PUBLICATION

Dissection of vertebrate hematopoiesis using zebrafish thrombopoietin

Authors
Svoboda, O., Stachura, D.L., Machonova, O., Pajer, P., Brynda, J., Zon, L.I., Traver, D., Bartunek, P.
ID
ZDB-PUB-140530-4
Date
2014
Source
Blood   124(2): 220-8 (Journal)
Registered Authors
Bartunek, Petr, Machonova, Olga, Svoboda, Ondrej, Traver, David, Zon, Leonard I.
Keywords
none
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Blood Platelets/physiology
  • Cell Differentiation
  • Cell Proliferation
  • Cells, Cultured
  • Embryo, Nonmammalian
  • Hematopoiesis/genetics*
  • Hematopoietic Stem Cells/physiology
  • Thrombopoietin/genetics*
  • Zebrafish/embryology
  • Zebrafish/physiology*
PubMed
24869937 Full text @ Blood
Abstract
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.
Genes / Markers
Figures
Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes