title: Absence of GPIbalpha is responsible for aberrant membrane development during megakaryocyte maturation: ultrastructural study using a transgenic model.
abstract: OBJECTIVE
  The glycoprotein Ib/IX/V complex (GPIb-IX-V) mediates platelet attachment to von Willebrand factor in exposed subendothelium. Molecular defects in the genes for GPIbalpha, GPIbbeta, and GPIX give rise to the Bernard-Soulier syndrome, in which thrombocytopenia and giant platelets suggest that this receptor also is involved in platelet production. To study how giant platelets are produced in vivo, we used a model of GPIbalpha-deficient mice (GPIbalpha(null)) and mice rescued with the human GPIbalpha transgene (GPIbalpha(null;hTg)).
  
  
  METHODS
  Using electron microscopy and immunogold labeling, we examined megakaryocytopoiesis in the bone marrow of these mice and developed a method to quantify the membranes of megakaryocytes (MK) and proplatelets by computer analysis.
  
  
  RESULTS
  Abnormal membrane development in the perinuclear zone was found in immature MK of GPIbalpha(null) mice. This led to a poorly developed demarcation membrane system and other ultrastructural changes. As a result, well-organized platelet territories were rarely seen within the cytoplasm of mature MK. Membrane quantification confirmed that MK of GPIbalpha(null) mice had a reduced internal membrane pool. Whereas these MK normally crossed the endothelial barrier, their migration was accompanied by the production of unusually large MK fragments or proplatelets in the vascular sinus with an approximately 50% decrease in internal membrane content compared to wild-type. In the rescued GPIbalpha(null;hTg) model, GPIbalpha was normally localized in MK, and there was a total correction of the ultrastructural defects.
  
  
  CONCLUSIONS
  GPIbalpha is essential for membrane development and distribution in maturing MK. Its absence leads to abnormal partitioning of the membrane systems and abnormal proplatelet production.
authors.affiliation: UMR 5533 CNRS, Laboratoire d'Hématologie, Hôpital Cardiologique, Avenue de Magellan, 33604 Pessac, France.
authors.forename: Christel
authors.initials: C
authors.lastname: Poujol
authors.forename: Jerry
authors.initials: J
authors.lastname: Ware
authors.forename: Bernhard
authors.initials: B
authors.lastname: Nieswandt
authors.forename: Alan T
authors.initials: AT
authors.lastname: Nurden
authors.forename: Paquita
authors.initials: P
authors.lastname: Nurden
mesh_terms: D000818:Animals; D001606:Bernard-Soulier Syndrome; D001792:Blood Platelets / Q000473:pathology; D001854:Bone Marrow Cells; D002454:Cell Differentiation; D002462:Cell Membrane / Q000235:genetics / Q000473:pathology*; D002465:Cell Movement; D004730:Endothelium, Vascular; D006801:Humans; D008533:Megakaryocytes / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure; D008565:Membrane Proteins / Q000235:genetics / Q000502:physiology; D051379:Mice; D008822:Mice, Transgenic; D008854:Microscopy, Electron; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000235:genetics / Q000502:physiology*; D010980:Platelet Membrane Glycoproteins
chemical_list: D008565:Membrane Proteins; D019038:Platelet Glycoprotein GPIb-IX Complex; D010980:Platelet Membrane Glycoproteins; C032193:glycoprotein receptor GPIb-IX