Thromb Haemost 2002; 88(03): 495-502
DOI: 10.1055/s-0037-1613243
Review Article
Schattauer GmbH

Effects of Enhanced P2X1 Receptor Ca2+ Influx on Functional Responses in Human Platelets

Michael G. Rolf
1   Department of Physiology, University of Cambridge, Cambridge, UK
,
Martyn P. Mahaut-Smith
1   Department of Physiology, University of Cambridge, Cambridge, UK
› Institutsangaben
Weitere Informationen

Publikationsverlauf

Received 17. April 2001

Accepted after resubmission 28. Mai 2002

Publikationsdatum:
08. Dezember 2017 (online)

Summary

G-protein-coupled P2Y1 and P2Y12 receptors play key roles in platelet activation, however the importance of ionotropic P2X1 receptors remains unclear. Platelet P2X1 responses are highly labile in vitro, but were greatly enhanced by increasing [Ca2+]o in the range 1–10 mM. The P2X1 agonist α,β-MeATP stimulated a shape change which saturated at peak [Ca2+]i of ≥ 400 nM, without evidence for aggregation. The maximal P2X1-evoked transmission decrease was 82% of that obtained via P2Y1 receptors. α., β-MeATP caused a disc to sphere transformation in virtually all platelets, but lacked the long processes produced by ADP. Following block of P2Y1 receptors with A3P5PS, co-stimulation with α., β-MeATP and ADP failed to induce aggregation despite the generation of peak [Ca2+]i responses similar to those stimulated via P2Y1 receptors. Therefore early, transient Ca2+ influx via P2X1 receptors can contribute to platelet activation by stimulating a significant morphological change, but does not readily synergise with P2Y12 receptors to support aggregation.

