Skip to main content
SpringerLink
Log in

Real-time imaging of apoptotic cell-membrane changes at the single-cell level in the beating murine heart

  • On the Market
  • Published:

From Nature Medicine

View current issue Submit your manuscript

An Erratum to this article was published on 01 January 2002

Abstract

We report a novel real-time imaging model to visualize apoptotic membrane changes of single cardiomyocytes in the injured heart of the living mouse, using fluorescent labeled annexin-V. Annexin-V binds to externalized phosphatidylserine (PS) of cells undergoing programmed cell death. With high-magnification (×100–160) real-time imaging, we visualized the binding of annexin-V to single cardiomyocytes. Kinetic studies at the single-cell level revealed that cardiomyocytes started to bind annexin-V within minutes after reperfusion, following an ischemic period of 30 minutes. The amount of bound annexin-V increased rapidly and reached a maximum within 20–25 minutes. Caspase inhibitors decreased the number of annexin-V–positive cardiomyocytes and slowed down the rate of PS exposure of cardiomyocytes that still bound annexin-V. This technology to study cell biology in the natural environment will enhance knowledge of intracellular signaling pathways relevant for cell-death regulation and strategies to manipulate these pathways for therapeutic effect.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1: Experimental setup.
Figure 2: In vivo single-cell imaging.
Figure 3: Kinetics of Anx-V-OG binding at the tissue level.
Figure 4: Kinetics of Anx-V-OG binding at the single-cell level.
Figure 5: Caspase inhibition decreases Anx-V-OG binding.

References

  1. Tempany, C.M. & McNeil, B.J. Advances in biomedical imaging. JAMA 285, 562–567 (2001).

    Article  CAS  Google Scholar 

  2. Plymale, D.R., Haskins, J.R. & De La Iglesia, F.A. Monitoring simultaneous subcellular events in vitro by means of coherent multiprobe fluorescence. Nature Med. 5, 351–355 (1999).

    Article  CAS  Google Scholar 

  3. Sweeney, T.J. et al. Visualizing the kinetics of tumor-cell clearance in living animals. Proc. Natl. Acad. Sci. USA 96, 12044–12049 (1999).

    Article  CAS  Google Scholar 

  4. Yang, M. et al. Whole-body and intravital optical imaging of angiogenesis in orthotopically implanted tumors. Proc. Natl. Acad. Sci. USA 98, 2616–2621 (2001).

    Article  CAS  Google Scholar 

  5. Dumont, E.A. et al. Cardiomyocyte death induced by myocardial ischemia and reperfusion: measurement with recombinant human annexin-V in a mouse model. Circulation 102, 1564–1568 (2000).

    Article  CAS  Google Scholar 

  6. Koopman, G. et al. Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood 84, 5–20 (1994).

    Google Scholar 

  7. Martin, S.J. et al. Early redistribution of plasma membrane phosphatidylserine is a general feature of apoptosis regardless of the initiating stimulus: inhibition by overexpression of Bcl-2 and Abl. J. Exp. Med. 182, 552–556 (1995).

    Article  Google Scholar 

  8. Goldstein, J.C., Waterhouse, N.J., Juin, P., Evan, G.I. & Green, D.R. The coordinate release of cytochrome c during apoptosis is rapid, complete and kinetically invariant. Nature Cell Biol. 2, 156–162 (2000).

    Article  CAS  Google Scholar 

  9. Xiang, J., Chao, D.T. & Korsmeyer, S.J. Bax-induced cell death may not require interleukin 1β converting enzyme-like proteases. Proc. Natl. Acad. Sci. USA 93, 14559–14563 (1996).

    Article  CAS  Google Scholar 

  10. Bell, J.I. Clinical research is dead; long live clinical research. Nature Med. 5, 477–478 (1999).

    Article  CAS  Google Scholar 

  11. Contag, P. R., Olomu, I. N., Stevenson, D. K. & Contag, C. H. Bioluminescent indicators in living mammals. Nature Med. 4, 245–247 (1998).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank N. Steinmetz and B. Armstrong for comments and suggestions. This study was supported from grants from the Dutch Heart Foundation (NHS 98.195 and NHS 2000-D035) and the Wynand-Pon Foundation. L.H. is a clinical research fellow for the Dutch Heart Foundation (NHS 2000-D035)

Author information

Authors and Affiliations

  1. Cardiovascular Research Institute Maastricht, Universiteitssingel 50, Maastricht, the Netherlands

    E.A. Dumont, C.P.M. Reutelingsperger, J.F.M. Smits, M.J.A.P. Daemen, P.A.F. Doevendans, H.J.J. Wellens & L. Hofstra

Authors
  1. E.A. Dumont
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. C.P.M. Reutelingsperger
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J.F.M. Smits
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M.J.A.P. Daemen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. P.A.F. Doevendans
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. H.J.J. Wellens
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. L. Hofstra
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to E.A. Dumont.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dumont, E., Reutelingsperger, C., Smits, J. et al. Real-time imaging of apoptotic cell-membrane changes at the single-cell level in the beating murine heart. Nat Med 7, 1352–1355 (2001). https://doi.org/10.1038/nm1201-1352

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm1201-1352

  • Springer Nature America, Inc.

This article is cited by

Search

Navigation