Physiol Rep. 2013 Oct;1(5):e00085. doi: 10.1002/phy2.85. Epub 2013 Oct 11.

Pericellular Ca(2+) recycling potentiates thrombin-evoked Ca(2+) signals in human platelets.

Physiological reports

Stewart O Sage, Nicholas Pugh, Richard W Farndale, Alan G S Harper

PMID: 24303163 PMCID: PMC3841026 DOI: 10.1002/phy2.85

Abstract

We have previously demonstrated that Na(+)/Ca(2+) exchangers (NCXs) potentiate Ca(2+) signaling evoked by thapsigargin in human platelets, via their ability to modulate the secretion of autocoids from dense granules. This link was confirmed in platelets stimulated with the physiological agonist, thrombin, and experiments were performed to examine how Ca(2+) removal by the NCX modulates platelet dense granule secretion. In cells loaded with the near-membrane indicator FFP-18, thrombin stimulation was observed to elicit an NCX-dependent accumulation of Ca(2+) in a pericellular region around the platelets. To test whether this pericellular Ca(2+) accumulation might be responsible for the influence of NCXs over platelet function, platelets were exposed to fast Ca(2+) chelators or had their glycocalyx removed. Both manipulations of the pericellular Ca(2+) rise reduced thrombin-evoked Ca(2+) signals and dense granule secretion. Blocking Ca(2+)-permeable ion channels had a similar effect, suggesting that Ca(2+) exported into the pericellular region is able to recycle back into the platelet cytosol. Single cell imaging with extracellular Fluo-4 indicated that thrombin-evoked rises in extracellular [Ca(2+)] occurred within the boundary described by the cell surface, suggesting their presence within the open canalicular system (OCS). FFP-18 fluorescence was similarly distributed. These data suggest that upon thrombin stimulation, NCX activity creates a rise in [Ca(2+)] within the pericellular region of the platelet from where it recycles back into the platelet cytosol, acting to both accelerate dense granule secretion and maintain the initial rise in cytosolic [Ca(2+)].

Keywords: Ca2+; FFP-18; nanojunction; open canalicular system; pericellular; platelets

