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J. Biol. Chem., Vol. 276, Issue 18, 15316-15325, May 4, 2001
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From In this report, the expression and
function of the platelet collagen receptor glycoprotein VI
(GPVI) were studied in human megakaryocytes during differentiation and
maturation of mobilized blood and cord blood derived
CD34+ cells. By flow cytometry, using an anti-GPVI
monoclonal antibody or convulxin, a GPVI-specific ligand, GPVI was
detected only on CD41+ cells including some
CD41+/CD34+ cells, suggesting expression at a
stage of differentiation similar to CD41. These results were confirmed
at the mRNA level using reverse transcription-polymerase chain
reaction. GPVI expression was low during megakaryocytic
differentiation but increased in the more mature megakaryocytes
(CD41high). As in platelets, megakaryocyte GPVI associates
with the Fc receptor
Expression and Function of the Collagen Receptor
GPVI during Megakaryocyte Maturation*
,,, and
INSERM E9907, Faculté Xavier Bichat, 75870 Paris Cedex 18, Paris, France, § INSERM U362,
Institut Gustave Roussy, Villejuif 94805, France, and ¶ Millenium
Pharmaceuticals Inc., Cambridge, Massachusetts 02139
chain (FcR). The FcR chain was detected
at the RNA and protein level at all stages of megakaryocyte maturation
preceding the expression of GPVI. The other collagen receptor,
2
1 integrin (CD49b/CD29), had a pattern
of expression similar to GPVI. Megakaryocytic GPVI was recognized as a
55-kDa protein by immunoblotting and ligand blotting, and thus it
presented a slightly lower apparent molecular mass than platelet GPVI
(58 kDa). Megakaryocytes began to adhere to immobilized convulxin via
GPVI after only 8-10 days of culture, at a time when megakaryocytes
were maturing. At this stage of maturation, they also adhered to
immobilized collagen by 21
integrin-dependent and -independent mechanisms. Convulxin induced a very similar pattern of protein tyrosine phosphorylation in
megakaryocytes and platelets including Syk, FcR, and
PLC
2. Our results showed that GPVI is
expressed early during megakaryocytic differentiation but functionally
allows megakaryocyte adherence to collagen only at late stages of
differentiation when its expression increases.
*
This work was supported by grants from the INSERM,
Association de Recherche Contre le Cancer Grant 9472, and University
Paris 7.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
To whom correspondence should be addressed. Tel.:
33-1-42-11-4233; Fax: 33-1-42-11-5240; E-mail: verpre@igr.fr.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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