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The role of Na+/H+ exchanger in serotonin secretion from porcine blood platelets

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This study was undertaken to evaluate whether a link exists between the activation of protein kinase C (PKC), operation of Na+/H+ exchanger (NHE), cell swelling and serotonin (5-HT) secretion in porcine platelets. Activation of platelets by thrombin or phorbol 12-myristate 13-acetate (PMA), a PKC activator, initiated a rapid rise in the activity of Na+/H+ exchanger and secretion of 5-HT. Both thrombin- and PMA-evoked activation of Na+/H+ exchanger was less pronounced in the presence of ethyl-isopropyl-amiloride (EIPA), an NHE inhibitor, and by GF 109203X, a PKC inhibitor. Monensin (simulating the action of NHE) caused a dose-dependent release of 5-HT that was not abolished by GF 109203X or EGTA. Lack of Na+ in the suspending medium reduced thrombin-, PMA-, and monensin-evoked 5-HT secretion. GF 109203X nearly completely inhibited 5-HT release induced by PMA-, partly that induced by thrombin, and had no effect on 5-HT release induced by monensin. EIPA partly inhibited 5-HT release induced by thrombin and nearly totally that evoked by PMA. Electronic cell sizing measurements showed an increase in mean platelet volume upon treatment of cells with monensin, PMA or thrombin. The PMA- and thrombin-evoked rise in mean platelet volume was strongly reduced in the presence of EIPA. As judged by optical swelling assay monensin and PMA produced a rapid rise in platelet volume. The swelling elicited by PMA was inhibited by EIPA and its kinetics was similar to that observed in the presence of monensin. Hypoosmotically evoked platelet swelling did not affect platelet aggregation but significantly potentiated thrombin-evoked release of 5-HT and ATP. Taken together, these results show that in porcine platelets PKC may promote 5-HT secretion through the activation of NHE. It is hypothesized that enhanced Na+/H+ antiport may result in a rise in cell membrane tension (due to cell swelling) which in turn facilitates fusion of secretory granules with the plasma membrane leading to 5-HT secretion.
Physical description
  • Department of Physical Chemistry, Medical University of Bialystok, Bialystok, Poland
  • Department of Physical Chemistry, Medical University of Bialystok, Bialystok, Poland
  • Department of Physical Chemistry, Medical University of Bialystok, Bialystok, Poland
  • Aviv A (1992) The roles of cell Ca2+, protein kinase C and the Na+-H+ antiport in the development of hypertension and insulin resistance. J Am Soc Nephrol 3: 1049-1063.
  • Apodaca G (2002) Modulation of membrane traffic by mechanical stimuli. Am J Physiol Renal Physiol 282: F179-F190.
  • Bednar RA, Gaul SL, Hamill TG, Egbertson MS, Shafer JA (1998) Identification of low molecular weight GP IIb/IIIa antagonists that bind preferentially to activated platelets. J Pharmacol Exp Ther 285: 1317-1326.
  • Berry S, Dawidcki DD, Agarval KC, Steiner M (1989) The role of microtubules in platelet secretory release. Biochim Biophys Acta 1012: 46-56.
  • Blaustein MP, Lederer WJ (1999) Sodium/calcium exchange: its physiological implications Physiol Rev 79: 763-854.
  • Borin M, Siffert W (1990) Stimulation by thrombin increases the cytosolic free Na+ concentration in human platelets. Studies with the novel fluorescent cytosolic Na+ indicator sodium-binding benzofuran isophtalate. J Biol Chem 265: 19543-19550.
  • Borin M, Siffert W (1991) Further characterization of the mechanisms mediating the rise in cytosolic free Na+ in thrombin-stimulated platelets. Evidence for inhibition of the Na+,K+-ATPase and for Na+ entry via a Ca2+ influx pathway. J Biol Chem 266: 13153-13160.
  • Connolly TM, Limbrid LE (1983) Removal of extraplatelet Na+ eliminates indomethacin-sensitive secretion from human platelets stimulated by epinephrine, ADP and thrombin. Proc Natl Acad Sci USA 80: 5320-5324.
  • Da Prada M, Cesura AM, Launay JM, Richards JG (1988) Platelets as a model for neurones? Experientia 44: 115-126.
  • Elzagallaai A, Rose S, Trifaro J-M (1998) Platelet secretion induced by phorbol esters stimulation is mediated through phosphorylatioin of MARCKS. Blood 95: 894-902.
