Imaging analyses of coagulation-dependent initiation of fibrinolysis on activated platelets and its modification by thrombin-activatable fibrinolysis inhibitor

Tomasz Brzoska; Yuko Suzuki; Hideto Sano; Seiichirou Suzuki; Martyna Tomczyk; Hiroki Tanaka; Tetsumei Urano

doi:10.1160/TH16-09-0722

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2017; 117(04): 682-690
DOI: 10.1160/TH16-09-0722

Coagulation and Fibrinolysis

Schattauer GmbH

Imaging analyses of coagulation-dependent initiation of fibrinolysis on activated platelets and its modification by thrombin-activatable fibrinolysis inhibitor

Tomasz Brzoska

¹Department of Medical Physiology, Hamamatsu University School of Medicine, Handa-yama, Higashi-ku, Hamamatsu, Japan

,

Yuko Suzuki

¹Department of Medical Physiology, Hamamatsu University School of Medicine, Handa-yama, Higashi-ku, Hamamatsu, Japan

,

Hideto Sano

¹Department of Medical Physiology, Hamamatsu University School of Medicine, Handa-yama, Higashi-ku, Hamamatsu, Japan

,

Seiichirou Suzuki

¹Department of Medical Physiology, Hamamatsu University School of Medicine, Handa-yama, Higashi-ku, Hamamatsu, Japan

,

Martyna Tomczyk

¹Department of Medical Physiology, Hamamatsu University School of Medicine, Handa-yama, Higashi-ku, Hamamatsu, Japan

,

Hiroki Tanaka

¹Department of Medical Physiology, Hamamatsu University School of Medicine, Handa-yama, Higashi-ku, Hamamatsu, Japan

,

Tetsumei Urano

¹Department of Medical Physiology, Hamamatsu University School of Medicine, Handa-yama, Higashi-ku, Hamamatsu, Japan

› Author Affiliations

Abstract

Summary

Using intravital confocal microscopy, we observed previously that the process of platelet phosphatidylserine (PS) exposure, fibrin formation and lysine binding site-dependent plasminogen (plg) accumulation took place only in the centre of thrombi, not at their periphery. These findings prompted us to analyse the spatiotemporal regulatory mechanisms underlying coagulation and fibrinolysis. We analysed the fibrin network formation and the subsequent lysis in an in vitro experiment using diluted platelet-rich plasma supplemented with fluorescently labelled coagulation and fibrinolytic factors, using confocal laser scanning microscopy. The structure of the fibrin network formed by supplemented tissue factor was uneven and denser at the sites of coagulation initiation regions (CIRs) on PS-exposed platelets. When tissue type plasminogen activator (tPA; 7.5 nM) was supplemented, labelled plg (50 nM) as well as tPA accumulated at CIRs, from where fibrinolysis started and gradually expanded to the peripheries. The lysis time at CIRs and their peripheries (50 µm from the CIR) were 27.9 ± 6.6 and 44.4 ± 9.7 minutes (mean ± SD, n=50 from five independent experiments) after the addition of tissue factor, respectively. Recombinant human soluble thrombomodulin (TMα; 2.0 nM) attenuated the CIR-dependent plg accumulation and strongly delayed fibrinolysis at CIRs. A carboxypeptidase inhibitor dose-dependently enhanced the CIR-de- pendent fibrinolysis initiation, and at 20 µM it completely abrogated the TMα-induced delay of fibrinolysis. Our findings are the first to directly present crosstalk between coagulation and fibrinolysis, which takes place on activated platelets’ surface and is further controlled by thrombin-activatable fibrinolysis inhibitor (TAFI).

Supplementary Material to this article is available online at www.thrombosis-online.com.

Keywords

Fibrinolysis - thrombin-activatable fibrinolysis inhibitor - coagulation initiation region - plasminogen - thrombomodulin

Full Text

References

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