Oestrogen Receptors in Chick Oviduct

Best‐Belpomme, Martin; Mešter, Ján; Weintraub, Hadassa; Baulieu, Étienne-Émile

doi:10.1111/j.1432-1033.1975.tb02329.x

Cited by 59 publications

(15 citation statements)

References 33 publications

(11 reference statements)

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“…The results obtained with these exchange techniques are in reasonable agreement with published data on oestrogen receptor site concentrations in chick oviduct cells [7,. Studies on the ontogeny of oestrogen receptor concentrations in the chiclk Mullerian duct have demonstrated the presence of 590 fmol of specifically bound oestradiol/mg DNA (810 sites/cell) in oviduct nuclei immediately prior to birth, and 22.50 fmol of cytoplasmic receptor/mg DNA (3100 sites/cell) in 4-day-old chickens [15,16].…”

Section: Discussionsupporting

confidence: 81%

Quantitative Estimates of Cytoplasmic and Nuclear Oestrogen Receptors in Chick Oviduct

Sutherland

Baulieu

1976

European Journal of Biochemistry

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[3H]Oestradiol exchange techniques were developed for the determination of specific oestrogen receptor site concentrations in the cytoplasm and nuclei of chick oviduct cells. Non-labelled, receptor-bound oestrogens were exchanged with [3H]oestradiol during a 24-h incubation at 20 'C, 2 h at 30 "C or 45 min at 37 "C. Both 'soluble' and 'insoluble' nuclear receptors were stable for at least 6 h at 30 "C and 37 "C but a proportion (approx. 30%) of cytoplasmic sites from withdrawn chickens were inactivated after 2 h at 20 "C.The magnum of 4-week-old immature chickens (weight = 15 mg) contained 0.20 pmol of oestrogen receptor which corresponds to 4275 receptor sites/cell, when it is assumed that all magnum cells have equal concentrations of receptor. In primarily stimulated chickens of similar age which had received 10 x 1 mg of oestradiol benzoate/day, the magnum weighed approximately 800 mg and contained 8.65 pmol of oestrogen receptor (4610 sites/cell). Withdrawal from primary oestrogenic stimulation for 3 -6 weeks resulted in a 110 mg magnum which contained 1.20 pmol of receptor (2225 sites/cell).Oviducts from immature and withdrawn chickens had the majority (73 -77 %) of their oestrogen receptor sites in the cytoplasmic fraction, while in primary stimulated chicken oviducts the majority (82%) of receptor sites were located in the nucleus.A single secondary injection of oestradiol, to oestrogen-withdrawn chickens, resulted in apparent translocation of cytoplasmic receptors to the nucleus during the first hour after injection. The magnitude of the decline in cytoplasmic receptor, and the concurrent increase in nuclear receptor concentration, was dose-dependent between 2 and 100 yg oestradiol/kg body weight. Larger doses of oestradiol up to 1 mg/kg did not increase the concentration of nuclear receptor above the maximum level seen at 100 pg oestradiol/kg. The initial rapid accumulation of nuclear receptor sites was followed by a period of progressive decline. By 15 h after an injection of 100 pg oestradiol/kg, the concentration of nuclear sites had reached pre-injection levels. During the same time period, the depleted cytoplasmic receptor levels were replenished such that they reached control values by 12 h and were about 150% of the pre-injection level at 24 h.The administration of oestrogenic compounds to the immature chicken induces growth, cytodifferentiation and the onset of egg-white protein synthesis in the magnum region of the oviduct [l]. Subsequent withdrawal from oestrogenic treatment results in cessation of protein synthesis and a quiescent, but partially differentiated oviduct which is capable of responding to secondary oestrogenic stimulation by the rapid onset of egg-white protein synthesis [2,3]. The response of the chick magnum to secondary oestrogenic stimulation offers a useful model for monitoring the early steps in oestrogenic hormone action, and may be useful in evaluating the role of changes in the concentration and subcellular distribution of specific oestrogen receptor molecules, in the ex...

