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Colorimetric and fluorescent TRAP assays for visualising and quantifying fish osteoclast activity
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Colorimetric and fluorescent TRAP assays for visualising and quantifying fish osteoclast activity
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Journal Article
Colorimetric and fluorescent TRAP assays for visualising and quantifying fish osteoclast activity
2022
Overview
Histochemical detection of tartrate-resistant acid phosphatase (TRAP) activity is a fundamental technique for visualizing osteoclastic bone resorption and assessing osteoclast activity status in tissues. This approach has mostly employed colorimetric detection, which has limited quantification of activity in situ and co-labelling with other skeletal markers. Here we report simple colorimetric and fluorescent TRAP assays in zebrafish and medaka, two important model organisms for investigating the pathogenesis of bone disorders. We show fluorescent TRAP staining, utilising the ELF97 substrate, is a rapid, robust and stable system to visualise and quantify osteoclast activity in zebrafish, and is compatible with other fluorescence stains, transgenic lines and antibody approaches. Using this approach, we show that TRAP activity is predominantly found around the base of the zebrafish pharyngeal teeth, where osteoclast activity state appears to be heterogeneous.
Publisher
PAGEPress Publications,PAGEPress Publications, Pavia, Italy
