Expression of Vitellogenin Gene by Androgens in Rasinbow Trout, Oncorhynchus mykiss

웅성호르몬에 의한 무지개송어의 vitellogenin 유전자 발현

  • 권혁추 (선문대학교 응용생물과학부) ;
  • 윤종만 (군산대학교 해양생명의학과) ;
  • 이종영 (게이오대학 의학부 분자생물학교실)
  • Published : 2000.03.01

Abstract

The effects of estrogen and androgens on Vg gene expression were examined in primary hepatocyte culture and livers of the immature male trout. Specific primers of Vg cDNA were designed with already reported Vg gene nucleotide sequences. PCR product was sequenced and verified with Vg cDNA of rainbow trout. Total RNA was extracted from the cultured hepatocytes and livers of steroid-treated rainbow trout and then it was analyzed by reverse transcriptase- polymerase chain reaction (RT-PCR) analysis. The Vg mRNA and Vg protein synthesis were increased in rainbow trout in vivo and in vitro with E$_2$ and methyltestosterone (MT) There were dose and time-related effects of E$_2$ and MT on vitellogesis. Androgens such as progesterone androsterone and testosterone also stimulated Vg mRNA expression in vitro. The results show that androgens as well as E$_2$ can induce expression of Vg mRNA in trout in vivo and in vitro.

자성 및 웅성스테로이드 호르몬들이 Vg 유전자발현에 영향을 미치는지를 미성숙 무지개송어의 배양간세포 막간을 이용하여 조사하였다. 이미 보고된 송어의 Vg gene의 염기배열을 참고로 Vg cDNA 단편(600 bp)을 증폭시킬 수 있는 primer들을 작성하였다. 이들 primer를 이용하여 증폭된 PCR 산물의 염기배열을 결정하여 송어의 Vg cDNA임을 확인하였고, RT-PCR법을 이용하여 배양간세포 그리고 E$_2$ 및 MT 처리된 송어의 간으로부터 Vg mRNA의 전사량 변화를 조사하였다. 호르몬 처리된 간세포 및 송어의 간에서 추출한 total RNA를 이용하여 RT-PCR법으로 분석한 결과 in vivo, in vitro 실험 모두에서 E$_2$ 또는 MT처리된 간세포 및 송어의 간으로부터 Vg mRNA와 Vg 단백질합성이 유도되었고, 이들의 증가 경향은 처리된 호르몬 농도 및 시간에 의존하고 있음이 밝혀졌다. 또한 progesterone, androsterone 그리고 testosterone 등의 웅성호르몬들도 Vg mRNA의 전사를 유도하고 있다는 것이 시사되었다. 이와 같은 결과로부터E$_2$ 뿐 아니라 웅성스테로이드들도 Vg mRNA의 발현을 유도하고 있음이 송어의 in vivo 또는 in vitro 실험에 의해서 확인되었다.

