Knockout and Knock-in Mouse Models to Study Purinergic Signaling

  • Robin M. H. Rumney
  • Dariusz C. GóreckiEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2041)


Purinergic signaling involves extracellular purines and pyrimidines acting upon specific cell surface purinoceptors classified into the P1, P2X, and P2Y families for nucleosides and nucleotides. This widespread signaling mechanism is active in all major tissues and influences a range of functions in health and disease. Orthologs to all but one of the human purinoceptors have been found in mouse, making this laboratory animal a useful model to study their function. Indeed, analyses of purinoceptors via knock-in or knockout approaches to produce gain or loss of function phenotypes have revealed several important therapeutic targets. None of the homozygous purinoceptor knockouts proved to be developmentally lethal, which suggest that either these receptors are not involved in key developmental processes or that the large number of receptors in each family allowed for functional compensation. Different models for the same purinoceptor often show compatible phenotypes but there have been examples of significant discrepancies. These revealed unexpected differences in the structure of human and mouse genes and emphasized the importance of the genetic background of different mouse strains. In this chapter, we provide an overview of the current knowledge and new trends in the modifications of purinoceptor genes in vivo. We discuss the resulting phenotypes, their applications and relative merits and limitations of mouse models available to study purinoceptor subtypes.

Key words

Knock-in Knockout Genetically modified animals Purinergic signaling Purinoceptor 



The authors would like to acknowledge the Polish Ministry of National Defence project “Kościuszko” no: 523/2017/DA and the EU COST Program (BM1406).


  1. 1.
    Drury AN, Szent-Györgyi A (1929) The physiological activity of adenine compounds with especial reference to their action upon the mammalian heart. J Physiol 68:213–237PubMedPubMedCentralCrossRefGoogle Scholar
  2. 2.
    Burnstock G (1972) Purinergic nerves. Pharmacol Rev 24:509–581PubMedGoogle Scholar
  3. 3.
    Webb TE, Simon J, Krishek BJ et al (1993) Cloning and functional expression of a brain G-protein-coupled ATP receptor. FEBS Lett 324:219–225PubMedCrossRefGoogle Scholar
  4. 4.
    Ralevic V, Burnstock G (1998) Receptors for purines and pyrimidines. Pharmacol Rev 50:413–492PubMedGoogle Scholar
  5. 5.
    Abbracchio MP (2006) International Union of Pharmacology. LVIII: Update on the P2Y G protein-coupled nucleotide receptors: from molecular mechanisms and pathophysiology to therapy. Pharmacol Rev 58:281–341PubMedPubMedCentralCrossRefGoogle Scholar
  6. 6.
    Khakh BS, Burnstock G, Kennedy CL et al (2001) International union of pharmacology. XXIV. Current status of the nomenclature and properties of P2X receptors and their subunits. Pharmacol Rev 53:107–118PubMedGoogle Scholar
  7. 7.
    Fredholm BB, IJzerman AP, Jacobson KA et al (2001) International Union of Pharmacology. XXV. Nomenclature and classification of adenosine receptors. Pharmacol Rev 53:527–552PubMedGoogle Scholar
  8. 8.
    Herbert JM, Savi P (2003) P2Y12, a new platelet ADP receptor, target of clopidogrel. Semin Vasc Med 3:113–122PubMedCrossRefGoogle Scholar
  9. 9.
    Fountain SJ, Parkinson K, Young MT et al (2007) An intracellular P2X receptor required for osmoregulation in Dictyostelium discoideum. Nature 448:200–203PubMedPubMedCentralCrossRefGoogle Scholar
  10. 10.
    Burnstock G (2007) Purine and pyrimidine receptors. Cell Mol Life Sci 64:1471–1483PubMedCrossRefGoogle Scholar
  11. 11.
    Evans MJ, Kaufman MH (1981) Establishment in culture of pluripotential cells from mouse embryos. Nature 292:154–156CrossRefGoogle Scholar
  12. 12.
    Bradley A, Evans M, Kaufman MH et al (1984) Formation of germ-line chimaeras from embryo-derived teratocarcinoma cell lines. Nature 309:255–256PubMedCrossRefGoogle Scholar
  13. 13.
    Mansour SL, Thomas KR, Capecchi MR (1988) Disruption of the proto-oncogene int-2 in mouse embryo-derived stem cells: a general strategy for targeting mutations to non-selectable genes. Nature 31:681–683Google Scholar
  14. 14.
    Li D, Qiu Z, Shao Y et al (2013) Heritable gene targeting in the mouse and rat using a CRISPR-Cas system. Nat Biotechnol 31:681–683CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Metzger D, Clifford J, Chiba H et al (1995) Conditional site-specific recombination in mammalian cells using a ligand-dependent chimeric Cre recombinase. Proc Natl Acad Sci U S A 92:6991–6995PubMedPubMedCentralCrossRefGoogle Scholar
  16. 16.
