Receptor-Estrogen Complex in the Nuclear Fraction of Rat Uterine Cells during the Estrous Cycle

Clark, James H.; Anderson, J.C.; Peck, Ernest J.

doi:10.1126/science.176.4034.528

Cited by 91 publications

(18 citation statements)

References 27 publications

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“…That the direct nuclear exchange only measures about half ( 1 -1.3 pmol of RnE/mg of DNA) of the nuclear receptor sites actually present is consistent with Mester and Baulieu (1975) but at variance with Clark et al (1972) who showed that RnE in adult cycling rats was 2-3 pmol/mg of DNA after injection of I O yg of E. This discrepancy might be due, however, to the difference in preparation of uteri. We utilized frozen uteri from adrenalectomized-ovariectomized rats, whereas Clark et al ( I 972) used fresh uteri from adult cycling rats.…”

Section: Discussionsupporting

confidence: 79%

Nuclear estradiol receptor in the adult rat uterus: a new exchange assay

Zava¹,

Harrington²,

McGuire³

1976

Biochemistry

109

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A protamine exchange assay has been developed to measure uterine nuclear estrogen receptor in mature rats exposed to estradiol (E). After ovariectomized-adrenalectomized mature rats are injected with E, estrogen receptor (RnE) is extracted from uterine nuclei with 0.6 M potassium chloride, diluted, and quantitatively precipitated with protamine sulfate. The precipitate is subjected to a ligand exchange with radiolabeled estradiol (E), with or without unlabeled diethylstilbesterol, to determine nonspecific binding. At 37 degrees C complete exchange of E for E in RnE is observed at 2.5 h; virtually no receptor degradation occurs up to at least 5 h. Exchange does not occur at 4 degrees C. Using the protamine assay, the depletion of cytoplasmic estrogen receptor (Rc) and the accumulation of RnE were studied at various doses of E at specific time points. Increasing doses of E result in a decrease of Rc with an equal increase of RnE. At the highest dose of E (10 mug) Rc is completely depleted within 10 min, by 6 h it is 25% replenished, and by 24 h returns to slightly above control levels. Within 10 min after the injection, RnE increases to 80-90% of the original cytoplasmic level of receptor (approximately 2-3 pmol/mg of DNA or approximately 1.5 pmol/100 mg of uterus). At 6 h RnE is 75% depleted and it is completely absent at 24 h. The protamine assay permits precise quantitative studies of nuclear estrogen receptor and avoids the problems of receptor degradation and excessive nonspecific binding often found in exchange reactions at elevated temperatures.

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Section: Discussionsupporting

confidence: 79%

Nuclear estradiol receptor in the adult rat uterus: a new exchange assay

Zava¹,

Harrington²,

McGuire³

1976

Biochemistry

109

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“…Erikkson [57] has shown that rat uterine "cytoplasmic" estrogen receptor levels increase during the first 5 days after OVX in adult rats and then gradually decline by 74% by 14 days after OVX. However, the levels reached during this period remain within the range observed during the rat estrous cycle [58] and are above the level necessary for a physiological response to E 2 in target tissues [59]. Similar uterine estrogen receptor levels are detected in adult mice after OVX, but OVX + ADX further reduces estrogen receptor levels [55,56].…”

Section: Discussionmentioning

confidence: 75%

Ovarian and Adrenal Contributions to Postnatal Growth and Differentiation of the Rat Uterus

Branham

Sheehan

1995

Biology of Reproduction

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We tested the hypothesis that ovarian and/or adrenal factors contribute to uterine growth, differentiation, and acquisition of estrogen responsiveness in the postnatal rat. In untreated rats, normalized uterine weight (5.2 mg/10 g BW on postnatal days [PND] 1-10) increased by about 35% on PND 11-19; PND 6 ovariectomy (OVX) eliminated this increase. Adrenalectomy (ADX) on PND 6 lowered normalized uterine weight only when combined with OVX and only on PND 16 and 19, demonstrating the presence of uterotropic adrenal products. OVX +/- ADX on PND 6 delayed uterine gland genesis by about 2 days but did not alter final gland numbers. There was no change in the normal pattern of luminal epithelium morphology. A uterotropic response to 17 beta-estradiol (E2) occurred on PND 10 in OVX rats and on PND 12 in OVX + ADX rats, but not until PND 14 in controls. We conclude that normal uterine growth is independent of the ovaries and adrenals prior to PND 10, partially dependent during PND 10-15, and completely dependent during PND 16-26. Additionally, a uterotropic response to exogenous E2 occurs concomitantly with, but independently of, the endogenous estrogen surge. Finally, while uterine gland genesis is slightly retarded by OVS +/- ADX, estrogens from these organs do not induce uterine differentiation.

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“…Oestrogen receptors appear in the hypothalamus after birth and increase in number between 14 and 26 days (Kato, Atsumi & Inaba, 1971, 1974Kulin & Reiter, 1972;Plapinger & McEwen, 1973). Rat uterine oestradiol binding capacity varies during the oestrous cycle, being highest at pro-oestrus in both the cytoplasm (Shain & Barnea, 1971) and nucleus (Clark, Anderson & Peck, 1972). Oestradiol binding in the rat corpus luteum varies during pregnancy, reaching a maximum around day 18 (Richards, 1974).…”

Section: Introductionmentioning

confidence: 96%

Cytoplasmic Oestrogen Receptor Complex of Female Ovine Pituitary: Changes Associated With the Reproductive State and Oestradiol Treatment

Wise

Payne

Karsch

et al. 1975

Journal of Endocrinology

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Binding of oestrone and oestradiol to a pituitary cytosol receptor was studied in ovariectomized sheep during the breeding season, during anoestrus and after oestradiol treatment during anoestrus. All sheep were ovariectomized 2 weeks before removal of the anterior pituitary. Oestrogen treatment consisted of the subcutaneous implantation of Silastic capsules containing oestradiol-17beta 3 weeks before ovariectomy. Association constants (KA) for oestrone and oestradiol and the concentration of binding sites for oestradiol varied with changes in reproductive state. Oestradiol bound more tightly and the concentration of available binding sites was lower during the breeding season than during anoestrus. The affinity of oestrone binding was less during the breeding season than during anoestrus, whilst the concentration of binding sites for this steroid did not change with changes in reproductive state. Treatment of anoestrous sheeep with oestradiol increased the KA for oestradiol and decreased the concentration of binding sites. However, no effect of oestradiol treatment was observed with regard to the KA or concentration of binding sites for oestrone. The results suggest that either the pituitary oestrogen cytoplasmic receptor undergoes a conformational change during the breeding season causing oestradiol to be more tightly bound or the onset of the breeding season is accompanied by the appearance of a new receptor which binds to oestradiol with greater affinity. Oestradiol treatment of anoestrous sheep mimics this change.

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Receptor-Estrogen Complex in the Nuclear Fraction of Rat Uterine Cells during the Estrous Cycle

Cited by 91 publications

References 27 publications

Nuclear estradiol receptor in the adult rat uterus: a new exchange assay

Nuclear estradiol receptor in the adult rat uterus: a new exchange assay

Ovarian and Adrenal Contributions to Postnatal Growth and Differentiation of the Rat Uterus

Cytoplasmic Oestrogen Receptor Complex of Female Ovine Pituitary: Changes Associated With the Reproductive State and Oestradiol Treatment

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