Venous Thrombosis and Thrombocyte Activity in Zebrafish Models of Quantitative and Qualitative Fibrinogen Disorders
Venous thrombosis occurs in patients with quantitative and qualitative fibrinogen disorders. Injury-induced thrombosis in zebrafish larvae has been used to model human coagulopathies. We aimed to determine whether zebrafish models of afibrinogenemia and dysfibrinogenemia have different thrombotic ph...
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doaj-825c2bfc9b4b4006ade52d17841a2c2e2021-01-12T00:03:17ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-01-012265565510.3390/ijms22020655Venous Thrombosis and Thrombocyte Activity in Zebrafish Models of Quantitative and Qualitative Fibrinogen DisordersRichard J. Fish0Cristina Freire1Corinne Di Sanza2Marguerite Neerman-Arbez3Department of Genetic Medicine and Development, Faculty of Medicine, University of Geneva, 1211 Geneva, SwitzerlandDepartment of Genetic Medicine and Development, Faculty of Medicine, University of Geneva, 1211 Geneva, SwitzerlandDepartment of Genetic Medicine and Development, Faculty of Medicine, University of Geneva, 1211 Geneva, SwitzerlandDepartment of Genetic Medicine and Development, Faculty of Medicine, University of Geneva, 1211 Geneva, SwitzerlandVenous thrombosis occurs in patients with quantitative and qualitative fibrinogen disorders. Injury-induced thrombosis in zebrafish larvae has been used to model human coagulopathies. We aimed to determine whether zebrafish models of afibrinogenemia and dysfibrinogenemia have different thrombotic phenotypes. Laser injuries were used to induce venous thrombosis and the time-to-occlusion (TTO) and the binding and aggregation of fluorescent <i>Tg(itga2b:EGFP)</i> thrombocytes measured. The <i>fga<sup>−/−</sup></i> larvae failed to support occlusive venous thrombosis and showed reduced thrombocyte binding and aggregation at injury sites. The <i>fga<sup>+/−</sup></i> larvae were largely unaffected. When genome editing zebrafish to produce fibrinogen Aα R28C, equivalent to the human Aα R35C dysfibrinogenemia mutation, we detected in-frame skipping of exon 2 in the fga mRNA, thereby encoding Aα<sup>Δ<i>19–56</i></sup>. This mutation is similar to Fibrinogen Montpellier II which causes hypodysfibrinogenemia. Aα<sup><i>+/</i>Δ<i>19–56</i></sup> fish had prolonged TTO and reduced thrombocyte activity, a dominant effect of the mutation. Finally, we used transgenic expression of fga R28C cDNA in fga knock-down or <i>fga<sup>−/−</sup></i> mutants to model thrombosis in dysfibrinogenemia. Aα R28C expression had similar effects on TTO and thrombocyte activity as Aα<sup><i>+/</i>Δ<i>19–56</i></sup>. We conclude that thrombosis assays in larval zebrafish can distinguish between quantitative and qualitative fibrinogen disorder models and may assist in anticipating a thrombotic phenotype of novel fibrinogen mutations.https://www.mdpi.com/1422-0067/22/2/655fibrinogenfibrinthrombocytesthrombosiszebrafish |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Richard J. Fish Cristina Freire Corinne Di Sanza Marguerite Neerman-Arbez |
spellingShingle |
Richard J. Fish Cristina Freire Corinne Di Sanza Marguerite Neerman-Arbez Venous Thrombosis and Thrombocyte Activity in Zebrafish Models of Quantitative and Qualitative Fibrinogen Disorders International Journal of Molecular Sciences fibrinogen fibrin thrombocytes thrombosis zebrafish |
author_facet |
Richard J. Fish Cristina Freire Corinne Di Sanza Marguerite Neerman-Arbez |
author_sort |
Richard J. Fish |
title |
Venous Thrombosis and Thrombocyte Activity in Zebrafish Models of Quantitative and Qualitative Fibrinogen Disorders |
title_short |
Venous Thrombosis and Thrombocyte Activity in Zebrafish Models of Quantitative and Qualitative Fibrinogen Disorders |
title_full |
Venous Thrombosis and Thrombocyte Activity in Zebrafish Models of Quantitative and Qualitative Fibrinogen Disorders |
title_fullStr |
Venous Thrombosis and Thrombocyte Activity in Zebrafish Models of Quantitative and Qualitative Fibrinogen Disorders |
title_full_unstemmed |
Venous Thrombosis and Thrombocyte Activity in Zebrafish Models of Quantitative and Qualitative Fibrinogen Disorders |
title_sort |
venous thrombosis and thrombocyte activity in zebrafish models of quantitative and qualitative fibrinogen disorders |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1661-6596 1422-0067 |
publishDate |
2021-01-01 |
description |
Venous thrombosis occurs in patients with quantitative and qualitative fibrinogen disorders. Injury-induced thrombosis in zebrafish larvae has been used to model human coagulopathies. We aimed to determine whether zebrafish models of afibrinogenemia and dysfibrinogenemia have different thrombotic phenotypes. Laser injuries were used to induce venous thrombosis and the time-to-occlusion (TTO) and the binding and aggregation of fluorescent <i>Tg(itga2b:EGFP)</i> thrombocytes measured. The <i>fga<sup>−/−</sup></i> larvae failed to support occlusive venous thrombosis and showed reduced thrombocyte binding and aggregation at injury sites. The <i>fga<sup>+/−</sup></i> larvae were largely unaffected. When genome editing zebrafish to produce fibrinogen Aα R28C, equivalent to the human Aα R35C dysfibrinogenemia mutation, we detected in-frame skipping of exon 2 in the fga mRNA, thereby encoding Aα<sup>Δ<i>19–56</i></sup>. This mutation is similar to Fibrinogen Montpellier II which causes hypodysfibrinogenemia. Aα<sup><i>+/</i>Δ<i>19–56</i></sup> fish had prolonged TTO and reduced thrombocyte activity, a dominant effect of the mutation. Finally, we used transgenic expression of fga R28C cDNA in fga knock-down or <i>fga<sup>−/−</sup></i> mutants to model thrombosis in dysfibrinogenemia. Aα R28C expression had similar effects on TTO and thrombocyte activity as Aα<sup><i>+/</i>Δ<i>19–56</i></sup>. We conclude that thrombosis assays in larval zebrafish can distinguish between quantitative and qualitative fibrinogen disorder models and may assist in anticipating a thrombotic phenotype of novel fibrinogen mutations. |
topic |
fibrinogen fibrin thrombocytes thrombosis zebrafish |
url |
https://www.mdpi.com/1422-0067/22/2/655 |
work_keys_str_mv |
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