  • Escolar G, White JG (1991) The platelet open canicular system: A final common pathway. Blood Cells 17: 467-485.
  • Flaumenhaft R (2003) Molecular basis of platelet granule secretion. Arterioscler Thromb Vasc Biol 23: 1152-1160.
  • Fritz M, Radmacher M, Gauub HE (1994) Granula motion and membrane spreading during activation of human platelets imaged by atomic force microscopy. Biophys J 66: 1328-1334.
  • Fuentealba J, Olivares R, Ales E, Tapia L, Rojo J, Arroyo G, Aldea M, Criado M, Gandia L, Garcia AG (2004) A choline-evoked [Ca2+]c signal causes catecholamine release and hyperpolarization of chromaffin cells. FASEB J 18: 1468-1470.
  • Harrison P, Cramer EM (1993) Platelet alpha-granules. Blood Rev 7: 52-62.
  • Hamill OP, Martinac B (2001) Molecular basis of mechanotransduction in living cells. Physiol Rev 81: 685-729.
  • Holmsen H (1981) Platelet secretion. in Haemostasis and Thrombosis: Basic Principles and Clinical Practice. Colman RW, Hirsh J, Marder VJ, Salzman EW, eds, pp 390-403. JB Lippincott Comp. Philadelphia.
  • Holmsen H, Dangelmaier CA (1989) Measurement of secretion of serotonin. Methods Enzymol 169: 195-205.
  • Joseph S, Siffert W, Gorter G, Akkerman JW (1990) Stimulation of human platelets by collagen occurs by a Na+/H+ exchanger independent mechanism. Biochim Biophys Acta 1054: 26-32.
  • Krishnamurti S, Patel Y, Morgan WA, Wheeler-Jones CP, Kakkar VV (1989) Na+/H+ exchange is not necessary for protein kinase C-mediated effects in platelets. FEBS Lett 252: 147-152.
  • Kroll MH, Schafer AL (1996) The analysis of ligand-receptor interactions in platelet activation. in Immunopharmacology of Platelets. pp 631-665. Academic Press Ltd.
  • Lang F, Busch GL, Ritter M, Vokl H, Waldegger S, Gulbins E, Haussinger D (1998) Functional significance of cell volume regulatory mechanisms. Physiol Rev 78: 247-306.
  • Lesch KP, Wolozin BL, Murphy DL, Riederer P (1993) Primary structure of the human platelet serotonin uptake site: identity with the brain serotonin transporter. J Neurochem 60: 2319-2322.
  • Lida H, Barron W, Page E (1988) Monensin turns on microtubule-associated translocation of secretory granules in cultured rat atrial myocytes. Circ Res 62: 1159-1170.
  • Le Quan Sang KH, Laude D, Devynck MA (1987) Platelet cytosolic free Ca2+ concentration and serotonin content in essential hypertension. J Hypertens 5: 237-240.
  • Livne AA, Sardet C, Pouyssegur J (1991) The Na+/H+ exchanger is phosphorylated in human platelets in response to activating agents. FEBS Lett 284: 219-222.
  • McNicol A, Nickolaychuk BR (1995) Inhibition of collagen-induced platelet activation by arachidonyl trifluoromethyl ketone. Biochem Pharmacol 50: 1795-1802.
  • McNicol A, Israels SJ (1999) Platelet dense granules: structure, function and implications for haemostasis. Thromb Res 95: 1-18.
  • Mollenhauer HH, Morre DJ, Rowe LD (1990) Alteration of intracellular traffic by monensin; mechanism specificity and relationship to toxicity. Biochim Biophys Acta 1031: 225-246.
  • Morris CE, Homann U (2001) Cell surface area regulation and membrane tension. J Membr Biol 179: 79-102.
  • Nishizuka Y (1986) Studies and perspectives of protein kinase C. Science 233: 305-312.
  • Putney L, Denker SP, Barber DL (2002) The changing face of the Na+/H+ exchanger, NHE1: structure, regulation, and cellular actions. Annu Rev Pharmacol Toxicol 42: 527-552.
  • Reed G, Fitzgerald M, Polgar J (2000) Molecular mechanisms of platelet exocytosis: insights into the "secrete" life of thrombocytes. Blood 15: 3334-3342.
  • Rink TJ, Sanchez A, Hallam TJ (1993) Diacylglycerol and phorbol ester stimulate secretion without raising cytoplasmic free calcium in human platelets. Nature 305: 317-319.
