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Ultrastructural demonstration of tubular inclusions coinciding with von
Willebrand factor in pig megakaryocytes
EM Cramer, J Breton-Gorius, JE Beesley and JF Martin
Service d'Hematologie et d'immunologie biologiques, CHU Bichat, Paris,
France.
The appearance of von Willebrand factor (vWF) in bone marrow megakaryocytes
was studied by standard electron microscopy (EM) and immuno-EM using an
original purification technique. Eighty percent pure megakaryocytes were
isolated from porcine rib bone marrow using Percoll gradients followed by
counterflow centrifugation. Activation was prevented by prostacyclin and
prefixation with low concentrations of glutaraldehyde. In early
megakaryoblasts, standard EM revealed the presence of tubular structures in
the small vesicles located in the Golgi area, in the small immature
alpha-granules and in the rare mature alpha-granules. Immunolabeling for
vWF was simultaneously observed in small vesicles and small alpha-granules,
mainly in the Golgi zone. In mature megakaryocytes, standard EM showed that
tubular structures were numerous, regularly spaced, and aligned in
parallel. Immunolabeling for vWF was intense, restricted to the
alpha-granules, and distributed in a similar manner to porcine platelets.
Gold particles were located eccentrically at one pole of the alpha-granule,
labeling only its periphery or outlining one side of an elongated granule.
Tubule profiles could be seen underlying the immunolabeling and were
usually located at one side of the granule. In conclusion, this study
demonstrates the presence of tubular structures in megakaryocyte alpha-
granules, their association with vWF, and the appearance of both in the
Golgi-associated vesicles.
Volume 71,
Issue 6,
pp. 1533-1538,
06/01/1988
Copyright © 1988 by The American Society of Hematology
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