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Multivesicular Bodies Are an Intermediate Stage in the Formation
of Platelet -Granules
Harry F.G. Heijnen,
Najet Debili,
William Vainchencker,
Janine Breton-Gorius,
Hans J. Geuze, and
Jan J. Sixma
From the Department of Hematology, University Hospital Utrecht, and
Department of Cell Biology, Medical Faculty, and Institute of
Biomembranes, Utrecht University, Utrecht, The Netherlands; INSERM U
362, Institut Gustave Roussy, Villejuif; and INSERM U 91, Hopital Henri
Mondor, Creteil, France.
We have used ultrathin cryosectioning and immunogold cytochemistry
to study the position of -granules in the endocytic and biosynthetic
pathways in megakaryocytes and platelets. Morphologically, we
distinguished three types of granules; so-called multivesicular bodies
type I (MVB I) with internal vesicles only, granules with internal
vesicles and an electron dense matrix (MVB II), and the -granules
with mainly a dense content and often internal membrane vesicles at
their periphery. The MVBs were prominent in cultured megakaryocytes and
the megakaryoblastic cell line CHRF-288, but were less numerous in bone
marrow megakaryocytes and platelets, whereas -granules were most
prominent in mature bone marrow megakaryocytes and in platelets. The
internalization kinetics of bovine serum albumin-gold particles and of
fibrinogen positioned the MVB subtypes and -granules sequentially in
the endocytic pathway. MVBs contained the secretory proteins von
Willebrand factor (vWF) and -thromboglobulin ( -TG), the platelet-specific membrane protein P-selectin,
and the lysosomal membrane protein CD63. Within the MVBs, endocytosed fibrinogen and endogenous -TG were restricted to the matrix, while vWF was predominantly associated with internal vesicles. CD63 was
also observed in association with internal membrane vesicles in the
-granules. These observations, and the gradual morphologic transition from granules containing vesicles to granules containing predominantly dense material, suggest that MVBs represent a
developmental stage in -granule maturation.
Blood, Vol. 91 No. 7 (April 1), 1998:
pp. 2313-2325
© 1998 by The American Society of Hematology.

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