ZFIN ID: ZDB-PERS-960805-341
Linney, Elwood
Email: elwood.linney@duke.edu
URL: http://glowfish.mc.duke.edu/
Affiliation: Linney Lab
Address: Laboratory of Molecular Development Department of Molecular Genetics and Microbiology Duke University Medical Center Box 3020 Durham, NC 27710 USA
Country: United States
Phone: (919) 684-6095
Fax: (919) 684-8735

This laboratory has focused its attention on retinoid signaling in the developing mouse embryo. This occurs through ligand-inducible transcription factors named retinoic acid receptors(RARs) or retinoid-X receptors(RXRs). These receptors can heterodimerize and bind to response elements associated with the transcriptional promoters of genes. We have used molecular, cellular and animal techniques to identify regions of active retinoic acid receptor activity in embryos, sources of retinoic acid in embryos, and the molecular structure of some of the genes encoding these receptors. In the mouse, we are currently using procedures that should allow us to identify genes that are regulated via these receptors.

In addition to this work, we have been involved in using magnetic resonance microscopy to image and render in three dimensions, developing mouse embryos. This has resulted in a CD ROM 'Digital Atlas of Mouse Embryology' (B.R. Smith, G.A. Johnson and E. Linney).

We recognized certain limitations in working with complex signaling in developing mouse embryos and have recently begun to work with an alternative vertebrate system, the zebrafish. This system allows us to obtain quite easily hundreds of developing embryos and to use simple injections to introduce proteins, mRNAs and DNAs into the embryos.

Because the zebrafish embryo does not markedly grow in overall size during embryonic development, it can be 'optically sectioned' completely throughout embryonic development using fluorescent markers and the scanning confocal laser microscope.

For the above reasons and more, we have begun to use the zebrafish system in our studies of retinoid signaling during embryonic development and are adapting methods and techniques which should allow us to 'see' gene expression in developing zebrafish embryos using fluorescent reporter molecules. Because of its small embryonic size, one can examine the pattern of a gene throughout development using the technique of whole mount in situ hybridization. We are adapting this technique to provide us with a fluorescent signal so that we can capture, in three dimensions, the localization of specific mRNAs and then use computer reconstruction and rendering techniques to construct the location of the mRNA in three dimensions.

With the above technologies, we are now addressing several different questions:

1) Are there localized regions of retinoic acid receptor activity in the developing zebrafish embryo?

2) Can we interfere and re-direct this signaling through introducing of in vitro synthesized mRNAs that would inhibit receptor activity?

3) Can we identify adaptor molecules which allow the receptor to communicate with the transciption complex using the yeast two-hybrid selection system?

4) Can we develop the zebrafish embryo as a biosensor for environmental estrogens using the above technology in combination with the creation of transgenic indicator zebrafish for estrogen receptor activity?

