Estrone Interaction with the Rat Uterus: In Vitro Response and Nuclear Uptake1

Ruh, Thomas S.; Katzenellenbogen, Benita S.; Katzenellenbogen, John A.; Gorski, Jack

doi:10.1210/endo-92-1-125

Cited by 100 publications

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“…Although E| can be converted into E2, the interconversion between these two estrogens strongly favors the oxidative direction, as has been shown previously [6] and has also been reported in human endo metrium [24. 27] and rat uterus [22], Significant concentrations of unconju gated 3H-estrogens were also observed in other fetal organs, which could be related to the presence of estrogen receptors in these tis sues [18]: unmetabolized 3H-E| is predomi nant in fetal lung, brain and ovary, while important quantities of 3H-E| arc converted into 3H-E2 in the fetal kidney, which is in agreement with previous studies in this labo ratory [17],…”

Section: Discussionsupporting

confidence: 90%

“…The loss of cytosol estrogen receptors could be due to the fact that E| could translocate estrogen receptors into the nucleus, as has been dem onstrated in previous in vitro studies in fetal uterus [26] and in other systems [21,22]. Furthermore, the nuclear transfer of Epre ceptor complex is also indirectly suggested by the subcellular uterine compartmentalization o f 3H-estrogens after in vivo injection of 3H-E!…”

Section: Discussionmentioning

confidence: 75%

“…Moreover, it has been sug gested that the conversion of estradiol to estrone may constitute a release mechanism for estradiol turnover in the target cells [9,24], for instance in the human endometrium during the secretory phase of the menstrual cycle, when the higher steroid 17[3-dehydrogenase activity reduces the intracellular con centration of estradiol by increasing the over all conversion to estrone and consequently modulates estrogenic action [10], However, it is difficult to explain all the biological effects of estrone on the basis of these mechanisms since estrone itself is able to induce several estrogenic responses [22].…”

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confidence: 99%

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Uptake, Receptor and Biological Response of Estrone in the Fetal Uterus of Guinea Pig

Lanzone¹,

Nguyen²,

Pasqualini³

1983

Horm Res

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In vivo and in situ subcutaneous injection of 30 µCi of ³H-estrone (³H-E₁) to guinea pig fetuses was carried out to study the dynamics of the selective uptake and retention of this estrogen in the uterus and other fetal organs. In the fetal uterus an increase in the uptake of radioactivity with time was found which was maximal 1 h after injection of ³H-E₁. The analysis of radioactive material showed that in the fetal uterus unmetabolized ³H-E₁ accounts for 65–75% of the total radioactivity between 10 and 60 min after the injection and its maximal concentration is 2.5–15 times and 10 times higher than the maximal concentrations found in other fetal organs and fetal plasma, respectively. Significant amounts of ³H-E₁ were also present in lung (33–60% of the total radioactivity in this tissue), brain (37–52%) and ovary (18–47%), while an important conversion of ³H-E₁ into ³H-estradiol was observed in the kidney (20–26 %). Estrogen sulfates, essentially estrone sulfate, represented the greater part of the fetal plasma radioactivity (30–49%). ³H-E₁-specific binding sites in the cytosol of the fetal uterus were found with physicochemical characteristics (specifically limited to natural and synthetic estrogens, K_d = 8.1 ± 1.2 × 10^-10M(SD), sedimentation coefficient 8S) similar to those of the typical estrogen receptor. The cytosol receptor-³H-E₁ complex has a dissociation rate constant (K_-1) of 16 ± 0.7 × 10^-4 s^-1 at 25 °C. The responsiveness of the fetal uterus to estrone treatment was studied after subcutaneous administration of estrone to pregnant guinea pigs (1 mg/kg/day for 3 consecutive days). Two biological responses were observed: an increase in fetal uterine weight (50–100% in relation to the nontreated animals) and a significant stimulation of progesterone receptor protein (10–16 times higher than in control animals). At the same time, there was a 50–70% decrease in the total estrogen receptor concentration.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 75%

mentioning

confidence: 99%

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Uptake, Receptor and Biological Response of Estrone in the Fetal Uterus of Guinea Pig

Lanzone¹,

Nguyen²,

Pasqualini³

1983

Horm Res

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“…Inhibition of about 80% of the radioactivity bound to the 8S complex was observed in the presence of approximately 41 pmol of AFP per uterine equivalent (22 (19), and that the binding affinity for estradiol of the 8S complex is 10-to 20-fold higher than that for AFP (see Table 1). Binding specificity: competitive studies It has been previously reported that the affinity of the 8S macromolecular complex in uterine cytosol from 10-day-old rats is higher for estradiol than for estrone, whereas the reverse occurs with the 4-5S complex (13).…”

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confidence: 98%

Alpha-fetoprotein: the major high-affinity estrogen binder in rat uterine cytosols.

Uriel¹,

Bouillon²,

Aussel³

et al. 1976

Proc. Natl. Acad. Sci. U.S.A.

