The progesterone receptor regulates the expression of TRPV4 channel

  • Carole Jung
  • César Fandos
  • Ivan M. Lorenzo
  • Cristina Plata
  • Jacqueline Fernandes
  • Gemma G. Gené
  • Esther Vázquez
  • Miguel A. Valverde
Molecular and Genomic Physiology


The transient receptor potential cationic channel TRPV4 contributes to different aspects of cell physiology via the generation of a Ca2+ signal and/or depolarization of the membrane potential. TRPV4 channel integrates distinct physical and chemical stimuli, including osmotic and mechanical stress, heat, acidic pH, endogenous ligands, and synthetic agonists such as 4α-phorbol 12,13-didecanoate (4αPDD). Although several regulatory sites controlling TRPV4 channel activity have been identified, very little is known about the regulation of TRPV4 expression, a situation common to other TRP channels. Here we show that TRPV4 expression is under the control of progesterone in both human airways and mammary gland epithelial cells, as well as in vascular smooth muscle cells. Exposure of human airways epithelial CFT1-LCFSN and mammary gland epithelial T47D cells to progesterone decreased TRPV4 mRNA and protein expression. Consequently, 4αPDD-induced cationic currents and Ca2+ signals were also diminished in progesterone-treated cells. The effect of progesterone was reverted by the progesterone receptor (PR) antagonist RU-486 or following transfection with small interference RNA (siRNA) against both PRA and PRB isoforms. Interestingly, TRPV4 expression and activity were increased in T47D mammary gland epithelial cells when PR was silenced with siRNA. Transcriptional regulation of −1.3 kB TRPV4 promoter-luciferase plasmids was also evaluated in vascular smooth muscle cells. TRPV4 promoter activity was reduced by coexpression with PR and further reduced in the presence of PG. Together, our data report the regulation of TRPV4 expression by progesterone, a process that requires a functional PR.


Epithelia Vascular smooth muscle Progesterone receptor Promoter Transcription Expression TRPV4 Calcium 



We would like to thank Dr. Albert Jordan (CSIC, Barcelona) for the human PRA and PRB expressing vectors. This work was supported by the Spanish Ministry of Science and Innovation (SAF2006-04973), Fondo de Investigación Sanitaria (Red HERACLES RD06/0009), Generalitat de Catalunya (SGR05-266), and Fundació la Marató de TV3 (061331,080430). The authors declare that they have no competing financial interests.

Supplementary material

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  1. 1.
    Andrade YN, Fernandes J, Vazquez E, Fernandez-Fernandez JM, Arniges M, Sanchez TM, Villalon M, Valverde MA (2005) TRPV4 channel is involved in the coupling of fluid viscosity changes to epithelial ciliary activity. J Cell Biol 168:869–874CrossRefPubMedGoogle Scholar
  2. 2.
    Arniges M, Fernandez-Fernandez JM, Albrecht N, Schaefer M, Valverde MA (2006) Human TRPV4 channel splice variants revealed a key role of ankyrin domains in multimerization and trafficking. J Biol Chem 281:1580–1586CrossRefPubMedGoogle Scholar
  3. 3.
    Arniges M, Vazquez E, Fernandez-Fernandez JM, Valverde MA (2004) Swelling-activated Ca2+ entry via TRPV4 channel is defective in cystic fibrosis airway epithelia. J Biol Chem 279:54062–54068CrossRefPubMedGoogle Scholar
  4. 4.
    Bagchi MK, Tsai SY, Tsai MJ, O'Malley BW (1990) Identification of a functional intermediate in receptor activation in progesterone-dependent cell-free transcription. Nature 345:547–550CrossRefPubMedGoogle Scholar
  5. 5.
    Bahamonde MI, Fernandez-Fernandez JM, Guix FX, Vazquez E, Valverde MA (2003) Plasma membrane voltage-dependent anion channel mediates antiestrogen-activated maxi Cl- currents in C1300 neuroblastoma cells. J Biol Chem 278:33284–33289CrossRefPubMedGoogle Scholar
  6. 6.
