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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Although follicle-stimulating hormone (FSH) and estrogens are known to be the main physiological stimuli for the development of the ovarian follicle in mammals, their growth-promoting activity has not been clearly established "in vitro". Furthermore, experimental evidence indicates that FSH and estradiol can independently inhibit granulosa cell proliferation. The present study was aimed at examining the effect of sex steroids in combination with FSH, on DNA synthesis in rat granulosa cells cultured in completely defined medium. Estradiol and FSH, when added separately, produced a significant inhibition of [3H] thymidine incorporation. In contrast, a combination of a low dose of FSH (20 ng/ml) with estradiol (100 ng/ml) produced a shift in the period of maximal DNA synthesis from 96 to 48 h after plating. Dose response studies showed that estradiol effects were produced at physiological intraovarian concentrations (1-100 ng/ml), whereas the effects of FSH were biphasic, with high doses (200 ng/ml) being inhibitory. A similar biphasic dose response curve was observed with increasing concentrations of a cAMP derivative in the presence of maximally effective doses of either an aromatizable steroid (androstenedione), insulin or insulin-like growth factor I. Non-aromatizable androgens (5alpha-dihydrotestosterone, 5alpha-androstane 3alpha-17beta diol and androsterone) showed a potency comparable to that of estradiol. The effect of 5alpha-dihydrotestosterone was completely blocked by a specific antiandrogen (hydroxy-flutamide), indicating that it was mediated by the androgen receptor. The effects of estradiol and androgens were not additive. The interaction between estradiol and FSH was further amplified in the presence of a maximally effective dose of insulin. Data presented herein indicate that both estrogens and androgens are able to elicit a mitogenic response in purified granulosa cells, cultured in a completely defined medium, provided the cells are stimulated by a physiological dose of FSH. These results suggest that, during follicular development, the stimulus for granulosa cell proliferation is given by the concerted action of steroid and peptide hormones acting through different signalling pathways.
J Steroid Biochem Mol Biol 1997 May
PMID:Concerted stimulation of rat granulosa cell deoxyribonucleic acid synthesis by sex steroids and follicle-stimulating hormone. 936 94

Conversion of Candida albicans from yeast to mycelial growth is believed to be associated with the organism's virulence. We investigated the role of mammalian hormones in initiating this transformation. Three clinical isolates of Candida albicans were tested for their ability to produce germ tubes under various conditions. Controlled hormonal conditions were provided by stripping rabbit serum with activated charcoal. Steroid compounds under investigation were added back to the stripped serum and yeast were inoculated into the test materials. Microscopic counts of germinated versus ungerminated cells were used as an indicator of morphogenic transformation. The percent of yeast cells germinating was profoundly reduced in stripped compared to unstripped serum. The addition of 1 microM estradiol, cholesterol or testosterone only slightly increased levels of germination above that seen in controls. Estradiol at concentrations 100 times less, however, proved a strong inducer of germination. Cholesterol did not synergize germination when combined with estradiol and the alpha isomer of estradiol had almost no activity as an inducer of morphogenic change in Candida albicans. We conclude that beta estradiol was a morphogenic inducer in three clinical isolates of Candida albicans but only at concentrations typical in vivo.
Cell Mol Life Sci 1997 Sep
PMID:Candida albicans morphogenesis is influenced by estrogen. 936 71

Cardiovascular effects of estrogens and particularly that of estradiol involve protection of the heart against ischemia. These effects were believed to be mainly indirect, mediated via changes in the blood and blood vessels. In the present paper a direct action of estradiol on the heart is demonstrated. Estradiol stimulates (p < 0.001) the Na,K-ATPase activity of cardiac sarcolemmal membranes by stimulating in an allosteric manner, the activation of the enzyme by potassium. The latter activation involves also an increase in affinity to potassium of the potassium binding sites on the enzyme molecule, but remains without any effect on the capacity and KD value of specific ouabain binding to the Na,K-ATPase. Estradiol is also antagonizing the depression of Na,K-ATPase activity that may be caused by ischemia and it is stimulating (p < 0.01) the ouabain-sensitive uptake of 86Rb into the heart cells. Our results indicate, that in addition to the known indirect effects of estradiol on the heart, the hormone also stimulates the activity and improves the kinetics of interaction of cardiac sarcolemmal Na,K-ATPase with ATP as well as with Na+ and K+ ions. This direct action may also account for the cardioprotective effects of estradiol.
Mol Cell Biochem 1997 Nov
PMID:Estradiol modulates the sodium pump in the heart sarcolemma. 940 52

