, Volume 15, Issue 3, pp 349–360 | Cite as

P2Y4 nucleotide receptor: a novel actor in post-natal cardiac development

  • Michael Horckmans
  • Bernard Robaye
  • Elvira Léon-Gόmez
  • Nicolas Lantz
  • Philippe Unger
  • Frédérique Dol-Gleizes
  • Sophie Clouet
  • Dorothée Cammarata
  • Paul Schaeffer
  • Pierre Savi
  • Christian Gachet
  • Jean-Luc Balligand
  • Chantal Dessy
  • Jean-Marie Boeynaems
  • Didier CommuniEmail author
Original Paper


Communication between endothelial cells and cardiomyocytes is critical for cardiac development and regeneration. However the mechanisms involved in these endothelial-cardiomyocyte interactions remain poorly understood. Nucleotides are released within the heart, especially under ischemia or pressure overload. The function of P2Y nucleotide receptors in cardiac development has never been investigated. Here we show that adult P2Y4-null mice display microcardia. P2Y4 nucleotide receptor is expressed in cardiac endothelial cells but not in cardiomyocytes. Loss of P2Y4 in cardiac endothelial cells strongly inhibits their growth, migration and PDGF-B secretion in response to UTP. Proliferation of microvessels and cardiomyocytes is reduced in P2Y4-null hearts early after birth, resulting in reduced heart growth. Our study uncovers mouse P2Y4 receptor as an essential regulator of cardiac endothelial cell function, and illustrates the involvement of endothelial-cardiomyocyte interactions in post-natal heart development. We also detected P2Y4 expression in human cardiac microvessels. P2Y4 receptor could constitute a therapeutic target to regulate cardiac remodelling and post-ischemic revascularisation.


Heart development Angiogenesis Nucleotides Endothelial cells P2Y receptors 



We thank Larissa di Pietrantonio, Olivier Vosters, Dominique Fokan, Hrag Esfahani and Steven Droogmans for help and technical assistance. The authors are grateful to Dr Andreas Markl for invaluable assistance in telemetry and helpful discussion. This work was supported by an Action de Recherche Concertée of the Communauté Française de Belgique, an Interuniversity Attraction Pole grant from the Politique Scientifique Fédérale (IAP-P6/30), Prime Minister’s Office, Federal Service for Science, Technology and Culture, by grants of the Fonds de la Recherche Scientifique Médicale (F.R.S.M.), the Fonds d’Encouragement à la Recherche (F.E.R.), the Fonds Emile DEFAY, the Fonds de la Recherche Scientifique Médicale of Belgium, the Walloon Region (Programme d’Excellence CIBLES), and the LifeSciHealth programme of the European Community (grant LSHB-2003-503337). D. Communi and C. Dessy are Senior Research Associates of the Fonds National de la Recherche Scientifique (F.N.R.S.). M. Horckmans is supported by the Fonds National de la Recherche Scientifique/FRIA, Belgium. Philippe Unger has received a grant from the “Fonds pour la Chirurgie Cardiaque”, Elvira Leon-Gomez is supported by a grant of the F.S.R. (Fonds Spécial de Recherche) of the U.C.L. (Université catholique de Louvain). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Conflict of interest



