Antibodies to estrogen receptor: Immunochemical similarity of estrophilin from various mammalian species

Greene, Geoffrey L.; Closs, Liselotte E.; Fleming, H. P.; DeSombre, Eugene R.; Jensen, Elwood V.

doi:10.1073/pnas.74.9.3681

Cited by 130 publications

(52 citation statements)

References 17 publications

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“…Similar observations were made with antibodies against the rabbit uterine progesterone receptor [7]. These results contrast with the limited shift to the 10-14-S region induced by antibodies against calf uterine oestrogen receptor [4]. An attempt to quantitate the heavy immunoglobulin-receptor complexes showed that they increased with IgG-G concentration, reached a plateau and then decreased.…”

Section: Discussioncontrasting

confidence: 41%

Antibodies against Progesterone Receptor from Chick Oviduct. Cross-Reactivity with Mammalian Progesterone Receptors

et al. 1982

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The highly purified, molybdate-stabilized '8 -9-S' form of the chick oviduct progesterone receptor complexed with [3H]progesterone (Specific activity z 10-12 nmol bound steroid/mg protein) was prepared according to the three-step method Eur. J. Biochem. 127,71-791 and 50 -90 pg samples were injected into a goat and a rabbit. Specific antibodies were observed as early as 2 weeks following the first booster injection and increased considerably 12 weeks later after further booster injections. Results were similar in the two animals, with higher titers observed in the goat. Antibodies cross-reacted with the 'transformed' 4-S form of the chick oviduct progesterone receptor. They also cross-reacted with molybdate-stabilized 8 -9-S progesterone receptors of mammalian species (obtained from MCF-7 human breast cancer cells and from rabbit and mouse uteri), suggesting some evolutionary conservation of steroid hormone receptors. No cross-reaction was observed with chick oviduct oestradiol receptor and with chick plasma transcortin (a non-receptor progesterone-binding protein).The chick oviduct progesterone receptor has been extensively studied, but reports of its purification (review in [I]) have not been followed by successful immunological studies [2]. Anti-receptor antibodies have been obtained against partially purified oestradiol receptor from calf uterus [3, 41, rat liver glucocorticosteroid receptor [5, 61, and progesterone receptor from rabbit uterus [7]. Some of these antibodies have been used for receptor immuno-assay [8] and immuno-cytological studies [9 -1 I]. We have recently achieved the purification to apparent homogeneity of a molybdate-stabilized '8-9-S' form of the chick oviduct progesterone receptor [12-141 and have obtained, after injection of the receptor into a goat and a rabbit, antibodies which cross-react with the progesterone receptor from mammalian species. MATERIALS AND METHODS ReagentsTritiated progesterone, ([2,4,6,7-3H]progesterone, 96 Ci/ mmol), [3H]dexamethasone (1 10 Ci/mmol) and t3H]ORG 2058 (45 Ci/mmol) were from the Radiochemical Centre, Amersham (England). The synthetic radioactive progestin [3H]-R 5020 (87 Ci/mmol) was from New England Nuclear. Their purity was verified by thin-layer chromatography. Non-radioactive steroids were obtained from Roussel-Uclaf, Romainville, except ORG 2058 (a gift from Organon, Oss, The Netherlands).They were > 95 % pure.All chemicals were reagent grade and obtained from Merck (Darmstadt). Dialysis tubing was from Thomas Lab Co. Philadelphia, PA.

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Section: Discussioncontrasting

confidence: 41%

Antibodies against Progesterone Receptor from Chick Oviduct. Cross-Reactivity with Mammalian Progesterone Receptors

et al. 1982

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“…Sucrose gradient sedimentation with [ 3 H]estradiol as ligand confirmed the presence of ER␣ in the 2-week-old mouse VP. In this system, ER␤ sediments in the 4 S region (fractions 11-20), whereas ER␣ sediments as an 8 S peak (fractions 1-10) (16). A specific 8S [ 3 H]estradiol binding peak was present in extracts from 2-week-old VP as well as from adult uterus (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Sedimentation studies were carried out as described in ref. 16. Cytosols were incubated for 3 h at 0°C with 10 nM [ 3 H]estradiol-17␤ (Amersham Pharmacia) in the presence or absence of 50-fold excess of cold estradiol, and the unbound steroid was removed with dextran-coated charcoal.…”

Section: Preparation Of Cytosol For Sucrose Density Gradient Centrifumentioning

confidence: 99%

Estrogen receptor α and imprinting of the neonatal mouse ventral prostate by estrogen

Omoto

Imamov

Warner

et al. 2005

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

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Exposure to estrogen in the neonatal period affects prostatic growth and leads to an increased incidence of prostatic intraepithelial neoplasia in later life. The effects of neonatal estrogen are clearly dependent on estrogen receptor (ER) α because they do not occur in ERα-knockout mice. Because ERα is expressed in the stroma, but not in the epithelium, of the adult ventral prostate, the concept of indirect estrogen effects through stromal signaling has been proposed. Here, we show that during the first 4 weeks of life, there are profound and rapid changes in the ER profile in the mouse ventral prostate. ERα is abundant in the stroma during week 1, but by week 2 it is exclusively epithelial, and then by week 4, ERα is lost and ERβ is dominant in the prostatic epithelium. The presence of ERα is associated with a high proliferation index, and ERβ is associated with quiescence. Branching morphogenesis was altered in ERα -/- , but not in ERβ -/- , mice. We conclude that imprinting and branching morphogenesis of the ventral prostate are mediated by estrogen acting directly on epithelial and stromal ERα during the first 2 weeks of life.

