A phenotypic screen in zebrafish identifies a novel small-molecule inducer of ectopic tail formation suggestive of alterations in non-canonical Wnt/PCP signaling
Gebruers, Evelien and Cordero-Maldonado, María Lorena and Gray, Alexander I and Clements, Carol and Harvey, Alan L and Edrada-Ebel, Ruangelie and de Witte, Peter A M and Crawford, Alexander D and Esguerra, Camila V (2013) A phenotypic screen in zebrafish identifies a novel small-molecule inducer of ectopic tail formation suggestive of alterations in non-canonical Wnt/PCP signaling. PLOS One, 8 (12). e83293. ISSN 1932-6203
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Abstract
Zebrafish have recently emerged as an attractive model for the in vivo bioassay-guided isolation and characterization of pharmacologically active small molecules of natural origin. We carried out a zebrafish-based phenotypic screen of over 3000 plant-derived secondary metabolite extracts with the goal of identifying novel small-molecule modulators of the BMP and Wnt signaling pathways. One of the bioactive plant extracts identified in this screen - Jasminum gilgianum, an Oleaceae species native to Papua New Guinea - induced ectopic tails during zebrafish embryonic development. As ectopic tail formation occurs when BMP or non-canonical Wnt signaling is inhibited during the tail protrusion process, we suspected a constituent of this extract to act as a modulator of these pathways. A bioassay-guided isolation was carried out on the basis of this zebrafish phenotype, identifying para-coumaric acid methyl ester (pCAME) as the active compound. We then performed an in-depth phenotypic analysis of pCAME-treated zebrafish embryos, including a tissue-specific marker analysis of the secondary tails. We found pCAME to synergize with the BMP-inhibitors dorsomorphin and LDN-193189 in inducing ectopic tails, and causing convergence-extension defects in compound-treated embryos. These results indicate that pCAME may interfere with non-canonical Wnt signaling. Inhibition of Jnk, a downstream target of Wnt/PCP signaling (via morpholino antisense knockdown and pharmacological inhibition with the kinase inhibitor SP600125) phenocopied pCAME-treated embryos. However, immunoblotting experiments revealed pCAME to not directly inhibit Jnk-mediated phosphorylation of c-Jun, suggesting additional targets of SP600125, and/or other pathways, as possibly being involved in the ectopic tail formation activity of pCAME. Further investigation of pCAME's mechanism of action will help determine this compound's pharmacological utility.
Creators(s): | Gebruers, Evelien, Cordero-Maldonado, María Lorena, Gray, Alexander I, Clements, Carol, Harvey, Alan L, Edrada-Ebel, Ruangelie, de Witte, Peter A M, Crawford, Alexander D and Esguerra, Camila V; | Item type: | Article |
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ID code: | 47962 |
Keywords: | phenotypic screen, zebrafish, novel small-molecule inducer, ectopic tail formation, non-canonical wnt/pcp, signalling, bmp signaling, embryos, notochords, phenotypes, plant signalling, tails, wnt signaling cascade, Therapeutics. Pharmacology, Pharmacology, Toxicology and Pharmaceutics(all) |
Subjects: | Medicine > Therapeutics. Pharmacology |
Department: | Faculty of Science > Strathclyde Institute of Pharmacy and Biomedical Sciences |
Depositing user: | Pure Administrator |
Date deposited: | 09 May 2014 10:22 |
Last modified: | 26 Feb 2021 01:43 |
URI: | https://strathprints.strath.ac.uk/id/eprint/47962 |
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