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Thromb Haemost 2001; 86(05): 1264-1271
DOI: 10.1055/s-0037-1616060
Review Article
Schattauer GmbH

The ATP-Gated P2X1 Ion Channel Acts as a Positive Regulator of Platelet Responses to Collagen

Cecile Oury   *
1   Center for Molecular and Vascular Biology, University of Leuven, Leuven, Belgium
,
Emese Toth-Zsamboki   *
1   Center for Molecular and Vascular Biology, University of Leuven, Leuven, Belgium
,
Chantal Thys
1   Center for Molecular and Vascular Biology, University of Leuven, Leuven, Belgium
,
Jan Tytgat
2   Laboratory of Toxicology, University of Leuven, Belgium
,
Jos Vermylen
1   Center for Molecular and Vascular Biology, University of Leuven, Leuven, Belgium
,
Marc F. Hoylaerts
1   Center for Molecular and Vascular Biology, University of Leuven, Leuven, Belgium
› Author AffiliationsETZ is involved in the Central and Eastern European Initiatives-KULeuven scholarships programme. C. O. is holder of a postdoctoral research mandate of the FWO. Financial support was obtained from the bilateral scientific and technological cooperation between Flanders and Hungary (BIL00/12) and from the FWO (project G 0376.01).
Further Information

Publication History

Received 01 March 2001

Accepted after resubmission 04 July 2001

Publication Date:
13 December 2017 (online)

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Summary

ATP is a potent agonist of the P2X1 ion channel, mediating a rapid, quickly desensitized influx of Ca2+. In hirudinized PRP, containing apyrase, the two stable selective P2X1 agonists, α,β-methylene ATP, and L-β, γ-methylene ATP induced extracellular Ca2+-dependent fast and reversible platelet shape change, leading to desensitization of the P2X1 ion channel. Preincubation with HPLC-purified ADP potently antagonized the subsequent α, β-methylene ATP- and L-β, γ-methylene ATP-evoked platelet shape change. Accordingly, upon heterologous expression of P2X1 in Xenopus oocytes, HPLC-purified ADP acted as an antagonist of the ATP-induced current, but was inactive itself. Since ATP and ADP are co-released from dense granules during platelet activation, we investigated whether the P2X1 ion channel is involved in the response of platelets to collagen. We found that platelet shape change and aggregation induced by low concentrations of collagen were strongly inhibited after selective desensitization of P2X1 with its agonists or by pretreating the platelets with a low concentration of ADP (0.5 μM), that antagonizes the P2X1 channel without desensitizing the P2Y1 receptor. Our data suggest that, during collagen-initiated platelet activation, the early secretion of ATP results in the activation of the P2X1 ion channel, which plays a role as a positive regulator of further platelet responses.

Keywords

Platelet receptor - ATP - collagen - shape change - aggregation

* considered as first authors.


 

