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Title: Estrogen- and progestin-receptor complexes and their interaction with rat liver cell nuclei during ontogenesis

Abstract

Steroid-receptor complexes (SRC) of estrogens and progestins have been isolated from the rat liver and purified 1500-2000-fold. Both in the state of the initial cytosol and purified 2000-fold, the SRC were characterized by gel filtration on Sephadex G-100 and by DEAE-cellulose chromatography. The purified SRC from the rat liver were used for binding to isolated liver cell nuclei from rats of different ages (1.5 months, 6 months, 12 months, 24 months). The maximum binding of SRC of progestins and estrogens from the rat liver is observed with homologous nuclei of 1.5-month-old rats. With age the binding of SRC by the nuclei decreases progressively and reaches a minimum by 24 months. The detected differences in the binding of SRC by the nuclei of cells of animals of different ages, in their opinion, may lie at the basis of changes in the functioning of the organism under the influence of hormones at different stages of ontogenesis.

Authors:
; ; ;
Publication Date:
Research Org.:
Sector of Gerontology, Minsk, USSR
OSTI Identifier:
5629373
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemistry (Engl. Transl.); (United States); Journal Volume: 51:5; Other Information: Translated from Biokhimiya; 51: 5, 775-781(May 1986)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; ESTROGENS; RECEPTORS; LIVER CELLS; CELL NUCLEI; ONTOGENESIS; RADIORECEPTOR ASSAY; PROGESTERONE; STEROIDS; AGE DEPENDENCE; BINDING ENERGY; CHROMATOGRAPHY; COMPLEXES; LABELLING; LIVER; RADIOACTIVITY; RATS; SCINTILLATION COUNTING; TRITIUM COMPOUNDS; ANIMAL CELLS; ANIMALS; BODY; CELL CONSTITUENTS; COUNTING TECHNIQUES; DIGESTIVE SYSTEM; ENERGY; GLANDS; HORMONES; ISOTOPE APPLICATIONS; KETONES; LABELLED COMPOUNDS; MAMMALS; MEMBRANE PROTEINS; ORGANIC COMPOUNDS; ORGANS; PREGNANES; PROTEINS; RODENTS; SEPARATION PROCESSES; SOMATIC CELLS; STEROID HORMONES; TRACER TECHNIQUES; VERTEBRATES; 550301* - Cytology- Tracer Techniques; 550201 - Biochemistry- Tracer Techniques

