Original Article | Published:

Animal Models

Establishment of a congenital amegakaryocytic thrombocytopenia model and a thrombocyte–specific reporter line in zebrafish

Leukemia volume 31, pages 12061216 (2017) | Download Citation

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Abstract

Mutations in the human myeloproliferative leukemia (MPL) protein gene are known to cause congenital amegakaryocytic thrombocytopenia (CAMT). The prognosis of this heritable disorder is poor and bone marrow transplantation is the only effective treatment. Here, by using the TALEN (transcription activator-like effector nuclease) technology, we created a zebrafish mpl mutant to model human CAMT. Disruption of zebrafish mpl lead to a severe reduction in thrombocytes and a high bleeding tendency, as well as deficiencies in adult hematopoietic stem/progenitor cells. We further demonstrated that thrombocytopenia in mpl mutant zebrafish was caused by impaired Tpo/Mpl/Jak2 signaling, resulting in reduced proliferation of thrombocyte precursors. These results indicate that mpl mutant zebrafish develop thrombocytopenia resembling the human CAMT. To utilize fully zebrafish to study thrombocyte biology and thrombocytopenia disorders, we generated a transgenic reporter line Tg(mpl:eGFP)smu4, in which green fluorescent protein (GFP) expression was driven by the mpl promoter. Detailed characterization of Tg(mpl:eGFP)smu4 fish confirmed that the thrombocyte lineage was specifically marked by GFP expression. In conclusion, we generated the first transmissible congenital thrombocytopenia zebrafish model mimicking human CAMT and a thrombocyte-specific transgenic line. Together with Tg(mpl:eGFP)smu4, mpl mutant zebrafish provide a useful tool for drug screening and study of thrombocytopoiesis.

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Acknowledgements

We thank Dr Koichi Kawakami and Dr Bo Zhang for providing pTol vector and TALEN reagents and protocol, and thank Dr Yu Zhang (Merck Millipore) for FACS using an imaging flow cytometry. This work was supported by the Talent Recruitment funding and Excellent Young Teacher funding (Yq2013025) of Guangdong Higher Education Institutes, Peal River S&T Nova Program of Guangzhou (2013J2200032) and Team Program of Guangdong Natural Science Foundation (2014A030312002).

Author information

Author notes

    • Q Lin
    •  & Y Zhang

    These authors contributed equally to this work.

Affiliations

  1. Key Laboratory of Zebrafish Modeling and Drug Screening for Human Diseases of Guangdong Higher Education Institutes, Department of Developmental Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, People’s Republic of China

    • Q Lin
    • , Y Zhang
    • , R Zhou
    • , Y Zheng
    • , L Zhao
    • , M Huang
    • , X Zhang
    •  & W Zhang
  2. Division of Haematology, Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, People’s Republic of China

    • A Y H Leung

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The authors declare no conflict of interest.

Corresponding author

Correspondence to Y Zhang.

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https://doi.org/10.1038/leu.2016.320

Supplementary Information accompanies this paper on the Leukemia website (http://www.nature.com/leu)

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