PUBLICATION
Establishment of the expression system for studying the function of active caspase-3 in zebrafish
- Authors
- Kim, H.Y., Kim, G.Y., Kim, S.S., Nam, M.K., and Rhim, H.
- ID
- ZDB-PUB-071219-24
- Date
- 2009
- Source
- Molecular biology reports 36(2): 405-413 (Journal)
- Registered Authors
- Keywords
- Apoptosis, Caspase-3, Microinjection, Zebrafish
- MeSH Terms
-
- Active Transport, Cell Nucleus
- Animals
- Caspase 3/genetics*
- Enzyme Activation
- Green Fluorescent Proteins/administration & dosage
- Microinjections
- Models, Animal*
- Plasmids/administration & dosage
- Transfection/methods*
- Zebrafish
- PubMed
- 18080784 Full text @ Mol. Biol. Rep.
Citation
Kim, H.Y., Kim, G.Y., Kim, S.S., Nam, M.K., and Rhim, H. (2009) Establishment of the expression system for studying the function of active caspase-3 in zebrafish. Molecular biology reports. 36(2):405-413.
Abstract
Caspase-3, a key molecule in apoptosis, has been extensively studied in cell culture system; however, it has been less well characterized in vivo because certain mediators are required for the proteolytic activation of effector caspases, including caspase-3. In this study, various forms of caspase-3 with the C-terminal GFP tag were inserted into the pCS2+ plasmid, and the expression patterns of caspase-3 proteins were characterized in a zebrafish model system using microinjection of nucleic acids into zebrafish embryos. We have verified that active caspase-3 was generated by its autocatalytic activity under the condition of caspase-2 prodomain (C2P)-caspase-3-GFP overexpression, indicating that the C2P domain is crucial for the activation of caspase-3. We also confirmed that the C2P domain plays an important role in regulating the nuclear localization of the C2P-caspase-3 chimeric protein. We used this expression system to establish an animal model system suitable for the investigation of the functional characteristics of caspase-3 in vivo. Thus, our study provides a useful and specific tool for investigating the molecular mechanisms by which active caspase-3 regulates apoptosis during embryonic development.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping