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Dissection of vertebrate hematopoiesis using zebrafish thrombopoietin
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SYSNO ASEP 0436400 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Dissection of vertebrate hematopoiesis using zebrafish thrombopoietin Author(s) Svoboda, Ondřej (UMG-J)
Stachura, D.L. (US)
Machoňová, Olga (UMG-J)
Pajer, Petr (UMG-J)
Brynda, Jiří (UMG-J) RID
Zon, L.I. (US)
Traver, D. (US)
Bartůněk, Petr (UMG-J) RIDSource Title Blood. - : American Society of Hematology - ISSN 0006-4971
Roč. 124, č. 2 (2014), s. 220-228Number of pages 9 s. Language eng - English Country US - United States Keywords Zebrafish ; hematopoiesis ; progenitors ; thrombopoietin ; erythropoietin Subject RIV EB - Genetics ; Molecular Biology R&D Projects GAP305/10/0953 GA ČR - Czech Science Foundation (CSF) UT WOS 000342618600015 DOI 10.1182/blood-2014-03-564682 Annotation In non-mammalian vertebrates, the functional units of hemostasis are thrombocytes. Thrombocytes are thought to arise from bi-potent thrombocytic/erythroid progenitors (TEPs). TEPs have been experimentally demonstrated in avian models of hematopoiesis, and mammals possess functional equivalents known as megakaryocyte/erythroid progenitors (MEPs). However, the presence of TEPs in teleosts has only been speculated. To identify and prospectively isolate TEPs, we identified, cloned, and generated recombinant zebrafish thrombopoietin (Tpo). Tpo mRNA expanded itga2b:GFP+ (cd41:GFP+) thrombocytes as well as hematopoietic stem and progenitor cells (HSPCs) in the zebrafish embryo. Utilizing Tpo in clonal methylcellulose assays, we describe for the first time the prospective isolation and characterization of TEPs from transgenic zebrafish. Combinatorial use of zebrafish Tpo, erythropoietin (Epo), and granulocyte colony stimulating factor (Gcsf) allowed the investigation of HSPCs responsible for erythro-, myelo-, and thrombo-poietic differentiation. Utilizing these assays allowed the visualization and differentiation of hematopoietic progenitors ex vivo in real-time with time-lapse and high-throughput microscopy, allowing analyses of their clonogenic and proliferative capacity. These studies indicate that the functional role of Tpo in the differentiation of thrombocytes from HSPCs is well conserved among vertebrate organisms, positing the zebrafish as an excellent model to investigate diseases caused by dysregulated erythro- and thrombo-poietic differentiation. Workplace Institute of Molecular Genetics Contact Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Year of Publishing 2015
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