scholar 

  4. 4

    Wissenbach, U., Bödding, M., Freichel, M. & Flockerzi, V. Trp12, a novel Trp related protein from kidney. FEBS Lett. 485, 127–134 (2000)

    CAS  Article  PubMed Central  Google Scholar 

  5. 5

    Liedtke, W. et al. Vanilloid receptor-related osmotically activated channel (VR-OAC), a candidate vertebrate osmoreceptor. Cell 103, 525–535 (2000)

    CAS  Article  PubMed Central  Google Scholar 

  6. 6

    Nilius, B., Prenen, J., Wissenbach, U., Bödding, M. & Droogmans, G. Differential activation of the volume-sensitive cation channel TRP12 (OTRPC4) and the volume-regulated anion currents in HEK-293 cells. Pflügers Arch. 443, 227–233 (2001)

    CAS  Article  PubMed Central  Google Scholar 

  7. 7

    Watanabe, H. et al. Heat-evoked activation of TRPV4 channels in an HEK293 cell expression system and in native mouse aorta endothelial cells. J. Biol. Chem. 277, 47044–47051 (2002)

    CAS  Article  PubMed Central  Google Scholar 

  8. 8

    Watanabe, H. et al. Activation of TRPV4 channels (hVRL-2/mTRP12) by phorbol derivatives. J. Biol. Chem. 277, 13569–13577 (2002)

    CAS  Article  PubMed Central  Google Scholar 

  9. 9

    Seegers, H. C., Gross, R. W. & Boyle, W. A. Calcium-independent phospholipase A2-derived arachidonic acid is essential for endothelium-dependent relaxation by acetylcholine. J. Pharmacol. Exp. Ther. 302, 918–923 (2002)

    CAS  Article  PubMed Central  Google Scholar 

  10. 10

    Tan, J. Z., Kaley, G. & Gurtner, G. H. Nitric oxide and prostaglandins mediate vasodilation to 5,6-EET in rabbit lung. Adv. Exp. Med. Biol. 407, 561–566 (1997)

    CAS  Article  PubMed Central  Google Scholar 

  11. 11

    Fuloria, M., Smith, T. K. & Aschner, J. L. Role of 5,6-epoxyeicosatrienoic acid in the regulation of newborn piglet pulmonary vascular tone. Am. J. Physiol. Lung Cell Mol. Physiol. 283, L383–L389 (2002)

    CAS  Article  PubMed Central  Google Scholar 

  12. 12

    Jarai, Z. et al. Cannabinoid-induced mesenteric vasodilation through an endothelial site distinct from CB1 or CB2 receptors. Proc. Natl Acad. Sci. USA 96, 14136–14141 (1999)

    ADS  CAS  Article  PubMed Central  Google Scholar 

  13. 13

    Voets, T. et al. Molecular determinants of permeation through the cation channel TRPV4. J. Biol. Chem. 277, 33704–33710 (2002)

    CAS  Article  PubMed Central  Google Scholar 

  14. 14

    Ueda, N. Endocannabinoid hydrolases. Prostaglandins Other Lipid Mediat. 68–69, 521–534 (2002)

    Article  PubMed Central  Google Scholar 

  15. 15

    Fleming, I. Cytochrome P450 enzymes in vascular homeostasis. Circ. Res. 89, 753–762 (2001)

    CAS  Article  PubMed Central  Google Scholar 

  16. 16

    Roman, R. J. P-450 metabolites of arachidonic acid in the control of cardiovascular function. Physiol. Rev. 82, 131–185 (2002)

    CAS  Article  PubMed Central  Google Scholar 

  17. 17

    Randall, M. D. & Kendall, D. A. Endocannabinoids: A new class of vasoactive substances. Trends Pharmacol. Sci. 19, 55–58 (1998)

    CAS  Article  PubMed Central  Google Scholar 

  18. 18

    Zygmunt, P. M. et al. Vanilloid receptors on sensory nerves mediate the vasodilator action of anandamide. Nature 400, 452–457 (1999)

    ADS  CAS  Article  Google Scholar 

  19. 19

    Randall, M. D. & Kendall, D. A. Anandamide and endothelium-derived hyperpolarizing factor act via a common vasorelaxant mechanism in rat mesentery. Eur. J. Pharmacol. 346, 51–53 (1998)

    CAS  Article  PubMed Central  Google Scholar 

  20. 20

    Rzigalinski, B. A., Willoughby, K. A., Hoffman, S. W., Falck, J. R. & Ellis, E. F. Calcium influx factor, further evidence it is 5,6-epoxyeicosatrienoic acid. J. Biol. Chem. 274, 175–182 (1999)

    CAS  Article  Google Scholar 

  21. 21

    Xie, Q., Zhang, Y., Zhai, C. & Bonanno, J. A. Calcium influx factor from cytochrome P-450 metabolism and secretion-like coupling mechanisms for capacitative calcium entry in corneal endothelial cells. J. Biol. Chem. 277, 16559–16566 (2002)

    CAS  Article  Google Scholar 

  22. 22

    Graier, W. F., Simecek, S. & Sturek, M. Cytochrome P450 mono-oxygenase-regulated signalling of Ca2+ entry in human and bovine endothelial cells. J. Physiol. (Lond.) 482, 259–274 (1995)

    CAS  Article  Google Scholar 

  23. 23

    Wilson, R. I. & Nicoll, R. A. Endocannabinoid signaling in the brain. Science 296, 678–682 (2002)

    ADS  CAS  Article  PubMed Central  Google Scholar 

  24. 24

    Suh, S. H. et al. Characterisation of explanted endothelial cells from mouse aorta: electrophysiology and Ca2+ signalling. Pflügers Arch. 438, 612–620 (1999)

    CAS  PubMed  PubMed Central  Google Scholar 

  25. 25

    Freichel, M. et al. Lack of an endothelial store-operated Ca2+ current impairs agonist-dependent vasorelaxation in TRP4-/- mice. Nature Cell Biol. 3, 121–127 (2001)

    CAS  Article  Google Scholar 

  26. 26

    Grainger, J. & Boachie Ansah, G. Anandamide-induced relaxation of sheep coronary arteries: The role of the vascular endothelium, arachidonic acid metabolites and potassium channels. Br. J. Pharmacol. 134, 1003–1012 (2001)

    CAS  Article  PubMed Central  Google Scholar 

  27. 27

    Williams, D. A. & Fay, F. S. Intracellular calibration of the fluorescent calcium indicator Fura-2. Cell Calcium 11, 75–83 (1990)

    CAS  Article  PubMed Central  Google Scholar 

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Acknowledgements

We thank V. Flockerzi and C. D. Benham for comments, and V. Flockerzi and U. Wissenbach for providing the mTRP12 clone (mTRPV4). This work was supported by the Belgian Federal Government, the Flemish Government and the Onderzoeksraad KU Leuven (Interuniversity Poles of Attraction Program, IUAP). T.V. is a postdoctoral Fellow of the Fund for Scientific Research–Flanders (Belgium) (FWO–Vlaanderen).

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Correspondence to Bernd Nilius.

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Watanabe, H., Vriens, J., Prenen, J. et al. Anandamide and arachidonic acid use epoxyeicosatrienoic acids to activate TRPV4 channels. Nature 424, 434–438 (2003). https://doi.org/10.1038/nature01807

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