Protocol development for discovery of angiogenesis inhibitors via automated methods using zebrafish
Autoři:
Antonio Mauro aff001; Robin Ng aff001; Jamie Yuanjun Li aff001; Rui Guan aff001; Youdong Wang aff001; Krishna Kumar Singh aff001; Xiao-Yan Wen aff001
Působiště autorů:
Zebrafish Centre for Advanced Drug Discovery, Keenan Research Center, Li Ka Shing Knowledge Institute, St. Michael’s Hospital, Unity Health Toronto, Toronto, Ontario, Canada
aff001; Institute of Medical Science, University of Toronto, Toronto, Ontario, Canada
aff002; Cardiovascular Sciences Collaborative Program, University of Toronto, Toronto, Ontario, Canada
aff003; Department of Pharmacology and Toxicology, University of Toronto, Toronto, Ontario, Canada
aff004; Department of Surgery, University of Toronto, Toronto, Ontario, Canada
aff005; Department of Medical Biophysics, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Ontario, Canada
aff006; Department of Medicine, University of Toronto, Toronto, Ontario, Canada
aff007; Department of Physiology, University of Toronto, Toronto, Ontario, Canada
aff008
Vyšlo v časopise:
PLoS ONE 14(11)
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pone.0221796
Souhrn
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.
Klíčová slova:
Embryos – Library screening – Drug discovery – Zebrafish – Computer software – Drug screening – Robots – Angiogenesis
Zdroje
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