PUBLICATION

Venous Thrombosis and Thrombocyte Activity in Zebrafish Models of Quantitative and Qualitative Fibrinogen Disorders

Authors
Fish, R.J., Freire, C., Di Sanza, C., Neerman-Arbez, M.
ID
ZDB-PUB-210115-1
Date
2021
Source
International Journal of Molecular Sciences   22(2): (Journal)
Registered Authors
di Sanza, Corinne, Fish, Richard, Neerman-Arbez, Marguerite
Keywords
fibrin, fibrinogen, thrombocytes, thrombosis, zebrafish
MeSH Terms
  • Animals
  • Base Sequence
  • Biomarkers*
  • Blood Coagulation
  • Blood Platelets/metabolism*
  • Disease Models, Animal
  • Exons
  • Fibrinogen/chemistry
  • Fibrinogen/genetics
  • Fibrinogen/metabolism*
  • Gene Editing
  • Gene Expression
  • Plasmids/genetics
  • Platelet Activation
  • RNA, Guide, Kinetoplastida
  • Sequence Deletion
  • Venous Thrombosis/blood*
  • Venous Thrombosis/diagnosis
  • Venous Thrombosis/etiology*
  • Zebrafish
PubMed
33440782 Full text @ Int. J. Mol. Sci.
Abstract
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 Tg(itga2b:EGFP) thrombocytes measured. The fga-/- larvae failed to support occlusive venous thrombosis and showed reduced thrombocyte binding and aggregation at injury sites. The fga+/- 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 AαΔ19-56. This mutation is similar to Fibrinogen Montpellier II which causes hypodysfibrinogenemia. Aα+/Δ19-56 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 fga-/- mutants to model thrombosis in dysfibrinogenemia. Aα R28C expression had similar effects on TTO and thrombocyte activity as Aα+/Δ19-56. 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.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping