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

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.
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.

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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

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  1. E.A. Dumont
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Correspondence to E.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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