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Blood, Vol. 93 No. 9 (May 1), 1999:
pp. 2876-2883
Expression of a Functional N-Methyl-D-Aspartate-Type
Glutamate Receptor by Bone Marrow Megakaryocytes
Paul G. Genever,
David J.P. Wilkinson,
Amanda J. Patton,
Nicky M. Peet,
Ying Hong,
Anthony Mathur,
Jorge D. Erusalimsky, and
Tim M. Skerry
From the Department of Biology, University of York, York; and The
Wolfson Institute for Biomedical Research, University College London,
Cruciform Project Laboratories, Rayne Institute, London, UK.
Better understanding of hemostasis will be possible by the
identification of new lineage-specific stimuli that regulate platelet formation. We describe a novel functional megakaryocyte receptor that
belongs to a family of ionotropic glutamate receptors of the
N-methyl-D-aspartate (NMDA) subtype responsible for
synaptic neurotransmission in the central nervous system (CNS).
Northern blotting and reverse-transcriptase polymerase chain reaction
(RT-PCR) studies identified expression of NMDAR1 and NMDAR2D type
subunit mRNA in rat marrow, human megakaryocytes, and MEG-01 clonal
megakaryoblastic cells. Immunohistochemistry and in vivo
autoradiographic binding of the NMDA receptor-specific antagonist
MK-801 confirmed that megakaryocytes expressed open channel-forming
NMDA receptors in vivo. Western blots indicated that megakaryocyte
NMDAR1 was either unglycosylated or only glycosylated to low levels,
and of identical size to CNS-type NMDAR1 after deglycosylation with
endoglycosidase F/peptide-N-glycosidase F. In functional
studies, we demonstrated that NMDA receptor activity was necessary for
phorbol myristate acetate (PMA)-induced differentiation of
megakaryoblastic cells; NMDA receptor blockade by specific antagonists
significantly inhibited PMA-mediated increases in cell size, CD41
expression, and adhesion of MEG-01 cells. These results provide
evidence for a novel pathway by which megakaryocytopoiesis and
platelet production may be regulated.
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