    Dymecki SM (1996) A modular set of Flp, FRT and lacZ fusion vectors for manipulating genes by site-specific recombination. Gene 171:197–201PubMedCrossRefGoogle Scholar
  17. 17.
    Chaiyachati BH, Kaundal R, Zhao J et al (2012) LoxP-FRT Trap (LOFT): a simple and flexible system for conventional and reversible gene targeting. BMC Biol 10:96PubMedPubMedCentralCrossRefGoogle Scholar
  18. 18.
    Geurts AM, Cost GJ, Freyvert Y et al (2009) Knockout rats via embryo microinjection of zinc-finger nucleases. Science 325:433PubMedPubMedCentralCrossRefGoogle Scholar
  19. 19.
    Kitada K, Keng VW, Takeda J et al (2009) Generating mutant rats using the Sleeping Beauty transposon system. Methods 49:236–242PubMedCrossRefGoogle Scholar
  20. 20.
    Furushima K, Jang CW, Chen DW et al (2012) Insertional mutagenesis by a hybrid piggyBac and Sleeping Beauty transposon in the rat. Genetics 192:1235–1248PubMedPubMedCentralCrossRefGoogle Scholar
  21. 21.
    Carter M, Shieh J, Carter M et al (2010) Making and using transgenic organisms. In: Guide to research techniques in neuroscience. Elsevier B.V., Amsterdam, pp 243–262CrossRefGoogle Scholar
  22. 22.
    Bouabe H, Okkenhaug K (2013) Gene targeting in mice: a review. Methods Mol Biol 1064:315–336PubMedPubMedCentralCrossRefGoogle Scholar
  23. 23.
    Eisener-Dorman AF, Lawrence DA, Bolivar VJ (2009) Cautionary insights on knockout mouse studies: the gene or not the gene? Brain Behav Immun 23:318–324PubMedCrossRefGoogle Scholar
  24. 24.
    Guan C, Ye C, Yang X et al (2010) A review of current large-scale mouse knockout efforts. Genesis 48:73–85PubMedGoogle Scholar
  25. 25.
    Matherne GP, Linden J, Byford AM et al (1997) Transgenic A1 adenosine receptor overexpression increases myocardial resistance to ischemia. Proc Natl Acad Sci U S A 94:6541–6546PubMedPubMedCentralCrossRefGoogle Scholar
  26. 26.
    Johansson B, Halldner L, Dunwiddie TV et al (2001) Hyperalgesia, anxiety, and decreased hypoxic neuroprotection in mice lacking the adenosine A1 receptor. Proc Natl Acad Sci U S A 98:9407–9412PubMedPubMedCentralCrossRefGoogle Scholar
  27. 27.
    Sun D, Samuelson LC, Yang T et al (2001) Mediation of tubuloglomerular feedback by adenosine: evidence from mice lacking adenosine 1 receptors. Proc Natl Acad Sci U S A 98:9983–9988PubMedPubMedCentralCrossRefGoogle Scholar
  28. 28.
    Funakoshi H, Chan TO, Good JC et al (2006) Regulated overexpression of the A1-adenosine receptor in mice results in adverse but reversible changes in cardiac morphology and function. Circulation 114:2240–2250PubMedCrossRefGoogle Scholar
  29. 29.
    Serchov T, Clement HW, Schwarz MK et al (2015) Increased signaling via adenosine A1 receptors, sleep deprivation, imipramine, and ketamine inhibit depressive-like behavior via induction of homer1a. Neuron 87:549–562PubMedPubMedCentralCrossRefGoogle Scholar
  30. 30.
    Ledent C, Vaugeois JM, Schiffmann SN et al (1997) Aggressiveness, hypoalgesia and high blood pressure in mice lacking the adenosine A2a receptor. Nature 388:674–678PubMedCrossRefGoogle Scholar
  31. 31.
    Chen JF, Huang Z, Ma J et al (1999) A2A adenosine receptor deficiency attenuates brain injury induced by transient focal ischemia in mice. J Neurosci 19:9192–9200PubMedPubMedCentralCrossRefGoogle Scholar
  32. 32.
    Giménez-Llort L, Schiffmann SN, Shmidt T et al (2007) Working memory deficits in transgenic rats overexpressing human adenosine A2A receptors in the brain. Neurobiol Learn Mem 87:42–56PubMedCrossRefGoogle Scholar
  33. 33.
    Boknik P, Drzewiecki K, Eskandar J et al (2018) Phenotyping of mice with heart specific overexpression of A2A-adenosine receptors: evidence for cardioprotective effects of A2A-adenosine receptors. Front Pharmacol 9:13PubMedPubMedCentralCrossRefGoogle Scholar
  34. 34.
    Yang D, Zhang Y, Nguyen HG et al (2006) The A2B adenosine receptor protects against inflammation and excessive vascular adhesion. J Clin Invest 116:1913–1923PubMedPubMedCentralCrossRefGoogle Scholar
  35. 35.