 
  • References

  • 1 Kunapuli SP, Daniel JL. P2 receptor subtypes in the cardiovascular system. Biochem J 1998; 336: 513-23.
  • 2 Gachet C. ADP receptors of platelets and their inhibition. Thromb Haemost 2001; 86: 222-32.
  • 3a Hollopeter G, Jantzen HM, Vincent D, Li G, England L, Ramakrishnan V, Yang RB, Nurden P, Nurden A, Julius D, Conley PB. Identification of the platelet ADP receptor targeted by antithrombotic drugs. Nature 2001; 409: 202-7.
  • 3b Foster CJ, Prosser DM, Agans JM, Zhai Y, Smith MD, Lachowicz JE, Zhang FL, Gustafson E, Monsma FJJ, Wiekowski MT, Abbondanzo SJ, Cook DN, Bayne ML, Lira SA, Chintala MS. Molecular identification and characterization of the platelet ADP receptor targeted by thienopyridine antithrombotic drugs. J Clin Invest 2001; 107: 1591-8.
  • 4 Takasaki J, Kamohara M, Saito T, Matsumoto M, Matsumoto S, Ohishi T, Soga T, Matsushime H, Furuichi K. Molecular cloning of the platelet P2TAC ADP receptor: pharmacological comparison with another ADP receptor, the P2Y1 receptor. Mol Pharmacol 2001; 60: 432-9.
  • 5 Hechler B, Leon C, Vial C, Vigne P, Frelin C, Cazenave JP, Gachet C. The P2Y1 receptor is necessary for adenosine 5’-diphosphate-induced platelet aggregation. Blood 1998; 92: 152-9.
  • 6 Savi P, Beauverger P, Labouret C, Delfaud M, Salel V, Kaghad M, Herbert JM. Role of P2Y1 purinoceptor in ADP-induced platelet activation. FEBS Lett 1998; 422: 291-5.
  • 7 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.
  • 8 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.
  • 9 Savi P, Bornia J, Salel V, Delfaud M, Herbert JM. Characterization of P2X1 purinoreceptors on rat platelets: effect of clopidogrel. Br J Haematol 1997; 98: 880-6.
  • 10 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.
  • 11 Greco NJ, Tonon G, Chen W, Luo X, Dalal R, Jamieson GA. Novel structurally altered P2X1 receptor is preferentially activated by adenosine diphosphate in platelets and megakaryocytic cells. Blood 2001; 98: 100-7.
  • 12 Oury C, Toth-Zsamboki E, Vermylen J, Hoylaerts MF, Greco NJ, Tonon G, Luo X, Dalal R, Jamieson GA. Does the P2X1del variant lacking 17 amino acids in its extracellular domain represent a relevant functionalion channel in platelets?. Blood 2002; 99: 2275-7.
  • 13 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.
  • 14 Oury C, Toth-Zsamboki E, Thys C, Tytgat J, Vermylen J, Hoylaerts MF. The ATP-gated P2X1 ion channel acts as a positive regulator of platelet responses to collagen. Thromb Haemost 2001; 86: 1264-71.
  • 15 Vial C, Rolf MG, Mahaut-Smith MP, Evans RJ. A study of P2X1 receptor function in murine megakaryocytes and human platelets reveals synergy with P2Y receptors. Br J Pharmacol 2002; 135: 363-72.
  • 16 Gaarder A, Jonsen J, Laland S, Hellem A, Owren PA. Adenosine diphosphate in red cells as a factor in the adhesiveness of human blood platelets. Nature 1961; 192: 531-2.
  • 17 Born GVR. Aggregation of blood platelets by adenosine diphosphate and its reversal. Nature 1962; 194: 927-9.
  • 18 Leon C, Hechler B, Vial C, Leray C, Cazenave JP, 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.
  • 19 Sage SO, Rink TJ. The kinetics of changes in intracellular calcium concentration in fura-2-loaded human platelets. J Biol Chem 1987; 262: 16364-9.
  • 20 Mahaut-Smith MP, Sage SO, Rink TJ. Receptor-activated single channels in intact human platelets. J Biol Chem 1990; 265: 10479-83.
  • 21 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.
  • 22 Sage SO, Yamoah EH, Heemskerk JW. The roles of P2X1 and P2TAC receptors in ADP-evoked calcium signalling in human platelets. Cell Calcium 2000; 28: 119-26.
  • 23 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.
  • 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.
  • 25 Macfarlane DE, Mills DC. The effects of ATP on platelets: evidence against the central role of released ADP in primary aggregation. Blood 1975; 46: 309-20.
  • 26 Paul BZ, Daniel JL, Kunapuli SP. Platelet shape change is mediated by both calcium-dependent and -independent signaling pathways. Role of p160 Rho-associated coiled-coil-containing protein kinase in platelet shape change. J Biol Chem 1999; 274: 28293-300.
  • 27 Grynkiewicz G, Poenie M, Tsien RY. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 1985; 260: 3440-50.
  • 28 Valera S, Hussy N, Evans RJ, Adami N, North RA, Surprenant A, Buell G. A new class of ligand-gated ion channel defined by P2X receptor for extracellular ATP. Nature 1994; 371: 516-9.
  • 29 Hallam TJ, Rink TJ. Responses to adenosine diphosphate in human platelets loaded with the fluorescent calcium indicator quin2. J Physiol 1985; 368: 131-46.
  • 30 Sage SO, Rink TJ, Mahaut-Smith MP. Resting and ADP-evoked changes in cytosolic free sodium concentration in human platelets loaded with the indicator SBFI. J Physiol 1991; 441: 559-73.
  • 31 Mahaut-Smith MP, Sage SO, Rink TJ. Rapid ADP-evoked currents in human platelets recorded with the nystatin permeabilized patch technique. J Biol Chem 1992; 267: 3060-5.
