scholarly journals 17β-Estradiol Stimulates Arachidonate Release from Human Amnion-Like WISH Cells through a Rapid Mechanism Involving a Membrane Receptor

Endocrinology ◽  
2003 ◽  
Vol 144 (8) ◽  
pp. 3359-3367 ◽  
Author(s):  
Sara Fiorini ◽  
Maria E. Ferretti ◽  
Carla Biondi ◽  
Barbara Pavan ◽  
Laura Lunghi ◽  
...  
Keyword(s):  
Cell Permeabilization ◽  
Human Amnion ◽  
Permeabilized Cells ◽  
Wistar Institute ◽  
N Terminus ◽  
Dependent Cell ◽  
17Β Estradiol ◽  
Arachidonate Release ◽  
Estradiol 17Β ◽  
Membrane Binding Sites

Abstract 17β-Estradiol (17β-E2) greatly and dose-dependently stimulates [3H]arachidonic acid (AA) release from the human amnion-like Wistar Institute Susan Hayflick (WISH) cells. This action is abolished by the phospholipase A2 inhibitor AACOCF3, significantly reduced by the estrogen receptor (ER) antagonist ICI 182,780, and uninfluenced by cycloheximide. The estradiol-BSA conjugate E2coBSA, which binds putative membrane ERs and is unable to enter the cell, also highly stimulates [3H]AA release from WISH cells, although to a lesser extent compared with 17β-E2. The fluorescent conjugate E2coBSA-FITC specifically binds to the surface of a subset of intact WISH cells, and labeling intensity appears dose and time dependent. Cell permeabilization results in a dense intracellular staining, mainly in the peripheral cytoplasm. H-150, an antibody against the N terminus of human ERβ, also labels the plasma membrane of intact WISH cells and the cytoplasm of permeabilized cells. Almost no labeling is observed using ER-21, an antibody against the N terminus of human ERα. RT-PCR evidences the presence of mRNA for ERβ, not for ERα. Our data suggest that 17β-E2 stimulates [3H]AA release from WISH cells through an apparently nongenomic pathway and interaction with membrane binding sites. These last are, at least in part, similar if not identical to ERβ.

scholarly journals A Basic Cluster in the N Terminus of Yellow Fever Virus NS2A Contributes to Infectious Particle Production

2015 ◽  
Vol 89 (9) ◽  
pp. 4951-4965 ◽  
Author(s):  
Stephanie Voßmann ◽  
Janett Wieseler ◽  
Romy Kerber ◽  
Beate Mareike Kümmerer
Keyword(s):  
Yellow Fever ◽  
Particle Production ◽  
Yellow Fever Virus ◽  
Fever Virus ◽  
Physical Interaction ◽  
Site Mutation ◽  
Virion Assembly ◽  
Permeabilized Cells ◽  
Basic Cluster ◽  
N Terminus

ABSTRACTThe flavivirus NS2A protein is involved in the assembly of infectious particles. To further understand its role in this process, a charged-to-alanine scanning analysis was performed on NS2A encoded by an infectious cDNA clone of yellow fever virus (YFV). Fifteen mutants containing single, double, or triple charged-to-alanine changes were tested. Five of them did not produce infectious particles, whereas efficient RNA replication was detectable for two of the five NS2A mutants (R22A-K23A-R24A and R99A-E100A-R101A mutants). Prolonged cultivation of transfected cells resulted in the recovery of pseudorevertants. Besides suppressor mutants in NS2A, a compensating second-site mutation in NS3 (D343G) arose for the NS2A R22A-K23A-R24A mutant. We found this NS3 mutation previously to be suppressive for the NS2Aα cleavage site Q189S mutant, also deficient in virion assembly. In this study, the subsequently suggested interaction between NS2A and NS3 was proven by coimmunoprecipitation analyses. Using selectively permeabilized cells, we could demonstrate that the regions encompassing R22A-K23A-R24A and Q189S in NS2A are localized to the cytoplasm, where NS3 is also known to reside. However, the defect in particle production observed for the NS2A R22A-K23A-R24A and Q189S mutants was not due to a defect in physical interaction between NS2A and NS3, as the NS2A mutations did not interrupt NS3 interaction. In fact, a region just upstream of R22-K23-R24 was mapped to be critical for NS2A-NS3 interaction. Taken together, these data support a complex interplay between YFV NS2A and NS3 in virion assembly and identify a basic cluster in the NS2A N terminus to be critical in this process.IMPORTANCEDespite an available vaccine, yellow fever remains endemic in tropical areas of South America and Africa. To control the disease, antiviral drugs are required, and an understanding of the determinants of virion assembly is central to their development. In this study, we identified a basic cluster of amino acids in the N terminus of YFV NS2A which inhibited virion assembly upon mutation. The defect was rescued by a spontaneously occurring mutation in NS3. Our study proves an interaction between NS2A and NS3, which, remarkably, was maintained for the NS2A mutant in the presence and absence of the NS3 mutation. This suggests a role for other viral and/or cellular proteins in virion assembly. Residues important for YFV virion production reported here only partially coincided with those reported for other flaviviruses, suggesting that the determinants for particle production are virus specific. Reconstruction of a YFV encoding tagged NS2A paves the way to identify further NS2A interaction partners.