Studies suggest that the steroid, dehydroepiandrosterone (DHEA) can exert effects directly, in addition to its indirect role serving as a precursor for other steroids such as androgens and estrogens. Because DHEA is one of the most abundant adrenal steroids secreted in man, we investigated the functional activity of DHEA on the classic estrogen response element (ERE) in the presence of the estrogen receptor (ER) in transiently transfected cells. GT1-7 hypothalamic neuronal cells, devoid of the estrogen receptor, were transiently transfected with the estrogen receptor expression plasmid (HEGO) and the estrogen response element luciferase (ERELUC) reporter vector. As expected, a dose-response stimulation of luciferase activity was observed in cells treated with estradiol. Concentrations of estradiol from 10(-10)-10(-6) M resulted in a 136-195 percent increase in luciferase activity compared with control. A dose-response stimulation was also observed in the cells treated with DHEA. A maximum stimulation of 177 percent increase in luciferase activity compared with control was observed with DHEA at a concentration of 10(-5) M. Both the estradiol and DHEA stimulation of ERE luciferase activity was inhibited by the estrogen receptor antagonist, ICI 182,780. The aromatase inhibitor, formestane in combination with estradiol or DHEA had no effect on luciferase activity, suggesting that the effect of DHEA is independent of its conversion to estradiol. Estradiol levels, as measured by ELISA, were appropriately elevated in the estradiol-treated cells but were not significantly different from the control cells in the DHEA-treated cells. These studies suggest a functional in vitro role of DHEA in activating the ERE in the presence of the classic ER.
J Steroid Biochem Mol Biol 1997 Aug
PMID:Dehydroepiandrosterone stimulates the estrogen response element. 944 50

Estradiol (E2) and progesterone (P) play different roles in generating the preovulatory surge release of luteinizing hormone-releasing hormone (LH-RH) and luteinizing hormone (LH). Results of our previous studies suggest that at least some of these steroid-specific effects may be mediated by beta-endorphinergic neurons. However, it is also possible that E2 and P differentially regulate responsiveness to opioids by altering mu-opioid receptor gene expression. To test this hypothesis, we used quantitative in situ hybridization histochemistry (ISHH) to measure the effects of E2 and P on mu-opioid receptor mRNA levels in cells of the preoptic area (POA) and arcuate nucleus (Arc). We examined several groups of animals in the morning and afternoon on the day of LH surge release: (1) 1-week ovariectomized (OVX) rats with or without E2 treatment sacrificed between 09:00 and 09:30 h (48 h after E2 capsules inserted); (2) OVX with or without E2 treatment sacrificed between 15:30 and 16:00 h; and (3) OVX with both E2 and P treatment sacrificed between 15:30 and 16:00 h (approximately 54 h after E2 and 6 h after P administration). We found that E2 had no effect on morning or afternoon levels of mu-opioid receptor mRNA levels in either the POA or Arc. In contrast, P treatment increased afternoon levels of mu-opioid receptor mRNA in both regions. These findings indicate that differential effects of E2 and P on LH-RH release may be mediated by steroid-specific effects on mu-opioid receptor gene expression in neurons of the POA and/or Arc.
Brain Res Mol Brain Res 1997 Dec 01
PMID:Progesterone increases levels of mu-opioid receptor mRNA in the preoptic area and arcuate nucleus of ovariectomized, estradiol-treated female rats. 945 Jun 74

An anti-estradiol antibody with improved specificity is searched for by combining steroid analog binding studies, mutant antibodies obtained from a phage-display library and structural modeling. Three-dimensional models for the anti-estradiol antibody 57-2 were constructed by comparative model building. Estradiol and analogs were docked into the combining site and molecular dynamics simulation was used to further refine this area of the protein. Cross-reactivities measured against 36 steroid analogs were used to help in the docking process and to evaluate the models. The roles of a number of residues were assessed by characterization of cross-reactivity mutants obtained from a phage display library. The cross-reactivity data and the results observed for mutants are explained by the structural model, in which the estradiol D-ring inserts deeply into the binding site and interacts with the antibody through at least one specific hydrogen bond. The binding data strongly suggest that this hydrogen bond connects the estradiol 17-hydroxyl group with the side chain of Gln H35. As expected for the binding of a small aromatic molecule, the antibody binding site contains many aromatic residues, e.g. Trp H50, H95 and L96 and Tyr L32, L49 and Phe L91.
Mol Immunol
PMID:Structural analysis of an anti-estradiol antibody. 956 68

