title: Thrombin Generation in Zebrafish Blood.
abstract: To better understand hypercoagulability as an underlying cause for thrombosis, the leading cause of death in the Western world, new assays to study ex vivo coagulation are essential. The zebrafish is generally accepted as a good model for human hemostasis and thrombosis, as the hemostatic system proved to be similar to that in man. Their small size however, has been a hurdle for more widespread use in hemostasis related research. In this study we developed a method that enables the measurement of thrombin generation in a single drop of non-anticoagulated zebrafish blood. Pre-treatment of the fish with inhibitors of FXa and thrombin, resulted in a dose dependent diminishing of thrombin generation, demonstrating the validity of the assay. In order to establish the relationship between whole blood thrombin generation and fibrin formation, we visualized the resulting fibrin network by scanning electron microscopy. Taken together, in this study we developed a fast and reliable method to measure thrombin generation in whole blood collected from a single zebrafish. Given the similarities between coagulation pathways of zebrafish and mammals, zebrafish may be an ideal animal model to determine the effect of novel therapeutics on thrombin generation. Additionally, because of the ease with which gene functions can be silenced, zebrafish may serve as a model organism for mechanistical research in thrombosis and hemostasis.
authors.affiliation: Cardiovascular Research Institute Maastricht, Maastricht University Medical Centre, Maastricht, The Netherlands.|Synapse BV, Maastricht, The Netherlands.
authors.forename: Evelien
authors.initials: E
authors.lastname: Schurgers
authors.affiliation: Cardiovascular Research Institute Maastricht, Maastricht University Medical Centre, Maastricht, The Netherlands.|Synapse BV, Maastricht, The Netherlands.
authors.forename: Martijn
authors.initials: M
authors.lastname: Moorlag
authors.affiliation: Cardiovascular Research Institute Maastricht, Maastricht University Medical Centre, Maastricht, The Netherlands.|Synapse BV, Maastricht, The Netherlands.
authors.forename: Coenraad
authors.initials: C
authors.lastname: Hemker
authors.affiliation: Cardiovascular Research Institute Maastricht, Maastricht University Medical Centre, Maastricht, The Netherlands.|Synapse BV, Maastricht, The Netherlands.
authors.forename: Theo
authors.initials: T
authors.lastname: Lindhout
authors.affiliation: Cardiovascular Research Institute Maastricht, Maastricht University Medical Centre, Maastricht, The Netherlands.|Synapse BV, Maastricht, The Netherlands.
authors.forename: Hilde
authors.initials: H
authors.lastname: Kelchtermans
authors.affiliation: Cardiovascular Research Institute Maastricht, Maastricht University Medical Centre, Maastricht, The Netherlands.|Synapse BV, Maastricht, The Netherlands.
authors.forename: Bas
authors.initials: B
authors.lastname: de Laat
mesh_terms: D000818:Animals; D001777:Blood Coagulation; D001780:Blood Coagulation Tests / Q000379:methods*; D015951:Factor Xa / Q000378:metabolism; D005260:Female; D005337:Fibrin / Q000378:metabolism / Q000648:ultrastructure; D029941:Fish Proteins / Q000097:blood / Q000378:metabolism; D008297:Male; D013917:Thrombin / Q000032:analysis / Q000378:metabolism*; D015027:Zebrafish / Q000097:blood* / Q000378:metabolism
chemical_list: D029941:Fish Proteins; D005337:Fibrin; D013917:Thrombin; D015951:Factor Xa