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Thromb Haemost 2012; 107(01): 158-166
DOI: 10.1160/TH11-04-0221
Animal Models
Schattauer GmbH

Developmental expression and organisation of fibrinogen genes in the zebrafish

Richard J. Fish
1   Department of Genetic Medicine and Development, University of Geneva Medical Centre, Geneva, Switzerland
,
Silja Vorjohann
1   Department of Genetic Medicine and Development, University of Geneva Medical Centre, Geneva, Switzerland
,
Frédérique Béna
2   Service of Genetic Medicine, University Hospitals of Geneva, Geneva, Switzerland
,
Alexandre Fort
1   Department of Genetic Medicine and Development, University of Geneva Medical Centre, Geneva, Switzerland
,
Marguerite Neerman-Arbez
1   Department of Genetic Medicine and Development, University of Geneva Medical Centre, Geneva, Switzerland
3   Service of Angiology and Haemostasis, University Hospital and Faculty of Medicine, Geneva, Switzerland
› Author Affiliations Financial support: This work was funded by the Swiss National Science Foundation, the Société Académique de Genève, the Dr Henri Dubois-Ferrière Dinu Lipatti Foundation, the Ernst and Lucie Schmidheiny Foundation and the Ernest Boninchi Foundation.
Further Information

Publication History

Received: 11 April 2011

Accepted after major revision: 03 October 2011

Publication Date:
29 November 2017 (online)

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Summary

The zebrafish is a model organism for studying vertebrate development and many human diseases. Orthologues of the majority of human coagulation factors are present in zebrafish, including fibrinogen. As a first step towards using zebrafish to model human fibrinogen disorders, we cloned the zebrafish fibrinogen cDNAs and made in situ hybridisations and quantitative reverse transcription-polymerase chain reactions (qRT-PCR) to detect zebrafish fibrinogen mRNAs. Prior to liver development or blood flow we detected zebrafish fibrinogen expression in the embryonic yolk syncytial layer and then in the early cells of the developing liver. While human fibrinogen is encoded by a three-gene, 50 kilobase (kb) cluster on chromosome 4 (FGB-FGA-FGG), recent genome assemblies showed that the zebrafish fgg gene appears distanced from fga and fgb, which we confirmed by in situ hybridisation. The zebrafish fibrinogen Bβ and γ protein chains are conserved at over 50% of amino acid positions, compared to the human polypeptides. The zebrafish Aα chain is less conserved and its C-terminal region is nearly 200 amino acids shorter than human Aα. We generated transgenic zebrafish which express a green fluorescent protein reporter gene under the control of a 1.6 kb regulatory region from zebrafish fgg. Transgenic embryos showed strong fluorescence in the developing liver, mimicking endogenous fibrinogen expression. This regulatory sequence can now be used for overexpression of transgenes in zebrafish hepatocytes. Our study is a proof-of-concept step towards using zebrafish to model human disease linked to fibrinogen gene mutations.

Keywords

Fibrinogen - zebrafish - gene cluster - transgenic - liver

 

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