Development of platelet inhibition by cAMP during megakaryocytopoiesis

doi:10.1074/jbc.M111390200

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Papers In Press, published online ahead of print May 7, 2002
J. Biol. Chem, 10.1074/jbc.M111390200
Submitted on November 29, 2001
Revised on May 7, 2002
Accepted on May 6, 2002

Development of platelet inhibition by cAMP during megakaryocytopoiesis

Els den Dekker, Gertie Gorter, Johan W. M. Heemskerk, and Jan-Willem N. Akkerman

Haematology, University Medical Center Utrecht, Utrecht 3508 GA

Corresponding Author: J.W.N.Akkerman{at}lab.azu.nl

Prostacyclin is a potent inhibitor of agonist-induced Ca[2+] increases in platelets, but in the megakaryocytic cell line MEG-01 this inhibition is absent. Using human megakaryocytic cell lines representing different stages in megakaryocyte (Mk) maturation and stem cells, immature and mature Mk’s, we show that the inhibition by prostacyclin develops at a late maturation stage, shortly before platelets are formed. This late appearance is not caused by insufficient cAMP formation, or absent protein kinase A (PKA) activity in immature cells. Instead, appearance of Ca2+ inhibition by prostacyclin is accompanied by a sharp increase in expression of the catalytic subunit of PKA (PKA-C), but not by changes in expression of the PKA-regulatory subunits Ialpha/beta, IIalpha and IIbeta. Overexpression of PKA-C in the megakaryocytic cell line CHRF-288-11 potentiates the Ca[2+] inhibition by prostacyclin. Thus, up-regulation of PKA-C appears a key step in the development of Ca[2+] inhibition by prostacyclin in platelets.

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