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.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.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.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

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

Files

Original bundle

Now showing 1 - 1 of 1
Loading...
Thumbnail Image
Name:
Matson et al. 2008.pdf
Size:
449.55 KB
Format:
Adobe Portable Document Format
Description:
Published version