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Section: Discussionsupporting

confidence: 81%

Quantitative Estimates of Cytoplasmic and Nuclear Oestrogen Receptors in Chick Oviduct

Sutherland

Baulieu

1976

European Journal of Biochemistry

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“…animal hormone receptors (15), but contrasts with others for which dissociation is relatively slow and strongly temperaturedependent (1,5,6,10 (Fig. 4) With the improved binding assay procedure that was used here, the concentration of specific NAA-binding sites, indicated by the point at which the line in a Scatchard plot intersects the abscissa, varied from about 30 to 50 pmol/g of tissue, fresh weight, in a number of different experiments (e.g.…”

Section: Resultsmentioning

confidence: 99%

Characterization of Naphthaleneacetic Acid Binding to Receptor Sites on Cellular Membranes of Maize Coleoptile Tissue

1977

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“…Nuclei from the oviducts of egg-laying hens were prepared according to Best-Belpomme et al (9). Estrogen receptor was extracted from purified nuclei with 500 mM KCl/50 mM Tris HCl, pH 8.7/10 mM EDTA/2 mM 2-mercaptoethanol/1 mM phenylmethylsulfonyl fluoride.…”

Section: Methodsmentioning

confidence: 99%

“…A second chromatographic step on a hydroxylapatite column was necessary to improve the DNA binding capacity of the estrogen-receptor complex. In some experiments estrogen-receptor was first "stripped" of its endogenous estrogen by a 10-min incubation at 370C with 5% charcoal/0.5% dextran (9).…”

Section: Methodsmentioning

confidence: 99%

Preferential binding of estrogen-receptor complex to a region containing the estrogen-dependent hypomethylation site preceding the chicken vitellogenin II gene.

Jost¹,

Seldran²,

Geiser³

1984

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

147

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DNA-cellulose competition binding assays were used to measure the ability of cloned DNA fragments of the chicken vitellogenin H gene to displace the estrogen-receptor complex from total chicken DNA coupled to cellulose. The DNA fragment that gave the highest competition is situated in the upstream region of the gene between nucleotides -458 and -725. This DNA fragment has four small clusters of A+T-rich sequences and contains the estrogen-dependent hypomethylation site. In vitro methylation of the Msp I site does not change the capacity of the DNA fragment to compete for estrogenreceptor complex, whereas cleavage of the C-C-G-G (Msp I site) results in a complete loss of competition of this fragment for estrogen-receptor complex. These results, combined with deoxyribonuclease I protection experiments, suggest that the most probable binding site for estrogen-receptor complex is.. This sequence has 73% homology with the core enhancer sequence of simian virus 40, .. .One mechanism in the regulation of specific gene expression by steroid hormones is the binding of the steroid receptor complex to the chromatin (1, 2). Given the large number of receptor molecules translocated to the nucleus and the relatively small number of genes that are stimulated, Yamamoto and Alberts (3, 4) proposed a two-site model of DNA binding in which the selective action of the steroid-receptor complex is achieved through interaction of a small number of highaffinity binding sites and a large number of low-affinity sites. This binding of steroid receptor to high-affinity binding sites could be, for example, responsible for the hypomethylation of a Msp I site situated at -611 base pairs (bp) in the upstream region of the vitellogenin II gene (5-7). Similarly, an interaction of the steroid-receptor complex with the DNA in this region of the gene may explain the presence of a deoxyribonuclease I-hypersensitive site situated approximately 700 bp upstream from the gene (8). The appearance of deoxyribonuclease I (DNase I) hypersensitivity at -700 bp and the hypomethylation of the Msp I site take place only in the presence of estrogen and are both observed in the estrogentreated liver and the oviduct. The common denominator of these two phenomena in liver and oviduct is the requirement of estrogen and therefore of nuclear receptor complex by which the effect of estrogen is expressed. We therefore formulate the working hypothesis that the steroid-receptor complex binds to the 5' upstream region of the gene where both hypomethylation and DNase I hypersensitivity are observed. We have tested this hypothesis and demonstrate that nuclear estrogen-receptor complex has a preferential binding to a DNA fragment situated upstream from the 5' end of the gene containing the Msp I hypomethylation site.The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact. MATERIALS AND METHODSPreparation of Nuclear R...

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Oestrogen Receptors in Chick Oviduct

Cited by 59 publications

References 33 publications

Quantitative Estimates of Cytoplasmic and Nuclear Oestrogen Receptors in Chick Oviduct

Quantitative Estimates of Cytoplasmic and Nuclear Oestrogen Receptors in Chick Oviduct

Characterization of Naphthaleneacetic Acid Binding to Receptor Sites on Cellular Membranes of Maize Coleoptile Tissue

Preferential binding of estrogen-receptor complex to a region containing the estrogen-dependent hypomethylation site preceding the chicken vitellogenin II gene.

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