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Uncoordinated expression of fibrinogen compared with thrombospondin and von
Willebrand factor in maturing human megakaryocytes
EM Cramer, N Debili, JF Martin, AM Gladwin, J Breton-Gorius, P Harrison, GF Savidge and W Vainchenker
Departement d'Hematologie, C.H.U. Bichat, Paris, France.
The localization of three known alpha-granule proteins, thrombospondin
(TSP), von Willebrand factor (vWF), and fibrinogen (Fg) has been studied in
human megakaryocytes (MK) by immunofluorescence and immunoelectron
microscopy. For this study, highly purified populations of MK were prepared
from human bone marrow either by counterflow centrifugal elutriation or by
cell culture from normal subjects and from two patients with
megakaryoblastic leukemia. In normal bone marrow immature MK, TSP, and vWF
were observed in the Golgi-associated vesicles and in small immature
alpha-granules; in mature MK, they were found in the matrix of the mature
large alpha-granules. Surprisingly, Fg was detected neither in the Golgi
area, nor in the small precursors of alpha-granules; it was only found in
the mature alpha-granules but this labeling was generally weaker than in
blood platelets. In order to confirm these differences between the
expression of Fg and vWF or TSP additional studies were performed on
cultured maturing MK: immunofluorescent and ultrastructural immunogold
labeling confirmed that vWF appeared early in the maturation while the same
immature MK were negative for Fg. In the late maturation stage, the three
proteins were detected in the alpha-granules. In order to know whether Fg
was lately synthesized or endocytosed from the outside medium, normal MK
were grown in the presence of either normal or afibrinogenemic plasma, and
normal serum. Fg was detected only in the alpha-granules of MK grown in
normal plasma. Similar results were observed with malignant MK, whose
maturation was independent of the culture conditions. In conclusion, this
study brings immunocytochemical evidence that vWF and TSP are synthesized
by immature MK, whereas Fg appears later in the MK alpha-granules and its
expression is dependent of the presence of an exogenous Fg source.
Volume 73,
Issue 5,
pp. 1123-1129,
04/01/1989
Copyright © 1989 by The American Society of Hematology
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