  • References

  • 1 Reimers H-J. Adenine nucleotides in blood platelets. In: The platelets. Physiology and pharmacology. Longenecker GL, ed. Alabama: Academic Press, Inc; 1985: 85-106.
    Google Scholar
  • 2 Mills DC. ADP receptors on platelets. Thromb Haemost 1996; 76: 835-56.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 3 Gachet C, Hechler B, Lèon C, Vial C, Leray C, Ohlmann P, Cazenave JP. Activation of ADP receptors and platelet function. Thromb Haemost 1997; 78: 271-5.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 4 Cattaneo M, Lombardi R, Zighetti ML, Gachet C, Ohlmann P, Cazenave JP, Mannucci PM. Deficiency of (33P)2MeS-ADP binding sites on platelets with secretion defect, normal granule stores and normal thromboxane A2 production. Evidence that ADP potentiates platelet secretion independently of the formation of large platelet aggregates and thromboxane A2 production. Thromb Haemost 1997; 77: 986-90.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 5 Cattaneo M, Canciana MT, Lecchi A, Kinlough-Rathbone RL, Packham MA, Mannucci PM, Mustard JF. Released adenosine diphosphate stabilizes thrombin-induced human platelet aggregates. Blood 1990; 75: 1081-6.
    CrossrefPubMedGoogle Scholar
  • 6 Trumel C, Payrastre B, Plantavid M, Hechler B, Vial C, Presek P, Martinson EA, Cazenave JP, Chap H, Gachet C. A key role of adenosine diphosphate in the irreversible platelet aggregation induced by the PAR1-activating peptide through the late activation of phosphoinositide 3-kinase. Blood 1999; 94: 4156-65.
    CrossrefPubMedGoogle Scholar
  • 7 Cattaneo M, Akkawat B, Lecchi A, Cimminiello C, Capitanio AM, Mannucci PM. Ticlopidine selectively inhibits human platelet responses to adenosine diphosphate. Thromb Haemost 1991; 66: 694-9.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 8 Cattaneo M, Gachet C. ADP receptors and clinical bleeding disorders. Arterioscler Thromb Vasc Biol 1999; 19: 2281-5.
    CrossrefPubMedGoogle Scholar
  • 9 Hollopeter G, Jantzen H-M, Vincent D, Li G, England L, Ramakrishnan V, Yang R-B, Nurden P, Nurden A, Julius DJ, Conley PB. Identification of the platelet ADP receptor targeted by antithrombotic drugs. Nature 2001; 409: 202-7.
    CrossrefPubMedGoogle Scholar
  • 10 Lèon C, Hechler B, Freund M, Eckly A, Vial C, Ohlmann P, Dierich A, LeMeur M, Cazenave J-P, Gachet C. Defective platelet aggregation and increased resistance to thrombosis in purinergic P2Y(1) receptor-null mice. J Clin Invest 1999; 104: 1731-7.
    CrossrefPubMedGoogle Scholar
  • 11 Fabre J-E, Nguyen M, Latour A, Keifer JA, Audoly LP, Coffman TM, Koller BH. Decreased platelet aggregation, increased bleeding time and resistance to thromboembolism in P2Y1-deficient mice. Nature med 1999; 5: 1199-202.
    CrossrefPubMedGoogle Scholar
  • 12 MacKenzie AB, Mahaut-Smith MP, Sage SO. Activation of receptor-operated cation channels via P2X1 not P2T purinoceptors in human platelets. J Biol Chem 1996; 271: 2879-81.
    CrossrefPubMedGoogle Scholar
  • 13 Jin J, Kunapuli SP. Coactivation of two different G protein-coupled receptors is essential for ADP-induced platelet aggregation. Proc Natl Acad Sci USA 1998; 95: 8070-4.
    CrossrefPubMedGoogle Scholar
  • 14 Jin J, Daniel JL, Kunapuli SP. Molecular basis for ADP-induced platelet activation. II. The P2Y1 receptor mediates ADP-induced intracellular calcium mobilization and shape change in platelets. J Biol Chem 1998; 273: 2030-4.
    CrossrefPubMedGoogle Scholar
  • 15 Takano S, Kimura J, Matsuoka I, Ono T. No requirement of P2X1 purinoceptors for platelet aggregation. Eur J Pharmacol 1999; 372: 305-9.
    CrossrefPubMedGoogle Scholar
  • 16 Savi P, Bornia J, Salel V, Delfaud M, Herbert J-M. Characterization of P2X1 purinoceptors on rat platelets: effect of clopidogrel. Br J Haematol 1997; 98: 880-6.
    CrossrefPubMedGoogle Scholar
  • 17 Mahaut-Smith MP, Ennion SJ, Rolf MG, Evans RJ. ADP is not an agonist at P2X1 receptors: evidence for separate receptors stimulated by ATP and ADP on human platelets. Br J Pharmacol 2000; 131: 108-14.
    CrossrefPubMedGoogle Scholar
  • 18 Lèon C, Hechler B, Vial C, Leray C, Cazenave J-P, Gachet C. The P2Y1 receptor is an ADP receptor antagonized by ATP and expressed in platelets and megakaryoblastic cells. FEBS lett 1997; 403: 26-30.
    CrossrefPubMedGoogle Scholar
  • 19 Fagura MS, Dainty IA, McKay GD, Kirk IP, Humphries RG, Robertson MJ, Dougall IG, Leff P. P2Y1 receptors in human platelets which are pharmacologically distinct from P2YADP-receptors. Br J Pharmacol 1998; 124: 157-64.
    CrossrefPubMedGoogle Scholar
