Secreted Dense Granule Adenine Nucleotides Promote Calcium Influx and the Maintenance of Elevated Cytosolic Calcium Levels in Stimulated Human Platelets

Bruce Lages; Harvey J. Weiss

doi:10.1055/s-0037-1614459

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1999; 81(02): 286-292
DOI: 10.1055/s-0037-1614459

Review Articles

Schattauer GmbH

Secreted Dense Granule Adenine Nucleotides Promote Calcium Influx and the Maintenance of Elevated Cytosolic Calcium Levels in Stimulated Human Platelets

Bruce Lages

¹From the Department of Medicine, Division of Hematology-Oncology, St. Luke’s-Roosevelt Hospital Center, and Columbia University College of Physicians and Surgeons, New York, NY, USA

,

Harvey J. Weiss

¹From the Department of Medicine, Division of Hematology-Oncology, St. Luke’s-Roosevelt Hospital Center, and Columbia University College of Physicians and Surgeons, New York, NY, USA

› Author Affiliations

Abstract

Summary

Evidence that secreted dense granule adenine nucleotides mediate part of the agonist-induced cytosolic calcium ([Ca²⁺]_i) responses in human platelets was obtained from comparisons of fura-2-loaded platelets from normal subjects and from patients with a form of platelet storage pool deficiency (SPD) in which the secretory dense granules and their contents are virtually absent. SPD platelets had normal initial [Ca²⁺]_i in creases induced by thrombin and the endoperoxide analog U46619, but a significantly enhanced decay of elevated [Ca²⁺]_i levels following the initial increases. With thrombin, this enhanced [Ca²⁺]_i decay was associated with decreased Ca²⁺ influx, as measured by Mn²⁺ quench of fura-2 fluorescence. Addition of micromolar concentrations of ADP, alone or together with ATP, after stimulation reversed the enhanced [Ca²⁺]_I decay and increased Mn²⁺ quench in SPD platelets, but had no effect on these responses in normal platelets, while addition of 100-fold higher concentrations of ATP or apyrase before stimulation increased [Ca²⁺]_I decay and decreased Mn²⁺ quench in normal platelets, but had little effect in SPD platelets. ATP and α,β-methylene ATP, a specific agonist for P2X1 receptors, at micromolar concentrations also increased Mn²⁺ quench, but to lesser extents than did ADP, in SPD platelets isolated and loaded with fura-2 in the presence of apyrase. Similar effects of ADP and excess ATP were seen in U46619-stimulated platelets, but decreased Ca²⁺ influx could not be measured directly in SPD platelets, presumably due to the very transient influx response seen with U46619. These results suggest that secreted dense granule ADP and ATP contribute to the maintenance of elevated [Ca²⁺]_i levels, but not to the initial [Ca²⁺]_i increases, in stimulated human platelets, most likely via a nucleotide-specific component of Ca²⁺ influx which may be mediated by interactions with both P2X₁ and P2Y₁ purinoceptors.

Full Text

References

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