DOI QR코드

DOI QR Code

Transgene chgH-rfp expression at developmental stages and reproductive status in marine medaka (Oryzias dancena)

  • Cho, Young Sun (Department of Marine Bio-Materials and Aquaculture, Pukyong National University) ;
  • Nam, Yoon Kwon (Department of Marine Bio-Materials and Aquaculture, Pukyong National University)
  • 투고 : 2016.08.05
  • 심사 : 2016.11.16
  • 발행 : 2016.11.30

초록

Background: The transgenic approach using estrogen-responsive regulator in fish has been given much attention as a potential means to detect and/or address estrogen-related aquatic pollutions. In order to address the development stage- and reproduction status-dependent expression patterns of the chgH-rfp transgene (red fluorescent protein transgene driven by choriogenin H promoter) in marine medaka Oryzias dancena, naturally occurring red fluorescent protein (RFP) signals under non-exposed conditions as well as the transgenically induced RFP signals under estrogen-exposed conditions were assayed. Results: Female transgenics begun to show naturally occurring RFP signals from the age of 7 weeks post hatching (WPH) without experimental estrogen exposure. Afterward, these RFP signals in female transgenics became robust with the progress of ovarian maturation. On the other hand, male transgenics did not show any naturally occurring RFP signal under non-exposed conditions irrespective of developmental stages and maturation statue. Upon exposures using estradiol-$17{\beta}$ (E2) and $17{\alpha}$-ethinylestradiol (EE2), RFP signals were significantly induced specifically in the livers of transgenic males. Conclusions: Male chgH-rfp transgenics were able to keep the "off" state of RFP expression during their entire life cycle unless exposed to exogenous estrogens. Owing to their tight regulation capability of estrogen-responsive transgene, transgenesis of chgH-rfp in male marine medaka could offer a useful model system for future ecotoxicogenomic studies regarding estrogenicity-related issues in aquatic and marine environments.

