Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1983 May 1;96(5):1234–1240. doi: 10.1083/jcb.96.5.1234

Role of sodium in ADP- and thrombin-induced megakaryocyte spreading

PMCID: PMC2112653  PMID: 6404909

Abstract

We investigated the role of sodium in megakaryocyte spreading induced by thrombin and ADP. We found that if extracellular sodium was replaced by lithium, potassium, or choline, spreading was inhibited. When extracellular sodium was present, amiloride or tetrodotoxin inhibited spreading. Using intracellular recording we found spreading to be associated with a permanent membrane depolarization. The extent and rate of thrombin-induced depolarization was reduced when lithium replaced sodium. Unspread cells had an average membrane potential of - 44.8 mV. Spread cells had an average membrane potential of -18.46 mV. When choline replaced sodium, or when in the presence of tetrodotoxin and amiloride, the spread cells repolarized, indicating that the depolarization is due to an increase in sodium permeability. Similar treatments did not change the membrane potential of unspread cells. Incubation of megakaryocytes with A23187 together with monensin or methylamine induced spreading. Methylamine occasionally caused spreading by itself, but neither ionophore alone caused spreading. These results indicate that megakaryocyte spreading induced by ADP and thrombin depends on an increase in sodium conductance.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Begg D. A., Rebhun L. I. pH regulates the polymerization of actin in the sea urchin egg cortex. J Cell Biol. 1979 Oct;83(1):241–248. doi: 10.1083/jcb.83.1.241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Catterall W. A. Neurotoxins that act on voltage-sensitive sodium channels in excitable membranes. Annu Rev Pharmacol Toxicol. 1980;20:15–43. doi: 10.1146/annurev.pa.20.040180.000311. [DOI] [PubMed] [Google Scholar]
  3. Connor C. G., Brady R. C., Brownstein B. L. Trifluoperazine inhibits spreading and migration of cells in culture. J Cell Physiol. 1981 Sep;108(3):299–307. doi: 10.1002/jcp.1041080303. [DOI] [PubMed] [Google Scholar]
  4. Dabrowska R., Hartshorne D. J. A Ca2+-and modulator-dependent myosin light chain kinase from non-muscle cells. Biochem Biophys Res Commun. 1978 Dec 29;85(4):1352–1359. doi: 10.1016/0006-291x(78)91152-x. [DOI] [PubMed] [Google Scholar]
  5. Daniel J. L., Holmsen H., Adelstein R. S. Thrombin-stimulated myosin phosphorylation in intact platelets and its possible involvement secretion. Thromb Haemost. 1977 Dec 15;38(4):984–989. [PubMed] [Google Scholar]
  6. Davis M. H., Pato C. N., Gruenstein E. Analysis of neurotoxin and mitogen-stimulated sodium transport in human fibroblasts. J Biol Chem. 1982 Apr 25;257(8):4356–4361. [PubMed] [Google Scholar]
  7. Deitmer J. W., Ellis D. Interactions between the regulation of the intracellular pH and sodium activity of sheep cardiac Purkinje fibres. J Physiol. 1980 Jul;304:471–488. doi: 10.1113/jphysiol.1980.sp013337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dunnett J., Nayler W. G. Effect of pH on calcium accumulation and release by isolated fragments of cardiac and skeletal muscle sarcoplasmic reticulum. Arch Biochem Biophys. 1979 Dec;198(2):434–438. doi: 10.1016/0003-9861(79)90517-4. [DOI] [PubMed] [Google Scholar]
  9. Fedorko M. E. The functional capacity of guinea pig megakaryocytes. I. Uptake of 3H-serotonin by megakaryocytes and their physiologic and morphologic response to stimuli for the platelet release reaction. Lab Invest. 1977 Mar;36(3):310–320. [PubMed] [Google Scholar]
  10. Feinberg H., Sandler W. C., Scorer M., Le Breton G. C., Grossman B., Born G. V. Movement of sodium into human platelets induced by ADP. Biochim Biophys Acta. 1977 Oct 17;470(2):317–324. doi: 10.1016/0005-2736(77)90109-2. [DOI] [PubMed] [Google Scholar]
  11. Feinstein M. B., Becker E. L., Fraser C. Thrombin, collagen and A23187 stimulated endogenous platelet arachidonate metabolism: differential inhibition by PGE1, local anesthetics and a serine-protease inhibitor. Prostaglandins. 1977;14(6):1075–1093. doi: 10.1016/0090-6980(77)90286-6. [DOI] [PubMed] [Google Scholar]
  12. Feinstein M. G., Fiekers J., Fraser C. An analysis of the mechanism of local anesthetic inhibition of platelet aggregation and secretion. J Pharmacol Exp Ther. 1976 Apr;197(1):215–228. [PubMed] [Google Scholar]
