Thromb Haemost 1987; 58(04): 971-977
DOI: 10.1055/s-0038-1646039
Original Article
Schattauer GmbH Stuttgart

Characterization of Na+/H+ Exchange in Platelets

Avinoam Livne
The Division of Cell Biology, The Hospital for Sick Children, Toronto, Ontario, Canada
,
Sergio Grinstein
The Division of Cell Biology, The Hospital for Sick Children, Toronto, Ontario, Canada
,
Aser Rothstein
The Division of Cell Biology, The Hospital for Sick Children, Toronto, Ontario, Canada
› Author Affiliations
Further Information

Publication History

Received 10 December 1986

Accepted after revision 03 July 1987

Publication Date:
29 June 2018 (online)

Summary

Acidification of the cytoplasm of human blood platelets leads to activation of Na+/H+ exchange. As a result, alkalinization occurs that is detectable by an intracellular fluorescent pH indicator. The activity of the exchanger can also be measured by the swelling of platelets suspended in Na-propionate medium using a Coulter Counter: the rapid entry of propionic acid leads to acidification and activation of Na+/H+ exchange with the parallel entry of Na+ and propionic acid leading to osmotic swelling. The Na+/H+ exchanger is sensitive to amiloride and to derivatives that are reported to be more specific inhibitors; it is specific for Na+ and Li+ with no measurable transfer of K+, Rb+ or Cs+; its Km for Na+ is 75 to 90 mM; it displays competitive behavior between Na+ and amiloride; its activity is decreased in cells loaded with Na+ by prolonged ouabain treatment; and it has a high temperature coefficient. These properties are in general similar to those of the exchanger in other cells. It is suggested that the Na+/H+ exchanger plays a role in platelet pH regulation.

 
  • References

  • 1 Mahnesmith RL, Aronson PS. The plasma membrane sodium-hydrogen exchanger and its role in physiological and pathophysiological processes. Circ Res 1985; 57: 773-788
  • 2 Aronson PS. Kinetic properties of the plasma membrane Na+/H+ exchanger. Ann Rev Physiol 1985; 47: 545-560
  • 3 Grinstein S, Rothstein A. Topical review; Mechanisms of regulation of the Na+/H+ exchanger. J Membr Biol 1986; 90: 1-12
  • 4 Siffert W, Miickenhoff K, Schied P. Evidence for a role of Na+/H+ exchange in platelets activated with calcium-ionophore A23187. Biochem Biophys Res Commun 1984; 125: 1123-1128
  • 5 Siffert W, Fox G, Miickenhoff K, Scheid P. Thrombin stimulates Na+/H+ exchange across the human platelet plasma membrane. FEBS Letters 1984; 172: 272-274
  • 6 Sweatt JD, Johnson SL, Cragoe EJ, Limbird LE. 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 1985; 260: 12910-12919
  • 7 Greenberg-Sepersky SM, Simons ER. Cation gradient dependence of the steps in thrombin stimulation of human platelets. J Biol Chem 1984; 259: 1502-1508
  • 8 Zavoico GB, Cragoe EJ, Feinstein MB. Regulation of intracellular pH in human platelets. Effects of thrombin, A 23187 and ionomvein and evidence for activation of Na+/H+ exchange and its inhibition by amiloride analogs. J Biol Chem 1986; 261: 12120-12127
  • 9 Livne A, Grinstein S, Rothstein A. Volume regulating behavior of human platelets. J Cell Physiol 1987; 131: 354-363
  • 10 Livne A, Balfe JW, Veitch R, Grinstein S, Rothstein A. Elevated platelet Na+/H+ exchange rates in essential hypertension: application of a novel test. Lancet 1987; i: 533-536
  • 11 Grinstein S, Cohen S, Rothstein A. Cytoplasmic pH regulation in thymic lymphocytes by an amiloride sensitive Na+/H+ antiport. J Gen Physiol 1984; 83: 341-369
  • 12 Grinstein S, Goetz JD, Furuya W, Rothstein A, Gelfand EW. Amiloride-sensitive Na+/H+ exchange in platelets and leukocytes: detection by electronic cell sizing. Am J Physiol 1984; 247: C293-8
  • 13 Again G, Livne A. Passive participation of fixed platelets in aggregation facilitated by covalently bound fibrinogen. Blood 1983; 61: 186-191
  • 14 Roos A, Boron WF. Intracellular pH. Physiol Rev 1981; 61: 296-434
  • 15 Thomas JA, Buchsbaum RH, Zimniak A, Rackcr FE. Intracellular pH measurements in Ehrlich ascites cells utilizing spectroscopic probes generated in situ. Biochemistry 1979; 18: 2210-2218
  • 16 Grinstein S, Furuya W. Characterization of the amiloride-sensitive Na+/H+ antiport of human neutrophils. Am J Physiol 1986; 250: C238-91
  • 17 Zhuang YX, Cragoe Jr EF, Sharkewitz T, Glaser L, Cassel D. Characterization of potent Na+/H+ exchange inhibitors from the amiloride series in A431 cells. Biochemistry 1984; 23: 4481-4488
  • 18 Feinberg H, Sandler WC, Scorer M, LeBrctton GC, Grossman B, Born G VR. Movement of sodium into human platelets induced by ADP. Biochim Biophys Acta 1977; 470: 317-324
  • 19 Parsegian A. Energy of an ion crossing a low dielectric membrane: solutions to four relevant electrostatic problems. Nature 1969; 221: 844-846
  • 20 Vignc P, Frclin C, Audinot M, Borsotto M, Cragoe Jr EJ, Lazdunski M. [|3H]ethylpropylamiloride. a radiolabelled diuretic for the analysis of the Na+/H+ exchange system. Its use with kidney cell membranes. EMBO J 1984; 3: 2647-2651
  • 21 Grinstein S, Cohen S, Goetz-Smith JD, Dixon SJ. Measurements of cytoplasmic pH and cellular volume for detection of Na+/H+ exchange in lymphocytes. Meth in Enzymol. 1987 (in press)
  • 22 Sandler WC, LeBreton GC, Feinberg H. Movement of sodium into human platelets. Biochim Biophys Acta 1980; 600: 448-455