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Evidence is presented that alpha-fetoprotein (AFP), a serum globulin, accounts mainly, if not entirely, for the high estrogen-binding properties of uterine cytosols from immature rats. By the use of specific immunoadsorbents to AFP and by competitive assays with unlabeled steroids and pure AFP, it has been demonstrated that in hypotonic cytosols AFP is present partly as free protein with a sedimentation coefficient of about 4-5 S and partly in association with some intracellular constituent(s) to form an 8S estrogen-binding entity. The (7,8) and in adult animals bearing primary liver cancer (9). In immature rats, serum AFP ranges from about 1 mg/ml (21-day-old animals) to several hundred ,ug/ml (25-day-old animals).Numerous studies, reviewed recently (10, 11), have demonstrated the presence of high-affinity hormone binders, called "receptor" proteins, in soluble homogenates of estrogen-responsive tissues. Rat-uterine cytoplasmic extracts (cytosols) incubated with tritiated estrogens and subjected to density gradient centrifugation have revealed two major macromolecular complexes with sedimentation coefficients close to 4-5 S and 8-9 S; these are currently referred to as the 4-5S and the 8S "receptors." The 8S complex predomiAbbreviation: AFP, alpha-fetoprotein. nates in hypotonic solutions, whereas in salt concentrations above 0.2 M the 4S complex is by far the major binding entity.The relatively high levels of serum AFP in immature rats prompted us to explore the contribution of AFP to the estrogen-binding capacity of uterine homogenates. The results obtained with specific anti-AFP immunoadsorbents (12,13) provided evidence that at low salt concentrations,'AFP ac-' counts for most of the estrogen-binding capacity associated with the 4-5S macromolecular complex. It was concluded that the 4-5S entity and AFP were identical and that the presence of AFP in uterine cytosols was probably due to serum contamination.We report here further observations which strongly suggest that the other macromolecular complex, the 8S entity, is also made up of AFP in reversible association with some intracellular constituent(s). Immunologic data and competitive assays with unlabeled estrogens and anti-estrogens have also enabled us to demonstrate significant changes in antigenicity and binding specificity of native serum AFP upon its transition to the 8S form and the reversion to the original properties of native AFP after the 8S --4-5S transformation in hypertonic solutions.MATERIAL AND METHODS Animals. Female Wistar rats were used in all experiments. Amniotic fluids were obtained by puncturing, the amniotic sacs of animals on the 17th to 19th day of pregnancy. The fluids were pooled, treated with charcoal (2 mg/ml) for 20 min at 370 with mild agitation, centrifuged at 5000 rpm (4000 X g), and stored at -80°. Uteri

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“…IP synthesis, which is detectable by 40 min (1), is preceded by synthesis of RNA required for IP synthesis [detectable by 10 min (2) 1; this synthesis can be induced by physiological concentrations of estrogens in vitro (3)(4)(5)(6) and can be inhibited by some anti-estrogens (7). However, the function of IP is unknown and little has been reported about its physical properties.…”

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confidence: 99%

Uterine Estrogen-Induced Protein: Physical and Immunological Comparison with Ovalbumin

Katzenellenbogen

Williams

1974

Proc. Natl. Acad. Sci. U.S.A.

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The physical and immunological properties of the estrogen-induced protein from rat uterus are compared with those of ovalbumin from chick oviduct. Both proteins are induced by estrogen and are under estrogenic regulation. Electrophoretic mobility on sodium dodecyl sulfate-polyacrylamide gels and elution behavior on Sephadex G-100 column chromatography indicate that the uterine protein has a molecular weight of about 42,000, similar to that of ovalbumin, and likewise is composed of only one subunit. On isoelectric focusing in polyacrylamide gels, both proteins focus at a pH of 4.6.The uterine protein, in addition, shows a component with an isoelectric point of 5.1. Despite considerable similarity in the physical properties of the uterine protein and ovalbumin, there is no significant crossreactivity of antiovalbumin gamma globulin with the uterine protein, implying that these two estrogen-regulated proteins are immunologically distinct.The induction of the synthesis of a specific uterine protein called "induced protein" (IP) is the earliest known biosynthetic tissue response after estrogen binding. IP synthesis, which is detectable by 40 min (1), is preceded by synthesis of RNA required for IP synthesis [detectable by 10 min (2) 1; this synthesis can be induced by physiological concentrations of estrogens in vitro (3-6) and can be inhibited by some anti-estrogens (7). However, the function of IP is unknown and little has been reported about its physical properties.The similarity we observed in the migration of IP and ovalbumin in several gel electrophoretic and protein separation procedures was intriguing because ovalbumin, from chick oviduct, is also a protein whose synthesis is induced by estrogen (8,9). If IP and ovalbumin were closely related, it might be suggestive of the function of IP and have evolutionary implications. We, therefore, compared the physical and immunological properties of IP and ovalbumin.This report shows that IP and ovalbumin have similar electrophoretic properties on polyacrylamide gels and have nearly identical molecular weights and similar isoelectric points as judged by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, Sephadex column chromatography, and isoelectric focusing. However, despite considerable similarity in their physical properties, the two proteins show no significant immunological crossreactivity, as monitored by immunoprecipitation techniques. (5). MATERIALS AND METHODS IncubationPolyacrylamide Gel Electrophoresis. Uterine supernatant fractions were separated on 6% polyacrylamide gels (9 X 0.7 cm) that were prepared and run in TBE buffer (66 mM Tris-20 mM boric acid-3 mM Na2 EDTA, pH 8.6 at 250) and processed as described (5). SDS-Polyacrylamide Gel Electrophoresis. Samples were run on 10% polyacrylamide-0.1% SDS, pH 7.2 gels (9 cm X 0.7 cm) by the method of Weber and Osborn (10). For preparation of the IP sample, gel slices are cut from TBEpolyacrylamide gels and mashed into small pieces. The proteins in the gel fragments from two (2.3 mm) gel ...

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Estrone Interaction with the Rat Uterus: In Vitro Response and Nuclear Uptake1

Cited by 100 publications

References 0 publications

Uptake, Receptor and Biological Response of Estrone in the Fetal Uterus of Guinea Pig

Uptake, Receptor and Biological Response of Estrone in the Fetal Uterus of Guinea Pig

Alpha-fetoprotein: the major high-affinity estrogen binder in rat uterine cytosols.

Uterine Estrogen-Induced Protein: Physical and Immunological Comparison with Ovalbumin

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