Development of the morpholino gene knockdown technique in Fundulus heteroclitus : a tool for studying molecular mechanisms in an established environmental model
Matson, Cole W.
Clark, Bryan W.
Jenny, Matthew J.
Fleming, Carrie R.
Hahn, Mark E.
Di Giulio, Richard T.
MetadataShow full item record
KeywordFundulus heteroclitus; Antisense morpholino oligonucleotide; CYP1A; Atlantic killifish; Genetic toxicology; Development; Teratogenesis
A significant challenge in environmental toxicology is that many genetic and genomic tools available in laboratory models are not developed for commonly used environmental models. The Atlantic killifish (Fundulus heteroclitus) is one of the most studied teleost environmental models, yet few genetic or genomic tools have been developed for use in this species. The advancement of genetic and evolutionary toxicology will require that many of the tools developed in laboratory models be transferred into species more applicable to environmental toxicology. Antisense morpholino oligonucleotide (MO) gene knockdown technology has been widely utilized to study development in zebrafish and has been proven to be a powerful tool in toxicological investigations through direct manipulation of molecular pathways. To expand the utility of killifish as an environmental model, MO gene knockdown technology was adapted for use in Fundulus. Morpholino microinjection methods were altered to overcome the significant differences between these two species. Morpholino efficacy and functional duration were evaluated with molecular and phenotypic methods. A cytochrome P450-1A (CYP1A) MO was used to confirm effectiveness of the methodology. For CYP1A MO-injected embryos, a 70% reduction in CYP1A activity, a 86% reduction in total CYP1A protein, a significant increase in β-naphthoflavone-induced teratogenicity, and estimates of functional duration (50% reduction in activity 10 dpf, and 86% reduction in total protein 12 dpf) conclusively demonstrated that MO technologies can be used effectively in killifish and will likely be just as informative as they have been in zebrafish.
Author Posting. © Elsevier B.V., 2008. This is the author's version of the work. It is posted here by permission of Elsevier B.V. for personal use, not for redistribution. The definitive version was published in Aquatic Toxicology 87 (2008): 289-295, doi:10.1016/j.aquatox.2008.02.010.
Showing items related by title, author, creator and subject.
Development and characterization of polyclonal antibodies against the aryl hydrocarbon receptor protein family (AHR1, AHR2, and AHR repressor) of Atlantic killifish Fundulus heteroclitus Merson, Rebeka R.; Franks, Diana G.; Karchner, Sibel I.; Hahn, Mark E. (2005-12-20)The aryl hydrocarbon receptor (AHR) and AHR repressor (AHRR) proteins regulate gene expression in response to some halogenated aromatic hydrocarbons and polycyclic aromatic hydrocarbons. The Atlantic killifish is a valuable ...
Role of DNA methylation of AHR1 and AHR2 promoters in differential sensitivity to PCBs in Atlantic Killifish, Fundulus heteroclitus Aluru, Neelakanteswar; Karchner, Sibel I.; Hahn, Mark E. (2010-10)Atlantic killifish (Fundulus heteroclitus) inhabiting the PCB-contaminated Superfund site in New Bedford Harbor (MA, USA) have evolved genetic resistance to the toxic effects of these compounds. They also lack induction ...
New cytochrome P450 1B1, 1C2 and 1D1 genes in the killifish Fundulus heteroclitus : Basal expression and response of five killifish CYP1s to the AHR agonist PCB126 Zanette, Juliano; Jenny, Matthew J.; Goldstone, Jared V.; Woodin, Bruce R.; Watka, Lauren A.; Bainy, Afonso C. D.; Stegeman, John J. (2009-04-30)Knowledge of the complement of cytochrome P450 (CYP) genes is essential to understanding detoxification and bioactivation mechanisms for organic contaminants.We cloned three new CYP1 genes, CYP1B1, CYP1C2 and CYP1D1, ...