    Belikoff BG, Hatfield S, Georgiev P et al (2011) A2B adenosine receptor blockade enhances macrophage-mediated bacterial phagocytosis and improves polymicrobial sepsis survival in mice. J Immunol 275:4429–4434Google Scholar
  36. 36.
    Salvatore CA, Tilley SL, Latour AM et al (2000) Disruption of the A(3) adenosine receptor gene in mice and its effect on stimulated inflammatory cells. J Biol Chem 275:4429–4434PubMedCrossRefGoogle Scholar
  37. 37.
    Zhao Z, Yaar R, Ladd D et al (2002) Overexpression of A3 adenosine receptors in smooth, cardiac, and skeletal muscle is lethal to embryos. Microvasc Res 63:61–69PubMedCrossRefGoogle Scholar
  38. 38.
    Brown R, Ollerstam A, Johansson B et al (2001) Abolished tubuloglomerular feedback and increased plasma renin in adenosine A1 receptor-deficient mice. Am J Physiol Regul Integr Comp Physiol 281:R1362–R1367PubMedGoogle Scholar
  39. 39.
    Lee HT, Xu H, Nasr SH et al (2004) A1 adenosine receptor knockout mice exhibit increased renal injury following ischemia and reperfusion. Am J Physiol Ren Physiol 286:F298–F306CrossRefGoogle Scholar
  40. 40.
    Kim M, Chen SWC, Park SW et al (2009) Kidney-specific reconstitution of the A1 adenosine receptor in A1 adenosine receptor knockout mice reduces renal ischemia-reperfusion injury. Kidney Int 75:809–823PubMedPubMedCentralCrossRefGoogle Scholar
  41. 41.
    Schweda F, Segerer F, Castrop H et al (2005) Blood pressure-dependent inhibition of renin secretion requires A1 adenosine receptors. Hypertension 46:780–786PubMedCrossRefGoogle Scholar
  42. 42.
    Koeppen M, Eckle T, Eltzschig HK (2009) Selective deletion of the A1 adenosine receptor abolishes heart-rate slowing effects of intravascular adenosine in vivo. PLoS One 4:e6784PubMedPubMedCentralCrossRefGoogle Scholar
  43. 43.
    Kochanek PM, Vagni VA, Janesko KL et al (2006) Adenosine A1 receptor knockout mice develop lethal status epilepticus after experimental traumatic brain injury. J Cereb Blood Flow Metab 26:565–575PubMedCrossRefGoogle Scholar
  44. 44.
    Zucchi R, Cerniway RJ, Ronca-Testoni S et al (2002) Effect of cardiac A1 adenosine receptor overexpression on sarcoplasmic reticulum function. Cardiovasc Res 53:326–333PubMedCrossRefGoogle Scholar
  45. 45.
    Chan TO, Funakoshi H, Song J et al (2008) Cardiac-restricted overexpression of the A2A-adenosine receptor in FVB mice transiently increases contractile performance and rescues the heart failure phenotype in mice overexpressing the A1-adenosine receptor. Clin Transl Sci 1:126–133PubMedPubMedCentralCrossRefGoogle Scholar
  46. 46.
    Chiodi V, Ferrante A, Ferraro L et al (2016) Striatal adenosine-cannabinoid receptor interactions in rats over-expressing adenosine A2A receptors. J Neurochem 136:907–917PubMedCrossRefGoogle Scholar
  47. 47.
    Domenici MR, Chiodi V, Averna M et al (2018) Neuronal adenosine A2Areceptor overexpression is neuroprotective towards 3-nitropropionic acid-induced striatal toxicity: a rat model of Huntington’s disease. Purinergic Signal 14:235–243PubMedPubMedCentralCrossRefGoogle Scholar
  48. 48.
    Johnston-Cox H, Koupenova M, Yang D et al (2012) The A2b adenosine receptor modulates glucose homeostasis and obesity. PLoS One 7:e40584PubMedPubMedCentralCrossRefGoogle Scholar
  49. 49.
    Eisenstein A, Carroll SH, Johnston-Cox H et al (2014) An adenosine receptor-Krüppel-like factor 4 protein axis inhibits adipogenesis. J Biol Chem 289:21071–21081PubMedPubMedCentralCrossRefGoogle Scholar
  50. 50.
    Koupenova M, Johnston-Cox H, Vezeridis A et al (2012) A2b adenosine receptor regulates hyperlipidemia and atherosclerosis. Circulation 125:354–363PubMedCrossRefGoogle Scholar
  51. 51.
    Zhong H, Shlykov SG, Molina JG et al (2003) Activation of murine lung mast cells by the adenosine A3 receptor. J Immunol 53:147–155Google Scholar
  52. 52.