Citation Formats

Konoplya, E.F., Luksha, G.L., Savateev, S.K., and Naumov, A.D. Estrogen- and progestin-receptor complexes and their interaction with rat liver cell nuclei during ontogenesis. United States: N. p., 1986. Web.
Konoplya, E.F., Luksha, G.L., Savateev, S.K., & Naumov, A.D. Estrogen- and progestin-receptor complexes and their interaction with rat liver cell nuclei during ontogenesis. United States.
Konoplya, E.F., Luksha, G.L., Savateev, S.K., and Naumov, A.D. Mon . "Estrogen- and progestin-receptor complexes and their interaction with rat liver cell nuclei during ontogenesis". United States. doi:.
@article{osti_5629373,
title = {Estrogen- and progestin-receptor complexes and their interaction with rat liver cell nuclei during ontogenesis},
author = {Konoplya, E.F. and Luksha, G.L. and Savateev, S.K. and Naumov, A.D.},
abstractNote = {Steroid-receptor complexes (SRC) of estrogens and progestins have been isolated from the rat liver and purified 1500-2000-fold. Both in the state of the initial cytosol and purified 2000-fold, the SRC were characterized by gel filtration on Sephadex G-100 and by DEAE-cellulose chromatography. The purified SRC from the rat liver were used for binding to isolated liver cell nuclei from rats of different ages (1.5 months, 6 months, 12 months, 24 months). The maximum binding of SRC of progestins and estrogens from the rat liver is observed with homologous nuclei of 1.5-month-old rats. With age the binding of SRC by the nuclei decreases progressively and reaches a minimum by 24 months. The detected differences in the binding of SRC by the nuclei of cells of animals of different ages, in their opinion, may lie at the basis of changes in the functioning of the organism under the influence of hormones at different stages of ontogenesis.},
doi = {},
journal = {Biochemistry (Engl. Transl.); (United States)},
number = ,
volume = 51:5,
place = {United States},
year = {Mon Nov 10 00:00:00 EST 1986},
month = {Mon Nov 10 00:00:00 EST 1986}
}
  • The accumulation of (/sup 3/H)estradiol-receptor complexes by liver nuclei after preliminary incubation of the hormone with rat liver cytosol was investigated. It was shown that the addition of a purified preparation of the unusual estrogen-binding protein (UEBP) isolated from the liver of males to the cytosol of females induces a dose-dependent inhibition of the subsequent accumulation of specifically bound (/sup 3/H)estradiol in the nuclei. The addition of 1.5 ..mu..M 2..cap alpha..-hydroxytestosterone, testosterone, 1-dehydrotestosterone, 5..cap alpha..-androstane-3..cap alpha..,17..beta..-diol, and 5..cap alpha..-dihydrotestosterone - a compound possessing pronounced affinity for the UEBP - to the liver cytosol of males leads to a 2-5-fold increasemore » in subsequent nuclear accumulation of the estrogen-receptor complexes. Androsterone, which does not interact with the UEBP, does not exhibit such an effect. When liver cytosol of females, lacking the UEBP, is used, the indicated steroids are ineffective with respect to estrogen reception. It was concluded that the UEBP of the liver of male rats is capable of modulating estrogen reception.« less
  • Studies on structurally related aromatic amines with different carcinogenic properties have shown that 2-acetylaminofluorene (2-AAF) and 2-acetylaminophenanthrene (AAP) inhibit the binding of 2,3,7,8-tetrachlorodibenzo-p-dioxin to the Ah receptor in vitro. The apparent inhibitor constants (K{sub i}) are 2.3 {mu}M for 2-AAF and 2.7 {mu}M for AAP. In contrast, 4-acetylaminofluorene, an isomer of 2-AAF, and trans-4-acetylaminostilbene do not bind to the rat hepatic cytosolic Ah receptor. Pretreating female Wistar rats with 2-AAF or AAP leads to the induction of the P-450 isoenzymes that are under the control of the Ah receptor. Ornithine decarboxylase activity is induced by all aromatic amines tested irrespectivemore » of their Ah receptor affinity. The aromatic amines used as model compounds do not inhibit the binding of 17-{beta}-estradiol to the 8S and 4S estrogen receptor of rat uterus or rat liver in a competition assay analyzed using sucrose density gradient centrifugation. On the other hand, the aromatic amines bind to varying extents to another estrogen-binding protein of rat liver whose function and identity is still unknown. The study demonstrates that structurally related aromatic amines in their unmetabolized form interact differentially with a cellular target protein, the Ah receptor, in vitro as well as in vivo. However, a relationship between these effects and the postulated promoting properties of 2-AAF remains to be established.« less
  • Effects of antiestrogen on progestin binding in uterine cell types were determined and compared to those of estrogen. Effects on uterine morphology were also studied. Immature rats were treated with four daily sc injections of 100 micrograms hydroxytamoxifen (TAM(OH)), 5 micrograms estradiol (E2), or oil. On day 5 the rats were injected iv with 1 microgram of the synthetic progestin (/sup 3/H)Org 2058, and 1 h later uteri were excised, weighed, and processed for thaw-mount autoradiography. Treatment with TAM(OH) or E2 resulted in uterine weight gain, which was greater in animals treated with E2. E2 treatment resulted in cellular hypertrophymore » in all tissue compartments, especially in the luminal epithelium and myometrium, but TAM(OH) treatment resulted in hypertrophy of only the luminal epithelium. Treatment with TAM(OH) or E2 changed the pattern and intensity of nuclear binding of (/sup 3/H)Org 2058 from that in oil-treated controls. E2 increased progestin binding in stroma and myometrium and decreased it in luminal epithelium. TAM(OH), similarly, decreased progestin binding in the luminal epithelium and increased it, albeit less than E2, in the myometrium, but left it unchanged in the stroma. The results indicate that E2 and TAM(OH) differentially effect progestin binding among the uterine tissue compartments.« less
  • The effects of estrogen on osteoclast survival and differentiation were studied using CD14-selected mononuclear osteoclast precursors from peripheral blood. Estradiol at {approx} 1 nM reduced RANKL-dependent osteoclast differentiation by 40-50%. Osteoclast differentiation was suppressed 14 days after addition of RANKL even when estradiol was withdrawn after 18 h. In CD14+ cells apoptosis was rare and was not augmented by RANKL or by 17-{beta}-estradiol. Estrogen receptor-{alpha} (ER{alpha}) expression was strongly down-regulated by RANKL, whether or not estradiol was present. Mature human osteoclasts thus cannot respond to estrogen via ER{alpha}. However, ER{alpha} was present in CD14+ osteoclast progenitors, and a scaffolding protein,more » BCAR1, which binds ER{alpha} in the presence of estrogen, was abundant. Immunoprecipitation showed rapid ({approx} 5 min) estrogen-dependent formation of ER{alpha}-BCAR1 complexes, which were increased by RANKL co-treatment. The RANKL-signaling intermediate Traf6, which regulates NF-{kappa}B activity, precipitated with this complex. Reduction of NF-{kappa}B nuclear localization occurred within 30 min of RANKL stimulation, and estradiol inhibited the phosphorylation of I{kappa}B in response to RANKL. Inhibition by estradiol was abolished by siRNA knockdown of BCAR1. We conclude that estrogen directly, but only partially, curtails human osteoclast formation. This effect requires BCAR1 and involves a non-genomic interaction with ER{alpha}.« less
  • The present study examined the number and distribution of progestin receptor cells in the 8-day-old male and female cortex and compared cortical labeling with that in the preoptic area and central hypothalamus. Eight-day postnatal mice (four males and four females), treated with estradiol, were each sc injected with 0.32 micrograms/100 g BW (125I)progestin. Brains were frozen 2 h after injection of (125I)progestin, sectioned, and processed for thaw-mount autoradiography. Cells with a nuclear concentration of radioactivity were localized in lamina VI of the lateral cortical regions of the male and female brain, while only a few cortical cells were seen inmore » laminae II, III, and V of the suprarhinal, lateral, and cingulate/paracingulate regions. Comparison of the number of labeled cells revealed that the female cortex contained significantly more labeled cells than the male at three of the four levels investigated. Similarly, the number of target cells was higher in the female medial preoptic nucleus, but not in the arcuate nucleus and ventromedial hypothalamic nucleus, while the distributions of labeled cells in the male and female preoptic/hypothalamic regions were comparable. Injection of unlabeled progesterone or R5020 1 h before (125I)progestin reduced the nuclear concentration of radioactivity in all target regions and verified the specificity of (125I)progestin for the progestin receptor. The results of these studies indicate that mouse 8-day-old cortex and preoptic area in the female animal have more progestin receptor cells than those in the male and demonstrate that progestin receptor cells are localized in a region of the cortex known to contain few estrogen target cells. These results further suggest that a sexual dimorphism in progestin cell number may result in a differential effect of progestin on the cortex and preoptic area of the mouse.« less