Expression of a functional N-methyl-D-aspartate-type glutamate receptor by bone marrow megakaryocytes

TY - JOUR

T1 - Expression of a functional N-methyl-D-aspartate-type glutamate receptor by bone marrow megakaryocytes

AU - Genever, P G

AU - Wilkinson, D J P

AU - Patton, A J

AU - Peet, N M

AU - Hong, Y

AU - Mathur, A

AU - Erusalimsky, J D

AU - Skerry, T M

PY - 1999/5/1

Y1 - 1999/5/1

N2 - 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, (C) 1999 by The American Society of Hematology.

AB - 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, (C) 1999 by The American Society of Hematology.

KW - LEUKEMIA-CELL LINE

KW - C-MPL LIGAND

KW - PROTEIN-KINASE

KW - NMDA-RECEPTOR

KW - HEL CELLS

KW - BLOOD-PLATELETS

KW - 5-HT2 RECEPTOR

KW - HIGH-AFFINITY

KW - ACETYLCHOLINESTERASE

KW - DIFFERENTIATION

M3 - Article

SN - 0006-4971

VL - 93

SP - 2876

EP - 2883

JO - Blood

JF - Blood

IS - 9

ER -