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Analysis of Apoptosis in Zebrafish Embryos by Whole-mount Immunofluorescence to Detect Activated Caspase 3
Published on: December 20, 2013
Caspase-3 induced apoptosis in transgenic zebrafish
Chiranjib Chakraborty1, Gargi Saha, Biplab Sarkar
1Biotechnology Department, Institute of Applied Medicines and Research, Delhi-Meerat Road, 9th Mile Stone, Ghaziabad, UP, India. drchiranjib@yahoo.com
Biotechnology Letters
|February 21, 2006
Summary
This study developed a transgenic zebrafish model to visualize apoptosis. Overexpressing mouse caspase-3 (CASP3) induced widespread cell death in various organs, offering a new tool for apoptosis research.
Area of Science:
- Developmental Biology
- Cell Biology
- Genetics
Background:
- Zebrafish are valuable model organisms for studying biological processes due to their genetic tractability.
- Apoptosis, or programmed cell death, is crucial for development and tissue homeostasis.
Purpose of the Study:
- To create a transgenic zebrafish model for visualizing and studying the induction and spread of apoptosis.
- To investigate the effects of overexpressing mouse caspase-3 (CASP3) in zebrafish.
Main Methods:
- Generated transgenic zebrafish expressing mouse CASP3 under the zebrafish beta-actin promoter.
- Utilized a chimeric transgene encoding CASP3 fused to green fluorescent protein (GFP) for apoptotic cell visualization.
- Observed the spatial and temporal dissemination of apoptosis in embryos and adult zebrafish.
Main Results:
- Successfully induced clonally derived apoptosis in transgenic zebrafish.
- Apoptosis originated in the thymus and spread to adjacent tissues like gill arches and retro-orbital soft tissue.
- Observed dissemination of apoptosis into abdominal organs, including the testis and kidney.
- Demonstrated that overexpressing CASP3 leads to extensive apoptosis in both embryonic and adult zebrafish.
Conclusions:
- The developed transgenic zebrafish model is effective for studying apoptosis.
- This model allows for the visualization and tracking of apoptosis induction and spread.
- Provides a platform for further research into caspase-3-mediated apoptosis in vivo.