  1. 1.
    Hsieh PC, Davis ME, Lisowski LK, Lee RT (2006) Endothelial-cardiomyocyte interactions in cardiac development and repair. Annu Rev Physiol 68:51–66PubMedCrossRefGoogle Scholar
  2. 2.
    Dutta AK, Sabirov RZ, Uramoto H, Okada YR (2004) Role of ATP-conductive anion channel in ATP release from neonatal rat cardiomyocytes in ischaemic or hypoxic conditions. J Physiol 559:799–812PubMedGoogle Scholar
  3. 3.
    Nishida M, Sato Y, Uemura A, Narita Y, Tozaki-Saitoh H, Nakaya M, Ide T, Suzuki K, Inoue K, Nagao T, Kurose H (2008) P2Y6 receptor-Galpha12/13 signalling in cardiomyocytes triggers pressure overload-induced cardiac fibrosis. EMBO J 27(23):3104–3115PubMedCrossRefGoogle Scholar
  4. 4.
    Cheung KK, Ryten M, Burnstock G (2003) Abundant and dynamic expression of G protein coupled P2Y receptors in mammalian development. Dev Dyn 228:254–266PubMedCrossRefGoogle Scholar
  5. 5.
    Communi D, Pirotton S, Parmentier M, Boeynaems JM (1995) Cloning and functional expression of a human uridine nucleotide receptor. J Biol Chem 270(52):30849–30852PubMedCrossRefGoogle Scholar
  6. 6.
    Bogdanov YD, Wildman SS, Clements MP, King BF, Burnstock G (1998) Molecular cloning and characterization of rat P2Y4 nucleotide receptor. Br J Pharmacol 124(3):428–430PubMedCrossRefGoogle Scholar
  7. 7.
    Webb TE, Henderson D, Roberts JA, Barnard EA (1998) Molecular cloning and characterization of the rat P2Y4 receptor. J Neurochem 71(4):1348–1357PubMedCrossRefGoogle Scholar
  8. 8.
    Lazarowski ER, Rochelle LG, O’Neal WK, Ribeiro CM, Grubb BR, Zhang V, Harden TK, Boucher RC (2001) Cloning and functional characterization of two murine uridine nucleotide receptors reveal a potential target for correcting ion transport deficiency in cystic fibrosis gallbladder. J Pharmacol Exp Ther 297(1):43–49PubMedGoogle Scholar
  9. 9.
    Lustig KD, Shiau AK, Brake AJ, Julius D (1993) Expression cloning of an ATP receptor from mouse neuroblastoma cells. Proc Natl Acad Sci USA 90(11):5113–5117PubMedCrossRefGoogle Scholar
  10. 10.
    Robaye B, Ghanem E, Wilkin F, Fokan D, Van Driessche W, Schurmans S, Boeynaems JM, Beauwens R (2003) Loss of nucleotide regulation of epithelial chloride transport in the jejunum of P2Y4-null mice. Mol Pharmacol 63(4):777–783PubMedCrossRefGoogle Scholar
  11. 11.
    Ghanem E, Robaye B, Leal T, Leipziger J, Van Driessche W, Beauwens R, Boeynaems JM (2005) The role of epithelial P2Y2 and P2Y4 receptors in the regulation of intestinal chloride secretion. Br J Pharmacol 146(3):364–369PubMedCrossRefGoogle Scholar
  12. 12.
    Matos JE, Sorensen MV, Geyti CS, Robaye B, Boeynaems JM, Leipziger J (2007) Distal colonic Na(+) absorption inhibited by luminal P2Y(2) receptors. Pflugers Arch Eur J Physiol 454:977–987CrossRefGoogle Scholar
  13. 13.
    Yitzhaki S, Shneyvays V, Jacobson KA, Shainberg A (2005) Involvement of uracil nucleotides in protection of cardiomyocytes from hypoxic stress. Biochem Pharmacol 69:1215–1223PubMedCrossRefGoogle Scholar
  14. 14.
    Yitzhaki S, Shainberg A, Cheporko Y, Vidne BA, Sagie A, Jacobson KA, Hochhauser E (2006) Uridine-5′-triphosphate (UTP) reduces infarct size and improves rat heart function after myocardial infarct. Biochem Pharmacol 72:949–955PubMedCrossRefGoogle Scholar
  15. 15.
    Vanderhaeghen P, Lu Q, Prakash N, Frisén J, Walsh CA, Frostig RD, Flanagan JG (2000) A mapping label required for normal scale of body representation in the cortex. Nat Neurosci 3:358–365PubMedCrossRefGoogle Scholar
  16. 16.
    Marelli-Berg FM, Peek E, Lidington EA, Stauss HJ, Lechler RI (2000) Isolation of endothelial cells from murine tissue. J Immunol Methods 244:205–215PubMedCrossRefGoogle Scholar
  17. 17.
    Alvarez BV, Johnson DE, Sowah D, Soliman D, Light PE, Xia Y, Karmazyn M, Casey JR (2007) Carbonic anhydrase inhibition prevents and reverts cardiomyocyte hypertrophy. J Physiol 579(1):127–145PubMedCrossRefGoogle Scholar
  18. 18.
