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Bioluminescence imaging of caspase-3 activity in mouse liver

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Abstract

Apoptosis is an essential process for the maintenance of liver physiology. The ability to noninvasively image apoptosis in livers would provide unique insights into its role in liver disease processes. In the present work, we established a stable mouse model by hydrodynamics methods to study the activity of caspase-3 and evaluate the effect of the apoptosis inhibitors in mouse livers under true physiological conditions by bioluminescence imaging. The reporter plasmid attB-ANLuc(DEVD)BCLuc that contains fragment of attB and ANLuc(DEVD)BCLuc was codelivered with the mouse-codon optimized φC31 (φC31o) integrase plasmids specifically to mouse liver by hydrodynamic injection procedure. Then, φC31o integrase mediated intramolecular recombination between wild-type attB and attP site in mice, and thus the reporter expression cassette attB-ANLuc(DEVD)BCLuc was integrated permanently into mouse liver chromosome. We used these mice to characterize in vivo activation of caspase-3 upon treatment with LPS/d-GalN. Our data show that liver apoptosis could be reflected by the activity of luciferase. The shRNA targeting caspase-3 protein or apoptosis inhibitors could effectively downregulate luciferase activity in vivo. Also, this model could be used to measure caspase-3 activation during inflammatory and infectious events in vivo as verified by infected with MHV-3. This model could be used for screening anti-apoptosis compounds target mouse livers.

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Acknowledgments

We thank Yusen Zhou for the MHV used in this work. This work was partially supported by Mega-projects of Science Research for the 12th Five-Year Plan (#2012ZX10004-502, 2011ZXJ092-031), the Natural Science Foundation of China (#81170387, #30901812, #30900824, #30972615).

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Authors and Affiliations

  1. Laboratory of Blood-borne Virus, Beijing Institute of Transfusion Medicine, 27(9) Tai Ping Road, 100850, Beijing, China

    Qiuxia Fu, Shaoduo Yan, Licui Wang, Yong Zhou, Shuaizheng Jia, Juan Du, Xiaohui Wang, Yuhua Zhang & Linsheng Zhan

  2. Ningxia Medical University, Yinchuan, China

    Xiangguo Duan

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  1. Qiuxia Fu
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  2. Xiangguo Duan
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Correspondence to Linsheng Zhan.

Additional information

Qiuxia Fu, Xiangguo Duan and Shaoduo Yan contributed equally to this study.

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Fu, Q., Duan, X., Yan, S. et al. Bioluminescence imaging of caspase-3 activity in mouse liver. Apoptosis 18, 998–1007 (2013). https://doi.org/10.1007/s10495-013-0849-z

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