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(293) Germ-line transgenic zebrafish as biosensors.
Linney, Elwood*,1, Yacisin, Kari1, Dobbs-McAuliffe, Betsy1, Donerly, Susan1, Zhao, Qingshun1, Christopher, Lassiter1, 1 Duke University Medical Center, Durham, NC, USA
ABSTRACT- Germ-line transgenic zebrafish as biosensors. Linney, E*, Yacisin, K, Dobbs-McAuliffe, B, Donerly, S, Zhao, Q, and Lassiter, CS, Duke University Medical Center, Durham, NC, USA. Our work is based upon two assumptions: 1)that environmental toxicants affect normal regulatory pathways in developing embryos and 2)that there is a conservation of genes and networks between different vertebrate species. We have produced transgenic lines of zebrafish which 1)express nuclear green fluorescent protein(GFP) and nuclear yellow fluorescent protein(YFP) in most of the cells of the developing embryo 2)transgenic lines of zebrafish which are biosensors for chemicals which affect the retinoic acid receptor mediated regulatory pathway and 3)transgenic lines which express cytoplasmic YFP in the developing nervous system. These lines allow us to follow the development of the nervous system and to visualize effects produced by challenge of the embryos with compounds which impact upon the developing nervous system. Embryos can be embedded live in agarose and fluorescent capture of developing embryos can be controlled by programming our computer to capture images at selected intervals. These images can then be used to generate time-lapse movies of fluorescent transgene expression during embryonic development. To complement these physical observations, through the NIEHS-funded toxicogenomics consortium we are analyzing gene expression changes during development with zebrafish microarrays.
Key words: biosensors, transgenic, zebrafish, fluorescence
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