In previous papers we provided evidence for a glucocorticoid (GC) responsive site in a highly purified rat liver plasma membrane (PM) fraction, which has proved to be osmotically active, 'right side-out' vesicles, free of CBG, glucocorticoid receptors (GR) and ATP (J. Steroid Biochem. Molec. Biol. 42 (1992) 737-756 and 757-771). This site, now called 'GC importer', mediates active transmembrane transport of corticosterone (B). Pronounced specificity, including stereo- and enantiomeric specificity, of ligand-GC importer interaction was demonstrated by competition assays using 54 different steroidal hormones and molecules. Important structural prerequisites for ligands with high specificity for the GC importer are plane C21-steroid hormones with 1-ene and/or 4-ene or 5alpha-reduced configuration, and/or OH-group(s) at C11beta>C17alpha>C21. Unexpectedly, other preferred ligands are C17alpha-ethynyl steroids like estrogens with an OH- or OCH3-group at C3 (EE2, mestranol) as well as progestins with C3-OH and 4-ene configuration (ethynodiol). C21-steroids with 11alpha-OH, 11-keto, 16alpha-CH3, 16beta-CH3, 16alpha-OH or 5beta-reduced configuration are low specificity ligands. The importer even displays different specificity for enantiomers (levonorgestrel>L-norgestrel). Altogether, the GC importer preferentially recognizes active GC and natural progestins which act as GC-antagonist (e.g. prednisolone>11beta-cortisol = B > or = progestins). Synthetic GC-agonists (e.g. dexamethasone, betamethasone, triamcinolone), most synthetic progestins, biologically inactive GC (e.g. 11alpha-cortisol, prednisone, cortisone, 11-dehydro-B), mineralocorticoids (aldosterone), natural estrogens (e.g. E1, E2, E3), DES and vitamin D3 derivatives do not interact with the GC importer. Osmotic shrinkage experiments revealed that interaction of high as well as low specificity ligands with the GC importer comprises reversible binding and transport through the PM. The ligand specificity profile of the GC importer and the GR exhibit pronounced differences, suggesting that both GC recognizing sites are different proteins. Performing immunoblotting, using specific mono- and polyclonal antibodies directed against the intracellular rat GR, of the PM pretreated with the membrane protein solubilizing detergent CHAPSO, we found that specific steroid binding to the PM site is not due to contamination with GR. Colchicine, daunorubicine, quinine, reserpine, verapamil and vinblastine, representatives of lipophilic xenobiotics which are known to be transported out of cells by the glycoprotein P170, did not compete with B for uptake into PM-vesicles, indicating that the GC importer is not a member of the ABC/mdr superfamily. The GC importer seems to be an additional link in the chain of steroid signal transduction and may be functionally involved in the action of natural GC-agonists and GC-antagonists.
J Steroid Biochem Mol Biol 1998 Jan
PMID:Glucocorticoid-recognizing and -effector sites in rat liver plasma membrane. Kinetics of corticosterone uptake by isolated membrane vesicles. III. Specificity and stereospecificity. 956 12

This study was undertaken to characterize the relationship between changes in steroid production, cell cycle activity (ie, cell proliferation) and apoptosis in antral and mural bovine granulosa cells cultured in vitro. This was done to select conditions promoting optimal estradiol production by bovine granulosa cells cultured in completely defined conditions. In the first experiment, antral granulosa cells were cultured over the entire 4 days of the culture period in the presence of either 0, 2, or 10 ng/ml of FSH (chronic conditions) or were maintained under minimal FSH support (0.5 ng/ml FSH) for the first 3 days of culture and then were challenged over the fourth day of culture with either 0, 2, or 10 ng/ml FSH (challenged conditions). Compared with cells exposed to constant FSH levels (chronic conditions), the FSH-induced production of estradiol was higher (P < 0.006) and that of progesterone was lower (P < 0.02) over the last 24 h of culture, when antral granulosa cells were maintained under minimal FSH support during the first 3 days of culture (challenged conditions). In the second experiment, dynamics of estradiol and progesterone productions, conversion of [14C]androstenedione into subsequent steroid metabolites, DNA content, cell cycle activity, and apoptosis (as assessed by flow cytometry) of antral and mural granulosa cells over the first 3 days of culture under minimal FSH support and in response to a challenge with FSH during the last 24 h of culture were evaluated. Estradiol production as well as the conversion of androstenedione into testosterone and estradiol were greater (P < 0.01) in antral than in mural granulosa cells cultured under challenged conditions. A higher proportion of mural than antral granulosa cells were in the proliferative state at the end of culture (P < 0.03). This may be related to the decreased ability of mural cells to produce estradiol. FSH suppressed (P < 0.05) the spontaneous onset of apoptosis in both cell types. These results suggest that functional differences between these two cell compartments need to be considered in studying bovine granulosa cells in vitro. Because of their large (400 to 600%) FSH-induced estradiol production, antral granulosa cells cultured under challenged conditions provide a model that can be used to examine substances for their ability to alter estradiol production and apoptosis in bovine granulosa cells.
Mol Reprod Dev 1998 Jun
PMID:Steroid production, cell proliferation, and apoptosis in cultured bovine antral and mural granulosa cells: development of an in vitro model to study estradiol production. 959 May 33

