Abstract
Many cells die during mammalian development and are engulfed by macrophages. In DNase II−/− embryos, the TUNEL-positive DNA of apoptotic cells is left undigested in macrophages, providing a system for studying programmed cell death during mouse development. Here, we showed that an Apaf-1-null mutation in the DNase II−/− embryos greatly reduced the number of macrophages carrying DNA at E11.5. However, at later stages of the embryogenesis, a significant number of macrophages carrying undigested DNA were present in Apaf-1−/− embryos, indicating that cells died and were engulfed in an Apaf-1-independent manner. In most tissues of the Apaf-1−/− embryos, no processed caspase-3 was detected, and the DNA of dead cells accumulated in the macrophages appeared intact. Many nonapoptotic dead cells were found in the tail of the Apaf-1−/− embryos, suggesting that the Apaf-1-independent programmed cell death occurred, and these dead cells were engulfed by macrophages. In contrast, active caspase-3 was detected in E14.5 thymus of Apaf-1−/− embryos. Treatment of fetal thymocytes with staurosporine, but not etoposide, induced processing of procaspases 3 and 9, indicating that the E14.5 thymocytes have the ability to undergo caspase-dependent apoptosis in an Apaf-1-independent manner. Thus, programmed cell death in mouse development, which normally proceeds in an efficient Apaf-1-depenent mechanism, appears to be backed up by Apaf-1-independent death systems.
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Abbreviations
- CAD:
-
caspase-activated DNase
- mAb:
-
monoclonal antibody
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Acknowledgements
We thank Dr. M Koike (Juntendo University) for his advice for the immunochemical staining, and M Fujii and M Harayama for secretarial assistance. This work was supported in part by Grants-in-Aid from the Ministry of Education, Science, Sports, and Culture in Japan, and by the Kyoto University Global COE Program (Center for Frontier Medicine). AN was a research assistant for the Osaka University Global COE Program (System Dynamics of Biological Function).
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Nagasaka, A., Kawane, K., Yoshida, H. et al. Apaf-1-independent programmed cell death in mouse development. Cell Death Differ 17, 931–941 (2010). https://doi.org/10.1038/cdd.2009.186
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DOI: https://doi.org/10.1038/cdd.2009.186
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