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Characterization of vasoactive intestinal peptide receptors on human
megakaryocytes and platelets
SK Park, TA Olson, N Ercal, M Summers and MS O'Dorisio
Department of Pediatrics, The Ohio State University, Columbus, USA.
Vasoactive intestinal peptide receptor I (VIPRI) expression was examined in
megakaryocytes using reverse transcriptase-polymerase chain reaction
(RT-PCR). VIPRI protein was characterized in platelet membranes using
covalent crosslinking techniques. Human megakaryocytes were isolated from
suspension cultures of cord blood and adult bone marrow mononuclear cells
using a murine monoclonal antibody to human platelet glycoprotein IIB/IIIA
(CD41) and immunomagnetic beads. RT-PCR primers were constructed for the
VIP, VIPRI, and VIPRII genes as well as for megakaryocyte specific genes,
c-mpl and platelet factor 4 (PF- 4). VIP, VIPRI, c-mpl, and PF-4 were
coexpressed in megakaryocyte mRNA. Southern blot analysis confirmed the
expression of VIPRI. 125I-VIP was covalently cross-linked to human platelet
membranes using the homobifunctional reagent disuccinimidyl suberate,
followed by polyacrylamide gel electrophoresis and autoradiography. A
125I-VIP- protein complex of Mr = 50,000 was identified. Labeling of the Mr
= 50,000 component was completely abolished by unlabeled VIP, but not by
peptide histidine methionine or growth hormone releasing factor, indicating
specific binding of VIP to the platelet membranes. Taken together, these
results suggest that VIP may have direct effects on megakaryocytopoiesis
and support our earlier observations of VIP modulation of platelet
aggregation.
Volume 87,
Issue 11,
pp. 4629-4635,
06/01/1996
Copyright © 1996 by The American Society of Hematology
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