Biochemical and Ultrastructural Studies Suggest that the Effects of Thapsigargin on Human Platelets Are Mediated by Changes in Intracellular Calcium but not by Intracellular Histamine

Satya P Saxena; Archibald McNicol; Allan B Becker; Lorne J Brandes; Ole Thastrup; Jon M Gerrard

doi:10.1055/s-0038-1646349

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1992; 68(06): 714-718
DOI: 10.1055/s-0038-1646349

Original Article

Schattauer GmbH Stuttgart

Biochemical and Ultrastructural Studies Suggest that the Effects of Thapsigargin on Human Platelets Are Mediated by Changes in Intracellular Calcium but not by Intracellular Histamine

Satya P Saxena

Manitoba Institute of Cell Biology, and Departments of Pediatrics, Medicine and Pharmacology, University of Manitoba, Winnipeg, Manitoba, Canada

,

Archibald McNicol

Manitoba Institute of Cell Biology, and Departments of Pediatrics, Medicine and Pharmacology, University of Manitoba, Winnipeg, Manitoba, Canada

,

Allan B Becker

Manitoba Institute of Cell Biology, and Departments of Pediatrics, Medicine and Pharmacology, University of Manitoba, Winnipeg, Manitoba, Canada

,

Lorne J Brandes

Manitoba Institute of Cell Biology, and Departments of Pediatrics, Medicine and Pharmacology, University of Manitoba, Winnipeg, Manitoba, Canada

,

Ole Thastrup

¹Department of Clinical Chemistry, University Hospital, Rigshospitalet, Denmark

,

Jon M Gerrard

Manitoba Institute of Cell Biology, and Departments of Pediatrics, Medicine and Pharmacology, University of Manitoba, Winnipeg, Manitoba, Canada

› Author Affiliations

Abstract

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Summary

The involvement of intracellular histamine in thapsigargin (Tg)-induced platelet aggregation was studied. Platelet aggregation induced by 0.25 and 0.5 µM Tg was not accompained by a rise in intracellular histamine but a significant (p <0.01) increase in the level of intracellular histamine was observed at 1 µM Tg. Preincubation of platelets with inhibitors of histamine metabolizing enzymes had little effect on intracellular histamine levels in platelets stimulated by 0.5 µM Tg. In addition, the inhibitors of histidine decarboxylase (HDC), α-methyl histidine (α-MH) and α-fluoromethyl histidine (α-FMH) failed to inhibit Tg-induced aggregation. The intracellular histamine receptor antagonist, N,N-diethyl-2-[4-(phenylmethyl)phenoxy] ethanamine. HC1 (DPPE), inhibited Tg-induced aggregation but with IC₅₀ values dependent on the concentration of agonist used. The inhibitory effects of DPPE on Tg-induced aggregation were not reversed by the addition of histamine to saponin-permeabilized platelets suggesting non-histamine mediated effects of DPPE on Tg-induced aggregation. Tg stimulated an increase in the cytosolic free calcium concentration which was unaffected by DPPE indicating that the effects of DPPE are also not due to the inhibition of mobilization of cytosolic calcium. The ultrastructural studies suggest that the major Tg-induced changes (pseudopod formation and granule centralization) are consistent with a primary role for Tg to mobilize calcium; DPPE had very little effect on these ultrastructural changes. The results indicate that the effects of Tg on human platelets are mediated by an increase in cytosolic calcium but not by intracellular histamine.

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References

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