An in vitro vitellogenin bioassay for estrogenic substances in the Rhynchocypris oxycephalus

버들치(Rhynchocypris oxycephalus)에서 에스트로겐류 물질에 의한 vitellogenin의 유도 및 단일클론항체 제작

  • Accepted : 2006.01.22
  • Published : 2006.03.29


Vitellogenin (Vtg), a phospholipoglycoprotein precursor of egg yolk is synthesized and secreted from the liver in response to estrogens in female fish. Vtg is normally undetectable in the blood of male fish, but can be induced by exposure to chemicals possessing estrogenic activity. Thus, the presence of Vtg in blood of male fish can serve as a useful biomarker for assessing previous exposure to estrogenic compounds. In the present study, Vtg was abnormally expressed in Rhynchocypris oxycephalus using estradiol benzoate ($E_2$). As the result, it was found that the level of Vtg in blood from R. oxycephalus was increased by treated quantity of $E_2$ with dose-effect manner. Monoclonal antibodies were generated against Vtg of R. oxycephalus. The hybridoma were screened with an enzyme immunoassay for the production of specific anti-Vtg antibodies. Five positive cell lines with a high specificity were selected. Monoclonal antibodies against vtg of R. oxycephalus that was developed in this study, may be a useful bio-indicator for the detection of estrogenic contamination in the aquatic ecosystem.


Grant : Estrogen류 화합물의 정량측정법 및 Rapid kit의 개발

Supported by : 과학고


  1. 김익수. 동물편 (담수어류). In: 교육부. 한국동식물도감. Vol 37. pp. 258-259, 교육부, 서울, 1997
  2. 류재천. 내분비계 장애물질의 개요와 검색법의 고찰. 한국농약과학회지 2002, 6, 135-156
  3. 이행석, 조은민, 류재천. Human Estrogen Recetptor Ligand Binding Domain (hER LBD)과 Co-activator로 구성된 효모 two-hybrid system을 이용한 내분비계장 애물질 검출계의 구축. Environmental Mutagens & Carcinogens 2002, 22, 175-182
  4. 최기철, 전상린, 김익수, 손영목. 원색 한국담수어도감. pp. 90-91, 향문사, 서울, 1990
  5. Carlson DB, Williams DE. Sex-specific vitellogenin production in immature rainbow trout, Environ Toxicol Chem. 1999, 18, 2361-2363<2361:SSVPII>2.3.CO;2
  6. Folmer LC, Gardner GR, Schreibman MP, Magliulo-Cepriano L, Mills LJ, Zaroogian G, Gutjahr-Gobell R, Haebler R, Horowitz DB, Denslow ND. Vitellogenin-induced pathology in male summer flounder (Paralichthys dentatus). Aquat Toxicol 2001, 51, 431-441
  7. Hennies M, Wiesmann M, Allner B, Sauerwein H. Vitellogenin in carp (Cyprinus carpio) and perch (Perca fluviatilis): purification, characterization and development of an ELISA for the detection of estrogenic effects. Sci Total Environ 2003, 309, 93-103
  8. Heppell SA, Denslow ND, Folmar LC, Sullivan CV. Universal assay for vitellogenin as a biomarker for environmental estrogens. Environ Health Perspect 1995, 103, 9-15
  9. Kendall, J. Principles and processes for evaluating endocrine disruption in wildlife. pp. 141-154, SETAC Press, Pensacola, 1998
  10. Pakdel F, Feon S, Le Gac F, Le Menn F, Valotaire Y. In vivo estrogen induction of hepatic estrogen receptor mRNA and correlation with vitellogenin mRNA in rainbow trout. Mol Cell Endocrinol 1991, 75, 205-212
  11. Smeets JM, Rankouhi TR, Nichols KM, Komen H, Kaminski NE, Giesy JP, van den Berg M. In vitro vitellogenin production by carp (Cyprinus carpio) hepatocytes as a screening method for determining (anti) estrogenic activity of xenobiotics. Toxicol Appl Pharmacol 1999, 157, 68-76
  12. Sumpter JP, Jobling S. Vitellogenesis as a biomarker for estrogenic contamination of the aquatic environment. Environ Health Perspect 1995, 103, 173-178
  13. Vaillant C, Le Guellec C, Pakdel F, Valotaire Y. Vitellogenin gene expression in primary culture of male rainbow trout hepatocytes. Gen Comp Endocrinol 1988, 70, 284-290
  14. Wallace RA, Bergink GW. Amphibian vitellogenin: Properties, hormonal regulation of hepatic synthesis and ovarian uptake, and conversion to yolk proteins. Amer Zool 1974, 14, 1159-1175