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)
Citation
Higuchi, A., Wakai, E., Tada, T., Koiwa, J., Adachi, Y., Shiromizu, T., Goto, H., Tanaka, T., Nishimura, Y. (2021) Generation of a Transgenic Zebrafish Line for In Vivo Assessment of Hepatic Apoptosis. Pharmaceuticals (Basel, Switzerland). 14(11):.
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
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping