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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.

dc.contributor.author Matson, Cole W
dc.contributor.author Clark, Bryan W
dc.contributor.author Jenny, Matthew J
dc.contributor.author Fleming, Carrie R
dc.contributor.author Hahn, Mark E
dc.contributor.author Di Giulio, Richard T
dc.coverage.spatial Netherlands
dc.date.accessioned 2013-04-30T14:14:58Z
dc.date.issued 2008-05-30
dc.identifier https://www.ncbi.nlm.nih.gov/pubmed/18378331
dc.identifier S0166-445X(08)00069-6
dc.identifier.issn 0166-445X
dc.identifier.uri https://hdl.handle.net/10161/6969
dc.description.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 beta-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.
dc.language eng
dc.publisher Elsevier BV
dc.relation.ispartof Aquat Toxicol
dc.relation.isversionof 10.1016/j.aquatox.2008.02.010
dc.subject Animals
dc.subject Cytochrome P-450 CYP1A1
dc.subject Embryo, Nonmammalian
dc.subject Fundulidae
dc.subject Heart Defects, Congenital
dc.subject Models, Biological
dc.subject Oligonucleotides, Antisense
dc.subject beta-Naphthoflavone
dc.title Development of the morpholino gene knockdown technique in Fundulus heteroclitus: a tool for studying molecular mechanisms in an established environmental model.
dc.type Journal article
duke.contributor.id Clark, Bryan W|0341357
duke.contributor.id Di Giulio, Richard T|0085052
pubs.author-url https://www.ncbi.nlm.nih.gov/pubmed/18378331
pubs.begin-page 289
pubs.end-page 295
pubs.issue 4
pubs.organisational-group Civil and Environmental Engineering
pubs.organisational-group Duke
pubs.organisational-group Environmental Sciences and Policy
pubs.organisational-group Marine Science and Conservation
pubs.organisational-group Nicholas School of the Environment
pubs.organisational-group Pratt School of Engineering
pubs.organisational-group Staff
pubs.publication-status Published
pubs.volume 87


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