We compared the effect of estradiol on activator protein-1 (AP-1) activity in estrogen receptor positive (ER alpha+) and estrogen receptor negative (ER alpha-) human breast cancer cell lines transiently transfected with the AP-1-responsive reporter plasmid AP-1-TK-CAT and an ER alpha expression vector. While estradiol increased AP-1 activity in the ER alpha+ cell lines MCF7, ZR75.1, and T47D, it decreased (MDA-MB231 and BT20 cells) or had no significant effect (MDA-MB435 cells) on AP-1-mediated transcription in ER alpha- cells. Estradiol also inhibited AP-1 activity in ER alpha-MDA-MB231 cells stably transfected with ER alpha and in which ER alpha levels are close to those found in MCF7. Use of ER alpha mutant expression vectors demonstrated that the DNA-binding domain of ER alpha was needed for stimulation or inhibition of AP-1 activity by estradiol but suggested that ER alpha binding to estrogen-responsive elements was not required for these effects. Changes in regulation paralleled quantitative and qualitative changes in protein binding to AP-1 sites, as demonstrated by gel shift assay: protein binding was greater and DNA/protein complexes migrated faster for ER alpha--than for ER alpha+ cells. In fact, by Northern blot, a high level of Fra-1 mRNA was found in BT20 and MDA-MB231 cells as compared with ER alpha+ cells, and MDA-MB435 cells showed an intermediary level of expression. The differential expression of Fra-1 in MCF7 and MDA-MB231 cells was confirmed at the protein level by supershift experiments. In addition, overexpression of Fra-1 in MCF7 cells decreased the positive effect of estradiol while inhibition of Fra-1 expression in MDA-MB231 cells, by transient transfection of the Fra-1 antisense expression vector, abolished the negative effect of the hormone. In conclusion, we demonstrated that ER alpha- breast cancer cell lines differ from ER+ cells by a high level of AP-1 DNA-binding activity due, at least in part, to high Fra-1 constitutive expression. High Fra-1 concentration is crucial for the negative regulation of AP-1 activity by estradiol and thus may take part in estradiol-induced inhibition of cell proliferation in ER alpha- breast cancer cells transfected with ER alpha expression construct.
Mol Endocrinol 1998 Jul
PMID:FRA-1 expression level modulates regulation of activator protein-1 activity by estradiol in breast cancer cells. 965 2

The nervous system is a target for sex steroid hormones which have profound actions on the growth, maturation, differentiation and functioning of brain cells. We found that some steroids, termed "neurosteroids", are synthesized within the brain by glial cells. The term "neurosteroids" designates their site of synthesis--the nervous system, either de novo from cholesterol or from steroid hormone precursors. The biological effects of steroid hormones are mediated by specific high-affinity intracellular receptors, which, after hormone binding, function as activated transcription factors. The presence of such receptors was shown in primary cultures of oligodendrocytes and astrocytes, derived from forebrains (CNS), and in Schwann cells, derived from sciatic nerves (PNS), of newborn rats. In glial cells of the CNS, progesterone-, glucocorticoid-, estrogen and androgen-receptors (PR, GR, ER, AR) were demonstrated and of these receptors, only PR was estrogen-inducible. In glial cells of the PNS, the presence of PR and ER was shown, but the PR in Schwann cell cultures was not inducible by estrogen treatment. Different effects of steroids on glial cell growth and differentiation during primary culture were observed. In particular, a striking increase of myelin-specific proteins such as myelin basic protein (MBP) and cyclic nucleotide phosphodiesterase (CNPase) was observed when oligodendrocytes, the myelinating glial cells of the CNS, were cultured in the presence of progesterone, as determined by indirect immunofluorescence staining and immunoblotting. Insulin also increases MBP and CNP-ase in oligodendrocytes and the combined treatment (insulin + progesterone) promotes a strong synergistic stimulation (14-fold increase) of myelin protein expression. Estradiol also increases MBP- and CNPase expression in oligodendrocytes, although to a lesser extent than progesterone. In the search for optimal stimulation of myelin-protein expression, several progesterone analogues were tested and the results are discussed.
J Steroid Biochem Mol Biol 1998 Apr
PMID:Steroid hormone receptors and steroid action in rat glial cells of the central and peripheral nervous system. 969 79


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