    Harrison G (2002) Effects of A3 adenosine receptor activation and gene knock-out in ischemic-reperfused mouse heart. Cardiovasc Res 53:147–155PubMedCrossRefGoogle Scholar
  53. 53.
    Cerniway RJ, Yang Z, Jacobson MA et al (2001) Targeted deletion of A(3) adenosine receptors improves tolerance to ischemia-reperfusion injury in mouse myocardium. Am J Physiol Heart Circ Physiol 281:H1751–H1758PubMedCrossRefGoogle Scholar
  54. 54.
    Lee HT, Ota-Setlik A, Xu H et al (2003) A3 adenosine receptor knockout mice are protected against ischemia- and myoglobinuria-induced renal failure. Am J Physiol Renal Physiol 284:F267–F273PubMedCrossRefGoogle Scholar
  55. 55.
    Yang T, Zollbrecht C, Winerdal ME et al (2016) Genetic abrogation of adenosine A3 receptor prevents uninephrectomy and high salt-induced hypertension. J Am Heart Assoc 5:e003868PubMedPubMedCentralGoogle Scholar
  56. 56.
    Hofer M, Pospíšil M, Dušek L et al (2014) Lack of adenosine A3 receptors causes defects in mouse peripheral blood parameters. Purinergic Signal 10:509–514PubMedPubMedCentralCrossRefGoogle Scholar
  57. 57.
    Hofer M, Pospíšil M, Dušek L et al (2013) Erythropoiesis-and thrombopoiesis-characterizing parameters in adenosine A3 receptor knock-out mice. Physiol Res 62:305–311PubMedGoogle Scholar
  58. 58.
    Chen Y, Corriden R, Inoue Y et al (2006) ATP release guides neutrophil chemotaxis via P2Y2 and A3 receptors. Science 314:1792–1795PubMedCrossRefGoogle Scholar
  59. 59.
    Joós G, Jákim J, Kiss B et al (2017) Involvement of adenosine A3 receptors in the chemotactic navigation of macrophages towards apoptotic cells. Immunol Lett 183:62–72PubMedCrossRefGoogle Scholar
  60. 60.
    Fedorova IM, Jacobson MA, Basile A et al (2003) Behavioral characterization of mice lacking the A3 adenosine receptor: sensitivity to hypoxic neurodegeneration. Cell Mol Neurobiol 23:431–447PubMedPubMedCentralCrossRefGoogle Scholar
  61. 61.
    Björklund O, Shang M, Tonazzini I et al (2008) Adenosine A1and A3receptors protect astrocytes from hypoxic damage. Eur J Pharmacol 596:6–13PubMedCrossRefGoogle Scholar
  62. 62.
    Rosito M, Deflorio C, Limatola C et al (2012) CXCL16 orchestrates adenosine A3 receptor and MCP-1/CCL2 activity to protect neurons from excitotoxic cell death in the CNS. J Neurosci 32:3154–3163PubMedPubMedCentralCrossRefGoogle Scholar
  63. 63.
    Björklund O, Halldner-Henriksson L, Yang J et al (2008) Decreased behavioral activation following caffeine, amphetamine and darkness in A3 adenosine receptor knock-out mice. Physiol Behav 95:668–676PubMedCrossRefGoogle Scholar
  64. 64.
    Little JW, Ford A, Symons-Liguori AM et al (2015) Endogenous adenosine A3receptor activation selectively alleviates persistent pain states. Brain 138:28–35PubMedCrossRefGoogle Scholar
  65. 65.
    Mulryan K, Gitterman DP, Lewis CJ et al (2000) Reduced vas deferens contraction and male infertility in mice lacking P2X1 receptors. Nature 183:2801–2809Google Scholar
  66. 66.
    Cockayne DA, Dunn PM, Zhong Y et al (2005) P2X2 knockout mice and P2X2/P2X3 double knockout mice reveal a role for the P2X2 receptor subunit in mediating multiple sensory effects of ATP. J Physiol 567:621–639PubMedPubMedCentralCrossRefGoogle Scholar
  67. 67.
    Cockayne DA, Hamilton SG, Zhu QM et al (2000) Urinary bladder hyporeflexia and reduced pain-related behaviour in P2X3-deficient mice. Nature 407:1011–1015PubMedCrossRefGoogle Scholar
  68. 68.
    Sim JA (2006) Altered hippocampal synaptic potentiation in P2X4 knock-out mice. J Neurosci 26:9006–9009PubMedPubMedCentralCrossRefGoogle Scholar
  69. 69.
    Yamamoto K, Sokabe T, Matsumoto T et al (2006) Impaired flow-dependent control of vascular tone and remodeling in P2X4-deficient mice. Nat Med 12:133–137PubMedCrossRefGoogle Scholar
  70. 70.
    Brône B, Moechars D, Marrannes R et al (2007) P2X currents in peritoneal macrophages of wild type and P2X4−/− mice. Immunol Lett 113:83–89PubMedCrossRefGoogle Scholar
  71. 71.
