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. 1974 Oct;144(1):99–106. doi: 10.1042/bj1440099

Oestradiol uptake and retention, and high-affinity binding sites in cultured rabbit uterus

Susan L Russell 1, Gordon H Thomas 1
PMCID: PMC1168469  PMID: 4376957

Abstract

Oestradiol uptake and turnover was examined in rabbit uterus maintained in organ culture for up to 3 days. Serum decreased the uptake of [3H]oestradiol, whereas insulin had no significant effect. During the first 24h of culture unoccupied high-affinity receptors for oestradiol were markedly depleted in the cytosol. Nuclear binding sites remained high during the first day of culture, and were still present after 3 days. The stability of nuclear-bound oestradiol was confirmed by examining the turnover of radioactivity during culture of uteri of rabbits injected with [3H]oestradiol 6h before death. Over half of the radioactivity was retained for as long as 3 days in tissue cultured in the absence of oestrogen. In tissue cultured for 24h with unlabelled oestrogen, there was a progressive increase in the displacement of [3H]oestradiol as the concentration of unlabelled hormone in the medium was increased from 0.1 to 5nm. Higher concentrations of oestradiol had little additional effect. The oestradiol involved in this displacement reaction was associated with macromolecules, characterized by Sephadex G-25 chromatography and sucrose-density-gradient ultracentrifugation of the 0.4m-KCl extract of the nuclear pellet.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson J. N., Clark J. H., Peck E. J., Jr The relationship between nuclear receptor-estrogen binding and uterotrophic responses. Biochem Biophys Res Commun. 1972 Sep 26;48(6):1460–1468. doi: 10.1016/0006-291x(72)90878-9. [DOI] [PubMed] [Google Scholar]
  2. Anderson J., Clark J. H., Peck E. J., Jr Oestrogen and nuclear binding sites. Determination of specific sites by ( 3 H)oestradiol exchange. Biochem J. 1972 Feb;126(3):561–567. doi: 10.1042/bj1260561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clark J. H., Anderson J. N., Peck E. J., Jr Estrogen receptor-anti-estrogen complex: atypical binding by uterine nuclei and effects on uterine growth. Steroids. 1973 Nov;22(5):707–718. doi: 10.1016/0039-128x(73)90118-9. [DOI] [PubMed] [Google Scholar]
  4. Clark J. H., Gorski J. Estrogen receptors: an evaluation of cytoplasmic-nuclear interactions in a cell-free system and a method for assay. Biochim Biophys Acta. 1969 Dec 30;192(3):508–515. doi: 10.1016/0304-4165(69)90400-0. [DOI] [PubMed] [Google Scholar]
  5. EAGLE H. Amino acid metabolism in mammalian cell cultures. Science. 1959 Aug 21;130(3373):432–437. doi: 10.1126/science.130.3373.432. [DOI] [PubMed] [Google Scholar]
  6. Giannopoulos G., Gorski J. Estrogen receptors. Quantitative studies on transfer of estradiol from cytoplasmic to nuclear binding sites. J Biol Chem. 1971 Apr 25;246(8):2524–2529. [PubMed] [Google Scholar]
  7. Gilman A. G. A protein binding assay for adenosine 3':5'-cyclic monophosphate. Proc Natl Acad Sci U S A. 1970 Sep;67(1):305–312. doi: 10.1073/pnas.67.1.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jensen E. V., Suzuki T., Kawashima T., Stumpf W. E., Jungblut P. W., DeSombre E. R. A two-step mechanism for the interaction of estradiol with rat uterus. Proc Natl Acad Sci U S A. 1968 Feb;59(2):632–638. doi: 10.1073/pnas.59.2.632. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. KAHN R. H. Effect of oestrogen and of vitamin A on vaginal cornification in tissue culture. Nature. 1954 Aug 14;174(4424):317–317. doi: 10.1038/174317a0. [DOI] [PubMed] [Google Scholar]