    Bidaux G, Roudbaraki M, Merle C, Crepin A, Delcourt P, Slomianny C, Thebault S, Bonnal JL, Benahmed M, Cabon F, Mauroy B, Prevarskaya N (2005) Evidence for specific TRPM8 expression in human prostate secretory epithelial cells: functional androgen receptor requirement. Endocr Relat Cancer 12:367–382CrossRefPubMedGoogle Scholar
  7. 7.
    Bolanz KA, Hediger MA, Landowski CP (2008) The role of TRPV6 in breast carcinogenesis. Mol Cancer Ther 7:271–279CrossRefPubMedGoogle Scholar
  8. 8.
    Burnstein KL, Cidlowski JA (1993) Multiple mechanisms for regulation of steroid hormone action. J Cell Biochem 51:130–134CrossRefPubMedGoogle Scholar
  9. 9.
    Camacho-Arroyo I, Ruiz A, Gamboa-Dominguez A, Perez-Palacios G, Cerbon MA (1994) Immunohistochemical localization of intracellular progesterone and glucocorticoid receptors in the rabbit lung. J Endocrinol 142:311–316CrossRefPubMedGoogle Scholar
  10. 10.
    Chang AS, Chang SM, Garcia RL, Schilling WP (1997) Concomitant and hormonally regulated expression of trp genes in bovine aortic endothelial cells. FEBS Lett 415:335–340CrossRefPubMedGoogle Scholar
  11. 11.
    Cheng KW, Cheng CK, Leung PC (2001) Differential role of PR-A and -B isoforms in transcription regulation of human GnRH receptor gene. Mol Endocrinol 15:2078–2092CrossRefPubMedGoogle Scholar
  12. 12.
    Coma M, Guix FX, Ill-Raga G, Uribesalgo I, Alameda F, Valverde MA, Munoz FJ (2008) Oxidative stress triggers the amyloidogenic pathway in human vascular smooth muscle cells. Neurobiol Aging 29:969–980CrossRefPubMedGoogle Scholar
  13. 13.
    D'hoedt D, Owsianik G, Prenen J, Cuajungco MP, Grimm C, Heller S, Voets T, Nilius B (2008) Stimulus-specific modulation of the cation channel TRPV4 by PACSIN 3. J Biol Chem 283:6272–6280CrossRefPubMedGoogle Scholar
  14. 14.
    Dijkema R, Schoonen WG, Teuwen R, van der Srtuik E, de Ries RJ, van der Kar BA, Olijve W (1998) Human progesterone receptor A and B isoforms in CHO cells. I. Stable transfection of receptor and receptor-responsive reporter genes: transcription modulation by (anti)progestagens. J Steroid Biochem Mol Biol 64:147–156CrossRefPubMedGoogle Scholar
  15. 15.
    Earley S, Heppner TJ, Nelson MT, Brayden JE (2005) TRPV4 forms a novel Ca2+ signaling complex with ryanodine receptors and BKCa channels. Circ Res 97:1270–1279CrossRefPubMedGoogle Scholar
  16. 16.
    Ehring GR, Kerschbaum HH, Eder C, Neben AL, Fanger CM, Khoury RM, Negulescu PA, Cahalan MD (1998) A nongenomic mechanism for progesterone-mediated immunosuppression: inhibition of K+ channels, Ca2+ signaling, and gene expression in T lymphocytes. J Exp Med 188:1593–1602CrossRefPubMedGoogle Scholar
  17. 17.
    Falkenstein E, Tillmann HC, Christ M, Feuring M, Wehling M (2000) Multiple actions of steroid hormones—a focus on rapid, nongenomic effects. Pharmacol Rev 52:513–556PubMedGoogle Scholar
  18. 18.
