Thromb Haemost 2001; 85(02): 303-308
DOI: 10.1055/s-0037-1615684
Review Article
Schattauer GmbH

Platelet Shape Change Evoked by Selective Activation of P2X1 Purinoceptors with α,β-Methylene ATP

Michael G. Rolf
1   Department of Physiology and Department of Plant Sciences, University of Cambridge, Cambridge, UK
,
Charles A. Brearley
2   Department of Plant Sciences, University of Cambridge, Cambridge, UK
,
Martyn P. Mahaut-Smith
1   Department of Physiology and Department of Plant Sciences, University of Cambridge, Cambridge, UK
› Author Affiliations
Further Information

Publication History

Received 24 January 2000

Accepted after resubmission 30 June 2000

Publication Date:
08 December 2017 (online)

Summary

Simultaneous measurements of [Ca2+]i and light transmission were used to examine the relationship between P2X1 receptor activation and functional platelet responses. The P2X1 agonist α,β-MeATP evoked a transient [Ca2+]i increase and a reversible decrease in light transmission; both responses required external Ca2+ and the nucleotidase apyrase. The transmission response was due to shape change only, verified by scanning electron microscopy and insensitivity to Reopro, a GPIIbIIIa antagonist. α,β-MeATP stimulated smaller shape changes than ADP, however P2X1 responses had a lifespan of <2 h following resuspension in saline and may be considerably larger in vivo. A peak [Ca2+]i increase of >50 nM was required for detectable shape change. Overlap of concentration-response relationships for α,β-MeATP-evoked [Ca2+]i and shape change suggests that other second messengers are not involved. Therefore, the physiological P2X1 agonist ATP can contribute to platelet activation, in contrast to its previously described inhibitory action at metabotropic platelet purinoceptors.