    Yang T, Shen JB, Yang R et al (2014) Novel protective role of endogenous cardiac myocyte P2X4 receptors in heart failure. Circ Heart Fail 7:510–518PubMedPubMedCentralCrossRefGoogle Scholar
  72. 72.
    Kim H, Walsh MC, Takegahara N et al (2017) The purinergic receptor P2X5 regulates inflammasome activity and hyper-multinucleation of murine osteoclasts. Sci Rep 7:196PubMedPubMedCentralCrossRefGoogle Scholar
  73. 73.
    de Baaij JHF, Kompatscher A, Viering DHHM et al (2016) P2X6 knockout mice exhibit normal electrolyte homeostasis. PLoS One 11:e0156803PubMedPubMedCentralCrossRefGoogle Scholar
  74. 74.
    Solle M, Labasi J, Perregaux DG et al (2001) Altered cytokine production in mice lacking P2X(7) receptors. J Biol Chem 276:125–132PubMedCrossRefGoogle Scholar
  75. 75.
    Chessell IP, Hatcher JP, Bountra C et al (2005) Disruption of the P2X7 purinoceptor gene abolishes chronic inflammatory and neuropathic pain. Pain 114:386–396PubMedCrossRefGoogle Scholar
  76. 76.
    Metzger MW, Walser SM, Aprile-Garcia F et al (2017) Genetically dissecting P2rx7 expression within the central nervous system using conditional humanized mice. Purinergic Signal 13:153–170PubMedCrossRefGoogle Scholar
  77. 77.
    Kaczmarek-Hajek K, Zhang J, Kopp R et al (2018) Re-evaluation of neuronal P2X7 expression using novel mouse models and a P2X7-specific nanobody. Elife 7:e36217PubMedPubMedCentralCrossRefGoogle Scholar
  78. 78.
    Lecut C, Frederix K, Johnson DM et al (2009) P2X1 ion channels promote neutrophil chemotaxis through rho kinase activation. J Immunol 183:2801–2809PubMedCrossRefGoogle Scholar
  79. 79.
    Lecut C, Faccinetto C, Delierneux C et al (2012) ATP-gated P2X1 ion channels protect against endotoxemia by dampening neutrophil activation. J Thromb Haemost 10:453–465PubMedCrossRefGoogle Scholar
  80. 80.
    Ren J, Bian X, DeVries M et al (2003) P2X2 subunits contribute to fast synaptic excitation in myenteric neurons of the mouse small intestine. J Physiol 552:809–821PubMedPubMedCentralCrossRefGoogle Scholar
  81. 81.
    Ryten M, Koshi R, Knight GE et al (2007) Abnormalities in neuromuscular junction structure and skeletal muscle function in mice lacking the P2X2 nucleotide receptor. Neuroscience 148:700–711PubMedCrossRefGoogle Scholar
  82. 82.
    Rong W, Gourine AV, Cockayne DA et al (2003) Pivotal role of nucleotide P2X2 receptor subunit of the ATP-gated ion channel mediating ventilatory responses to hypoxia. J Neurosci 23:11315–11321PubMedPubMedCentralCrossRefGoogle Scholar
  83. 83.
    Zhong Y, Dunn PM, Bardini M et al (2001) Changes in P2X receptor responses of sensory neurons from P2X3-deficient mice. Eur J Neurosci 14:1784–1792PubMedCrossRefGoogle Scholar
  84. 84.
    Bian X, Ren J, DeVries M et al (2003) Peristalsis is impaired in the small intestine of mice lacking the P2X3 subunit. J Physiol 551:309–322PubMedPubMedCentralCrossRefGoogle Scholar
  85. 85.
    McIlwrath SL, Davis BM, Bielefeldt K (2009) Deletion of P2X3 receptors blunts gastro-oesophageal sensation in mice. Neurogastroenterol Motil 21:890–e66PubMedPubMedCentralCrossRefGoogle Scholar
  86. 86.
    Eddy MC, Eschle BK, Barrows J et al (2009) Double P2X2/P2X3 purinergic receptor knockout mice do not taste NaCl or the artificial sweetener SC45647. Chem Senses 34:789–797PubMedPubMedCentralCrossRefGoogle Scholar
  87. 87.
    Sclafani A, Ackroff K (2014) Maltodextrin and fat preference deficits in “taste-blind” P2X2/P2X3 knockout mice. Chem Senses 39:507–514PubMedPubMedCentralCrossRefGoogle Scholar
  88. 88.
    Baxter AW, Choi SJ, Sim JA et al (2011) Role of P2X4 receptors in synaptic strengthening in mouse CA1 hippocampal neurons. Eur J Neurosci 34:213–220PubMedPubMedCentralCrossRefGoogle Scholar
  89. 89.
