Journal
EUROPEAN JOURNAL OF HISTOCHEMISTRY
Volume 66, Issue 2, Pages -Publisher
PAGEPRESS PUBL
DOI: 10.4081/ejh.2022.3369
Keywords
TRAP; tartrate-resistant acid phosphatase; osteoclast; ELF97; fracture; zebrafish; medaka
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In this study, we developed simple colorimetric and fluorescent TRAP assays in zebrafish and medaka, which can be used to assess osteoclast activity. Fluorescent TRAP staining using the ELF97 substrate is a rapid, stable system that is compatible with other fluorescence stains, transgenic lines, and antibody approaches.
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.
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