scholarly journals 17α-Estradiol Modulates IGF1 and Hepatic Gene Expression in a Sex-Specific Manner

2020 ◽  
Author(s):  
Silvana Sidhom ◽  
Augusto Schneider ◽  
Yimin Fang ◽  
Samuel McFadden ◽  
Justin Darcy ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Insulin Sensitivity ◽  
Growth Hormone Receptor ◽  
Health Benefits ◽  
Male Mice ◽  
Wild Type ◽  
Somatotropic Axis ◽  
Gh Secretion ◽  
17Β Estradiol ◽  
Estradiol 17Β

Abstract Aging is the greatest risk factor for most chronic diseases. The somatotropic axis is one of the most conserved biological pathways that regulates aging across species. 17α-Estradiol (17α-E2), a diastereomer of 17β-estradiol (17β-E2), was recently found to elicit health benefits, including improved insulin sensitivity and extend longevity exclusively in male mice. Given that 17β-E2 is known to modulate somatotropic signaling in females through actions in the pituitary and liver, we hypothesized that 17α-E2 may be modulating the somatotropic axis in males, thereby contributing to health benefits. Herein, we demonstrate that 17α-E2 increases hepatic insulin-like growth factor 1 (IGF1) production in male mice without inducing any changes in pulsatile growth hormone (GH) secretion. Using growth hormone receptor knockout (GHRKO) mice, we subsequently determined that the induction of hepatic IGF1 by 17α-E2 is dependent upon GH signaling in male mice, and that 17α-E2 elicits no effects on IGF1 production in female mice. We also determined that 17α-E2 failed to feminize the hepatic transcriptional profile in normal (N) male mice, as evidenced by a clear divergence between the sexes, regardless of treatment. Conversely, significant overlap in transcriptional profiles was observed between sexes in GHRKO mice, and this was unaffected by 17α-E2 treatment. Based on these findings, we propose that 17α-E2 acts as a pleiotropic pathway modulator in male mice by uncoupling IGF1 production from insulin sensitivity. In summary, 17α-E2 treatment upregulates IGF1 production in wild-type (and N) male mice in what appears to be a GH-dependent fashion, while no effects in female IGF1 production are observed following 17α-E2 treatment.

scholarly journals Modulating Effects of Cholecalciferol Treatment on Estrogen Deficiency-Induced Anxiety-Like Behavior of Adult Female Rats

Folia Medica ◽  
2017 ◽  
Vol 59 (2) ◽  
pp. 139-158 ◽  
Author(s):  
Julia Fedotova ◽  
Daria Zarembo ◽  
Jozef Dragasek ◽  
Martin Caprnda ◽  
Peter Kruzliak ◽  
...  
Keyword(s):  
Elevated Plus Maze ◽  
Open Field Test ◽  
Estrogen Deficiency ◽  
Female Rats ◽  
Ovx Rats ◽  
Plus Maze ◽  
17Β Estradiol ◽  
Experimental Rat Model ◽  
High Doses ◽  
Estradiol 17Β

AbstractBackground:Vitamin D can be one of the candidate substances that are used as additional supplementation in the treatment of anxiety-related disorders in women with estrogen imbalance.Materials and methods:The aim of the present study was to examine the effects of chronic cholecalciferol administration (1.0, 2.5 or 5.0 mg/kg/day, s.c.) on the anxiety-like behavior and monoamines levels in the rat hippocampus following ovariectomy in female rats. Cholecalciferol was given to ovariectomized (OVX) rats and OVX rats treated with 17β-estradiol (17β-E2, 0.5 μg/rat, s.c.). The anxiety-like behavior was assessed in the elevated plus maze (EPM) and the light-dark tests (LDT), locomotor and grooming activities were assessed in the open-field test (OFT).Results:Cholecalciferol in high doses alone or in combination with 17β-E2-induced anxiolytic-like effects in OVX and OVX rats treated with 17β-E2as evidenced in the EPM and LDT tests, and increased grooming activity in the OFT test. We found that DA and 5-HT levels increased while 5-HT turnover in the hippocampus decreased in these groups of OVX rats.Conclusion:Our results indicate that cholecalciferol in high doses has a marked anxiolytic-like effect due to an increase in the monoamines levels in the experimental rat model of estrogen deficiency.

