1. Protocol development for discovery of angiogenesis inhibitors via automated methods using zebrafish
- Author
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Krishna Kumar Singh, Robin Ng, Youdong Wang, Antonio Mauro, Rui Guan, Xiao-Yan Wen, and Jamie Yuanjun Li
- Subjects
0301 basic medicine ,Embryology ,Embryo, Nonmammalian ,Indoles ,Angiogenesis ,Physiology ,Drug Evaluation, Preclinical ,Angiogenesis Inhibitors ,Cardiovascular Physiology ,Green fluorescent protein ,Neovascularization ,Animals, Genetically Modified ,chemistry.chemical_compound ,0302 clinical medicine ,Drug Discovery ,Oximes ,Medicine and Health Sciences ,Zebrafish ,Multidisciplinary ,Drug discovery ,Eukaryota ,Equipment Design ,Animal Models ,Robotics ,Angiogenesis inhibitor ,Experimental Organism Systems ,Osteichthyes ,030220 oncology & carcinogenesis ,Vertebrates ,Medicine ,Engineering and Technology ,medicine.symptom ,Robots ,Algorithms ,Research Article ,Computer and Information Sciences ,Drug Research and Development ,Science ,High-throughput screening ,Neovascularization, Physiologic ,Computational biology ,Library Screening ,Biology ,Research and Analysis Methods ,Computer Software ,03 medical and health sciences ,Model Organisms ,medicine ,Animals ,Molecular Biology Techniques ,Molecular Biology ,Protocol (science) ,Automation, Laboratory ,Pharmacology ,Molecular Biology Assays and Analysis Techniques ,Drug Screening ,Dose-Response Relationship, Drug ,Mechanical Engineering ,Embryos ,Organisms ,Endothelial Cells ,Biology and Life Sciences ,biology.organism_classification ,High-Throughput Screening Assays ,030104 developmental biology ,Fish ,chemistry ,Animal Studies ,Indirubin ,Developmental Biology - 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(Flk1: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.
- Published
- 2019