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Published on: January 1, 2017
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Colorimetric and fluorescent TRAP assays for visualising and quantifying fish osteoclast activity
Lalith Prabha Ethiraj1, En Lei Samuel Fong2, Ranran Liu3
1Lee Kong Chian School of Medicine, Nanyang Technological University. ETHIRAJL001@e.ntu.edu.sg.
European Journal of Histochemistry : EJH
|March 25, 2022
Summary
Researchers developed new fluorescent assays for tartrate-resistant acid phosphatase (TRAP) in zebrafish and medaka. This method enhances visualization and quantification of osteoclast activity, crucial for studying bone disorders.
Area of Science:
- Biochemistry
- Developmental Biology
- Histology
Background:
- Tartrate-resistant acid phosphatase (TRAP) activity is key for assessing osteoclast function in bone resorption.
- Current colorimetric TRAP detection methods have limitations in quantification and co-staining.
- Zebrafish and medaka are valuable model organisms for bone disorder research.
Purpose of the Study:
- To develop simple and effective colorimetric and fluorescent TRAP assays for zebrafish and medaka.
- To establish a robust method for visualizing and quantifying osteoclast activity in vivo.
- To overcome limitations of existing TRAP detection techniques.
Main Methods:
- Development of novel colorimetric and fluorescent TRAP assays.
- Utilisation of the ELF97 substrate for fluorescent TRAP staining.
- Application of assays in zebrafish and medaka model organisms.
- Compatibility testing with fluorescence stains, transgenic lines, and antibody methods.
Main Results:
- Successful implementation of simple colorimetric and fluorescent TRAP assays in zebrafish and medaka.
- Fluorescent TRAP staining using ELF97 proved rapid, robust, and stable for visualizing osteoclast activity.
- The fluorescent method is compatible with various advanced biological techniques.
- TRAP activity was predominantly observed at the base of zebrafish pharyngeal teeth, showing heterogeneous activity states.
Conclusions:
- Fluorescent TRAP staining offers a superior method for visualizing and quantifying osteoclast activity in zebrafish.
- The developed assays are suitable for studying bone disorder pathogenesis in relevant model organisms.
- This technique facilitates detailed analysis of osteoclast heterogeneity in skeletal tissues.
