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Blood, 7 May 2009, Vol. 113, No. 19, pp. 4754-4762. Prepublished online as a Blood First Edition Paper on December 24, 2008; DOI 10.1182/blood-2008-06-162693. This Article Full Text Full Text (PDF) Supplemental Appendix, Tables, Figures, and Video All Versions of this Article: blood-2008-06-162693v1 113/19/4754    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by O'Connor, M. N. Articles by Ouwehand, W. H. Search for Related Content PubMed PubMed Citation Articles by O'Connor, M. N. Articles by Ouwehand, W. H. Related Collections Platelets and Thrombopoiesis Related Article in Blood Online Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  PLATELETS AND THROMBOPOIESIS Functional genomics in zebrafish permits rapid characterization of novel platelet membrane proteins Marie N. O'Connor1,*, Isabelle I. Salles2,*, Ana Cvejic1, Nicholas A. Watkins1,3, Adam Walker4, Stephen F. Garner1,3, Chris I. Jones5, Iain C. Macaulay1, Michael Steward4, Jaap-Jan Zwaginga6, Sarah L. Bray7, Frank Dudbridge7, Bernard de Bono8, Alison H. Goodall5, Hans Deckmyn2, Derek L. Stemple9, Willem H. Ouwehand1,3,9, on behalf of the Bloodomics Consortium 1 Department of Haematology, University of Cambridge, Cambridge, United Kingdom; 2 Laboratory for Thrombosis Research, Katholieke Universiteit Leuven, Campus Kortrijk, Kortrijk, Belgium; 3 National Health Service Blood and Transplant, Cambridge, United Kingdom; 4 Domantis Limited, Cambridge, United Kingdom; 5 Department of Cardiovascular Sciences, University of Leicester, Leicester, United Kingdom; 6 Department of Experimental Immunohaematology, Sanquin Research, Amsterdam, and the Department of Immunohaematology and Blood Transfusion of the Leiden University Medical Center, Leiden, The Netherlands; 7 Medical Research Council Biostatistics Unit, Institute of Public Health, Cambridge, United Kingdom; 8 European Bioinformatics Institute, Wellcome Trust Genome Campus, Hinxton, United Kingdom; and 9 Wellcome Trust Sanger Institute, Hinxton, United Kingdom In this study, we demonstrate the suitability of the vertebrate Danio rerio (zebrafish) for functional screening of novel platelet genes in vivo by reverse genetics. Comparative transcript analysis of platelets and their precursor cell, the megakaryocyte, together with nucleated blood cell elements, endothelial cells, and erythroblasts, identified novel platelet membrane proteins with hitherto unknown roles in thrombus formation. We determined the phenotype induced by antisense morpholino oligonucleotide (MO)–based knockdown of 5 of these genes in a laser-induced arterial thrombosis model. To validate the model, the genes for platelet glycoprotein (GP) IIb and the coagulation protein factor VIII were targeted. MO-injected fish showed normal thrombus initiation but severely impaired thrombus growth, consistent with the mouse knockout phenotypes, and concomitant knockdown of both resulted in spontaneous bleeding. Knockdown of 4 of the 5 novel platelet proteins altered arterial thrombosis, as demonstrated by modified kinetics of thrombus initiation and/or development. We identified a putative role for BAMBI and LRRC32 in promotion and DCBLD2 and ESAM in inhibition of thrombus formation. We conclude that phenotypic analysis of MO-injected zebrafish is a fast and powerful method for initial screening of novel platelet proteins for function in thrombosis. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? Related Article in Blood Online: Comparative genomics: fishing nets hemostatic catch Andrew S. Weyrich and Guy A. Zimmerman Blood 2009 113: 4479-4480. [Full Text] [PDF] This article has been cited by other articles: D. Q. Tran, J. Andersson, R. Wang, H. Ramsey, D. Unutmaz, and E. M. Shevach GARP (LRRC32) is essential for the surface expression of latent TGF-{beta} on platelets and activated FOXP3+ regulatory T cells PNAS, August 11, 2009; 106(32): 13445 - 13450. [Abstract] [Full Text] [PDF] A. S. Weyrich and G. A. Zimmerman Comparative genomics: fishing nets hemostatic catch Blood, May 7, 2009; 113(19): 4479 - 4480. [Full Text] [PDF] N. A. Watkins, A. Gusnanto, B. de Bono, S. De, D. Miranda-Saavedra, D. L. Hardie, W. G. J. Angenent, A. P. Attwood, P. D. Ellis, W. Erber, et al. A HaemAtlas: characterizing gene expression in differentiated human blood cells Blood, May 7, 2009; 113(19): e1 - e9. [Abstract] [Full Text] [PDF]

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