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

Nature Medicine volume 7, pages 1352–1355 (2001)Cite this article

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.

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Figure 1: Experimental setup.

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Figure 2: In vivo single-cell imaging.

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Figure 3: Kinetics of Anx-V-OG binding at the tissue level.

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Figure 4: Kinetics of Anx-V-OG binding at the single-cell level.

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Figure 5: Caspase inhibition decreases Anx-V-OG binding.

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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)

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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
  2. C.P.M. Reutelingsperger
  3. J.F.M. Smits
  4. M.J.A.P. Daemen
  5. P.A.F. Doevendans
  6. H.J.J. Wellens
  7. L. Hofstra

Corresponding author

Correspondence toE.A. Dumont.

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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

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