Abstract
Apoptosis plays a role in cardiomyocyte death in several cardiovascular disorders. Here, we show that primary postnatal cardiomyocytes did not die upon activation of the intrinsic (cytochrome c-dependent) apoptotic pathway. Release of cytochrome c from mitochondria to the cytosol occurred, but did not activate the effector phase of apoptosis. Myocardial cells did not express apoptotic protease-activating factor-1 (Apaf-1), the allosteric activator of caspase-9 acting downstream of cytochrome c release. Forced expression of Apaf-1 restored the competence to complete the cytochrome c-induced apoptotic program and this effect was prevented by overexpression of Bcl-XL. However, cardiomyocytes were able to enter the apoptotic program when it was initiated by activation of death receptors, as observed during serum deprivation and metabolic inhibition. Our results indicate that regulation of Apaf-1 expression may be a new regulatory mechanism developed in postmitotic cells in order to prevent irreversible commitment to die after release of cytochrome c.
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Abbreviations
- Ac-DEVD-afc:
-
acetyl-Asp[OMe]-Glu[OMe]-Val-Asp[OMe]-7-amino-4-trifluoromethylcoumarin
- PBS:
-
phosphate-buffered saline
- PFA:
-
paraformaldehyde
- TUNEL:
-
TdT-mediated dUTP nick end-labeling
- z-DEVD-fmk:
-
benzyloxycarbonyl-Asp[OMe]-Glu[OMe]-Val-Asp[OMe]-fluoromethylketone
- z-IETD-fmk:
-
benzyloxycarbonyl-Ile-Glu[OMe]-Thr-Asp[OMe]-fluoromethylketone
- z-VAD-fmk:
-
benzyloxycarbonyl-Val-Ala- Asp[OMe]-fluoromethylketone
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Acknowledgements
We thank Gabriel Nuñez for the pcDNA3-Apaf-1XL construct. The present work has been partially supported by grants from the Fundació Marató de TV3 to JX Comella and M Ballester, and the Fondo de Investigaciones Sanitarias (01/3023 and PI020116 to D Sanchis and PI020051 to JX Comella). D Sanchis has been partially supported by a postdoctoral Grant from the Generalitat de Catalunya and by the Fondo de Investigaciones Sanitarias (01/3023). M Mayorga is supported by a Young Researcher Award from the Generalitat de Catalunya to JX Comella. We thank Neil Goodman for proof-reading and editing the manuscript.
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Sanchis, D., Mayorga, M., Ballester, M. et al. Lack of Apaf-1 expression confers resistance to cytochrome c-driven apoptosis in cardiomyocytes. Cell Death Differ 10, 977–986 (2003). https://doi.org/10.1038/sj.cdd.4401267
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DOI: https://doi.org/10.1038/sj.cdd.4401267
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