    Fernandes J, Lorenzo IM, Andrade YN, Garcia-Elias A, Serra SA, Fernandez-Fernandez JM, Valverde MA (2008) IP3 sensitizes TRPV4 channel to the mechano- and osmotransducing messenger 5′-6′-epoxyeicosatrienoic acid. J Cell Biol 181:143–155CrossRefPubMedGoogle Scholar
  19. 19.
    Fernandez-Fernandez JM, Andrade YN, Arniges M, Fernandes J, Plata C, Rubio-Moscardo F, Vazquez E, Valverde MA (2008) Functional coupling of TRPV4 cationic channel and large conductance, calcium-dependent potassium channel in human bronchial epithelial cell lines. Pflugers Arch 457:149–159CrossRefPubMedGoogle Scholar
  20. 20.
    Fernandez-Fernandez JM, Nobles M, Currid A, Vazquez E, Valverde MA (2002) Maxi K+ channel mediates regulatory volume decrease response in a human bronchial epithelial cell line. Am J Physiol Cell Physiol 283:C1705–C1714PubMedGoogle Scholar
  21. 21.
    Gao X, Wu L, O'Neil RG (2003) Temperature-modulated diversity of TRPV4 channel gating: activation by physical stresses and phorbol ester derivatives through protein kinase C-dependent and -independent pathways. J Biol Chem 278:27129–27137CrossRefPubMedGoogle Scholar
  22. 22.
    Garcia-Elias A, Lorenzo IM, Vicente R, Valverde MA (2008) IP3 receptor binds to and sensitizes TRPV4 channel to osmotic stimuli via a calmodulin-binding site. J Biol Chem 283:31284–31288CrossRefPubMedGoogle Scholar
  23. 23.
    Graham JD, Clarke CL (2002) Expression and transcriptional activity of progesterone receptor A and progesterone receptor B in mammalian cells. Breast Cancer Res 4:187–190CrossRefPubMedGoogle Scholar
  24. 24.
    Hoenderop JG, Nilius B, Bindels RJ (2003) Epithelial calcium channels: from identification to function and regulation. Pflugers Arch 446:304–308PubMedGoogle Scholar
  25. 25.
    Irnaten M, Blanchard-Gutton N, Praetorius J, Harvey BJ (2009) Rapid effects of 17beta-estradiol on TRPV5 epithelial Ca(2+) channels in rat renal cells. Steroids 74:642–649CrossRefPubMedGoogle Scholar
  26. 26.
    Kitahara T, Li HS, Balaban CD (2005) Changes in transient receptor potential cation channel superfamily V (TRPV) mRNA expression in the mouse inner ear ganglia after kanamycin challenge. Hear Res 201:132–144CrossRefPubMedGoogle Scholar
  27. 27.
    Liedtke W, Friedman JM (2003) Abnormal osmotic regulation in trpv4−/− mice. Proc Natl Acad Sci U S A 100:13698–13703CrossRefPubMedGoogle Scholar
  28. 28.
    Lorenzo IM, Liedtke W, Sanderson MJ, Valverde MA (2008) TRPV4 channel participates in receptor-operated calcium entry and ciliary beat frequency regulation in mouse airway epithelial cells. Proc Natl Acad Sci U S A 105:12611–12616CrossRefPubMedGoogle Scholar
  29. 29.
    Mahmood T, Saridogan E, Smutna S, Habib AM, Djahanbakhch O (1998) The effect of ovarian steroids on epithelial ciliary beat frequency in the human Fallopian tube. Hum Reprod 13:2991–2994CrossRefPubMedGoogle Scholar
  30. 30.
    Mori T, Murata M, Yoshino T, Nakasono S, Saito F, Takeyama H, Matsunaga T (2009) A stable human progesterone receptor expressing HeLa reporter cell line as a tool in chemical evaluation at the different cell-cycle phases. Toxicol Lett 186:123–129CrossRefPubMedGoogle Scholar
  31. 31.