    Falk W, Goodwin RH, Leonard EJ (1980) A 48-well micro chemotaxis assembly for rapid and accurate measurement of leucocyte migration. J Immunol Methods 33:239–247PubMedGoogle Scholar
  19. 19.
    Colville-Nash PR, Alam CA, Appleton I, Brown JR, Seed MP, Willoughby DA (1995) The pharmacological modulation of angiogenesis in chronic granulomatous inflammation. J Pharmacol Exp Ther 274(3):1463–1472PubMedGoogle Scholar
  20. 20.
    O’Connell TD, Ishizaka S, Nakamura A, Swigart PM, Rodrigo MC, Simpson GL, Cotecchia S, Rokosh DG, Grossman W, Foster E, Simpson PC (2003) The alpha(1A/C)- and alpha(1B)-adrenergic receptors are required for physiological cardiac hypertrophy in the double-knockout mouse. J Clin Invest 111:1783–1791PubMedGoogle Scholar
  21. 21.
    Ikeda Y, Aihara K, Sato T, Akaike M, Yoshizumi M, Suzaki Y, Izawa Y, Fujimura M, Hashizume S, Kato M, Yagi S, Tamaki T, Kawano H, Matsumoto T, Azuma H, Kato S, Matsumoto T (2005) Androgen receptor gene knockout male mice exhibit impaired cardiac growth and exacerbation of angiotensin II-induced cardiac fibrosis. J Biol Chem 280(33):29661–29666PubMedCrossRefGoogle Scholar
  22. 22.
    Nebigil CG, Choi DS, Dierich A, Hickel P, Le Meur M, Messaddeq N, Launay JM, Maroteaux L (2000) Serotonin 2B receptor is required for heart development. Proc Natl Acad Sci USA 97:9508–9513PubMedCrossRefGoogle Scholar
  23. 23.
    Nahrendorf M, Spindler M, Hu K, Bauer L, Ritter O, Nordbeck P, Quaschning T, Hiller KH, Wallis J, Ertl G, Bauer WR, Neubauer S (2005) Creatine kinase knockout mice show left ventricular hypertrophy and dilatation, but unaltered remodeling post-myocardial infarction. Cardiovasc Res 65:419–427PubMedCrossRefGoogle Scholar
  24. 24.
    Ellmers LJ, Knowles JW, Kim HS, Smithies O, Maeda N, Cameron VA (2002) Ventricular expression of natriuretic peptides in Nrp1−/− mice with cardiac hypertrophy and fibrosis. Am J Physiol Heart Circ Physiol 283:707–714Google Scholar
  25. 25.
    Bellomo D, Headrick JP, Silins GU, Paterson CA, Thomas PS, Gartside M, Mould A, Cahill MM, Tonks ID, Grimmond SM, Townson S, Wells C, Little M, Cummings MC, Hayward NK, Kay GF (2000) Mice lacking the vascular endothelial growth factor-B gene (Vegfb) have smaller hearts, dysfunctional coronary vasculature, and impaired recovery from cardiac ischemia. Circ Res 86:e29–e35PubMedCrossRefGoogle Scholar
  26. 26.
    Van den Akker NM, Winkel LC, Nisancioglu MH, Maas S, Wisse LJ, Armulik A, Poelmann RE, Lie-Venema H, Betsholtz C, Gittenberger-de Groot AC (2008) PDGF-B signaling is important for murine cardiac development: its role in developing atrioventricular valves, coronaries, and cardiac innervation. Dev Dyn 237(2):494–503PubMedCrossRefGoogle Scholar
  27. 27.
    Nyström HC, Lindblom P, Wickman A, Andersson I, Norlin J, Fäldt J, Lindahl P, Skøtt O, Bjarnegård M, Fitzgerald SM, Caidahl K, Gan LM, Betsholz C, Bergström G (2006) Platelet-derived growth factor B retention is essential for development of normal structure and function of conduit vessels and capillaries. Cardiovasc Res 71(3):557–565PubMedCrossRefGoogle Scholar
  28. 28.
    Bjarnegård M, Enge M, Norlin J, Gustafsdottir S, Fredriksson S, Abramsson A, Takemoto M, Gustafsson E, Fässler R, Betsholtz C (2004) Endothelium-specific ablation of PDGFB leads to pericyte loss and glomerular, cardiac and placental abnormalities. Development 131(8):1847–1857PubMedCrossRefGoogle Scholar
  29. 29.
    Hudlicka O, Brown MD (1996) Postnatal growth of the heart and its blood vessels. J Vasc Res 33:266–287PubMedCrossRefGoogle Scholar
  30. 30.
    Poolman RA, Li JM, Durand B, Brooks G (1999) Altered expression of cell cycle proteins and prolonged duration of cardiac myocyte hyperplasia in p27KIP1 knockout mice. Circ Res 85:117–127PubMedCrossRefGoogle Scholar
  31. 31.
    Jackson T, Allard MF, Sreenan CM, Doss LK, Bishop SP, Swain JL (1990) The c-myc proto-oncogene regulates cardiac development in transgenic mice. Mol Cell Biol 10:3709–3716PubMedGoogle Scholar
  32. 32.