Aluru, N., Kuo, E., Helfrich, L.W., Karchner, S.I., Linney, E.A., Pais, J.E., Franks, D.G. (2015) Developmental exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin alters DNA methyltransferase (dnmt) expression in zebrafish (Danio rerio). Toxicology and applied pharmacology. 284(2):142-51
Hahn, M.E., McArthur, A.G., Karchner, S.I., Franks, D.G., Jenny, M.J., Timme-Laragy, A.R., Stegeman, J.J., Woodin, B.R., Cipriano, M.J., Linney, E. (2014) The Transcriptional Response to Oxidative Stress during Vertebrate Development: Effects of tert-Butylhydroquinone and 2,3,7,8-Tetrachlorodibenzo-p-Dioxin. PLoS One. 9:e113158
Roy, N.M., Arpie, B., Lugo, J., Linney, E., Levin, E.D., and Cerutti, D. (2012) Brief embryonic strychnine exposure in zebrafish causes long-term adult behavioral impairment with indications of embryonic synaptic changes. Neurotoxicology and teratology. 34(6):587-591
Yen, J., Donerly, S., Levin, E.D., and Linney, E.A. (2011) Differential acetylcholinesterase inhibition of chlorpyrifos, diazinon and parathion in larval zebrafish. Neurotoxicology and teratology. 33(6):735-41
Levin, E.D., Sledge, D., Roach, S., Petro, A., Donerly, S., and Linney, E. (2011) Persistent behavioral impairment caused by embryonic methylphenidate exposure in zebrafish. Neurotoxicology and teratology. 33(6):668-73
Sledge, D., Yen, J., Morton, T., Dishaw, L., Petro, A., Donerly, S., Linney, E., and Levin, E.D. (2011) Critical duration of exposure for developmental chlorpyrifos-induced neurobehavioral toxicity. Neurotoxicology and teratology. 33(6):742-51
Linney, E., Donerly, S., Mackey, L., and Dobbs-McAuliffe, B. (2011) The negative side of retinoic acid receptors. Neurotoxicology and teratology. 33(6):631-40
Linney, E., Perz-Edwards, A., and Kelley, B. (2011) Identification and characterization of a functional zebrafish smrt corepressor (ncor2). Gene. 486(1-2):31-6
Aschner, M., Levin, E.D., Suñol, C., Olopade, J.O., Helmcke, K.J., Avila, D.S., Sledge, D., Ali, R.H., Upchurch, L., Donerly, S., Linney, E., Forsby, A., Ponnoru, P., and Connor, J.R. (2010) Gene-Environment Interactions: Neurodegeneration in Non-Mammals and Mammals. Neurotoxicology. 31(5):582-588
Alexeyenko, A., Wassenberg, D.M., Lobenhofer, E.K., Yen, J., Linney, E., Sonnhammer, E.L., and Meyer, J.N. (2010) Dynamic zebrafish interactome reveals transcriptional mechanisms of dioxin toxicity. PLoS One. 5(5):e10465
Powers, C.M., Yen, J., Linney, E.A., Seidler, F.J., and Slotkin, T.A. (2010) Silver exposure in developing Zebrafish (Danio rerio): Persistent effects on larval behavior and survival. Neurotoxicology and teratology. 32(3):391-397
Eddins, D., Cerutti, D., Williams, P., Linney, E., and Levin, E.D. (2010) Zebrafish Provide a Sensitive Model of Persisting Neurobehavioral Effects of Developmental Chlorpyrifos Exposure: Comparison with Nicotine and Pilocarpine Effects and Relationship to Dopamine Deficits. Neurotoxicology and teratology. 32(1):99-108
Timme-Laragy, A.R., Van Tiem, L.A., Linney, E.A., and Di Giulio, R.T. (2009) Antioxidant responses and NRF2 in synergistic developmental toxicity of PAHs in zebrafish. Toxicological sciences : an official journal of the Society of Toxicology. 109(2):217-227
Hu, P., Tian, M., Bao, J., Xing, G., Gu, X., Gao, X., Linney, E., and Zhao, Q. (2008) Retinoid regulation of the zebrafish cyp26a1 promoter. Developmental Dynamics : an official publication of the American Association of Anatomists. 237(12):3798-3808
Malone, M.H., Sciaky, N., Stalheim, L., Hahn, K.M., Linney, E., and Johnson, G.L. (2007) Laser-scanning velocimetry: A confocal microscopy method for quantitative measurement of cardiovascular performance in zebrafish embryos and larvae. BMC Biotechnology. 7(1):40
Kim, M.J., Liu, I.H., Song, Y., Lee, J.A., Halfter, W., Balice-Gordon, R.J., Linney, E., and Cole, G.J. (2007) Agrin is Required for Posterior Development and Motor Axon Outgrowth and Branching in Embryonic Zebrafish. Glycobiology. 17(2):231-247
Lassiter, C.S., and Linney, E. (2007) Embryonic Expression And Steroid Regulation of Brain Aromatase cyp19a1b in Zebrafish (Danio Rerio). Zebrafish. 4(1):49-58
Zhao, Q., Dobbs-McAuliffe, B., and Linney, E. (2005) Expression of cyp26b1 during zebrafish early development. Gene expression patterns : GEP. 5(3):363-369
Linney, E., Upchurch, L., and Donerly, S. (2004) Zebrafish as a neurotoxicological model. Neurotoxicology and teratology. 26(6):709-718
Levin, E.D., Swain, H.A., Donerly, S., and Linney, E. (2004) Developmental chlorpyrifos effects on hatchling zebrafish swimming behavior. Neurotoxicology and teratology. 26(6):719-723
Linney, E., Dobbs-McAuliffe, B., Sajadi, H., and Malek, R.L. (2004) Microarray gene expression profiling during the segmentation phase of zebrafish development. Comparative biochemistry and physiology. Toxicology & pharmacology : CBP. 138(3):351-362
Dobbs-McAuliffe, B., Zhao, Q., and Linney, E. (2004) Feedback mechanisms regulate retinoic acid production and degradation in the zebrafish embryo. Mechanisms of Development. 121(4):339-350
Linney, E., and Udvadia, A.J. (2004) Construction and detection of fluorescent, germline transgenic zebrafish. Methods in molecular biology (Clifton, N.J.). 254:271-288
Udvadia, A.J. and Linney, E. (2003) Windows into development: historic, current, and future perspectives on transgenic zebrafish. Developmental Biology. 256(1):1-17
Levin, E.D., Chrysanthis, E., Yacisin, K., and Linney, E. (2003) Chlorpyrifos exposure of developing zebrafish: effects on survival and long-term effects on response latency and spatial discrimination. Neurotoxicology and teratology. 25(1):51-57
Lassiter, C.S., Kelley, B., and Linney, E. (2002) Genomic structure and embryonic expression of estrogen receptor beta a (ERbetaa) in zebrafish (Danio rerio). Gene. 299(1-2):141-151
Perz-Edwards, A., Hardison, N.L., and Linney, E. (2001) Retinoic acid-mediated gene expression in transgenic reporter zebrafish. Developmental Biology. 229(1):89-101
Linney, E., Hardison, N.L., Lonze, B.E., Lyons, S., and DiNapoli, L. (1999) Transgene expression in zebrafish: a comparison of retroviral-vector and DNA-injection approaches. Developmental Biology. 213(1):207-216