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

  • Miroslaw Rybaltowski
  • Yuko Suzuki
  • Hideo Mogami
  • Iwona Chlebinska
  • Tomasz Brzoska
  • Aki Tanaka
  • Fumiaki Banno
  • Toshiyuki Miyata
  • Tetsumei Urano
Cardiovascular Physiology

DOI: 10.1007/s00424-011-0958-x

Cite this article as:
Rybaltowski, M., Suzuki, Y., Mogami, H. et al. Pflugers Arch - Eur J Physiol (2011) 461: 623. doi:10.1007/s00424-011-0958-x

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.

Keywords

Von Willebrand factor (VWF)A disintegrin-like and metalloprotease with thrombospondin type 1 motif 13 (ADAMTS13)Thrombotic thrombocytopenic purpura (TTP)Intravital confocal fluorescent microscopy

Supplementary material

424_2011_958_MOESM1_ESM.doc (24 kb)
ESM 1(DOC 24 kb)
424_2011_958_Fig8_ESM.jpg (42 kb)
Supplementary Fig. 1

(JPG 41.9 kb)

424_2011_958_MOESM2_ESM.tif (2.9 mb)
High resolution image file (TIFF 2.86 mb)
424_2011_958_Fig9_ESM.jpg (51 kb)
Supplementary Fig. 2

(JPG 50.5 kb)

424_2011_958_MOESM3_ESM.tif (1.5 mb)
High resolution image file (TIFF 1.48 mb)
424_2011_958_MOESM4_ESM.mov (1.3 mb)
Supplemental movie(MOV 1.31mb)

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Miroslaw Rybaltowski
    • 1
  • Yuko Suzuki
    • 1
  • Hideo Mogami
    • 1
  • Iwona Chlebinska
    • 1
  • Tomasz Brzoska
    • 1
  • Aki Tanaka
    • 1
  • Fumiaki Banno
    • 2
  • Toshiyuki Miyata
    • 2
  • Tetsumei Urano
    • 1
  1. 1.Department of PhysiologyHamamatsu University School of MedicineHamamatsuJapan
  2. 2.Department of Molecular PathogenesisNational Cerebral and Cardiovascular Center, Research InstituteOsakaJapan