  • 20 Rolf MG, Brearley CA, Mahaut-Smith MP. Platelet shape change evoked by selective activation of P2X1 purinoceptors with α,β-methylene ATP. Thromb Haemost 2001; 85: 303-8.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 21 Watson SP. Collagen receptor signaling in platelets and megakaryocytes. Thromb Haemost 1999; 82: 365-76.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 22 Deckmyn H, Stanssens P, Hoet B, Declerck PJ, Lauwereys M, Gansemans Y, Tornai I, Vermylen J. An echistatin-like Arg-Gly-Asp (RGD)-containing sequence in the heavy chain CDR3 of a murine monoclonal antibody that inhibits human platelet glycoprotein IIb/IIIa function. Br J Haematol 1994; 87: 562-71.
    CrossrefPubMedGoogle Scholar
  • 23 Oury C, Toth-Zsamboki E, Van Geet C, Thys C, Wei L, Nilius B, Vermylen J, Hoylaerts MF. A natural dominant negative P2X1 receptor due to deletion of a single amino acid residue. J Biol Chem 2000; 275: 22611-4.
    CrossrefPubMedGoogle Scholar
  • 24 Evans RJ, Lewis C, Buell G, Valera S, North RA, Surprenant A. Pharmacological characterization of heterologously expressed ATP-gated cation channels (P2X purinoceptors). Mol Pharmacol 1995; 48: 178-83.
    PubMedGoogle Scholar
  • 25 Werner P, Seward EP, Buell GN, North RA. Domains of P2X receptors involved in desensitization. Proc Natl Acad Sci USA 1996; 93: 15485-90.
    CrossrefPubMedGoogle Scholar
  • 26 Storey RF, Sanderson HM, White AE, May JA, Cameron KE, Heptinstall S. The central role of the P2T receptor in amplification of human platelet activation, aggregation, secretion, and procoagulant activity. Br J Haematol 2000; 110: 925-34.
    CrossrefPubMedGoogle Scholar
  • 27 Ohlmann P, Eckly A, Freund M, Cazenave J-P, Offermans S, Gachet C. ADP induces partial platelet aggregation without shape change and potentiates collagen-induced aggregation in the absence of Gαq. Blood 2000; 96: 2134-9.
    PubMedGoogle Scholar
  • 28 Ingall AH, Dixon J, Bailey A, Coombs ME, Cox D, Mc Inally JJ, Hunt SF, Kindon ND, Teobald BJ, Willis PA, Humphries RG, Leff P, Clegg JA, Smith JA, Tomlinson W. Antagonists of the platelet P2T receptor: a novel approach to antithrombotic therapy. J Med Chem 1999; 42: 213-20.
    CrossrefPubMedGoogle Scholar
  • 29 Malmgren R. ATP secretion occurs as an initial response in collagen-induced platelet activation. Thromb Res 1986; 43: 445-53.
    CrossrefPubMedGoogle Scholar
  • 30 Bauer M, Retzer M, Wilde JI, Maschberger P, Essler M, Aepfelbacher M, Watson SP, Siess W. Dichotomous regulation of myosin phosphorylation and shape change by Rho-kinase and calcium in intact human platelets. Blood 1999; 94: 1665-72.
    PubMedGoogle Scholar
  • 31 Cusack NJ, Hourani SMO. Adenosine 5′-diphosphate antagonists and human platelets: no evidence that aggregation and inhibition of stimulated adenylate cyclase are mediated by different receptors. Br J Pharmacol 1982; 76: 221-7.
    CrossrefPubMedGoogle Scholar
  • 32 Hall DA, Hourani SMO. Effects of analogues of adenine nucleotides on increases in intracellular calcium mediated by P2T-purinoceptors on human blood platelets. Br J Pharmacol 1993; 108: 728-33.
    CrossrefPubMedGoogle Scholar
  • 33 Baurand A, Eckly A, Bari N, Lèon C, Hechler B, Cazenave J-P, Gachet C. Desensitization of the platelet aggregation response to ADP: Differential down-regulation of the P2Y1 and P2cyc receptors. Thromb Haemost 2000; 84: 484-91.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 34 Wagner WR, Hubell JA. ADP receptor antagonists and converting enzyme systems reduce platelet deposition onto collagen. Thromb Haemost 1992; 67: 461-7.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 35 Weiss HJ, Witte LD, Kaplan KL, Lages BA, Chernoff A, Nossel HL, Goodman DS, Baumgartner HR. Heterogeneity in storage pool deficiency: studies on granule-bound substances in 18 patients including variants deficient in α-granules, platelet factor 4, β-thromboglobulin, and platelet-derived growth factor. Blood 1979; 54: 1296-319.
    PubMedGoogle Scholar
  • 36 Beigi R, Kobatake E, Aizawa M, Dubyak GR. Detection of local ATP release from activated platelets using cell surface-attached firely luciferase. Am J Physiol 1999; 276: C267-C278.
    CrossrefPubMedGoogle Scholar
  • 37 Sage SO, Yamoah EH, Heemskerk JWM. The roles of P2X1 and P2TAC receptors in ADP-evoked calcium signaling in human platelets. Cell Calcium 2000; 28: 119-26.
    CrossrefPubMedGoogle Scholar
  • 38 Gratacap MP, Hérault JP, Viala C, Ragab A, Savi P, Herbert JM, Chap H, Plantavid M, Payrastre B. FcγRIIA requires a Gi-dependent pathway for an efficient stimulation of phosphoinositide 3-kinase, calcium mobilization, and platelet aggregation. Blood 2000; 96: 3439-46.
    PubMedGoogle Scholar