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

Abstract

To increase the facility and throughput of scoring phenotypic traits in embryonic zebrafish, we developed an automated micro-well assay for heart rate using automated fluorescence microscopy of transgenic embryos expressing green fluorescent protein in myocardium. The assay measures heart rates efficiently and accurately over a large linear dynamic range, and it rapidly characterizes dose dependence and kinetics of small molecule-induced changes in heart rate. This is the first high-throughput micro-well assay for organ function in an intact vertebrate.

Myoscape controls cardiac calcium cycling and contractility via regulation of L-type calcium channel surface expression

“…To evaluate whether Myoscape deficiency interferes with key steps of zebrafish heart development, we examined the expression of cardiac chamber-specific proteins and messenger RNAs (mRNAs) by immunostainings and antisense RNA in situ hybridization, respectively, as well as the cardiac structure of Myoscape morphants by histology2930. While minimal residual Myoscape expression cannot be excluded, knockdown of Myoscape did not interfere with crucial steps of cardiogenesis, such as heart tube looping, chamber demarcation and the differentiation of ventricular and atrial cardiomyocytes (Supplementary Fig.…”

NADPH Oxidase 4 Induces Cardiac Arrhythmic Phenotype in Zebrafish

“…Zebrafish Strains and Studies-Transgenic strain myl7:GFP zebrafish were maintained as described previously (7,11). GFP expression is driven by a cardiac-specific promoter of myl7 (also known as cardiac myosin light chain 2).…”

Illuminating cardiac development: Advances in imaging add new dimensions to the utility of zebrafish genetics

“…Furthermore, it will be fruitful to employ high-speed imaging to the characterization of small molecules influencing cardiac function. Using automated microscopy, automated videography, and automated computational analysis, Burns et al have created a high-throughput assay that sensitively and accurately monitors heart rate in Tg(cmlc2:gfp) embryos [57]. This strategy can be used to rapidly screen large libraries of small molecules for their effects on contractility.…”