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H. Choi, C. Joo; The Effect of Lithium During Zebrafish Eye Development . Invest. Ophthalmol. Vis. Sci. 2003;44(13):651.
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Purpose: The Wnt/beta-catenin signaling plays multiple roles during embryonic development. Gsk3-beta(glycogen synthase kinase3-beta) plays a key role in cell-fate decisions by negatively regulating beta-catenin. It has been reported that lithium inhibit the Gsk3-beta activity thus mimicking Wnt signaling in zebrafish embryos. The purpose of this study is to search of differentially regulated gene(s) by lithium during zebrafish eye development. Methods: Treatment of zebrafish embryos with lithium after 4~6 hours post-fertilization leads to eyeless or small eyes. Total RNA was isolated from lithium treated embryos and untreated as a control. These RNA were used for cDNA synthesis followed by subtraction. Results: Approximately 700 clones from the cDNA subtraction libraries were blotted onto nitrocellulose membrane and hybridized with DIG-labeled cDNA from each group as probes. More than putative 70 individual clone were isolated and then sequenced. Among them the transcriptional levels of Type I cytokeratin, Type II cytokeratin and Ke3 were decreased in lithium treated embryos compared to untreated control group as determined by Northern analysis. This result is consistent with previous report that Type I cytokeratin and Type II cytokeratin were expressed in zebrafish eye. Conclusions: Our finding suggests that lithium may affect the expression of gene(s) responsible for eye formation during zebrafish early development.
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