PUBLICATION

Generation of a Transgenic Zebrafish Line for In Vivo Assessment of Hepatic Apoptosis

Authors
Higuchi, A., Wakai, E., Tada, T., Koiwa, J., Adachi, Y., Shiromizu, T., Goto, H., Tanaka, T., Nishimura, Y.
ID
ZDB-PUB-211129-66
Date
2021
Source
Pharmaceuticals (Basel, Switzerland)   14(11): (Journal)
Registered Authors
Nishimura, Yuhei, Tanaka, Toshio
Keywords
Förster resonance energy transfer, apoptosis, caspase, drug-induced liver injury, hepatoprotectant, in vivo fluorescence imaging, liver, zebrafish
MeSH Terms
none
PubMed
34832899 Full text @ Pharmaceuticals (Basel)
Abstract
Hepatic apoptosis is involved in a variety of pathophysiologic conditions in the liver, including hepatitis, steatosis, and drug-induced liver injury. The development of easy-to-perform and reliable in vivo assays would thus greatly enhance the efforts to understand liver diseases and identify associated genes and potential drugs. In this study, we developed a transgenic zebrafish line that was suitable for the assessment of caspase 3 activity in the liver by using in vivo fluorescence imaging. The larvae of transgenic zebrafish dominantly expressed Casper3GR in the liver under control of the promoter of the phosphoenolpyruvate carboxykinase 1 gene. Casper3GR is composed of two fluorescent proteins, tagGFP and tagRFP, which are connected via a peptide linker that can be cleaved by activated caspase 3. Under tagGFP excitation conditions in zebrafish that were exposed to the well-characterized hepatotoxicant isoniazid, we detected increased and decreased fluorescence associated with tagGFP and tagRFP, respectively. This result suggests that isoniazid activates caspase 3 in the zebrafish liver, which digests the linker between tagGFP and tagRFP, resulting in a reduction in the Förster resonance energy transfer to tagRFP upon tagGFP excitation. We also detected isoniazid-induced inhibition of caspase 3 activity in zebrafish that were treated with the hepatoprotectants ursodeoxycholic acid and obeticholic acid. The transgenic zebrafish that were developed in this study could be a powerful tool for identifying both hepatotoxic and hepatoprotective drugs, as well as for analyzing the effects of the genes of interest to hepatic apoptosis.
Genes / Markers
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping