PMID: 6403084May 1, 1983Paper

Characterization of the megakaryocyte secretory response: studies of continuously monitored release of endogenous ATP.

Blood
J L Miller

Abstract

Megakaryocytes share a number of structural and chemical properties with their progeny, blood platelets. With the availability of highly purified preparations of megakaryocytes isolated from guinea pig bone marrow, it is now also possible to study functional aspects of these cells. The present work reports the first study of the release of endogenously stored materials in megakaryocytes. Guinea pig megakaryocytes isolated to 75%-90% purity were exposed to thrombin or calcium ionophore (A23187) and the release of ATP was continuously monitored with the luciferin-luciferase reaction. Both maximal extent and initial rate of release were studied. Thrombin-induced release was half-maximal at thrombin concentrations of 0.2-0.5 NIH U/ml. At 4 U/ml thrombin, maximal release was 538 +/- 147 nmole ATP/10(9) megakaryocytes. A23187 induced half-maximal responses at concentrations of 7-8 microM. ATP release by ionophore showed a nearly absolute requirement for extracellular calcium, with release by thrombin showing only a partial calcium dependence. Following overnight culture, the response to thrombin was unchanged, whereas ATP release in response to ionophore was consistently increased (p less than 0.01). By comparison of maximally releas...Continue Reading

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