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Platelet and megakaryocyte dense granules contain glycoproteins Ib and
IIb-IIIa
T Youssefian, JM Masse, F Rendu, J Guichard and EM Cramer
INSERM U.91, Hopital Henri Mondor, Creteil, France.
Platelets contain two main types of secretory organelles, the dense
granules and the alpha-granules. P-selectin, a specific receptor for
leukocytes that is present in the alpha-granule membrane, has also been
demonstrated to be associated with the dense granule limiting membrane,
showing that a relationship exists between these two types of secretory
granules. We have previously shown that the plasma membrane receptors
glycoproteins (Gp) IIb-IIIa and Ib are also present in the alpha- granule
membrane. To document further the composition of the dense granule
membrane, we have used immunoelectron microscopy in the present work to
determine if the dense granule membrane also contains these glycoproteins.
First, the cytochemical method of Richards and Da Prada (J Histochem
Cytochem 25:1322, 1977), which specifically enhances dense body electron
density, was combined with immunogold-labeled anti-Gp IIb- IIIa or anti-Gp
Ib antibody. A consistent and reproducible labeling for Gp IIb-IIIa, but
less for Gp Ib, was found in the membrane of platelet dense granules.
Subsequently, double immunogold labeling was performed on frozen thin
sections of resting platelets using antibodies directed against the dense
body components granulophysin or P-selectin, followed by anti-Gp IIb-IIIa
or anti-Gp Ib. Consistent labeling for Gp IIb-IIIa and weaker labeling for
Gp Ib were detected in dense bodies. The possibility that the
granulophysin-positive structures could be lysosomes was excluded by the
presence of P-selectin. Immunogold labeling of isolated dense granule
fractions confirmed these results. Identical findings were made on human
cultured megakaryocytes using double immunolabeling. In conclusion, this
study demonstrates the presence of Gp IIb-IIIa and Gp Ib on the dense
granule membrane. This observation provides additional evidence of
similarities between the alpha-granule and dense granule membranes and
raises the possibility of a dual mechanism responsible for the formation of
dense granules similar to that of alpha-granules, ie, endogenous synthesis
as well as endocytosis from the plasma membrane.
Volume 89,
Issue 11,
pp. 4047-4057,
06/01/1997
Copyright © 1997 by The American Society of Hematology
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