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Zebrafish Kit ligands cooperate with erythropoietin to promote erythroid cell expansion
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SYSNO ASEP 0539675 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Zebrafish Kit ligands cooperate with erythropoietin to promote erythroid cell expansion Author(s) Oltová, Jana (UMG-J)
Svoboda, Ondřej (UMG-J)
Machoňová, Olga (UMG-J)
Svatoňová, Petra (UMG-J)
Traver, D. (US)
Kolář, Michal (UMG-J) RID, ORCID
Bartůněk, Petr (UMG-J) RIDNumber of authors 7 Source Title Blood Advances. - : Elsevier - ISSN 2473-9529
Roč. 4, č. 23 (2020), s. 5915-5924Number of pages 10 s. Publication form Online - E Language eng - English Country US - United States Keywords c-kit ; progenitor cells ; terminal differentiation ; glucocorticoid-receptor ; clonal analysis ; growth-factor ; gene-product ; hematopoiesis ; expression ; chicken Subject RIV EB - Genetics ; Molecular Biology OECD category Cell biology R&D Projects GA16-21024S GA ČR - Czech Science Foundation (CSF) NV19-07-00412 GA MZd - Ministry of Health (MZ) Method of publishing Open access Institutional support UMG-J - RVO:68378050 UT WOS 000596950900003 DOI 10.1182/bloodadvances.2020001700 Annotation Kit ligand (Kitlg) is pleiotropic cytokine with a prominent role in vertebrate erythropoiesis. Although the role of Kitlg in this process has not been reported in Danio rerio (zebrafish), in the present study we show that its function is evolutionarily conserved. Zebrafish possess 2 copies of Kitlg genes (Kitlga and Kitlgb) as a result of whole-genome duplication. To determine the role of each ligand in zebrafish, we performed a series of ex vivo and in vivo gain- and loss-of-function experiments. First, we tested the biological activity of recombinant Kitlg proteins in suspension culture from zebrafish whole-kidney marrow, and we demonstrate that Kitlga is necessary for expansion of erythroid progenitors ex vivo. To further address the role of kitlga and kitlgb in hematopoietic development in vivo, we performed gain-of-function experiments in zebrafish embryos, showing that both ligands cooperate with erythropoietin (Epo) to promote erythroid cell expansion. Finally, using the kita mutant (kita(b5/b5) or sparse), we show that the Kita receptor is crucial for Kitlga/b cooperation with Epo in erythroid cells. In summary, using optimized suspension culture conditions with recombinant cytokines (Epo, Kitlga), we report, for the first time, ex vivo suspension cultures of zebrafish hematopoietic progenitor cells that can serve as an indispensable tool to study normal and aberrant hematopoiesis in zebrafish. Furthermore, we conclude that, although partial functional diversification of Kit ligands has been described in other processes, in erythroid development, both paralogs play a similar role, and their function is evolutionarily conserved. Workplace Institute of Molecular Genetics Contact Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Year of Publishing 2021 Electronic address https://ashpublications.org/bloodadvances/article/4/23/5915/474371/Zebrafish-Kit-ligands-cooperate-with
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