ZFIN ID: ZDB-PUB-121004-22
In situ hybridization assay-based small molecule screening in zebrafish
Jing, L., Durand, E.M., Ezzio, C., Pagliuca, S.M., and Zon, L.I.
In vitro biochemical and cell-based small-molecule screens have been widely used to identify compounds that target specific signaling pathways, but the identified compounds frequently fail at the animal testing stage, largely due to the in vivo absorption, metabolism, and toxicity properties of the chemicals. Zebrafish has recently emerged as a vertebrate whole-organism model for small-molecule screening. The in vivo bioactivity and specificity of compounds are examined from the very beginning of zebrafish screens. In addition, zebrafish is suitable for large-scale chemical screens, similar to many cellular assays. This protocol describes an approach for in situ hybridization (ISH)-based chemical screening in zebrafish, which, in principle, can be used to screen any gene product. The described protocol has been used to identify small molecules affecting specific molecular pathways and biological processes. It can also be adapted to zebrafish screens with different readouts.