Skip to main content
SpringerLink
Log in

Clot Retraction

  • Protocol
  • First Online:
Platelets and Megakaryocytes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 788))

Abstract

The study of clot retraction in vitro has been adopted as a simple and reproducible approach to assess platelet function. Plasma clots should retract away from the sides of a glass tube within a few hours allowing the rapid characterization of outside-in signaling through platelet integrin αIIbβ3. In this chapter, we describe the role of platelets in fibrin clot retraction and provide a detailed description of the methods used to assess this process.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Abbreviations

AMPK α2:

Adenosine monophosphate (AMP)-activated protein kinase α2

PLCγ2:

Phospholipase Cγ2

PP2:

4-Amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo-d-3,4-pyrimidine

PRP:

Platelet-rich plasma

RBC:

Red blood cell

References

  1. Farndale, R. W., Sixma, J. J., Barnes, M. J., and de Groot, P. G. (2004) The role of collagen in thrombosis and hemostasis. J Thromb Haemost 2, 561–73.

    Article  PubMed  CAS  Google Scholar 

  2. Jackson, S. P., Nesbitt, W. S., and Kulkarni, S. (2003) Signaling events underlying thrombus formation. J Thromb Haemost 1, 1602–12.

    Article  PubMed  CAS  Google Scholar 

  3. Gibbins, J., Asselin, J., Farndale, R., Barnes, M., Law, C. L., and Watson, S. P. (1996) Tyrosine phosphorylation of the Fc receptor gamma-chain in collagen-stimulated platelets. J Biol Chem 271, 18095–9.

    Article  PubMed  CAS  Google Scholar 

  4. Coughlin, S. R. (2000) Thrombin signalling and protease-activated receptors. Nature 407, 258–64.

    Article  PubMed  CAS  Google Scholar 

  5. Shattil, S. J., and Newman, P. J. (2004) Integrins: dynamic scaffolds for adhesion and signaling in platelets. Blood 104, 1606–15.

    Article  PubMed  CAS  Google Scholar 

  6. Law, D. A., DeGuzman, F. R., Heiser, P., Ministri-Madrid, K., Killeen, N., and Phillips, D. R. (1999) Integrin cytoplasmic tyrosine motif is required for outside-in alphaIIbbeta3 signalling and platelet function. Nature 401, 808–11.

    Article  PubMed  CAS  Google Scholar 

  7. Shattil, S. J., Kashiwagi, H., and Pampori, N. (1998) Integrin signaling: the platelet paradigm. Blood 91, 2645–57.

    PubMed  CAS  Google Scholar 

  8. Weyrich, A. S., Denis, M. M., Schwertz, H., Tolley, N. D., Foulks, J., Spencer, E., Kraiss, L. W., Albertine, K. H., McIntyre, T. M., and Zimmerman, G. A. (2007) mTOR-dependent synthesis of Bcl-3 controls the retraction of fibrin clots by activated human platelets. Blood 109, 1975–83.

    Article  PubMed  CAS  Google Scholar 

  9. Berndt, M. C., and Andrews, R. K. (2007) Full clot retraction: running on mTOR. Blood 109, 1791–2.

    Article  CAS  Google Scholar 

  10. Suzuki-Inoue, K., Hughes, C. E., Inoue, O., Kaneko, M., Cuyun-Lira, O., Takafuta, T., Watson, S. P., and Ozaki, Y. (2007) Involvement of Src kinases and PLCgamma2 in clot retraction. Thromb Res 120, 251–8.

    Article  PubMed  CAS  Google Scholar 

  11. Schoenwaelder, S. M., Jackson, S. P., Yuan, Y., Teasdale, M. S., Salem, H. H., and Mitchell, C. A. (1994) Tyrosine kinases regulate the cytoskeletal attachment of integrin alpha IIb beta 3 (platelet glycoprotein IIb/IIIa) and the cellular retraction of fibrin polymers. J Biol Chem 269, 32479–87.

    PubMed  CAS  Google Scholar 

  12. Randriamboavonjy, V., Isaak, J., Fromel, T., Viollet, B., Fisslthaler, B., Preissner, K. T., and Fleming, I. AMPK alpha2 subunit is involved in platelet signaling, clot retraction, and thrombus stability. Blood 116, 2134–40.

    Google Scholar 

  13. Osdoit, S., and Rosa, J. P. (2001) Fibrin clot retraction by human platelets correlates with alpha(IIb)beta(3) integrin-dependent protein tyrosine dephosphorylation. J Biol Chem 276, 6703–10.

    Article  PubMed  CAS  Google Scholar 

  14. Seiffert, D., Pedicord, D. L., Kieras, C. J., He, B., Stern, A. M., and Billheimer, J. T. (2002) Regulation of clot retraction by glycoprotein IIb/IIIa antagonists. Thromb Res 108, 181–9.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute for Cardiovascular and Metabolic Research, School of Biological Sciences, The University of Reading, Reading, UK

    Katherine L. Tucker, Tanya Sage & Jonathan M. Gibbins

Authors
  1. Katherine L. Tucker
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tanya Sage
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jonathan M. Gibbins
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Katherine L. Tucker .

Editor information

Editors and Affiliations

  1. School of Biological Sciences, Inst. Cardiovascular & Metabolic Res., University of Reading, Reading, RG6 6UB, Berkshire, United Kingdom

    Jonathan M. Gibbins

  2. , Dept. Cell Physiology & Pharmacology, University of Leicester, University Road, Leicester, LE1 9HN, United Kingdom

    Martyn P. Mahaut-Smith

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Tucker, K.L., Sage, T., Gibbins, J.M. (2012). Clot Retraction. In: Gibbins, J., Mahaut-Smith, M. (eds) Platelets and Megakaryocytes. Methods in Molecular Biology, vol 788. Springer, New York, NY. https://doi.org/10.1007/978-1-61779-307-3_8

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-307-3_8

  • Published:

  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-61779-306-6

  • Online ISBN: 978-1-61779-307-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation