Development and Validation of an Automated High-Throughput System for Zebrafish In Vivo Screenings

Abstract

The zebrafish is a vertebrate model compatible with the paradigms of drug discovery. The small size and transparency of zebrafish embryos make them amenable for the automation necessary in high-throughput screenings. We have developed an automated high-throughput platform for in vivo chemical screenings on zebrafish embryos that includes automated methods for embryo dispensation, compound delivery, incubation, imaging and analysis of the results. At present, two different assays to detect cardiotoxic compounds and angiogenesis inhibitors can be automatically run in the platform, showing the versatility of the system. A validation of these two assays with known positive and negative compounds, as well as a screening for the detection of unknown anti-angiogenic compounds, have been successfully carried out in the system developed. We present a totally automated platform that allows for high-throughput screenings in a vertebrate organism.

Kctd10 regulates heart morphogenesis by repressing the transcriptional activity of Tbx5a in zebrafish

“…Adrenaline, digilanid and atropine were provided by Peking University Hospital. The embryos of 24 hpf derived from the incross of kctd10 þ / À zebrafish were treated with adrenaline (250 mM), atropine (100 mM) and digilanid (10 mM), respectively [53][54][55] . After incubating at 28°C for 24 h, the larvae were anesthetized with MS-222 (ethyl-3-aminobenzoate methanesulfonate salt, Sigma-Aldrich Inc.) at 28°C.…”

A Chemical Screening Procedure for Glucocorticoid Signaling with a Zebrafish Larva Luciferase Reporter System

“…At the same time, it allows for a substantial degree of automation, e.g. automated drug application by pipetting robots or automated distribution of embryos by embryo sorting devices 24,25 . The simple readout does not require automated screening microscopes or sophisticated image analysis software, yet provides a rich set of data on temporal and quantitative aspects of the studied signaling pathway.…”

G 2 /M cell cycle arrest on HT-29 cancer cells and toxicity assessment of triphenylphosphanegold(I) carbonimidothioates, Ph 3 PAu[SC(OR) = NPh], R = Me, Et, and iPr, during zebrafish development

“…The use of zebrafish (Danio rerio) has emerged as a powerful model system to monitor angiogenesis in vivo [15][16][17]. Zebrafish have been used as a model organism for research purposes for more than 20 years and therefore handling methods, optimum breeding techniques, husbandry, and maintenance are well established [18].…”

“…Recent research has shown that most of the genes found in humans have at least one zebrafish orthologue [19]. The above ensures zebrafish as being highly useful for the study of complex biological processes such as cell 6 senescence and apoptosis, aging, cardiovascular system development, and drug toxicity response [15][16][17][18][19][20][21]. There are a few special characteristics of zebrafish that are worth highlighting.…”