    Brutsaert DL (2003) Cardiac endothelial-myocardial signaling: its role in cardiac growth, contractile performance, and rhythmicity. Physiol Rev 83:59–115PubMedGoogle Scholar
  33. 33.
    Tirziu D, Chorianopoulos E, Moodie KL, Palac RT, Zhuang ZW, Tjwa M, Roncal C, Eriksson U, Fu Q, Elfenbein A, Hall AE, Carmeliet P, Moons L, Simons M (2007) Myocardial hypertrophy in the absence of external stimuli is induced by angiogenesis in mice. J Clin Invest 117:3188–3197PubMedCrossRefGoogle Scholar
  34. 34.
    Shiojima I, Sato K, Izumiya Y, Schiekofer S, Ito M, Liao R, Colucci WS, Walsh K (2005) Disruption of coordinated cardiac hypertrophy and angiogenesis contributes to the transition to heart failure. J Clin Invest 115(8):2059–2064CrossRefGoogle Scholar
  35. 35.
    Satterwhite CM, Farrelly AM, Bradley ME (1999) Chemotactic, mitogenic, and angiogenic actions of UTP on vascular endothelial cells. Am J Physiol 276:1091–1097Google Scholar
  36. 36.
    Kaczmarek E, Erb L, Koziak K, Jarzyna R, Wink MR, Guckelberger O, Blusztajn JK, Trinkaus-Randall V, Weisman GA, Robson SC (2005) Modulation of endothelial cell migration by extracellular nucleotides. Involvement of focal adhesion kinase and phosphatidylinositol 3-kinase-mediated pathways. Thromb Haemost 93(4):735–742PubMedGoogle Scholar
  37. 37.
    Gerasimovskaya EV, Woodward HN, Tucker DA, Stenmark KR (2008) Extracellular ATP is a pro-angiogenic factor for pulmonary artery vasa vasorum endothelial cells. Angiogenesis 11:169–182PubMedCrossRefGoogle Scholar
  38. 38.
    Lyubchenko T, Woodward H, Veo KD, Burns N, Nijmeh H, Liubchenko GA, Stenmark KR, Gerasimovskaya EV (2011) P2Y1 and P2Y13 receptors mediate Ca2+ signaling and proliferative responses in pulmonary artery vasa vasorum endothelial cells. Am J Physiol Cell Physiol 300:C266–C275PubMedCrossRefGoogle Scholar
  39. 39.
    Guns PJ, Korda A, Crauwels HM, Van Assche T, Robaye B, Boeynaems JM, Bult H (2005) Pharmacological characterization of nucleotide P2Y receptors on endothelial cells of the mouse aorta. Br J Pharmacol 146(2):288–295PubMedCrossRefGoogle Scholar
  40. 40.
    Goepfert C, Sundberg C, Sevigny J, Enjyoji K, Hoshi T, Csizmadia E, Robson E (2001) Disordered cellular migration and angiogenesis in cd39-null mice. Circulation 104(25):3109–3115PubMedCrossRefGoogle Scholar
  41. 41.
    Wihlborg AK, Balogh J, Wang L, Borna C, Dou Y, Joshi BV, Lazarowski E, Jacobson KA, Arner A, Erlinge D (2006) Positive inotropic effects by uridine triphosphate (UTP) and uridine diphosphate (UDP) via P2Y2 and P2Y6 receptors on cardiomyocytes and release of UTP in man during myocardial infarction. Circ Res 98:970–976PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Michael Horckmans
    • 1
  • Bernard Robaye
    • 2
  • Elvira Léon-Gόmez
    • 3
  • Nicolas Lantz
    • 4
  • Philippe Unger
    • 5
  • Frédérique Dol-Gleizes
    • 6
  • Sophie Clouet
    • 1
  • Dorothée Cammarata
    • 2
  • Paul Schaeffer
    • 6
  • Pierre Savi
    • 6
  • Christian Gachet
    • 4
  • Jean-Luc Balligand
    • 3
  • Chantal Dessy
    • 3
  • Jean-Marie Boeynaems
    • 1
    • 7
  • Didier Communi
    • 1
    Email author
  1. 1.Institute of Interdisciplinary Research, IRIBHMUniversité Libre de BruxellesBrusselsBelgium
  2. 2.Institute of Interdisciplinary Research, IRIBHMUniversité Libre de BruxellesGosseliesBelgium
  3. 3.Unit of Pharmacology and TherapeuticsUniversité catholique de LouvainBrusselsBelgium
  4. 4.Department Biologie et pharmacologie des plaquettes sanguines: hémostase, thrombose, transfusionUMR_S949 INSERM-Université de Strasbourg, Etablissement Français du Sang-AlsaceStrasbourgFrance
  5. 5.Department of Cardiology, Erasme HospitalUniversité Libre de BruxellesBrusselsBelgium
  6. 6.Département Thrombose and AngiogenèseService de Pharmacologie, Sanofi-AventisToulouseFrance
  7. 7.Department of Laboratory Medicine, Erasme HospitalUniversité Libre de BruxellesBrusselsBelgium

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