Abstract
Programmed cell death, which is required for the development and homeostasis of metazoans, includes mechanisms such as apoptosis, autophagic cell death, and necrotic (or type III) death. Members of the Bcl2 family regulate apoptosis, among which Bax and Bak act as a mitochondrial gateway. Although embryonic fibroblasts from Bax/Bak double-knockout (DKO) mice are resistant to apoptosis, we previously demonstrated that these cells die through an autophagy-dependent mechanism in response to various types of cellular stressors. To determine the physiological role of autophagy-dependent cell death, we generated Atg5/Bax/Bak triple-knockout (TKO) mice, in which autophagy is greatly suppressed compared with DKO mice. Embryonic fibroblasts and thymocytes from TKO mice underwent autophagy much less frequently, and their viability was much higher than DKO cells in the presence of certain cellular stressors, providing genetic evidence that DKO cells undergo Atg5-dependent death. Compared with wild-type embryos, the loss of interdigital webs was significantly delayed in DKO embryos and was even further delayed in TKO embryos. Brain malformation is a distinct feature observed in DKO embryos on the 129 genetic background, but not in those on a B6 background, whereas such malformations appeared in TKO embryos even on a B6 background. Taken together, our data suggest that Atg5-dependent cell death contributes to the embryonic development of DKO mice, implying that autophagy compensates for the deficiency in apoptosis.
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Acknowledgements
We are grateful to Dr N. Mizushima for providing us with Atg5+/− mice. This study was supported in part by Grant-in-Aid for Grant-in-Aid for Scientific Research (S) (22229002), Grant-in-Aid for Scientific Research (A) (17H01533), Grant-in-Aid for challenging Exploratory Research (16K15230), Grant-in-Aid for Scientific Research on Innovative Areas (15H01554, 26110001, 26110005), Grant-in-Aid for Encouragement of Young Scientists (B) (15K19004) from the MEXT of Japan, by the Project for Cancer Research and Therapeutic Evolution (P-CREATE) and by the Project for Psychiatric and Neurological Disorders from the Japan Agency for Medical Research and development, AMED. This study was also supported by the Joint Usage/Research Program of Medical Research Institute, Tokyo Medical and Dental University, and by a grant from the Secom Science and Technology Foundation and the Takeda Science Foundation.
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SA performed EM analyses. MT, TY, HS, YN, YM, IY, and SS performed other experiments and analyzed the results. YT and SS designed and conducted the research, and wrote the paper.
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Arakawa, S., Tsujioka, M., Yoshida, T. et al. Role of Atg5-dependent cell death in the embryonic development of Bax/Bak double-knockout mice. Cell Death Differ 24, 1598–1608 (2017). https://doi.org/10.1038/cdd.2017.84
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DOI: https://doi.org/10.1038/cdd.2017.84
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