  • Roberts DE, McNicol A, Bose R (2004) Mechanism of collagen activation in human platelets. J Biol Chem 279: 19421-19430.
  • Rosskopf D (1999) Sodium-hydrogen exchange and platelet function. J Thromb Thrombolysis 8: 15-23.
  • Rosskopf D, Morgenstern E, Scholz W, Osswald U, Siffert W (1991) Rapid determination of the elevated Na+/H+ exchange in platelets of patients with essential hypertension using an optical swelling assay. J Hypertens 9: 231-238.
  • Samson J, Stelmach H, Tomasiak M (2001) The importance of Na+/H+ exchanger for the generation of procoagulant activity by porcine blood platelets. Platelets 12: 436-442.
  • Siess W (1989) Molecular mechanisms of platelet activation. Physiol Rev 69: 58-178.
  • Siffert W, Dusing R (1996) Na+/H+ exchange in hypertension and in diabetes mellitus - facts and hypotheses. Basic Res Cardiol 91: 179-190.
  • Siffert W, Siffert G, Scheid P, Akkerman JWN (1989) Activation of Na+/H+ exchange and Ca2+ mobilization start simultaneously in thrombin-stimulated platelets. Biochem J 258: 521-527.
  • Stelmach H, Rusak T, Tomasiak M (2002) The involvement of the Na+/H+ exchanger in the formation of microvesicles by porcine platelets. Haematologia 32: 239-252.
  • Storey RF, Wilcox RG, Heptinstall S (1998) Differential effects of glycoprotein IIb/IIIa antagonists on platelet microaggregate and macroaggregate formation and effect of anticoagulant on antagonist potency. Circulation 98: 1616-2161.
  • Strbak V, Greer MA (2000) Regulation of hormone secretion by acute cell volume changes: Ca(2+)-independent hormone secretion. Cell Physiol Biochem 10: 393-402.
  • Straub SG, Daniel S, Sharp GWG (2002) Hyposmotic shock stimulates insulin secretion by two distinct mechanisms. Studies the βHC9 cell. Am J Physiol Endocrinol Metab 282: E1070-E1076.
  • Sweatt JD, Johnson SL, Cragoe EJ, Limbird LE (1985) Inhibitors of Na+/H+ exchange block stimulus-provoked arachidonic acid release in human platelets. Selective effects on platelet activation by epinephrine, ADP and lower concentrations of thrombin. J Biol Chem 260: 12910-12919.
  • Tomasiak M, Stelmach H, Rusak T, Wysocka J (2004) Nitric oxide and platelet energy metabolism. Acta Biochim Polon 51: 789-803.
  • Toullec D, Pianetti P, Coste H, Bellevergue P, Grand-Perret T, Ajakane M, Baudet V, Boissin P, Boursier E, Loriolle F, Duhamel L, Charon D, Kirilovsky J (1991) The bisindolylmaleimide, GF 109203X is a potent and selective inhibitor of protein kinase C. J Biol Chem 266: 15771-15781.
  • Turetta L, Donella-Deana A, Folda A, Bulato C, Deana R (2004) Characterisation of serotonin efflux induced by cytosolic Ca2+ and Na+ concentration increase in human platelets. Cell Physiol Biochem 14: 377-386.
  • Wehner F, Tinel H (1998) Role of Na+ conductance, Na+/H+ exchange, and Na+-K+-2Cl- exchange symport in the regulatory volume increase of rat hepatocytes. J Physiol 506: 127-142.
  • Weiner CP (1987) The role of serotonin in the genesis of hypertension in preeclampsia. Am J Obstet Gynecol 156: 885-888.
  • White JG (1999) Platelet secretory process. Blood 93: 2422-2425.
  • White JG, Clawson CC (1980) The surface-connected canalicular system of blood platelets - a fenestrated membrane system. Am J Pathol 101: 353-364.
  • White JG, Escolar G (1991) The blood platelet open canalicular system: A two way street. Eur J Cell Biol 56: 233-242.
  • Yoashioka A, Shirakawa R, Nishioka H, Tabuchi A, Higashi T, Ozaki H, Yamamoto A, Kita T, Horiuchi H (2001) Identification of protein kinase Cα as an essential, but not sufficient, cytosolic factor for Ca2+-induced α- and dense-core granule secretion in platelets. J Biol Chem 276: 39379-39385.
  • Zimmerberg J, Vogel SS, Chernomordic LV (1993) Mechanisms of membrane fusion. Annu Rev Biophys Biomol Struct 22: 433-466.
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