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Blood, Vol. 92 No. 6 (September 15), 1998:
pp. 2012-2023
Ultrastructural Analysis of Bone Marrow Hematopoiesis in Mice
Transgenic for the Thymidine Kinase Gene Driven by the
 IIb Promoter
Christel Poujol,
Diana Tronik-Le Roux,
Philippe Tropel,
Valérie Roullot,
Alan Nurden,
Gérard Marguerie, and
Paquita Nurden
From UMR 5533 CNRS, Hôpital Cardiologique, Pessac, France; and
the Laboratoire de Transgénèse et de Différenciation
Cellulaire du CEA, Grenoble, France.
Transgenic mice have been generated with expression of the herpes
virus thymidine kinase gene directed by a 2.7-kb fragment of the
 IIb murine promoter of the gene encoding the
IIb-subunit of the platelet integrin
IIb 3 (Tropel et al, Blood
90:2995, 1997). Administration of ganciclovir (GCV) to these mice
resulted not only in an acute cessation of platelet production due to
the depletion of the megakaryocytic lineage, but also a decrease in erythrocyte and leukocyte numbers. Immunogold staining on ultrathin frozen sections and electron microscopy has now shown that the remaining population of immature hematopoietic cells contain a high
proportion of Sca-1+ and CD34+ cells, with
CD45R+ cells of the lymphopoietic lineage being
maintained. Stromal cells were also preserved. Blood thrombopoietin
levels were high. At 4 days of the recovery phase, Sca-1 and CD34
antigen expression decreased with intense proliferation of cells of the
three lineages, with megakaryocyte (MK) progenitors being identified by
their positivity for glycoprotein IIb-IIIa. These results suggest that transcriptional activity for the IIb gene promoter was
present on pluripotent hematopoietic stem cells. At 6 to 8 days after cessation of GCV, numerous mature MK were observed, some of them with deformed shapes crossing the endothelial barrier through thin
apertures. Proplatelet production was visualized in the vascular sinus.
After 15 days, circulating platelet levels had increased to
approximately 65% of normal. Transgenic IIb-tk mice constitute a
valuable model to study in vivo megakaryocytopoiesis.
© 1998 by The American Society of Hematology.
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