Abstract
p35, a viral inhibitor of caspase, prevents cell death induced by various stimuli. We established an experimental system to study the involvement of caspases in cell death, using primary cultured cells from p35 transgenic mice in which the p35 open reading frame (ORF) had been disrupted by the insertion of a DNA segment flanked by loxP sites, the Cre recognition sites. In this system, p35 expression can be initiated by Cre recombinase. Cardiomyocytes, which are highly sensitive to hypoxic stress, were infected with an adenovirus carrying the cre gene (AxCANCre). Expression of p35 by infection with AxCANCre resulted in inhibition of caspase-3 activation and resistance to hypoxia-induced cell death. Hypoxia-induced cytochrome c release was also attenuated in p35-expressing cardiomyocytes. Our transgenic mice can be used as an experimental model for studying the involvement of caspases in various degenerative diseases as well as programmed cell death both in vitro and in vivo. Cell Death and Differentiation (2000) 7, 485–492
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Abbreviations
- PBS:
-
phosphate-buffered saline
- FBS:
-
fetal bovine serum
- RT–PCR:
-
reverse transcription PCR
- DMEM:
-
Dulbecco's modified Eagle's medium
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Acknowledgements
We are grateful to I Saito at University of Tokyo for providing us recombinant Adenovirus AxCANCre and AxCALacZ. This work was supported in part by grants from the Japanese Ministry of Education, Science, Sports and Culture to H Okano and M Miura, from Human Frontier Science Program to H Okano. This work was also supported in part to H Okano by Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Corporation. S Hisahara is a research fellow of the Japan Society for the Promotion of Science.
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Araki, T., Shibata, M., Takano, R. et al. Conditional expression of anti-apoptotic protein p35 by Cre-mediated DNA recombination in cardiomyocytes from loxP-p35-transgenic mice. Cell Death Differ 7, 485–492 (2000). https://doi.org/10.1038/sj.cdd.4400674
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DOI: https://doi.org/10.1038/sj.cdd.4400674
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