Abstract
Their optical clarity as larvae and embryos, small size, and high fecundity make zebrafish ideal for whole animal high throughput screening. A high-throughput drug discovery platform (HTP) has been built to perform fully automated screens of compound libraries with zebrafish embryos. A Tg(kdrl:EGFP) line, marking endothelial cell cytoplasm, was used in this work to help develop protocols and functional algorithms for the system, with the intent of screening for angiogenesis inhibitors. Indirubin 3’ Monoxime (I3M), a known angiogenesis inhibitor, was used at various concentrations to validate the protocols. Consistent with previous studies, a dose dependant inhibitory effect of I3M on angiogenesis was confirmed. The methods and protocols developed here could significantly increase the throughput of drug screens, while limiting human errors. These methods are expected to facilitate the discovery of novel anti-angiogenesis compounds and can be adapted for many other applications in which samples have a good fluorescent signal.
Highlights
The Zebrafish (Danio rerio) is a tropical fresh water fish belonging to the minnow family
The high-throughput drug discovery platform (HTP) is designed to work with 96-well plates and can be modified to work with 384-well plates
This manuscript exhibits a functional high throughput drug discovery protocol to be used with zebrafish models
Summary
The Zebrafish (Danio rerio) is a tropical fresh water fish belonging to the minnow family. The zebrafish has recently gained popularity as a well-managed vertebrate model for human disease. One of the main reasons for this popularity is its unique combination of optical clarity as embryos and larvae and its embryological manipulability. The optical clarity of zebrafish embryos and larvae allows for the extensive examination of the onset and progression of a pathological process in vivo and in real time. The breeding habits and large number of offspring per mating cycle (100–200 eggs per week) are additional positives for the zebrafish.
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