Keywords

References

  1. Proc. Natl. Acad. Sci v.85 Induction of vitellogenin in primary monolayer cultures of cockerel hepatocytes Boehm K.D.;R.L.;J. Ilan
  2. Gen. Comp. Endocrinol. v.84 Effects of 17β-estradiol and carp gonadotropin on European silver female eel (Anguilla anguilla L.) employing a homologous radioimmunoassay for vitellogenin Burzawa-Gerard E.;A. Dumas-Vidal
  3. Gen. Comp. Endocrinol. v.88 Hormoal control of in vitro vitellogenin synthesis in Rana esculenta liver: Effects of Mammalian and Amphibian growth hormone Carnevali O.;G. Mosconi;K. Yamamoto;T. Kobayashi;S. Kikuyama;A.M. Polzonetti-Magni
  4. Mol. Cell. Endocrinol. v.114 MUltihormonal control of vitellogenin mRNA expression in the liver of frog, Rana esculenta Carnevali O.;MG. Sabbieti;G. Mosconi;A.M. Polzonetti-Magni
  5. Comp. Biochem. Physiol. v.55B Natural Occurrence, and experimental induction by estradiol-17β, of a lipophosphoprotein (Vitellogenin) in flounder (Platichthys flesus, L.) Emmersen B.K.;I.M. Petersen
  6. J. Endocrinol. v.83 Effect of estradiol-17β on serum calcium and vitellogenin levels in rainbow trout Eliott J.A.K.;N.R. Bromage;C. Whitehead
  7. Comp. Biochem. Physiol. v.81B Sexual differences in the in vitro induction of vitellogenesis in the turtle: Role of the pituitary and growth hormone Ho S.M.;L.J. Wangh;I.P. Callard
  8. Gen. Comp. Endocrinol. v.37 Induction of vitellogenin synthesis in goldfish by massive doses of androgens Hori S.H.;T. Kodama;K. Tanahashi
  9. Comp. Biochem. Physiol. v.104B Vitellogenin induction by estradiol-17β in primary hepatocyte culture in the rainbow trout, Oncorhynchus mykiss Kwon H.C.;S. Hayashi;Y. Mugiya
  10. Gen. Comp. Endocrinol. v.93 Involvement of growth hormone and prolactin in the induction of vitellogenin synthesis in primary hepatocyte culture in the eel Kwon H.C.;Y. Mugiya
  11. Bull. Fac. Fish. Hokkaido Univ. v.41 Enzyme-linked immunosorbent assay (ELISA) of vitellogenin in white spotted charr, Salvelinus leucomaenis Kwon H.C.;A. Hara;Y. Mugiya;J. Yamada
  12. Korean J. Animal Reprod. v.20 Induction of vitellogenin synthesis by androgens in cultured hepatocytes of the eel, Anguilla japonica Kwon H.;H.Y. Park
  13. Nature v.227 Cleavage of structural roteins during the assembly of the head of bacteriophage T4 Laemmli U.K.
  14. Steroids v.35 Effect of androgen mediated by the estrogen receptor of fish liver: vitellogenin accumulation Le Menn F.;H. Rochefort;M. Garcia
  15. Vitro Cell. Dev. Biol. v.22 Estradiol-17β stimulation of vitellogenin synthesis in primary culture of male rainbow trout hepatocytes Maitre J.-L.;Y. Valotaire;C. Guguen-Guillouzo
  16. Mol. Cell. Endocrinol. v.141 Regulation of the vitellogenin gene B1 promoter after transfer into hepatocytes in primary cultures Marilley D.;D. Robyr;C. Schild-poulter;W. Wahli
  17. Fish Physiology v.XIA Vitellogenesis and oocyte assembly Mommsen T.P.;P.J. Walsh
  18. Mol. Cell. Endocrinol. v.66 Estradiol receptor in the lizard liver (Podarcis s. sicula);Seasonal changes and estradiol and growth hormone dependence Paolucci M.
  19. Chem. Biol. Interact. v.100 Effects of estrogen and nonylphenol on the post -transcriptional regulation of vitellogenin gene expression Ren L.;SK. Lewis;JJ. Lech
  20. A laboratory manual Molecular cloning Sambrook J.;E.F. Fritsch;T. Maniatis
  21. Gen. Comp. Endocrinol. v.52 A comparation of the female reproductive cycles of autumn-spawning and winter-spawning strains of rainbow trout (Salmo gairdneri Richardson) Scott A.P.;J.P. Sumpter
  22. Environ. Health Perspect v.7 Vitellogenesis as a biomarker for estrogenic contamination of the aquatic environment Sumpter JP.;S. Jobling
  23. Gen. Comp. Endocrinol. v.43 Streroid-induced synthesis of vitellogenin in the catfish Heteroppneustes fossilis (Bloch) Sundararaj B.I.;P. Nath
  24. Gen. Comp. Endocrinol. v.48 Role of testosterone, estradiol-17β, and cortisol during vitellogenin synthesis in the catfish, Heteropneustes fossilis(Block) Sundararaj B.I.;S.V. Goswami;V.J. Lamb
  25. Rec. Prog. Horm. Res. v.35 Vitellogenesis : A versatile model for hormonal regulation of gene expression Tata J.R.;D.F. Smith
  26. Mol. Cell Endocrinol. v.30 Rapid estrogen metabolism and vitellogenin gene expression in Xenopus hepatocyte cultures Tenniswood M.P.R.;P.F. Searle;A.P. Wolffe;J.R. Tata
  27. Gen. Comp. Endocrinol. v.53 Changes and serum concentrations of steriod hormones, thyroxine, and vitellogenin druing spawning migration of the chum salmon, Oncorhynchus Ueda H.;O. Hiroi;A. Hara;K. Yamauchi;Y. Nagahama
  28. Gen. Comp. Endocrinol. v.70 Vitellogenin gene expression in primary culture of male rainbow trout hepatocytes Vaillant C.;C. Le Guellec;F. Pakdel;Y. Valotaire
  29. Gen. Comp. Endocrinol. v.46 Estrone and estradiol particitation during exogenous vitellogenesis in the rainbow trout, Salmo gairdneri Van Bohemen Ch. G.;J.G.D. Lambert;H.J.Th.;P.G.W.J. van Oordt
  30. Developmental Biology v.1 Vitellogenin and oocyte growth in nonmammalian vertebrates Wallace R.A.;L. Browder(ed.)
  31. Dev. Biol. v.89 Glucocorticoids act together with estrogens and thyroid hormones in regulating the synthesis and secretion of Xenopus vitellogenin, serum albumin, and fibrinogen Wangh L.J.
  32. Proc. Natl. Acad. Sci. USA v.72 Synthesis of vitellogenin in cultures of male and female frog liver regulated by estradiol treatment in vitro Wangh L.J.;A.J. Knowland
  33. Dev. Biol. v.89 Thyroid hormones are corequisites for estradiol-17β in vitro induction of Xenopus vitellogenin Wangh L.J.;W. Schneider