High-throughput assay for small molecules that modulate zebrafish embryonic heart rate

Area of Science:

• Cardiovascular research
• Zebrafish models
• High-throughput screening

Background:

• Assessing organ function in vertebrate embryos is crucial for drug discovery.
• Current methods for scoring phenotypic traits in zebrafish embryos can be labor-intensive.
• There is a need for automated, high-throughput assays for organ function analysis.

Purpose of the Study:

• To develop an automated micro-well assay for measuring heart rate in embryonic zebrafish.
• To increase the efficiency and throughput of phenotypic trait scoring in zebrafish.
• To establish a high-throughput method for assessing small molecule effects on organ function.

Main Methods:

• Utilized transgenic zebrafish embryos expressing green fluorescent protein in the myocardium.
• Developed an automated micro-well assay combined with automated fluorescence microscopy.
• Measured heart rates by analyzing fluorescence signals from the heart.

Main Results:

• The assay efficiently and accurately measures heart rates over a large linear dynamic range.
• The assay rapidly characterizes dose dependence and kinetics of small molecule-induced heart rate changes.
• This represents the first high-throughput micro-well assay for organ function in an intact vertebrate.

Conclusions:

• The developed assay significantly improves the facility and throughput of heart rate assessment in zebrafish embryos.
• This automated method is suitable for high-throughput screening of compounds affecting cardiovascular function.
• This assay provides a novel platform for studying organ function in a whole vertebrate model.