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“…The sucrose gradient centrifugation experiments show that, in low-salt sucrose gradients, the native ER␤ present in cytosol from rat mammary glands sediments in the 4S region whereas ER␣ sediments as an 8S peak as expected (22). This has been observed in several tissues (prostate, lung, uterus) in which ER␣ and ER␤ are present (unpublished data).…”

Section: Discussionmentioning

confidence: 71%

“…Sedimentation studies were carried out as described previously (22). Breast tissue cytosols were incubated for 3 h at 0°C with 10 nM [ 3 H]estradiol-17␤ (Amersham) in the presence or absence of 50-fold excess of cold estradiol, and the unbound steroid was removed with dextrancoated charcoal.…”

Section: Preparation Of Cytosol For Sucrose Density Gradient Centrifumentioning

confidence: 99%

Estrogen receptors α and β in the rodent mammary gland

Saji

Jensen

Nilsson

et al. 2000

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

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323

155

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An obligatory role for estrogen in growth, development, and functions of the mammary gland is well established, but the roles of the two estrogen receptors remain unclear. With the use of specific antibodies, it was found that both estrogen receptors, ER␣ and ER␤, are expressed in the rat mammary gland but the presence and cellular distribution of the two receptors are distinct. In prepubertal rats, ER␣ was detected in 40% of the epithelial cell nuclei. This decreased to 30% at puberty and continued to decrease throughout pregnancy to a low of 5% at day 14. During lactation there was a large induction of ER␣ with up to 70% of the nuclei positive at day 21. Approximately 60 -70% of epithelial cells expressed ER␤ at all stages of breast development. Cells coexpressing ER␣ and ER␤ were rare during pregnancy, a proliferative phase, but they represented up to 60% of the epithelial cells during lactation, a postproliferative phase. Western blot analysis and sucrose gradient centrifugation confirmed this pattern of expression. During pregnancy, the proliferating cell nuclear antigen was not expressed in ER␣-positive cells but was observed in 3-7% of ER␤-containing cells. Because more than 90% of ER␤-bearing cells do not proliferate, and 55-70% of the dividing cells have neither ER␣ nor ER␤, it is clear that the presence of these receptors in epithelial cells is not a prerequisite for estrogen-mediated proliferation. E strogens are potent mitogens in the mammary gland, where they are obligatory for normal development as well as for induction and progression of mammary carcinoma (1). The female mammary gland undergoes a surge of cell division during puberty, and throughout adult life there is cyclical proliferation and involution during estrous cycles (2). Despite the clear role for estrogen in proliferation of the mammary epithelium, the role of estrogen receptors in this process remains unclear. During pubertal growth and during the estrous cycle, the majority of proliferating cells, both in terminal end buds and in ducts, do not contain estrogen receptor ␣ (ER␣). This has been observed in both rodent (3) and human (4, 5) mammary glands. Progesterone receptor (PR) is localized in those cells that contain ER␣, and induction of PR by estrogen occurs at much lower plasma levels of estrogen than are required for cell proliferation (5). These observations have led to the concept (6) of two distinct types of responses to estrogen in the breast: (i) an indirect action of estradiol in the mammary epithelium by inducing estrogen receptor-containing stromal cells to produce growth factors that stimulate epithelial cells to divide and (ii) a direct effect on ER␣-containing cells that occurs at low estrogen concentrations and results in induction of PR.Dependence of epithelium on stroma for estrogen-mediated induction of PR has been demonstrated in experiments in which epithelial and stromal components were cultured together (7-9). Stroma from responsive (mature) glands can confer estrogen sensitivity on epithelium from insensitiv...

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Antibodies to estrogen receptor: Immunochemical similarity of estrophilin from various mammalian species

Cited by 130 publications

References 17 publications

Antibodies against Progesterone Receptor from Chick Oviduct. Cross-Reactivity with Mammalian Progesterone Receptors

Antibodies against Progesterone Receptor from Chick Oviduct. Cross-Reactivity with Mammalian Progesterone Receptors

Estrogen receptor α and imprinting of the neonatal mouse ventral prostate by estrogen

Estrogen receptors α and β in the rodent mammary gland

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