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{"project":"2_test","denotations":[{"id":"17022820-11714745-8600395","span":{"begin":697,"end":699},"obj":"11714745"}],"text":"Marked differences in rebleeding time in B6 and A/J mice\nThe bleeding time, an indicator of hemostatic activity, was determined after prewarming the tail, clipping a 5 mm segment, placing the tail in warm saline and measuring the time for the bleeding to stop. We found this procedure was consistent and reproducible in different mouse strains and was not gender dependent. Bleeding time was not different in A/J mice compared to B6 mice (Figure 2A). In order to determine if specific pathways were altered in the B6 and A/J strains, mice with known alterations in hemostasis, Lepob mice, or thrombosis, Plg-/- and PAI-/- mice, were also tested. The Lepob mice have impaired platelet aggregation [26] and were evaluated as a reference for platelet function. In the Lepob mice (Figure 2A) there was nearly a 10-fold increase in bleeding time compared to B6 mice. Bleeding time was also significantly (P = 0.0001) increased by 66% in the Plg-/- mice compared to WT mice littermates. Bleeding time was not different in PAI-1-/- mice compared to B6 mice. In contrast to the Lepob mice with a platelet defect and 10-fold increase in bleeding time, there was no difference in the bleeding time between B6 and A/J mice and suggested platelet aggregation is not altered in these two strains.\nFigure 2 Bleeding and rebleeding time in B6, A/J, Lepob, Plg-/- and PAI-1-/- mice. Panel A, B: Bleeding time (seconds) and Panel C, D: Rebleeding time (seconds). To determine statistical differences, values from B6 and A/J and from WT and Plg-/- littermates were compared with a t-test, and values from B6, PAI-1-/- and Lepob were compared by a one-way ANOVA and a Newman-Keuls post-test. Symbols indicate a significant difference (*P \u003c 0.05, **P \u003c 0.01, ***P \u003c 0.001) between B6 or WT mice. Values are the mean ± SEM of 10–34 mice per genotype. Rebleeding time, an indicator of thrombus stability, was measured as the time between the cessation of bleeding and the start of the second bleeding (Figure 2B). Rebleeding times were significantly higher in A/J (2.6-fold, P \u003c 0.001) mice than the B6 mice. In contrast, the rebleeding time was nearly 8.5-fold less in the Lepob mice than the B6 mice. To determine how alterations in the fibrinolytic system may affect rebleeding time, Plg-/- and PAI-1-/- mice were also tested (Figure 2B). Rebleeding time was not different in Plg-/- mice compared to WT littermates, but PAI-1-/- mice had a significantly (P \u003c 0.05) reduced rebleeding time, which was 1.8-fold lower than for the B6 mice. Although a Plg deficiency did not result in a prolonged rebleeding time as anticipated, a PAI-1 deficiency resulted in a shortened time. The markedly increased rebleeding time in the A/J may suggest differences in the regulation of the Plg network."}

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PMC:1617083 / 14472-17239 JSON TXT