    Ulmann L, Levavasseur F, Avignone E et al (2013) Involvement of P2X4 receptors in hippocampal microglial activation after status epilepticus. Glia 61:1306–1319PubMedCrossRefGoogle Scholar
  90. 90.
    Khoja S, Huynh N, Asatryan L et al (2018) Reduced expression of purinergic P2X4 receptors increases voluntary ethanol intake in C57BL/6J mice. Alcohol 68:63–70PubMedCrossRefGoogle Scholar
  91. 91.
    Wyatt LR, Finn DA, Khoja S et al (2014) Contribution of P2X4 receptors to ethanol intake in male C57BL/6 mice. Neurochem Res 39:1127–1139PubMedPubMedCentralCrossRefGoogle Scholar
  92. 92.
    Wyatt LR, Godar SC, Khoja S et al (2013) Sociocommunicative and sensorimotor impairments in male P2X4-deficient mice. Neuropsychopharmacology 38:1993–2002PubMedPubMedCentralCrossRefGoogle Scholar
  93. 93.
    Ulmann L, Hatcher JP, Hughes JP et al (2008) Up-regulation of P2X4 receptors in spinal microglia after peripheral nerve injury mediates BDNF release and neuropathic pain. J Neurosci 28:11263–11268PubMedPubMedCentralCrossRefGoogle Scholar
  94. 94.
    Ulmann L, Hirbec H, Rassendren F (2010) P2X4 receptors mediate PGE2 release by tissue-resident macrophages and initiate inflammatory pain. EMBO J 29:2290–2300PubMedPubMedCentralCrossRefGoogle Scholar
  95. 95.
    Tsuda M, Kuboyama K, Inoue T et al (2009) Behavioral phenotypes of mice lacking purinergic P2X4 receptors in acute and chronic pain assays. Mol Pain 5:28PubMedPubMedCentralCrossRefGoogle Scholar
  96. 96.
    Yan Z, Khadra A, Li S et al (2010) Experimental characterization and mathematical modeling of P2X7 receptor channel gating. J Neurosci 30:14213–14224PubMedPubMedCentralCrossRefGoogle Scholar
  97. 97.
    Di Virgilio F, Dal Ben D, Sarti AC et al (2017) The P2X7 receptor in infection and inflammation. Immunity 47(1):15–31PubMedCrossRefGoogle Scholar
  98. 98.
    Young CNJ, Gorecki DC (2017) P2RX7 purinoceptor as a therapeutic target – the second coming? Front Chem 6:248CrossRefGoogle Scholar
  99. 99.
    Di Virgilio F, Schmalzing G, Markwardt F (2018) The elusive P2X7 macropore. Trends Cell Biol 28:392–404PubMedCrossRefGoogle Scholar
  100. 100.
    Adriouch S, Dox C, Welge V et al (2002) Cutting edge: a natural P451L mutation in the cytoplasmic domain impairs the function of the mouse P2X7 receptor. J Immunol 169:4108–4112PubMedCrossRefGoogle Scholar
  101. 101.
    Nicke A, Kuan YH, Masin M et al (2009) A functional P2X7 splice variant with an alternative transmembrane domain 1 escapes gene inactivation in P2X7 knock-out mice. J Biol Chem 284:25813–25822PubMedPubMedCentralCrossRefGoogle Scholar
  102. 102.
    Ke HZ, Qi H, Weidema AF et al (2003) Deletion of the P2X 7 nucleotide receptor reveals its regulatory roles in bone formation and resorption. Mol Endocrinol 17:1356–1367PubMedCrossRefGoogle Scholar
  103. 103.
    Li J, Liu D, Ke HZ et al (2005) The P2X7 nucleotide receptor mediates skeletal mechanotransduction. J Biol Chem 13:243–253Google Scholar
  104. 104.
    Gartland A, Buckley KA, Hipskind RA et al (2003) Multinucleated osteoclast formation in vivo and in vitro by P2X7 receptor-deficient mice. Crit Rev Eukaryot Gene Expr 13:243–253PubMedGoogle Scholar
  105. 105.
    Syberg S, Schwarz P, Petersen S et al (2012) Association between P2X7 receptor polymorphisms and bone status in mice. J Osteoporos 2012:637986PubMedPubMedCentralGoogle Scholar
  106. 106.
    Syberg S, Petersen S, Beck Jensen JE et al (2012) Genetic background strongly influences the bone phenotype of P2X7 receptor knockout mice. J Osteoporos 2012:391097PubMedPubMedCentralGoogle Scholar
  107. 107.
    Grygorowicz T, Dąbrowska-Bouta B, Strużyńska L (2018) Administration of an antagonist of P2X7 receptor to EAE rats prevents a decrease of expression of claudin-5 in cerebral capillaries. Purinergic Signal 14(4):385–393PubMedPubMedCentralCrossRefGoogle Scholar
  108. 108.
