title: Characterization of estrogen-responsive transgenic marine medaka Oryzias dancena germlines harboring red fluorescent protein gene under the control by endogenous choriogenin H promoter.
abstract: Transgenic marine medaka (Oryzias dancena) germlines were generated by the microinjection of the red fluorescent protein (RFP) reporter gene (rfp) driven by the endogenous choriogenin H gene (chgH) promoter. The selected transgenic lines contained multiple copies of the transgene (3-42 copies per cell) in their genomes. Although all the founders were mosaic, the transgene was stably transmitted from the F1 generation to all subsequent generations following a Mendelian pattern. Different transgenic lines showed different responsiveness to estradiol-17β (E2) exposure at the mRNA and protein levels, and the expression efficiency was dependent upon the transgene copy number. The induction of RFP was significantly affected by the developmental stage of transgenic larvae: later-stage larvae (older than 7 days post-hatching) showed higher sensitivity to E2 exposure than earlier-stage larvae. The response of transgenic expression to E2 was fairly dependent upon the E2 dose (200-3,200 ng/L) and exposure period (1-7 days), according to both a microscopic examination of RFP intensity and a qRT-PCR assay. The transgenic marine medaka showed similar transgenic responses to E2 under freshwater, brackish, and seawater conditions. In addition to E2, the transgenic RFP signal was also successfully induced during 1-week exposure to various other natural (1 μg/L estrone and 10 μg/L estriol) and synthetic (xeno)estrogens (0.1 μg/L 17α-ethynylestradiol, 1 μg/L diethylstilbestrol, and 10 mg/L bisphenol A). The efficiency of transgene expression varied greatly among the chemicals tested. The results of this study suggest that the chgH-rfp transgenic marine medaka species will be useful in the in vivo detection of waterborne estrogens under a wide range of salinity conditions.
authors.affiliation: Institute of Marine Living Modified Organisms, Pukyong National University, Busan 608-737, South Korea.
authors.forename: Young Sun
authors.initials: YS
authors.lastname: Cho
authors.forename: Dong Soo
authors.initials: DS
authors.lastname: Kim
authors.forename: Yoon Kwon
authors.initials: YK
authors.lastname: Nam
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D004305:Dose-Response Relationship, Drug; D004527:Egg Proteins / Q000235:genetics*; D004784:Environmental Monitoring / Q000379:methods; D004958:Estradiol / Q000494:pharmacology; D004967:Estrogens / Q000378:metabolism / Q000494:pharmacology*; D004970:Estrone / Q000494:pharmacology; D004997:Ethinyl Estradiol / Q000494:pharmacology; D005618:Fresh Water; D018628:Gene Dosage; D005786:Gene Expression Regulation / Q000187:drug effects; D005854:Germ Cells; D007814:Larva / Q000187:drug effects / Q000254:growth & development; D008164:Luminescent Proteins / Q000235:genetics*; D009990:Oryzias / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011498:Protein Precursors / Q000235:genetics*; D012623:Seawater; D014874:Water Pollutants, Chemical / Q000032:analysis; D000097573:Red Fluorescent Protein
chemical_list: D004527:Egg Proteins; D004958:Estradiol; D004967:Estrogens; D004970:Estrone; D004997:Ethinyl Estradiol; D008164:Luminescent Proteins; D011498:Protein Precursors; D014874:Water Pollutants, Chemical; D000097573:Red Fluorescent Protein; C104950:choriogenin H