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    Development of the morpholino gene knockdown technique in Fundulus heteroclitus : a tool for studying molecular mechanisms in an established environmental model

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    Fundulus morpholino Manuscript FINAL Draft.pdf (691.5Kb)
    Date
    2008-02-13
    Author
    Matson, Cole W.  Concept link
    Clark, Bryan W.  Concept link
    Jenny, Matthew J.  Concept link
    Fleming, Carrie R.  Concept link
    Hahn, Mark E.  Concept link
    Di Giulio, Richard T.  Concept link
    Metadata
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    Citable URI
    https://hdl.handle.net/1912/2271
    As published
    https://doi.org/10.1016/j.aquatox.2008.02.010
    Keyword
     Fundulus heteroclitus; Antisense morpholino oligonucleotide; CYP1A; Atlantic killifish; Genetic toxicology; Development; Teratogenesis 
    Abstract
    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.
    Description
    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.
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    • Biology
    Suggested Citation
    Preprint: Matson, Cole W., Clark, Bryan W., Jenny, Matthew J., Fleming, Carrie R., Hahn, Mark E., Di Giulio, Richard T., "Development of the morpholino gene knockdown technique in Fundulus heteroclitus : a tool for studying molecular mechanisms in an established environmental model", 2008-02-13, https://doi.org/10.1016/j.aquatox.2008.02.010, https://hdl.handle.net/1912/2271
     

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