    Nadal A, Diaz M, Valverde MA (2001) The estrogen trinity: membrane, cytosolic, and nuclear effects. News Physiol Sci 16:251–255PubMedGoogle Scholar
  32. 32.
    Nilius B, Vriens J, Prenen J, Droogmans G, Voets T (2004) TRPV4 calcium entry channel: a paradigm for gating diversity. Am J Physiol Cell Physiol 286:C195–C205CrossRefPubMedGoogle Scholar
  33. 33.
    Nutu M, Weijdegard B, Thomas P, Bergh C, Thurin-Kjellberg A, Pang Y, Billig H, Larsson DG (2007) Membrane progesterone receptor gamma: tissue distribution and expression in ciliated cells in the fallopian tube. Mol Reprod Dev 74:843–850CrossRefPubMedGoogle Scholar
  34. 34.
    Orshal JM, Khalil RA (2004) Gender, sex hormones, and vascular tone. Am J Physiol Regul Integr Comp Physiol 286:R233–R249PubMedGoogle Scholar
  35. 35.
    Paltieli Y, Eibschitz I, Ziskind G, Ohel G, Silbermann M, Weichselbaum A (2000) High progesterone levels and ciliary dysfunction—a possible cause of ectopic pregnancy. J Assist Reprod Genet 17:103–106CrossRefPubMedGoogle Scholar
  36. 36.
    Paria BC, Bair AM, Xue J, Yu Y, Malik AB, Tiruppathi C (2006) Ca2+ influx induced by protease-activated receptor-1 activates a feed-forward mechanism of TRPC1 expression via nuclear factor-kappaB activation in endothelial cells. J Biol Chem 281:20715–20727CrossRefPubMedGoogle Scholar
  37. 37.
    Pasanen S, Ylikomi T, Syvala H, Tuohimaa P (1997) Distribution of progesterone receptor in chicken: novel target organs for progesterone and estrogen action. Mol Cell Endocrinol 135:79–91CrossRefPubMedGoogle Scholar
  38. 38.
    Senti M, Fernandez-Fernandez JM, Tomas M, Vazquez E, Elosua R, Marrugat J, Valverde MA (2005) Protective effect of the KCNMB1 E65K genetic polymorphism against diastolic hypertension in aging women and its relevance to cardiovascular risk. Circ Res 97:1360–1365CrossRefPubMedGoogle Scholar
  39. 39.
    Shirasaki H, Watanabe K, Kanaizumi E, Konno N, Sato J, Narita S, Himi T (2004) Expression and localization of steroid receptors in human nasal mucosa. Acta Otolaryngol 124:958–963CrossRefPubMedGoogle Scholar
  40. 40.
    Strotmann R, Harteneck C, Nunnenmacher K, Schultz G, Plant TD (2000) OTRPC4, a nonselective cation channel that confers sensitivity to extracellular osmolarity. Nat Cell Biol 2:695–702CrossRefPubMedGoogle Scholar
  41. 41.
    Strotmann R, Schultz G, Plant TD (2003) Ca2+-dependent potentiation of the nonselective cation channel TRPV4 is mediated by a C-terminal calmodulin binding site. J Biol Chem 278:26541–26549CrossRefPubMedGoogle Scholar
  42. 42.
    Suzuki M, Mizuno A, Kodaira K, Imai M (2003) Impaired pressure sensation in mice lacking TRPV4. J Biol Chem 278:22664–22668CrossRefPubMedGoogle Scholar
  43. 43.
    Tait AS, Butts CL, Sternberg EM (2008) The role of glucocorticoids and progestins in inflammatory, autoimmune, and infectious disease. J Leukoc Biol 84:924–931CrossRefPubMedGoogle Scholar
  44. 44.
    Teilmann SC, Byskov AG, Pedersen PA, Wheatley DN, Pazour GJ, Christensen ST (2005) Localization of transient receptor potential ion channels in primary and motile cilia of the female murine reproductive organs. Mol Reprod Dev 71:444–452CrossRefPubMedGoogle Scholar
  45. 45.
