Abstract
Although proteases of the caspase family are essential mediators of apoptosis in nucleated cells, in anucleate cells their presence and potential functions are almost completely unknown. Human erythrocytes are a major cell population that does not contain a cell nucleus or other organelles. However, during senescence they undergo certain morphological alterations resembling apoptosis. In the present study, we found that mature erythrocytes contain considerable amounts of caspase-3 and -8, whereas essential components of the mitochondrial apoptotic cascade such as caspase-9, Apaf-1 and cytochrome c were missing. Strikingly, although caspases of erythrocytes were functionally active in vitro, they failed to become activated in intact erythrocytes either during prolonged storage or in response to various proapoptotic stimuli. Following an increase of cytosolic calcium, instead the cysteine protease calpain but not caspases became activated and mediated fodrin cleavage and other morphological alterations such as cell shrinkage. Our results therefore suggest that erythrocytes do not have a functional death system. In addition, because of the presence of procaspases and the absence of a cell nucleus and mitochondria erythrocytes may be an attractive system to dissect the role of certain apoptosis-regulatory pathways. Cell Death and Differentiation (2001) 8, 1197–1206
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Abbreviations
- Ac-DEVD-AMC:
-
N-acetyl-Asp-Glu-Val-Asp-aminomethylcoumarin
- Apaf-1:
-
apoptotic protease-activating factor-1
- CD95L:
-
CD95 ligand
- CHX:
-
cycloheximide
- RBC:
-
red blood cell
- TNF:
-
tumor necrosis factor
- z-VAD-fmk:
-
benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone
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Acknowledgements
The authors wish to thank Dr. V Dixit for kindly providing the bacterial expression plasmid for caspase-6. This work was supported in part by grants from the Federal Ministry of Education, Science, Research and Technology (Fö. 01KS9602) and the Interdisciplinary Center of Clinical Research (IZKF) of the Universities of Tübingen and Münster. S Wesselborg acknowledges a fellowship from the Federal Ministry of Education, Science, Research and Technology, and C Berg from the Fortune Program of the University of Tübingen.
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Berg, C., Engels, I., Rothbart, A. et al. Human mature red blood cells express caspase-3 and caspase-8, but are devoid of mitochondrial regulators of apoptosis. Cell Death Differ 8, 1197–1206 (2001). https://doi.org/10.1038/sj.cdd.4400905
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DOI: https://doi.org/10.1038/sj.cdd.4400905
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