  • 32 Clifford EE, Parker K, Humphreys BD, Kertesy SB, Dubyak GR. The P2X1 receptor, an adenosine triphosphate-gated cation channel, is expressed in human platelets but not in human blood leukocytes. Blood 1998; 91: 3172-81.
  • 33 Mahaut-Smith MP, Rink TJ, Collins SC, Sage SO. Voltage-gated potassium channels and the control of membrane potential in human platelets. J Physiol 1990; 428: 723-35.
  • 34 Daniel JL, Molish IR, Rigmaiden M, Stewart G. Evidence for a role of myosin phosphorylation in the initiation of the platelet shape change response. J Biol Chem 1984; 259: 9826-31.
  • 35 Nachmias VT, Kavaler J, Jacubowitz S. Reversible association of myosin with the platelet cytoskeleton. Nature 1985; 313: 70-2.
  • 36 Hathaway DR, Adelstein RS. Human platelet myosin light chain kinase requires the calcium-binding protein calmodulin for activity. Proc Natl Acad Sci USA 1979; 76: 1653-7.
  • 37 Fanger CM, Ghanshani S, Logsdon NJ, Rauer H, Kalman K, Zhou J, Beckingham K, Chandy KG, Cahalan MD, Aiyar J. Calmodulin mediates calcium-dependent activation of the intermediate conductance KCa channel, IKCa1 . J Biol Chem 1999; 274: 5746-54.
  • 38 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.
  • 39 Sage SO, MacKenzie AB, Jenner S, Mahaut-Smith MP. Purinoceptorevoked calcium signalling in human platelets. Prostaglandins Leukot Essent Fatty Acids 1997; 57: 435-8.
  • 40 Sargeant P, Farndale RW, Sage SO. Calcium store depletion in dimethyl BAPTA-loaded human platelets increases protein tyrosine phosphorylation in the absence of a rise in cytosolic calcium. Exp Physiol 1994; 79: 269-72.
  • 41 Rosado JA, Sage SO. Role of the ERK pathway in the activation of store-mediated calcium entry in human platelets. J Biol Chem 2001; 276: 15659-65.
  • 42 Heemskerk JW, Hoyland J, Mason WT, Sage SO. Spiking in cytosolic calcium concentration in single fibrinogen-bound fura-2-loaded human platelets. Biochem J 1992; 283: 379-83.
  • 43 Hussain JF, Mahaut-Smith MP. Reversible and irreversible intracellular Ca2+ spiking in single isolated human platelets. J Physiol 1999; 514: 713-8.
  • 44 Hallam TJ, Rink TJ. Agonists stimulate divalent cation channels in the plasma membrane of human platelets. FEBS Lett 1985; 186: 175-9.
  • 45 Nishio H, Ikegami Y, Nakata Y, Segawa T. Fluorescence digital image analysis of thrombin and ADP induced rise in intracellular Ca2+ concentration of single blood platelets. Neurochem Int 1992; 21: 75-81.
  • 46 Heemskerk JW, Vis P, Feijge MA, Hoyland J, Mason WT, Sage SO. Roles of phospholipase C and Ca2+-ATPase in calcium responses of single, fibrinogen-bound platelets. J Biol Chem 1993; 268: 356-63.
  • 47 Mahaut-Smith MP. Calcium-activated potassium channels in human platelets. J Physiol 1995; 484: 15-24.
  • 48 Somasundaram B, Mahaut-Smith MP. Three cation influx currents activated by purinergic receptor stimulation in rat megakaryocytes. J Physiol 1994; 480: 225-31.
  • 49 Somasundaram B, Norman JC, Mahaut-Smith MP. Primaquine, an inhibitor of vesicular transport, blocks the calcium-release-activated current in rat megakaryocytes. Biochem J 1995; 309: 725-9.
  • 50 Trumel C, Payrastre B, Plantavid M, Hechler B, Viala 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.
  • 51 Dangelmaier C, Jin J, Smith JB, Kunapuli SP. Potentiation of thromboxane A2-induced platelet secretion by Gi signalling through the phosphoinositide-3 kinase pathway. Thromb Haemost 2001; 85: 341-8.
  • 52 Kauffenstein G, Bergmeier W, Eckly A, Ohlmann P, Leon C, Cazenave JP, Nieswandt B, Gachet C. The P2Y12 receptor induces platelet aggregation through weak activation of the alphaIIbbeta3 integrin – a phosphoinositide 3-kinase-dependent mechanism. FEBS Lett 2001; 505: 281-90.
  • 53 Fabre JE, 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. Nat Med 1999; 05: 1199-202.
  • 54 Leon C, Hechler B, Freund M, Eckly A, Vial C, Ohlmann P, Dierich A, LeMeur M, Cazenave JP, Gachet C. Defective platelet aggregation and increased resistance to thrombosis in purinergic P2Y1 receptor-null mice. J Clin Invest 1999; 104: 1731-7.
  • 55 Jarvis GE, Humphries RG, Robertson MJ, Leff P. ADP can induce aggregation of human platelets via both P2Y1 and P2T receptors. Br J Pharmacol 2000; 129: 275-82.
  • 56 Wildman SS, Brown SG, King BF, Burnstock G. Selectivity of diadenosine polyphosphates for rat P2X receptor subunits. Eur J Pharmacol 1999; 367: 119-23.
  • 57 Lewis CJ, Gitterman DP, Schluter H, Evans RJ. Effects of diadenosine polyphosphates (ApnAs) and adenosine polyphospho guanosines (ApnGs) on rat mesenteric artery P2X receptor ion channels. Br J Pharmacol 2000; 129: 124-30.
  • 58 Savi P, Pflieger AM, Herbert JM. cAMP is not an important messenger for ADP-induced platelet aggregation. Blood Coagul Fibrinolysis 1996; 07: 249-52.
  • 59 Daniel JL, Dangelmaier C, Jin J, Kim YB, Kunapuli SP. Role of intracellular signaling events in ADP-induced platelet aggregation. Thromb Haemost 1999; 82: 1322-6.
  • 60 Mahaut-Smith MP, Hussain JF, Mason MJ. Depolarization-evoked Ca2+ release in a non-excitable cell, the rat megakaryocyte. J Physiol 1999; 515: 385-90.