Targeted mutagenesis of zebrafish antithrombin III triggers disseminated intravascular coagulation and thrombosis, revealing insight into function.

Autor:	Liu Y; Department of Pediatrics and., Kretz CA; Life Sciences Institute, University of Michigan, Ann Arbor, MI;, Maeder ML; Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, MA;, Richter CE; Department of Pediatrics and., Tsao P; Department of Pediatrics and., Vo AH; Department of Pediatrics and., Huarng MC; Department of Pediatrics and., Rode T; Department of Pediatrics and., Hu Z; Department of Pediatrics and., Mehra R; Department of Pathology, University of Michigan, Ann Arbor, MI;, Olson ST; Center for Molecular Biology of Oral Diseases, University of Illinois, Chicago, IL;, Joung JK; Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, MA; Department of Pathology, Harvard Medical School, Boston, MA; and., Shavit JA; Department of Pediatrics and Cellular and Molecular Biology Program, University of Michigan, Ann Arbor, MI.
Jazyk:	angličtina
Zdroj:	Blood [Blood] 2014 Jul 03; Vol. 124 (1), pp. 142-50. Date of Electronic Publication: 2014 Apr 29.
DOI:	10.1182/blood-2014-03-561027
Abstrakt:	Pathologic blood clotting is a leading cause of morbidity and mortality in the developed world, underlying deep vein thrombosis, myocardial infarction, and stroke. Genetic predisposition to thrombosis is still poorly understood, and we hypothesize that there are many additional risk alleles and modifying factors remaining to be discovered. Mammalian models have contributed to our understanding of thrombosis, but are low throughput and costly. We have turned to the zebrafish, a tool for high-throughput genetic analysis. Using zinc finger nucleases, we show that disruption of the zebrafish antithrombin III (at3) locus results in spontaneous venous thrombosis in larvae. Although homozygous mutants survive into early adulthood, they eventually succumb to massive intracardiac thrombosis. Characterization of null fish revealed disseminated intravascular coagulation in larvae secondary to unopposed thrombin activity and fibrinogen consumption, which could be rescued by both human and zebrafish at3 complementary DNAs. Mutation of the human AT3-reactive center loop abolished the ability to rescue, but the heparin-binding site was dispensable. These results demonstrate overall conservation of AT3 function in zebrafish, but reveal developmental variances in the ability to tolerate excessive clot formation. The accessibility of early zebrafish development will provide unique methods for dissection of the underlying mechanisms of thrombosis. (© 2014 by The American Society of Hematology.)
Databáze:	MEDLINE
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