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Bioluminescence imaging of caspase-3 activity in mouse liver
Bioluminescence imaging of caspase-3 activity in mouse liver
- Source :
- Apoptosis. 18:998-1007
- Publication Year :
- 2013
- Publisher :
- Springer Science and Business Media LLC, 2013.
-
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.
- Subjects :
- Cancer Research
Clinical Biochemistry
Pharmaceutical Science
Apoptosis
Caspase 3
Small hairpin RNA
Mice
Genes, Reporter
In vivo
Animals
Bioluminescence imaging
Luciferase
Luciferases
Pharmacology
Mice, Inbred BALB C
biology
Biochemistry (medical)
Cell Biology
Molecular biology
Integrase
Liver
Luminescent Measurements
biology.protein
Female
Expression cassette
Subjects
Details
- ISSN :
- 1573675X and 13608185
- Volume :
- 18
- Database :
- OpenAIRE
- Journal :
- Apoptosis
- Accession number :
- edsair.doi.dedup.....0824f4f74fcbcf7c8a14f704c8e44696
- Full Text :
- https://doi.org/10.1007/s10495-013-0849-z