  13. Gonnella P. A., Nachmias V. T. Platelet activation and microfilament bundling. J Cell Biol. 1981 Apr;89(1):146–151. doi: 10.1083/jcb.89.1.146. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hathaway D. R., Adelstein R. S. Human platelet myosin light chain kinase requires the calcium-binding protein calmodulin for activity. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1653–1657. doi: 10.1073/pnas.76.4.1653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Horne W. C., Norman N. E., Schwartz D. B., Simons E. R. Changes in cytoplasmic pH and in membrane potential in thrombin-stimulated human platelets. Eur J Biochem. 1981 Nov;120(2):295–302. doi: 10.1111/j.1432-1033.1981.tb05703.x. [DOI] [PubMed] [Google Scholar]
  16. Käser-Glanzmann R., Jakäbovä M., George J. N., Lüscher E. F. Stimulation of calcium uptake in platelet membrane vesicles by adenosine 3',5'-cyclic monophosphate and protein kinase. Biochim Biophys Acta. 1977 May 2;466(3):429–440. doi: 10.1016/0005-2736(77)90336-4. [DOI] [PubMed] [Google Scholar]
  17. Leven R. M., Nachmias V. T. Cultured megakaryocytes: changes in the cytoskeleton after ADP-induced spreading. J Cell Biol. 1982 Feb;92(2):313–323. doi: 10.1083/jcb.92.2.313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Meissner G. Calcium transport and monovalent cation and proton fluxes in sarcoplasmic reticulum vesicles. J Biol Chem. 1981 Jan 25;256(2):636–643. [PubMed] [Google Scholar]
  19. Menashi S., Davis C., Crawford N. Calcium uptake associated with an intracellular membrane fraction prepared from human blood platelets by high-voltage, free-flow electrophoresis. FEBS Lett. 1982 Apr 19;140(2):298–302. doi: 10.1016/0014-5793(82)80918-6. [DOI] [PubMed] [Google Scholar]
  20. Miller J. L., Sheridan J. D., White J. G. Electrical responses by guinea pig megakaryocytes. Nature. 1978 Apr 13;272(5654):643–645. doi: 10.1038/272643a0. [DOI] [PubMed] [Google Scholar]
  21. Moolenaar W. H., Boonstra J., van der Saag P. T., de Laat S. W. Sodium/proton exchange in mouse neuroblastoma cells. J Biol Chem. 1981 Dec 25;256(24):12883–12887. [PubMed] [Google Scholar]
  22. Mukherjee C., Lynn W. S. Role of ions and extracellular protein in leukocyte motility and membrane ruffling. Am J Pathol. 1978 Nov;93(2):369–381. [PMC free article] [PubMed] [Google Scholar]
  23. Munson R., Jr, Westermark B., Glaser L. Tetrodotoxin-sensitive sodium channels in normal human fibroblasts and normal human glia-like cells. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6425–6429. doi: 10.1073/pnas.76.12.6425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Nachmias V. T. Cytoskeleton of human platelets at rest and after spreading. J Cell Biol. 1980 Sep;86(3):795–802. doi: 10.1083/jcb.86.3.795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Pressman B. C. Biological applications of ionophores. Annu Rev Biochem. 1976;45:501–530. doi: 10.1146/annurev.bi.45.070176.002441. [DOI] [PubMed] [Google Scholar]
  26. Rothenberg P., Reuss L., Glaser L. Serum and epidermal growth factor transiently depolarize quiescent BSC-1 epithelial cells. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7783–7787. doi: 10.1073/pnas.79.24.7783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Scholey J. M., Taylor K. A., Kendrick-Jones J. Regulation of non-muscle myosin assembly by calmodulin-dependent light chain kinase. Nature. 1980 Sep 18;287(5779):233–235. doi: 10.1038/287233a0. [DOI] [PubMed] [Google Scholar]
  28. Sha'afi R. I., Naccache P. H., Alobaidi T., Molski T. F., Volpi M. Effect of arachidonic acid and the chemotactic factor F-Met-Leu-Phe on cation transport in rabbit neutrophils. J Cell Physiol. 1981 Feb;106(2):215–223. doi: 10.1002/jcp.1041060207. [DOI] [PubMed] [Google Scholar]
  29. Shen S. S., Steinhardt R. A. Intracellular pH and the sodium requirement at fertilisation. Nature. 1979 Nov 1;282(5734):87–89. doi: 10.1038/282087a0. [DOI] [PubMed] [Google Scholar]
  30. Shoshan V., MacLennan D. H., Wood D. S. A proton gradient controls a calcium-release channel in sarcoplasmic reticulum. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4828–4832. doi: 10.1073/pnas.78.8.4828. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Thomas R. C. The role of bicarbonate, chloride and sodium ions in the regulation of intracellular pH in snail neurones. J Physiol. 1977 Dec;273(1):317–338. doi: 10.1113/jphysiol.1977.sp012096. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tilney L. G., Kiehart D. P., Sardet C., Tilney M. Polymerization of actin. IV. Role of Ca++ and H+ in the assembly of actin and in membrane fusion in the acrosomal reaction of echinoderm sperm. J Cell Biol. 1978 May;77(2):536–550. doi: 10.1083/jcb.77.2.536. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. White J. G., Rao G. H., Gerrard J. M. Effects of the lonophore A23187 on blood platelets I. Influence on aggregation and secretion. Am J Pathol. 1974 Nov;77(2):135–149. [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES