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Real‐time measurement of non‐lethal platelet thromboenbolic responses in the anaethetized mouse
Author(s) -
Tymvios Charalambos M,
Jones Sarah,
Moore Christopher,
Emerson Michael
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.2_supplement.38
Subject(s) - platelet , in vivo , thrombin , platelet activation , pharmacology , chemistry , bleeding time , endothelium , in vitro , medicine , immunology , platelet aggregation , biochemistry , biology , microbiology and biotechnology
In vitro platelet aggregation is a poor predictor of platelet responses in vivo where the vascular endothelium plays an important role in the platelet response. Intravenous injection of platelet agonists into conscious mice has previously been shown to induce thromboembolic mortality that is largely mediated through platelets. This technique is limited scientifically and inflicts considerable pain and suffering in experimental animals. We therefore developed methodology for assessing in vivo platelet function non‐invasively and in real‐time. Platelets were isolated from anaesthetised donor mice, radiolabelled with 111 Indium Oxine and re‐infused into anaesthetised mice. Circulating platelets were monitored with a miniaturised crystal scintillation probe placed over the pulmonary vascular. Intravenous injection of ADP (0.4 – 400 μg/Kg), thrombin (100–1000 IU/Kg) and collagen (25 – 100 μg/Kg) induced dose‐dependent changes in platelet counts that consisted of a rapid increase in the pulmonary probe as platelets aggregated and became trapped in the pulmonary vascular. NO inhibitors (L‐NAME, L‐NMMA) were infused to inhibit NO from platelets and the vascular endothelium and significantly enhanced platelet responses. We therefore suggest that our model will be useful in determining the effects of platelet and endothelial modulators on platelet thromboembolic responses.