Fibrinolysis pathway is an important mechanism for dissolution of fibrin clot by the action of plasmin which is formed from plasminogen, a zymogen via the action of plasminogen activators, i.e. tissue plasminogen activator and urinary plasminogen activator. The regulation of fibrinolysis system in vivo is maintained by plasminogen activators and natural inhibitors i.e. α2-antiplasmin, α2-macroglobulin, Thrombin-activatable fibrinolysis inhibitor (TAFI) and plasminogen activator inhibitor 1 and 2 (PAI-1and PAI-2). There are several fibrinolytic assays developed for human plasma but there are no reports describing fibrinolytic assay using zebrafish plasma. In this study, a fibrinolytic assay via using small amount of zebrafish plasma was developed. This assay was performed under different conditions; one by the addition of exogenous tissue plasminogen activator alone to the pooled zebrafish plasma along with calcium chloride and thromboplastin, second Dade ACTIN was used instead of tissue plasminogen activator and third Dade ACTIN along with thromboplastin was used. Epsilon amino caproic acid (EACA), a synthetic antifibrinolytic agent was used at different concentrations to inhibit fibrinolysis successfully. Similar experiments were performed on human plasma as well to check the applicability of the assay to humans and positive results were obtained. Furthermore, knockdown of tissue plasminogen activator and plasminogen genes was performed and the prolongation of peak time, the time taken for the maximal formation of fibrin was observed, similar to the EACA inhibition. In conclusion, a fibrinolysis assay using miniscule amount of plasma was developed and applied to study knockdown of fibrinolytic pathway genes. The assay developed here may have clinical utility.
| Identifer | oai:union.ndltd.org:unt.edu/info:ark/67531/metadc1833569 |
| Date | 08 1900 |
| Creators | Gill, Jaspreet Kaur |
| Contributors | Jagadeeswaran, Pudur, Root, Dough, Goven, Arthur James, 1950- |
| Publisher | University of North Texas |
| Source Sets | University of North Texas |
| Language | English |
| Detected Language | English |
| Type | Thesis or Dissertation |
| Format | vii, 22 pages : illustrations (some color), Text |
| Rights | Public, Gill, Jaspreet Kaur, Copyright, Copyright is held by the author, unless otherwise noted. All rights Reserved. |
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