    Fabre JE, Nguyen M, Latour A et al (1999) Decreased platelet aggregation, increased bleeding time and resistance to thromboembolism in P2Y1-deficient mice. Nat Med 5:1199–1202PubMedCrossRefGoogle Scholar
  109. 109.
    Léon C, Hechler B, Freund M et al (1999) Defective platelet aggregation and increased resistance to thrombosis in purinergic P2Y1receptor-null mice. J Clin Invest 104:1731–1737PubMedPubMedCentralCrossRefGoogle Scholar
  110. 110.
    Hwang SJ, Blair PJ, Durnin L et al (2012) P2Y1 purinoreceptors are fundamental to inhibitory motor control of murine colonic excitability and transit. J Physiol 590:1957–1972PubMedPubMedCentralCrossRefGoogle Scholar
  111. 111.
    Homolya L, Watt WC, Lazarowski ER et al (1999) Nucleotide-regulated calcium signaling in lung fibroblasts and epithelial cells from normal and P2Y2 receptor (−/−) mice. J Biol Chem 274:26454–26460PubMedCrossRefGoogle Scholar
  112. 112.
    Robaye B, Ghanem E, Wilkin F et al (2003) Loss of nucleotide regulation of epithelial chloride transport in the jejunum of P2Y4-null mice. Mol Pharmacol 63:777–783PubMedCrossRefGoogle Scholar
  113. 113.
    Bar I, Guns P-J, Metallo J et al (2008) Knockout mice reveal a role for P2Y6 receptor in macrophages, endothelial cells, and vascular smooth muscle cells. Mol Pharmacol 74:777–784PubMedCrossRefGoogle Scholar
  114. 114.
    Giannattasio G, Ohta S, Boyce JR et al (2011) The P2Y6 receptor inhibits effector T cell activation in allergic pulmonary inflammation. J Immunol 187:1486–1495PubMedPubMedCentralCrossRefGoogle Scholar
  115. 115.
    Garcia RA, Yan M, Search D et al (2014) P2Y6receptor potentiates pro-inflammatory responses in macrophages and exhibits differential roles in atherosclerotic lesion development. PLoS One 9:e111385PubMedPubMedCentralCrossRefGoogle Scholar
  116. 116.
    Li R, Tan B, Yan Y et al (2014) Extracellular UDP and P2Y6 function as a danger signal to protect mice from vesicular stomatitis virus infection through an increase in IFN-β production. J Immunol 193:4515–4526PubMedCrossRefGoogle Scholar
  117. 117.
    Foster CJ, Prosser DM, Agans JM et al (2001) Molecular identification and characterization of the platelet ADP receptor targeted by thienopyridine antithrombotic drugs. J Clin Invest 107:1591–1598PubMedPubMedCentralCrossRefGoogle Scholar
  118. 118.
    André P, Delaney SM, LaRocca T et al (2003) P2Y12regulates platelet adhesion/activation, thrombus growth, and thrombus stability in injured arteries. J Clin Invest 112:398–406PubMedPubMedCentralCrossRefGoogle Scholar
  119. 119.
    Fabre AC, Malaval C, Ben Addi A et al (2010) P2Y13 receptor is critical for reverse cholesterol transport. Hepatology 52:1477–1483PubMedCrossRefGoogle Scholar
  120. 120.
    Bassil AK, Bourdu S, Townson KA et al (2009) UDP-glucose modulates gastric function through P2Y14 receptor-dependent and -independent mechanisms. Am J Physiol Gastrointest Liver Physiol 296:G923–G930PubMedCrossRefGoogle Scholar
  121. 121.
    Meister J, Le Duc D, Ricken A et al (2014) The G protein-coupled receptor P2Y14 influences insulin release and smooth muscle function in mice. J Biol Chem 289:23353–23366PubMedPubMedCentralCrossRefGoogle Scholar
  122. 122.
    Gil V, Martínez-Cutillas M, Mañé N et al (2013) P2Y1 knockout mice lack purinergic neuromuscular transmission in the antrum and cecum. Neurogastroenterol Motil 25:e170–e182PubMedCrossRefGoogle Scholar
  123. 123.
    Matos JE, Robaye B, Boeynaems JM et al (2005) K+ secretion activated by luminal P2Y2 and P2Y4 receptors in mouse colon. J Physiol 564:269–279PubMedPubMedCentralCrossRefGoogle Scholar
  124. 124.
    Horckmans M, Robaye B, Léon-Gómez E et al (2012) P2Y4 nucleotide receptor: a novel actor in post-natal cardiac development. Angiogenesis 15:349–360PubMedCrossRefGoogle Scholar
  125. 125.
    Horckmans M, Leon-Gomez E, Robaye B et al (2012) Gene deletion of P2Y4 receptor lowers exercise capacity and reduces myocardial hypertrophy with swimming exercise. AJP Heart Circ Physiol 303:H835–H843CrossRefGoogle Scholar
  126. 126.