scholarly journals 17β-Estradiol Modulates Prostaglandin E2 Release from Human Amnion-Derived WISH Cells1

2001 ◽  
Vol 64 (6) ◽  
pp. 1677-1681 ◽  
Author(s):  
Barbara Pavan ◽  
Carla Biondi ◽  
Maria Enrica Ferretti ◽  
Laura Lunghi ◽  
Guglielmo Paganetto
Keyword(s):  
Prostaglandin E2 ◽  
Human Amnion ◽  
17Β Estradiol

ATP-dependent transport of organic anions into isolated basolateral membrane vesicles from rat intestine

2004 ◽  
Vol 287 (4) ◽  
pp. G749-G756 ◽  
Author(s):  
Takahiro Shoji ◽  
Hiroshi Suzuki ◽  
Hiroyuki Kusuhara ◽  
Yuka Watanabe ◽  
Shingo Sakamoto ◽  
...  
Keyword(s):  
Rank Order ◽  
Basolateral Membrane ◽  
Membrane Vesicles ◽  
Organic Anions ◽  
Dependent Manner ◽  
Basolateral Membrane Vesicles ◽  
High K ◽  
17Β Estradiol ◽  
Dependent Transport ◽  
Estradiol 17Β

The mechanism for the cellular extrusion of organic anions across the intestinal basolateral membrane was examined using isolated membrane vesicles from rat jejunum, ileum, and colon. It was found that 17β-estradiol 17β-d-glucuronide (E217βG) is taken up in an ATP-dependent manner into the basolateral membrane vesicles (BLMVs) but not into the brush-border or microsomal counterparts. The ATP-dependent uptake of E217βG into BLMVs from jejunum and ileum was described by a single component with a Km value of 23.5 and 8.31 μM, respectively, whereas that into the BLMVs from colon was described by assuming the presence of high ( Km = 0.82 μM)- and low-affinity ( Km = 35.4 μM) components. Taurocholate, 6-hydroxy-5,7-dimethyl-2-methylamino-4-(3-pyridylmethyl) benzothiazole glucuronide and taurolithocholate sulfate, but not leukotriene C4, were significantly taken up by the BLMVs. In addition to such substrate specificity, the inhibitor sensitivity of the ATP-dependent transport in BLMVs was similar to that of rat multidrug resistance-associated protein 3 (Mrp3), which is located on the basolateral membrane of enterocytes. Together with the fact that the rank order of the extent of the expression of Mrp3 (jejunum < ileum << colon) is in parallel with that of the extent of the transport of ligands, these results suggest that the ATP-dependent uptake of organic anions into isolated intestinal BLMVs is at least partly mediated by Mrp3.

scholarly journals Macromolecular prodrugs. XIII. Hydrosoluble conjugates of 17β-estradiol and estradiol-17β-valerate with polyaspartamide polymer

2011 ◽  
Vol 61 (4) ◽  
pp. 465-472 ◽  
Author(s):  
Marijana Končič ◽  
Branka Zorc ◽  
Predrag Novak
Keyword(s):  
Drug Delivery ◽  
Water Solubility ◽  
The Other ◽  
Amino Groups ◽  
Chemically Modified ◽  
Drug Conjugates ◽  
17Β Estradiol ◽  
Macromolecular Prodrugs ◽  
Polymer Drug Conjugates ◽  
Estradiol 17Β

Macromolecular prodrugs. XIII. Hydrosoluble conjugates of 17β-estradiol and estradiol-17β-valerate with polyaspartamide polymerTwo hydrosoluble conjugates of 17β-estradiol (ED) and estradiol-17β-valerate (EV) with polyaspartamide polymer were prepared and characterized. ED and EV were first chemically modified and bound to poly[α,β-(N-2-hydroxyethyl-DL-aspartamide)]-poly[α,β-(N-2-aminoethyl-DL-aspartamide)] (PAHA), a hydrosoluble polyaspartamide-type copolymer bearing both hydroxyl and amino groups. ED was first converted to 17-hemisuccinate (EDS) and then bound to PAHA. In the resulting conjugate PAHA-EDS, the estradiol moiety was linked to the polymer through a 2-aminoethylhemisuccinamide spacer. On the other hand, EV was first converted to estradiol-17β-valerate-3-(benzotriazole-1-carboxylate), which readily reacted with amino groups in PAHA affording the polymer-drug conjugate PAHA-EV. In the prepared conjugate PAHA-EV, the estradiol moiety was covalently bound to the polyaspartamide backbone by carbamate linkage, through an ethylenediamine spacer. The polymer-drug conjugates were designed and prepared with the aim to increase water-solubility, bioavailability and to improve drug delivery of the lipophilic estrogen hormone.

