Zebrafish: a preclinical model for drug screening.
The zebrafish embryo has become an important vertebrate model for assessing drug effects. It is well suited for studies in genetics, embryology, development, and cell biology. Zebrafish embryos exhibit unique characteristics, including ease of maintenance and drug administration, short reproductive cycle, and transparency that permits visual assessment of developing cells and organs. Because of these advantages, zebrafish bioassays are cheaper and faster than mouse assays, and are suitable for large-scale drug screening. Here we describe the use of zebrafish bioassays for assessing toxicity, angiogenesis, and apoptosis. Using 18 chemicals, we demonstrated that toxic response, teratogenic effects, and LC(50) in zebrafish are comparable to results in mice. The effects of compounds on various organs, including the heart, brain, intestine, pancreas, cartilage, liver, and kidney, were observed in the transparent animals without complicated processing, demonstrating the efficiency of toxicity assays using zebrafish embryos. Using endogenous alkaline phosphatase staining and a whole-animal enzyme assay, we demonstrated that SU5416 and flavopiridol, compounds shown to have antiangiogenic effects in mammals, inhibit blood vessel growth in zebrafish, and this bioassay is suitable for high-throughput screening using a 96-well microplate reader. We also demonstrated that in vivo acridine orange staining can be used to visualize apoptotic events in embryos treated with brefeldin A, neomycin, or caspase inhibitors. After in vivo staining, acridine orange can be extracted and quantitated using a fluorescence microplate reader, providing a screening system for agents that modulate apoptosis.
Parng
Chuenlei
C
Phylonix, Cambridge, MA 02139, USA. chuenlei@phylonix.com
Seng
Wen Lin
WL
Semino
Carlos
C
McGrath
Patricia
P
D000165:Acridine Orange; D000469:Alkaline Phosphatase / Q000378:metabolism; D020533:Angiogenesis Inhibitors / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001681:Biological Assay; D004353:Drug Evaluation, Preclinical; D004625:Embryo, Nonmammalian; D005419:Flavonoids / Q000494:pharmacology; D005456:Fluorescent Dyes; D007211:Indoles / Q000494:pharmacology; D007928:Lethal Dose 50; D051379:Mice; D010880:Piperidines / Q000494:pharmacology; D011758:Pyrroles / Q000494:pharmacology; D018675:Toxicity Tests; D015027:Zebrafish / Q000235:genetics / Q000502:physiology*
D020533:Angiogenesis Inhibitors; D005419:Flavonoids; D005456:Fluorescent Dyes; D007211:Indoles; D010880:Piperidines; D011758:Pyrroles; C077990:alvocidib; C116890:Semaxinib; D000469:Alkaline Phosphatase; D000165:Acridine Orange