Effects of Calcium Ionophore on Vitellogenin Production in the Culture of Hepatocytes in the Rinbow Trout, Oncorhynchus mykess

무지개송어(Oncorhynchus mykiss) 배양 간세포에서 Vitellogenin 합성에 미치는 Calcium ionophore의 영향

  • Published : 1998.05.01

Abstract

Effects of A23187 on estradiol$-17^{\beta}$-induced vitellogenin (VTG) induction were electrophore-tically examined in primary hepatocyte cultures in rainbow trout. hepatocytes were predultured for 2 days and then estradiol-$17^{\beta}$(E2, $2{\times}10^{-6}$M) and calcium ionophore (A23187, $10^{-7)$~$10^{-5}$ M) were added to the incubation medium. The hepatocytes were cultured for 7 more days. In addition, effects of A23187 on $E_2$-primed VTG production were investigated for 7 days. The addition of A23187 ($10^{-7)$~$10^{-5}$M) to the incubation medium specifically reduced VTG production by hepatocytes in a concentration-dependent way. The addition of A23187 significantly reduced the rate of $E_2$-primed VTG production to 18% of the control (E2 only) on Day 7. However, $E_2$-primed VTG production was reduced to 47% of the control by withdrawal of $E_2$ from the incubation medium. Therefore, these results suggest that intracellualr sequestered calcium could regulate VTG synthesis at the translational and/or post-translational stage.

Vitellogenin(VTG) 합성에 미치는 A23187의 영향을 무지개송어의 배양 간세포를 이용하여 실험을 행하였다. 간세포는 2일간 배양한 후, Estradiol-$17^{\beta}$($E_2$, $2{\times}10^{-6}$M) 및 A23187 ($10^{-7)$~$10^{-5}$M)을 첨가하여 7일간 배양하였다. 그리고, $E_2$에 의한 VTG 합성시의 A23187이 미치는 영향에 대해서도 조사하였다. A23187이 미치는 영향에 대해서도 조사하였다. A23187 ($10^{-7)$~$10^{-5}$M)의 첨가에 의해 간세포에서의 VTG 합성은 농도의 증가에 따라 감소하였다.$E_2$에 의해 합성된 VTG는 A 23187 ($10^{-5}$M)의 첨가에 의해 대조군($E_2$만의 첨가)의 약 18%로 유의하게 감소하였다. 그러나,$E_2$제거로는 대조군의 약 47% 밖에 감소되지 않았다. 이러한 결과로 보아, 세포내의 저장 Ca은 번역 단계 또는 번역 후 단계를 조절함으로써, VTG 합성을 조절하는 것으로 추정된다.

Keywords

References

  1. Am. J. Physiol. v.251 Ionomycin acts as an ionophore to release TRH-regulated $Ca^{2+}$ stores from GH₄C₁cells Albert, P. R.;A. H. Tashjian Jr.
  2. Biochem. J. v.232 The $Ca^{2+}$ mobilizing actions of vasopressin and anbiotensin differ from those of the $^{\alpha}$-adrenergic agonist phenylephrine in the perfused rat liver Altin, J. G.;F. L. Bygrave
  3. J. Biol. Chem. v.258 no.23 Identification of a $Ca^{2+}$ requirement for protein synthesis in eukaryotic cells Brostrom, C. O.;S. B. Bocckino;M. A. Brostrom
  4. Mol. Pharmacol. v.29 Regulation of protein synthesis in isolated hepatocytes by calcium-mobilizing hormones Brostrom, C. O.;S. B. Bocckino;M. A. Brostrom;E. M. Galuska
  5. J. Biol. Chem. v.264 Inhibition of translational initiation in eukaryotic cells by calcium ionophore Brostrom, C. O.;K. V. Chin;W. L. Wong;C. Cade;M. A. Brostrom
  6. J. Biol. Chem. v.262 Calcium-dependent regulation of protein synthesis at translational initiation in eukaryotic cells Chin, K. -V.;C. Cade;C. O. Brostrom;E. M. Galuska;M. A. Brostrom
  7. Int. J. Biochem. v.20 Regulation of protein synthesis in intact rat liver by calcium mobilizing agents Chin, K. -V.;C. Cade;M. A. Brostrom;C. O. Brostrom
  8. Eur. J. Biochem. v.151 The role of protein phosphorylation in the hormonal control of enzyme activity Cohen, P.
  9. J. Biol. Chem. v.262 Regulated phosphorylation and low abundance of hera cell initiation factor eIF-4F suggest a role in translational control Duncan, R.;S. C. Milburn;J. W. B. Hershey
  10. J. Biol. Chem. v.264 A novel calcium-dependent phosphorylation of a ribosome-associated protein Fawell, E. H.;I. J. Boyer;M. A. Brostrom;C. O. Brostrom
  11. Reprod. Nutr. Develop. v.28 no.4A Biochemical analysis of vitellogenin from rainbow trout (Salmo gairdneri) : Fatty acid composition of phospholipids Fremont, L.;A. Riazi
  12. Comp. Biochem. Physiol. v.115C The in vitro effects of forskolin, IBMX and cyanoketone on meiotic maturation in follicle-enclosed catfish (Clarias batrachus) oocytes Haider, S.;S. K. Chaube
  13. Bull. Jpn. Soc. Sci. Fish. v.52 Primary culture of the eel hepatocytes in the serum free medium Hayashi, S.;Z. Ooshiro
  14. Comp. Biochem. Physiol. v.115A Relationship between vitellogenin and its related egg yolk proteins in sakhalin taimen(Hucho perryi) Hiramatsu, N.;A. Hara
  15. Am. J. Physiol. v.263 Reguration of protein synthesis by modulation of intracellular calcium in rat liver Kimball, S. R.;L. S. Jefferson
  16. Comp. Biochem. Physiol. v.104B Vitellogenin induction by estradiol-17β in primary hepatocyte culture in the rainbow trout, Oncorhynchus mykiss Kwon, H.;C. Hayashi;Y. Mugiya
  17. Nature v.227 Cleavage of structural proteins during assembly of the head of bateriophage T₄ Laemmli, U. K.
  18. J. Exp. Zool. v.272 Proteolytic cleavage of vitellogenin and yolk proteins during vitellogenin uptake and oocyte maturation in barfin flounder(Verasper moseri) Matsubara, T.;K. Sawano
  19. Biochemistry v.34 Vitellogenin and lipovitellin : Zinc proteins of Xenopus laevis oocytes Montorzi, M.;K. H. Falchuk;B. L. Vallee
  20. Biochem. v.235 Initiation of protein synthesis in mammalian cells Pain, V. M.
  21. Prog. Nucleic Acids Res. Mol. Biol. v.33 Regulation of protein synthesis by phosphorylation of ribosomal protein S6 and aminoacyl-tRNA synthesis. Traugh, J. A.;A. M. Pendergast
  22. Developmental Biology v.1 Vitellogenin and oocyte growth in nonmammalian vertebrates Wallace, R. A.;L. Browder(ed.)
  23. Gen. Comp. Endocrinol. v.105 Effects of extracellular calcium concentrations and calcium antagonists on vitellogenin induction by estradiol-17β in primary hepatocyte culture in the rainbow trout Oncorhynchus mykiss Yeo, I.-K.;Y. Mugiya
  24. Fish. Sci. v.64 no.3 Effects of calcium agonists on vitellogenin induction by estradiol-17β in primary hepatocyte culture in the rainbow trout Oncorhynchus mykiss Yeo, I. K.;Y. Mugiya