 
  • References

  • 1 Born GVR. Aggregation of blood platelets by adenosine diphosphate and its reversal. Nature 1962; 194: 927-8.
  • 2 Hourani SMO, Hall DA. Receptors for ADP on human blood-platelets. Trends In Pharmacological Sciences 1994; 15: 103-8.
  • 3 Kunapuli SP. Multiple P2 receptor subtypes on platelets: a new interpretation of their function. Trends In Pharmacological Sciences 1998; 19: 391-4.
  • 4 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 Letters 1997; 403: 26-30.
  • 5 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.
  • 6 Cooper DM, Rodbell M. ADP is a potent inhibitor of human platelet plasma membrane adenylate cyclase. Nature 1979; 282: 517-8.
  • 7 Cusack NJ, Hourani SM. 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.
  • 8 Ohlmann P, Laugwitz KL, Nurnberg B, Spicher K, Schultz G, Cazenave JP, Gachet C. The human platelet ADP receptor activates Gi2 proteins. Biochem J 1995; 312: 775-9.
  • 9 Jin J, Kunapuli SP. Coactivation of two different G protein-coupled receptors is essential for ADP-induced platelet aggregation. Proc Natl Acad Sci. U. S. A. 1998; 95: 8070-4.
  • 10 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.
  • 11 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.
  • 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.
  • 13 Sage SO, Mackenzie AB, Jenner S, Mahaut Smith MP. Purinoceptor-evoked calcium signalling in human platelets. Prostaglandins Leukot. Essent. Fatty Acids. 1997; 57: 435-8.
  • 14 Vial C, Hechler B, Leon C, Cazenave JP, Gachet C. Presence of P2X1 purinoceptors in human platelets and megakaryoblastic cell lines. Thromb Haemost 1997; 78: 1500-4.
  • 15 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.
  • 16 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.
  • 17 Siess W. Molecular mechanisms of platelet activation. Physiol Rev 1989; 69: 58-178.
  • 18 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.
  • 19 Holmsen H. Platelet metabolism and activation. Semin Hematol 1985; 22: 219-40.
  • 20 Sillen LG, Martel AE. Stability constants of metal-ion complexes. 1974
  • 21 McLean FC, Hastings AB. Clinical Estimation and Significance of Calcium-Ion Concentrations in the Blood. Amer J. Med Sci 1935; 189: 601-13.
  • 22 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.
  • 23 Grynkiewicz G, Poenie M, Tsien RY. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 1985; 260: 3440-50.
  • 24 Brearley CA, Hanke DE. Inositol phosphates in the duckweed Spirodela polyrhiza L. Biochem J 1996; 314: 215-25.
  • 25 Ohlmann P, Eckly A, Freund M, Cazenave JP, Offermanns S, Gachet C. ADP induces partial aggregation without shape change and potentiates collagen-induced aggregation in the absence of G q. Blood 2000; 96: 2134-9.
  • 26 Daniel JL, Dangelmaier C, Jin J, Ashby B, Smith JB, Kunapuli SP. Molecular basis for ADP-induced platelet activation. I. Evidence for three distinct ADP receptors on human platelets. J Biol Chem 1998; 273: 2024-9.
  • 27 Nicholas RA, Watt WC, Lazarowski ER, Li Q, Harden TK. Uridine nucleotide selectivity of three phospholipase C-activating P2 receptors: Identification of a UDP-selective, a UTP-selective, and an ATP- and UTP-specific receptor. Molecular Pharmacology 1996; 50: 224-9.
  • 28 Rolf MG, Lockwood AJ, Mahaut Smith MP, Sage SO. Properties of the human platelet ADP receptor which releases calcium from intracellular stores. J Physiol Lond 1998; 506: P: 68P-68P.
  • 29 Boyer JL, Romero Avila T, Schachter JB, Harden TK. Identification of competitive antagonists of the P2Y1 receptor. Molecular Pharmacology 1996; 50: 1323-9.
  • 30 Stoop R, Surprenant A, North RA. Different sensitivities to pH of ATP-induced currents at four cloned P2X receptors. J Neurophysiol 1997; 78: 1837-40.
  • 31 Coller BS. A new murine monoclonal antibody reports an activation-dependent change in the conformation and/or microenvironment of the platelet glycoprotein IIb/IIIa complex. J Clin Invest 1985; 76: 101-8.
  • 32 Humphries RG, Tomlinson W, Clegg JA, Ingall AH, Kindon ND, Leff P. Pharmacological profile of the novel P2T-purinoceptor antagonist, FPL 67085 in vitro and in the anaesthetized rat in vivo. Br J Pharmacol 1995; 115: 1110-6.
  • 33 Eberhard DA, Holz RW. Intracellular Ca2+ activates phospholipase C. TINS 1988; 11: 517-20.
  • 34 Bezprozvanny I, Ehrlich BE. The inositol 1,4,5-trisphosphate (InsP3) receptor. J Membr Biol 1995; 145: 205-16.
  • 35 Hallam TJ, Rink TJ. Responses to adenosine diphosphate in human platelets loaded with the fluorescent calcium indicator quin2. J Physiol Lond 1985; 368: 131-46.
  • 36 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.
  • 37 Rink TJ, Smith SW, Tsien RY. Cytoplasmic free Ca2+ in human platelets: Ca2+ thresholds and Ca- independent activation for shape-change and secretion. FEBS Lett. 1982; 148: 21-6.
  • 38 Paul BZS, Daniel JL, Kunapuli SP. Platelet Shape Change Is Mediated by both Calcium-dependent and -independent Signaling Pathways. J Biol Chem 1999; 40: 28293-300.
  • 39 Klages B, Brandt U, Simon MI, Schultz G, Offermanns S. Activation of G12/G13 results in shape change and Rho/Rho-kinase-mediated myosin light chain phosphorylation in mouse platelets. J Cell Biol 1999; 144: 745-54.
  • 40 Beigi R, Kobatake E, Aizawa M, Dubyak GR. Detection of local ATP release from activated platelets using cell surface-attached firefly luciferase. Am J Physiol 1999; 276: C267-78.