    Teilmann SC, Clement CA, Thorup J, Byskov AG, Christensen ST (2006) Expression and localization of the progesterone receptor in mouse and human reproductive organs. J Endocrinol 191:525–535CrossRefPubMedGoogle Scholar
  46. 46.
    Uotinen N, Puustinen R, Pasanen S, Manninen T, Kivineva M, Syvala H, Tuohimaa P, Ylikomi T (1999) Distribution of progesterone receptor in female mouse tissues. Gen Comp Endocrinol 115:429–441CrossRefPubMedGoogle Scholar
  47. 47.
    Vallejo G, Ballare C, Baranao JL, Beato M, Saragueta P (2005) Progestin activation of nongenomic pathways via cross talk of progesterone receptor with estrogen receptor beta induces proliferation of endometrial stromal cells. Mol Endocrinol 19:3023–3037CrossRefPubMedGoogle Scholar
  48. 48.
    Valverde MA, Rojas P, Amigo J, Cosmelli D, Orio P, Bahamonde MI, Mann GE, Vergara C, Latorre R (1999) Acute activation of Maxi-K channels (hSlo) by estradiol binding to the β subunit. Science 285:1929–1931CrossRefPubMedGoogle Scholar
  49. 49.
    Vazquez E, Nobles M, Valverde MA (2001) Defective regulatory volume decrease in human cystic fibrosis tracheal cells because of altered regulation of intermediate conductance Ca2+-dependent potassium channels. Proc Natl Acad Sci U S A 98:5329–5334CrossRefPubMedGoogle Scholar
  50. 50.
    Vazquez E, Valverde MA (2006) A review of TRP channels splicing. Semin Cell Dev Biol 17:607–617CrossRefPubMedGoogle Scholar
  51. 51.
    Vriens J, Owsianik G, Janssens A, Voets T, Nilius B (2007) Determinants of 4 alpha-phorbol sensitivity in transmembrane domains 3 and 4 of the cation channel TRPV4. J Biol Chem 282:12796–12803CrossRefPubMedGoogle Scholar
  52. 52.
    Watanabe H, Davis JB, Smart D, Jerman JC, Smith GD, Hayes P, Vriens J, Cairns W, Wissenbach U, Prenen J, Flockerzi V, Droogmans G, Benham CD, Nilius B (2002) Activation of TRPV4 channels (hVRL-2/mTRP12) by phorbol derivatives. J Biol Chem 277:13569–13577CrossRefPubMedGoogle Scholar
  53. 53.
    Wessel T, Schuchter U, Walt H (2004) Ciliary motility in bovine oviducts for sensing rapid non-genomic reactions upon exposure to progesterone. Horm Metab Res 36:136–141CrossRefPubMedGoogle Scholar
  54. 54.
    Wu D, Huang W, Richardson PM, Priestley JV, Liu M (2008) TRPC4 in rat dorsal root ganglion neurons is increased after nerve injury and is necessary for neurite outgrowth. J Biol Chem 283:416–426CrossRefPubMedGoogle Scholar
  55. 55.
    Yankaskas JR, Conrad M, Kovai D, Lazarowsky E, Paradiso AM, Rinehart CA, Sarkadi B, Schlegel R, Boucher RC (1993) Papilloma virus immortalized tracheal epithelial cells retain a well-differentiated phenotype. Am J Physiol 264:C1219–C1230PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Carole Jung
    • 1
  • César Fandos
    • 1
  • Ivan M. Lorenzo
    • 1
  • Cristina Plata
    • 1
  • Jacqueline Fernandes
    • 1
  • Gemma G. Gené
    • 1
  • Esther Vázquez
    • 1
    • 2
  • Miguel A. Valverde
    • 1
  1. 1.Laboratory of Molecular Physiology and Channelopathies, Department of Experimental and Health SciencesUniversitat Pompeu Fabra, Edifici PRBBBarcelonaSpain
  2. 2.CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN)BarcelonaSpain

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