References

1. Neuroreport. 1998 Oct 26;9(15):3403-7 - PubMed
2. Biochim Biophys Acta. 1993 Apr 16;1176(3):215-21 - PubMed
3. Biochim Biophys Acta. 1989 Jun 6;981(2):367-70 - PubMed
4. Biochim Biophys Acta. 1988 Mar 3;938(3):497-500 - PubMed
5. Biochem J. 1991 Dec 15;280 ( Pt 3):783-9 - PubMed
6. Cell Calcium. 2007 Oct-Nov;42(4-5):379-87 - PubMed
7. Cell Calcium. 1992 Mar;13(3):183-92 - PubMed
8. Cell Calcium. 2007 Dec;42(6):606-17 - PubMed
9. Thromb Haemost. 1982 Feb 26;47(1):22-6 - PubMed
10. Biochem Pharmacol. 2007 Dec 3;74(11):1596-607 - PubMed
11. Eur J Biochem. 2002 Mar;269(5):1543-52 - PubMed
12. J Gen Physiol. 1986 Aug;88(2):149-65 - PubMed
13. Exp Physiol. 2009 May;94(5):509-19 - PubMed
14. Cell Calcium. 2003 May-Jun;33(5-6):311-21 - PubMed
15. J Biol Chem. 2004 Oct 22;279(43):44250-7 - PubMed
16. Thromb Haemost. 1999 Feb;81(2):286-92 - PubMed
17. J Biol Chem. 1987 Dec 5;262(34):16364-9 - PubMed
18. Cell Calcium. 1998 May;23(5):269-79 - PubMed
19. Nature. 1987 Jan 29-Feb 4;325(6103):456-8 - PubMed
20. Thromb Haemost. 1985 Oct 30;54(3):645-9 - PubMed
21. Biochem Biophys Res Commun. 1988 Jun 16;153(2):848-54 - PubMed
22. Biophys J. 2010 Aug 4;99(3):755-64 - PubMed
23. Biochim Biophys Acta. 1987 Jun 15;929(1):88-102 - PubMed
24. Cell Calcium. 2009 May;45(5):413-20 - PubMed
25. Blood. 2009 Jul 2;114(1):e10-9 - PubMed
26. Arterioscler Thromb Vasc Biol. 1996 Feb;16(2):230-5 - PubMed
27. Cold Spring Harb Perspect Biol. 2010 Nov;2(11):a004051 - PubMed
28. J Thromb Haemost. 2010 Jul;8(7):1604-13 - PubMed
29. Cell Calcium. 2007 Oct-Nov;42(4-5):373-8 - PubMed
30. J Thromb Haemost. 2011 Mar;9(3):540-51 - PubMed
31. Blood. 2010 Aug 19;116(7):1147-56 - PubMed
32. Physiol Rev. 2006 Jan;86(1):155-203 - PubMed
33. Blood. 2008 Nov 15;112(10):4069-79 - PubMed
34. Blood. 2001 May 1;97(9):2648-56 - PubMed
35. Am J Pathol. 1972 Feb;66(2):295-312 - PubMed
36. Br J Pharmacol. 2012 Feb;165(4):922-36 - PubMed
37. Am J Physiol. 1998 Aug;275(2):C423-30 - PubMed
38. Cell Calcium. 1998 Oct;24(4):275-83 - PubMed
39. Proc Natl Acad Sci U S A. 1996 May 28;93(11):5368-73 - PubMed
40. Stroke. 2008 Jun;39(6):1774-8 - PubMed
41. Cell Calcium. 1989 Oct;10(7):491-8 - PubMed
42. J Physiol. 1999 Jul 15;518 ( Pt 2):463-7 - PubMed
43. J Histochem Cytochem. 1975 Feb;23(2):103-6 - PubMed
44. Physiol Rev. 1982 Jan;62(1):185-261 - PubMed
45. Annu Rev Med. 1979;30:119-34 - PubMed
46. J Biol Chem. 1985 Mar 25;260(6):3440-50 - PubMed
47. Biochim Biophys Acta. 1986 May 29;886(3):406-10 - PubMed
48. J Biol Chem. 2004 May 7;279(19):19421-30 - PubMed
49. Biochem J. 1990 Oct 15;271(2):515-22 - PubMed
50. J Physiol. 1995 Apr 1;484 ( Pt 1):15-24 - PubMed
51. J Physiol. 1997 Jun 15;501 ( Pt 3):485-95 - PubMed
52. FEBS Lett. 1985 Jul 8;186(2):175-9 - PubMed
53. Biophys Chem. 2004 Dec 20;112(2-3):131-40 - PubMed
54. J Thromb Haemost. 2007 May;5(5):910-7 - PubMed
55. J Biol Chem. 1985 Jul 25;260(15):8657-60 - PubMed
56. J Biol Chem. 1982 Dec 10;257(23):14000-5 - PubMed
57. J Physiol. 2001 Sep 15;535(Pt 3):625-35 - PubMed
58. Neuron. 1996 Jan;16(1):89-101 - PubMed
59. Cell Calcium. 1989 Feb-Mar;10(2):101-13 - PubMed
60. Thromb Haemost. 1987 Jun 3;57(3):337-40 - PubMed
61. Blood. 2012 Oct 11;120(15):e73-82 - PubMed
62. J Histochem Cytochem. 1980 Nov;28(11):1183-8 - PubMed
63. J Cell Biol. 2003 Mar 31;160(7):1151-61 - PubMed
64. Physiol Rev. 1999 Jul;79(3):763-854 - PubMed
65. J Physiol. 1990 Sep;428:723-35 - PubMed
66. FEBS Lett. 1987 Dec 10;225(1-2):72-6 - PubMed
67. J Thromb Haemost. 2010 Dec;8(12):2766-74 - PubMed
68. J Biol Chem. 1984 Oct 25;259(20):12571-5 - PubMed
69. J Thromb Haemost. 2009 Feb;7(2):330-8 - PubMed

Publication Types

• Journal Article

Grant support

• PG/07/100/23759/British Heart Foundation/United Kingdom
• RG/09/003/27122/British Heart Foundation/United Kingdom