  10. King R. J., Gordon J. The association of [6,7-3H]oestradiol with a nuclear protein. J Endocrinol. 1967 Dec;39(4):533–542. doi: 10.1677/joe.0.0390533. [DOI] [PubMed] [Google Scholar]
  11. Kohorn E. I., Tchao R. Conversion of proliferative endometrium to secretory endometrium by progesterone in organ culture. J Endocrinol. 1969 Nov;45(3):401–405. doi: 10.1677/joe.0.0450401. [DOI] [PubMed] [Google Scholar]
  12. LEATHEM J. H. Some biochmical aspects of the uterus. Ann N Y Acad Sci. 1959 Jan 9;75:463–471. doi: 10.1111/j.1749-6632.1959.tb44568.x. [DOI] [PubMed] [Google Scholar]
  13. LOSTROH A. J. SYNTHETIC ACTIVITY IN MOUSE ORGANS MAINTAINED ON CHEMICALLY-DEFINED MEDIA. I. EFFECT OF INSULIN ON PROTEIN SYNTHESIS IN UTERI FROM OVARIECTOMIZED MICE. Acta Endocrinol (Copenh) 1964 Oct;47:331–337. doi: 10.1530/acta.0.0470331. [DOI] [PubMed] [Google Scholar]
  14. MUELLER G. C., HERRANEN A. M., JERVELL K. F. Studies on the mechanism of action of estrogens. Recent Prog Horm Res. 1958;14:95–139. [PubMed] [Google Scholar]
  15. Méster J., Rbertson D. M., Feherty P., Kellie A. E. Determination of high-affinity oestrogen receptor sites in uterine supernatant preparations. Biochem J. 1970 Dec;120(4):831–836. doi: 10.1042/bj1200831. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. NICOLETTE J. A., GORSKI J. EFFECT OF ESTRADIOL ON GLUCOSE-U-C-14 METABOLISM IN THE RAT UTERUS. Arch Biochem Biophys. 1964 Aug;107:279–283. doi: 10.1016/0003-9861(64)90330-3. [DOI] [PubMed] [Google Scholar]
  17. Puca G. A., Bresciani F. Receptor molecule for oestrogens from rat uterus. Nature. 1968 Jun 8;218(5145):967–969. doi: 10.1038/218967a0. [DOI] [PubMed] [Google Scholar]
  18. Russell S. L., Thomas G. H. The interconversion of oestrone and oestradiol-17 and the concentration of oestradiol-17 receptors in cultured rabbit uterus. J Endocrinol. 1973 Feb;56(2):203–211. doi: 10.1677/joe.0.0560203. [DOI] [PubMed] [Google Scholar]
  19. Shyamala G. Estradiol receptors in mouse mammary tumors: absence of the transfer of bound estradiol from the cytoplasm to the nucleus. Biochem Biophys Res Commun. 1972 Feb 25;46(4):1623–1630. doi: 10.1016/0006-291x(72)90795-4. [DOI] [PubMed] [Google Scholar]
  20. Szego C. M., Davis J. S. Adenosine 3',5'-monophosphate in rat uterus: acute elevation by estrogen. Proc Natl Acad Sci U S A. 1967 Oct;58(4):1711–1718. doi: 10.1073/pnas.58.4.1711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Toft D., Shyamala G., Gorski J. A receptor molecule for estrogens: studies using a cell-free system. Proc Natl Acad Sci U S A. 1967 Jun;57(6):1740–1743. doi: 10.1073/pnas.57.6.1740. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Williams D., Gorski J. A new assessment of subcellular distribution of bound estrogen in the uterus. Biochem Biophys Res Commun. 1971 Oct 1;45(1):258–264. doi: 10.1016/0006-291x(71)90078-7. [DOI] [PubMed] [Google Scholar]
  23. Wyss R. H., Karsznia R., Heinrichs W. L., Herrmann W. L. Inhibition of uterine receptor binding of estradiol by anti-estrogens (clomiphene and CL-868). J Clin Endocrinol Metab. 1968 Dec;28(12):1824–1828. doi: 10.1210/jcem-28-12-1824. [DOI] [PubMed] [Google Scholar]

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