Re-establishment of VWF-dependent Weibel-Palade bodies in VWD endothelial cells.
Type 3 von Willebrand disease (VWD) is a severe hemorrhagic defect in humans. We now identify the homozygous mutation in the Chapel Hill strain of canine type 3 VWD that results in premature termination of von Willebrand factor (VWF) protein synthesis. We cultured endothelium from VWD and normal dogs to study intracellular VWF trafficking and Weibel-Palade body formation. Weibel-Palade bodies could not be identified in the canine VWD aortic endothelial cells (VWD-AECs) by P-selectin, VWFpp, or VWF immunostaining and confocal microscopy. We demonstrate the reestablishment of Weibel-Palade bodies that recruit endogenous P-selectin by expressing wild-type VWF in VWD-AECs. Expression of mutant VWF proteins confirmed that VWF multimerization is not necessary for Weibel-Palade body creation. Although the VWF propeptide is required for the formation of Weibel-Palade bodies, it cannot independently induce the formation of the granule. These VWF-null endothelial cells provide a unique opportunity to examine the biogenesis of Weibel-Palade bodies in endothelium from a canine model of type 3 VWD.
Haberichter
Sandra L
SL
Department of Pediatrics, Medical College of Wisconsin, Milwaukee 53226, USA. shaberic@mail.mcw.edu
Merricks
Elizabeth P
EP
Fahs
Scot A
SA
Christopherson
Pamela A
PA
Nichols
Timothy C
TC
Montgomery
Robert R
RR
D000595:Amino Acid Sequence; D000818:Animals; D001011:Aorta / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D001483:Base Sequence; D002478:Cells, Cultured; D004285:Dogs; D042783:Endothelial Cells / Q000378:metabolism* / Q000473:pathology*; D016368:Frameshift Mutation / Q000235:genetics; D005786:Gene Expression Regulation; D006801:Humans; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D020302:Weibel-Palade Bodies / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D014842:von Willebrand Diseases / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D014841:von Willebrand Factor / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D014841:von Willebrand Factor