키워드

참고문헌

  1. Bogers R, Mutsaerds E, Druke J, De Roode DF, Murk AJ, Van Der Burg B, Legler J. Estrogenic endpoints in fish early life-stage tests: luciferase and vitellogenin induction in estrogen-responsive transgenic zebrafish. Environ Toxicol Chem. 2006;25:241-7. https://doi.org/10.1897/05-234R.1
  2. Chen X, Li VWT, Yu RMK, Cheng SH. Choriogenin mRNA as a sensitive molecular biomarker for estrogenic chemicals in developing brackish medaka (Oryzias melastigma). Ecotoxicol Environ Saf. 2008;71:200-8. https://doi.org/10.1016/j.ecoenv.2007.10.005
  3. Chen H, Hu J, Yang J, Wang Y, Xu H, Jiang Q, Gong Y, Gu Y, Song H. Generation of a fluorescent transgenic zebrafish for detection of environmental estrogens. Aquat Toxicol. 2010;96:53-61. https://doi.org/10.1016/j.aquatox.2009.09.015
  4. Cho YS, Lee SY, Kim DS, Nam YK. Spawning performance, embryonic development and early viability under different salinity conditions in a euryhaline medaka species, Oryzias dancena. Korean J Ichthyol. 2010;22:25-33.
  5. Cho YS, Lee SY, Kim YK, Kim DS, Nam YK. Functional ability of cytoskeletal $\beta$-actin regulator to drive constitutive and ubiquitous expression of a fluorescent reporter throughout the life cycle of transgenic marine medaka Oryzias dancena. Transgenic Res. 2011;20:1333-55. https://doi.org/10.1007/s11248-011-9501-2
  6. Cho YS, Kim DS, Nam YK. Characterization of estrogen-responsive transgenic marine medaka Oryzias dancena germlines harboring red fluorescent protein gene under the control by endogenous choriogenin H promoter. Transgenic Res. 2013;22:501-17. https://doi.org/10.1007/s11248-012-9650-y
  7. Hong L, Fujita T, Wada T, Amano H, Hiramatsu N, Zhang X, Todo T, Hara A. Choriogenin and vitellogenin in red lip mullet (Chelon haematocheilus): purification, characterization, and evaluation as potential biomarkers for detecting estrogenic activity. Comp Biochem Physiol C Toxicol Pharmacol. 2009;149:9-17. https://doi.org/10.1016/j.cbpc.2008.05.017
  8. Kinoshita M, Murata K, Naruse K, Tanaka M. Reproduction of medaka. In: Kinoshita M, Murata K, Naruse K, Tanaka M, editors. Medaka: biology, management, and experimental protocols. Iowa: Wiley-Blackwell; 2009. p. 67-99.
  9. Kumar A, Crawford K, Flick R, Klevitsky R, Lorenz JN, Bove KE, Robbins J, Lessard JL. Transgenic overexpression of cardiac actin in the mouse heart suggests coregulation of cardiac, skeletal and vascular actin expression. Transgenic Res. 2004;13:531-40. https://doi.org/10.1007/s11248-004-2823-6
  10. Kurauchi K, Nakaguchi Y, Tsutsumi M, Hori H, Kurihara R, Hashimoto S, Ohnuma R, Yamamoto Y, Matsuoka S, Kawai S, Hirata T, Kinoshita M. In vivo visual reporter system for detection of estrogen-like substances by transgenic medaka. Environ Sci Technol. 2005;39:2762-68. https://doi.org/10.1021/es0486465
  11. Kurauchi K, Hirata T, Kinoshita M. Characteristics of ChgH-GFP transgenic medaka lines, an in vivo estrogenic compound detection system. Mar Pollut Bull. 2008;57:441-4. https://doi.org/10.1016/j.marpolbul.2008.03.016
  12. Lee SY, Kim DS, Nam YK. Gene structure and estrogen responsive mRNA expression of a novel choriogenin H isoform from a marine medaka (Oryzias dancena). Fish Aquat Sci. 2012;15:221-31.
  13. Lee SY, Kim DS, Nam YK. Molecular characterization of fast skeletal musclespecific myosin light chain 2 gene (mlc2f) in marine medaka Oryzias dancena. Genes Genom. 2013;35:289-303. https://doi.org/10.1007/s13258-013-0071-y
  14. Nam YK, Cho YS, Kim DS. Expression patterns of the chgH:rfp transgene in response to 17a-ethinylestradiol (EE2) exposure in marine medaka Oryzias dancena. Fish Aquat Sci. 2015;18:65-71.
  15. Salam MA, Sawada T, Ohya T, Ninomiya K, Hayashi S. Detection of environmental estrogenicity using transgenic medaka hatchlings (Oryzias latipes) expressing the GFP tagged choriogenin L gene. J Environ Sci Health, Part A. 2008;43:272-7. https://doi.org/10.1080/10934520701792761
  16. Scholz S, Kurauchi K, Kinoshita M, Oshima Y, Ozato K, Schirmer K, Wakamatsu Y. Analysis of estrogenic effects by quantification of green fluorescent protein in juvenile fish of a transgenic medaka. Environ Toxicol Chem. 2005;24:2553-61. https://doi.org/10.1897/04-525R.1
  17. Song HY. Cytogenetic and reproductive characteristics of interspecific hybrids between marine medaka Oryzias dancena and Javanese medaka O. javanicus. Ph.D. Dissertation Pukyong Natl Univ. Busan. 2010. p. 133.
  18. Song HY, Nam YK, Bang IC, Kim DS. Embryogenesis and early ontogenesis of a marine medaka, Oryzias dancena. Korean J Ichthyol. 2009;21:227-38.
  19. Song HY, Nam YK, Bang IC, Kim DS. Hybridization between marine medaka Oryzias dancena and Javanese medaka Oryzias javanicus. Kor J Fish Aquat Sci. 2010;43:462-73.
  20. Stegeman JJ, Goldstone JV, Hahn ME. Perspectives on zebrafish as a model in environmental toxicology. Fish Physiol. 2010;29:367-439.
  21. Ueno T, Yasumasu S, Hayashi S, Iuchi I. Identification of choriogenin cis-regulatory elements and production of estrogen-inducible, liver-specific transgenic medaka. Mech Dev. 2004;121:803-15. https://doi.org/10.1016/j.mod.2004.03.027
  22. Zeng Z, Shan T, Tong Y, Lam SH, Gong Z. Development of estrogen-responsive transgenic medaka for environmental monitoring of endocrine disrupters. Environ Sci Technol. 2005;39:9001-8. https://doi.org/10.1021/es050728l