title: A high-content screening assay in transgenic zebrafish identifies two novel activators of fgf signaling.
abstract: Zebrafish have become an invaluable vertebrate animal model to interrogate small molecule libraries for modulators of complex biological pathways and phenotypes. We have recently described the implementation of a quantitative, high-content imaging assay in multi-well plates to analyze the effects of small molecules on Fibroblast Growth Factor (FGF) signaling in vivo. Here we have evaluated the capability of the assay to identify compounds that hyperactivate FGF signaling from a test cassette of agents with known biological activities. Using a transgenic zebrafish reporter line for FGF activity, we screened 1040 compounds from an annotated library of known bioactive agents, including FDA-approved drugs. The assay identified two molecules, 8-hydroxyquinoline sulfate and pyrithione zinc, that enhanced FGF signaling in specific areas of the brain. Subsequent studies revealed that both compounds specifically expanded FGF target gene expression. Furthermore, treatment of early stage embryos with either compound resulted in dorsalized phenotypes characteristic of hyperactivation of FGF signaling in early development. Documented activities for both agents included activation of extracellular signal-related kinase (ERK), consistent with FGF hyperactivation. To conclude, we demonstrate the power of automated quantitative high-content imaging to identify small molecule modulators of FGF.
authors.affiliation: Department of Developmental Biology, University of Pittsburgh, School of Medicine, PA 15213, USA.
authors.forename: Manush
authors.initials: M
authors.lastname: Saydmohammed
authors.forename: Laura L
authors.initials: LL
authors.lastname: Vollmer
authors.forename: Ezenwa Obi
authors.initials: EO
authors.lastname: Onuoha
authors.forename: Andreas
authors.initials: A
authors.lastname: Vogt
authors.forename: Michael
authors.initials: M
authors.lastname: Tsang
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D004625:Embryo, Nonmammalian / Q000187:drug effects*; D005346:Fibroblast Growth Factors / Q000378:metabolism*; D057166:High-Throughput Screening Assays; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*; D015027:Zebrafish / Q000196:embryology / Q000235:genetics* / Q000378:metabolism*
chemical_list: D004364:Pharmaceutical Preparations; D005346:Fibroblast Growth Factors