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In vivo imaging analysis of the interaction between unusually large von Willebrand factor multimers and platelets on the surface of vascular wall

  • Cardiovascular Physiology
  • Published:
Pflügers Archiv - European Journal of Physiology Aims and scope Submit manuscript

Abstract

To elucidate how unusually large von Willebrand factor (UL-VWF) multimers facilitate thrombus formation, their behavior was analyzed together with that of platelets in living mice deficient in the gene encoding the protease that cleaves UL-VWF, a disintegrin-like and metalloprotease with thrombospondin type 1 motif 13 (ADAMTS13−/−). By crossing ADAMTS13−/− mice with green fluorescent protein-expressing transgenic mice (GFP mice), GFP-ADAMTS13−/− mice were obtained. The dynamics of GFP-expressing platelets were monitored employing intravital confocal fluorescent microscopy. Administration of a vasopressin derivative, DDAVP, a secretagogue of VWF increased the number of platelets adhered to vascular endothelial cells (VECs) on mesentery at sites recognized by an anti-VWF antibody. Some of these platelets were interconnected and aligned as beads on a string. They reached their maximum length at 5 min and were longer in GFP-ADAMTS13−/− mice than in GFP mice (5.3 ± 4.3, N = 6 vs 2.9 ± 2.1 μm, N = 4) (mean±SE). Focal injury of VECs by topical application of FeCl3 developed longer (25, 3–50 vs 10, 2–25 μm, P < 0.01) (mean, 10th–90th percentile) and more stable (1.3, 0.3–6.3 vs 0.3, 0.2–1.3 s, P < 0.01) connected platelets in GFP-ADAMTS13−/− mice than in GFP mice. This study revealed that ADAMTS13 cleaves platelet-bound UL-VWF multimers, both during their secretion from VECs and after their adherence to injured vascular walls in veins. UL-VWF multimers either being secreted from VECs or circulating in plasma of ADAMTS13−/− mice appeared to facilitate the accumulation of longer and more stable VWF strings with more associated platelets on injured vascular walls.

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Acknowledgments

This work was supported in part by Grants-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (JSPS) (to T.U., Y.S., H.M., F.B., and T.M.), the Ministry of Health, Labor, and Welfare of Japan (to T.M.), the Japan Society for the Promotion of Science (to T.M.), the Program for Promotion of Fundamental Studies in Health Sciences of the National Institute of Biochemical Innovation of Japan (to T.M.), the Smoking Research Foundation (to T.U.), and a scholarship to M.R. from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

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The authors declare that there are no conflicts of interest associated with study.

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Authors and Affiliations

  1. Department of Physiology, Hamamatsu University School of Medicine, 1-20-1, Handa-yama, Higashi-ku, Hamamatsu, 431–3192, Japan

    Miroslaw Rybaltowski, Yuko Suzuki, Hideo Mogami, Iwona Chlebinska, Tomasz Brzoska, Aki Tanaka & Tetsumei Urano

  2. Department of Molecular Pathogenesis, National Cerebral and Cardiovascular Center, Research Institute, 5-7-1 Fujishiro-dai, Suita, Osaka, 565-8565, Japan

    Fumiaki Banno & Toshiyuki Miyata

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  1. Miroslaw Rybaltowski
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Correspondence to Tetsumei Urano.

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Rybaltowski, M., Suzuki, Y., Mogami, H. et al. In vivo imaging analysis of the interaction between unusually large von Willebrand factor multimers and platelets on the surface of vascular wall. Pflugers Arch - Eur J Physiol 461, 623–633 (2011). https://doi.org/10.1007/s00424-011-0958-x

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