    Kauffenstein G, Tamareille S, Prunier F et al (2016) Central role of P2Y6 UDP receptor in arteriolar myogenic tone. Arterioscler Thromb Vasc Biol 36:1598–1606PubMedPubMedCentralCrossRefGoogle Scholar
  127. 127.
    Clouet S, Di Pietrantonio L, Daskalopoulos EP et al (2016) Loss of mouse P2Y6nucleotide receptor is associated with physiological macrocardia and amplified pathological cardiac hypertrophy. J Biol Chem 291:15841–15852PubMedPubMedCentralCrossRefGoogle Scholar
  128. 128.
    Placet M, Arguin G, Molle CM et al (2018) The G protein-coupled P2Y6 receptor promotes colorectal cancer tumorigenesis by inhibiting apoptosis. Biochim Biophys Acta Mol basis Dis 1864:1539–1551PubMedCrossRefGoogle Scholar
  129. 129.
    Liverani E, Rico MC, Yaratha L et al (2014) LPS-induced systemic inflammation is more severe in P2Y12 null mice. J Leukoc Biol 95:313–323PubMedPubMedCentralCrossRefGoogle Scholar
  130. 130.
    Li D, Wang Y, Zhang L et al (2012) Roles of purinergic receptor P2Y, G protein-coupled 12 in the development of atherosclerosis in apolipoprotein E-deficient mice. Arterioscler Thromb Vasc Biol 32:e81–e89PubMedGoogle Scholar
  131. 131.
    Harada K, Matsumoto Y, Umemura K (2011) Adenosine diphosphate receptor P2Y12-mediated migration of host smooth muscle-like cells and leukocytes in the development of transplant arteriosclerosis. Transplantation 92:148–154PubMedCrossRefGoogle Scholar
  132. 132.
    Ben Addi A, Cammarata D, Conley PB et al (2010) Role of the P2Y12 receptor in the modulation of murine dendritic cell function by ADP. J Immunol 185:5900–5906PubMedCrossRefGoogle Scholar
  133. 133.
    Haynes SE, Hollopeter G, Yang G et al (2006) The P2Y12 receptor regulates microglial activation by extracellular nucleotides. Nat Neurosci 9:1512–1519PubMedCrossRefGoogle Scholar
  134. 134.
    Gu N, Eyo UB, Murugan M et al (2016) Microglial P2Y12 receptors regulate microglial activation and surveillance during neuropathic pain. Brain Behav Immun 55:82–92PubMedCrossRefGoogle Scholar
  135. 135.
    Su X, Floyd DH, Hughes A et al (2012) The ADP receptor P2RY12 regulates osteoclast function and pathologic bone remodeling. J Clin Invest 122:3579–3592PubMedPubMedCentralCrossRefGoogle Scholar
  136. 136.
    Wang Y, Sun Y, Li D et al (2013) Platelet P2Y12 is involved in murine pulmonary metastasis. PLoS One 8:e80780PubMedPubMedCentralCrossRefGoogle Scholar
  137. 137.
    Wang N, Robaye B, Agrawal A et al (2012) Reduced bone turnover in mice lacking the P2Y13 receptor of ADP. Mol Endocrinol 26:142–152PubMedCrossRefGoogle Scholar
  138. 138.
    Wang N, Rumney RMH, Yang L et al (2013) The P2Y13 receptor regulates extracellular ATP metabolism and the osteogenic response to mechanical loading. J Bone Miner Res 28:1446–1456PubMedCrossRefGoogle Scholar
  139. 139.
    Wang N, Robaye B, Gossiel F et al (2014) The P2Y13 receptor regulates phosphate metabolism and FGF-23 secretion with effects on skeletal development. FASEB J 28:2249–2259PubMedCrossRefGoogle Scholar
  140. 140.
    Kafkafi N, Benjamini Y, Sakov A et al (2005) Genotype-environment interactions in mouse behavior: a way out of the problem. Proc Natl Acad Sci U S A 102:4619–4624PubMedPubMedCentralCrossRefGoogle Scholar
  141. 141.
    Richter SH, Garner JP, Würbel H (2009) Environmental standardization: cure or cause of poor reproducibility in animal experiments? Nat Methods 6:257–261PubMedCrossRefGoogle Scholar
  142. 142.
    Karp NA, Melvin D, Mott RF (2012) Robust and sensitive analysis of mouse knockout phenotypes. PLoS One 7:e52410PubMedPubMedCentralCrossRefGoogle Scholar
  143. 143.
    Ellenbroek B, Youn J (2016) Rodent models in neuroscience research: is it a rat race? Dis Model Mech 9:1079–1087PubMedPubMedCentralCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.School of Pharmacy and Biomedical SciencesUniversity of PortsmouthPortsmouthUK
  2. 2.Military Institute of Hygiene and EpidemiologyWarsawPoland

Personalised recommendations