Uptake of 17β-estradiol and its conjugates by isolated rabbit liver nuclei

1983 ◽  
Vol 61 (7) ◽  
pp. 784-789 ◽  
Author(s):  
J. H. Tong ◽  
E. I. Seper ◽  
D. S. Layne ◽  
D. G. Williamson
Keyword(s):  
Diffusion Process ◽  
Thin Layer ◽  
Thin Layer Chromatography ◽  
Substrate Concentration ◽  
Rabbit Liver ◽  
17Β Estradiol ◽  
Liver Nuclei ◽  
Layer Chromatography ◽  
Estradiol 17Β ◽  
Uptake And Metabolism

The present study compares the uptake and metabolism of 17β-estradiol, 17β-estradiol 3-glucoside, and 17β-estradiol 3-glucuronide by a highly purified preparation of rabbit liver nuclei. The uptake of the three estrogens was rapid and equilibration was reached within 60 s. The order of uptake was 17β-estradiol (64 fmol∙mg protein−1) > 17β-estradiol 3-glucoside (10 fmol∙mg protein−1) > 17β-estradiol 3-glucuronide (6.5 fmol∙mg protein−1). Thin-layer chromatography of the estrogens taken up by rabbit liver nuclei indicated the presence of a β-glucosidase activity associated with the nuclear preparation. The apparent Km value of this enzyme for 17β-estradiol 3-glucoside (3.5 μM) was about 10-fold higher when compared with the cytosolic enzyme. The uptake of the three estrogens was linearly proportional to the substrate concentration from 1 to 100 nM. No competition for uptake was observed among the steroids and the presence of diethylstilbestrol did not reduce the uptake of the steroids. These findings suggest that 17β-estradiol, 17β-estradiol 3-glucoside, and 17β-estradiol 3-glucuronide are taken up by nuclei by a nonsaturable diffusion process. The effect of cytosol on the uptake of estrogens by purified nuclei was also investigated. It was observed that cytosol reduced the uptake of 17β-estradiol but had little effect on that of its conjugates.

Substitution of Trp1242 of TM17 alters substrate specificity of human multidrug resistance protein 3

2003 ◽  
Vol 284 (2) ◽  
pp. G280-G289 ◽  
Author(s):  
Curtis J. Oleschuk ◽  
Roger G. Deeley ◽  
Susan P. C. Cole
Keyword(s):  
Multidrug Resistance ◽  
Substrate Specificity ◽  
Multidrug Resistance Protein ◽  
Wild Type ◽  
Multidrug Resistance Protein 3 ◽  
17Β Estradiol ◽  
Resistance Protein ◽  
Estradiol 17Β ◽  
Taurocholate Transport

Multidrug resistance protein 3 (MRP3) is an ATP-dependent transporter of 17β-estradiol 17β(d-glucuronide) (E217βG), leukotriene C4 (LTC4), methotrexate, and the bile salts taurocholate and glycocholate. In the present study, the role of a highly conserved Trp residue at position 1242 on MRP3 transport function was examined by expressing wild-type MRP3 and Ala-, Cys-, Phe-, Tyr-, and Pro-substituted mutants in human embryonic kidney 293T cells. Four MRP3-Trp1242 mutants showed significantly increased E217βG uptake, whereas transport by the Pro mutant was undetectable. Similarly, the Pro mutant did not transport LTC4. By comparison, LTC4transport by the Ala, Cys, Phe, and Tyr mutants was reduced by ∼35%. The Ala, Cys, Phe, and Tyr mutants all showed greatly reduced methotrexate and leucovorin transport, except the Tyr mutant, which transported leucovorin at levels comparable with wild-type MRP3. In contrast, the MRP3-Trp1242 substitutions did not significantly affect taurocholate transport or taurocholate and glycocholate inhibition of E217βG uptake. Thus Trp1242 substitutions markedly alter the substrate specificity of MRP3 but leave bile salt binding and transport intact.

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