Estrogens are known to exert significant structural and functional effects in the hippocampus of adult rodents. In particular, 17 beta-estradiol can improve, impair, or have no effect on hippocampus-dependent learning and memory depending on dose and time of administration. The effects of other forms of estrogen, such as estrone and 17 alpha-estradiol, on hippocampus-dependent learning have not been as thoroughly investigated. Therefore, the purpose of this study was to investigate the effects of 17 beta-estradiol, estrone, and 17 alpha-estradiol at three different doses on two different tasks: hippocampus-dependent contextual fear conditioning and hippocampus-independent cued fear conditioning. Adult ovariectomized female rats were injected with one of the estrogens at one of the three doses 30 mins before conditioning to assess the rapid effects of these estrogens on acquisition. Twenty-four hours later memory for the context was examined and 1 h later memory for the cue (tone) was assessed. Levels of synaptophysin were examined in the dorsal hippocampus of rats to identify a potential synaptic correlate of hormonal effects on contextual fear conditioning. Low 17 beta-estradiol and 17 alpha-estradiol enhanced, whereas high 17 beta-estradiol and 17 alpha-estradiol impaired, contextual fear conditioning. Only the middle dose of estrone severely impaired contextual fear conditioning. Estrogens did not alter performance in the hippocampus-independent cued task. Synaptophysin expression was increased by estrone (at a middle and high dose) and 17 beta-estradiol (at a middle dose) in the CA3 region of the hippocampus and was not correlated with cognition. The results of this study indicate that estradiol can positively or negatively influence hippocampus-dependent learning and memory, whereas estrone impairs hippocampus-dependent learning and memory in a dose-dependent manner. These results have important therapeutic implications, as estrone, a main component of a widely used hormone replacement therapy, was shown to have either a negative effect or no effect on learning and memory. It may be possible to use 17 alpha-estradiol and lower doses of estrogens as potential alternatives in hormone replacement therapies. Neuropsychopharmacology (2010) 35, 547-559; doi: 10.1038/npp.2009.161; published online 21 October 2009
We show here that baseline separation of dansylated estrone, 17β-estradiol, and 17α-estradiol can be done, contrary to previous reports, within a short run time on a single RP-LC analytical column packed with particles bonded with phenyl-hexyl stationary phase. The chromatographic method coupled with isotope dilution tandem MS offers a simple assay enabling the simultaneous analysis of these analytes. The method employs 13C-labeled estrogens as internal standards to eliminate potential matrix effects arising from the use of deuterated estrogens. The assay also offers adequate accuracy and sensitivity to be useful for biological samples. The practical applicability of the validated method is demonstrated by the quantitative analyses of in vivo samples obtained from rats treated with Premarin®. Figure Quantification of estrogens from rat samples by LC–MS/MS
The natural manure-borne hormones, 17α-estradiol (17α-E2), 17β-estradiol (17β-E2), and estrone (E1), are routinely detected in surface water near agricultural land and wastewater treatment facilities. Once in the stream network, hormones may enter the sediment bed where they are subject to anaerobic conditions. This study focuses on the difference in anaerobic transformation rates and formation of metabolites from 17α-E2, 17β-E2, and E1 (applied at ∼3.66 μmol kg(-1) of sediment on a dry weight basis) under nitrate- and sulfate-reducing conditions. Sediment extracts were analyzed using negative electrospray ionization tandem mass spectrometry. Under both redox conditions, degradation was stereospecific and followed similar trends in half-lives, 17β-E2 < 17α-E2 < E1, with degradation considerably slower under sulfate-reducing conditions. Both E2 isomers were predominantly converted to E1; however, isomeric conversion also occurred with peak concentrations of ∼1.7 mol % of 17β-E2 formed in 17α-E2 amended sediments and peak concentrations of ∼2.4 mol % of 17α-E2 formed from 17β-E2. In E1-amended systems, E1 transformed to E2 with preferential formation of the more potent 17β isomer up to ∼30 mol % suggesting that isomer interconversion is through E1. Sediments, therefore, may serve as both a sink and a source of the more estrogenic compound E2. Transformation of amended hormones in autoclaved sediments was markedly slower than in nonautoclaved sediments. Results support the inclusion of isomer-specific behavior and the potential for reversible transformation and interconversion in anaerobic sediments in modeling fate in stream networks and developing risk management strategies.
In estradiol-primed nonreceptive ovariectomized rats, activation of G protein-coupled estrogen receptor 1 (GPER) in the arcuate nucleus of the hypothalamus (ARH) rapidly facilitates sexual receptivity (lordosis). Estradiol priming activates ARH β-endorphin (β-END) neurons that then activate medial preoptic (MPN) μ-opioid receptors (MOP) to inhibit lordosis. ARH infusion of non-esterified 17β-estradiol (E2) 47.5 h after 17β-estradiol benzoate (2 μg EB) priming deactivates MPN MOP and rapidly facilitates lordosis within 30 min via activation of GPER. Since it was unclear where GPERs were located in the neuron, we tested the hypothesis that GPER signaling is initiated at the plasma membrane. Membrane impermeable estradiol (17β-estradiol conjugated to biotin; E-Biotin) infused into the ARH of EB primed rats facilitated lordosis within 30 min, and MPN MOP was deactivated. These actions were blocked by pretreating with GPER antagonist, G-15. Further, we used cell fractionation and western blot techniques to demonstrate that GPER is expressed both in plasma membrane and cytosolic ARH fractions. In previous studies, the orphanin FQ/nociceptin-opioid receptor-like receptor-1 (OFQ/N-ORL-1) system mediated estradiol-only facilitation of lordosis. Therefore, we tested whether the OFQ/N-ORL-1 system mediates E-Biotin-GPER facilitation of lordosis. Pretreatment of UFP-101, an ORL-1 selective antagonist, blocked the facilitation of lordosis and deactivation of MPN MOP by ARH infusion of E-Biotin. Double-label immunohistochemistry revealed that GPER is expressed within approximately 70% of OFQ/N neurons. These data indicate that membrane GPER mediates the E2/E-Biotin facilitation of lordosis by inducing OFQ/N neurotransmission, which inhibits β-END neurotransmission to reduce MPN MOP activation.
The application of manure and biosolids onto agricultural land has increased the risk of estrogenic exposure to aquatic systems. Both αE2 and βE2 have been routinely detected in surface and ground waters with higher concentrations reported near concentrated animal feeding operations and agricultural fields. Although movement through the soil to a water body is highly dependent on hormone–soil interactions, to date, only the interaction of βE2 with soils has been characterized despite αE2 often being the more common form excreted by livestock such as beef cattle and dairy. In predicting the transport of estradiol, sorption characteristics for the stereoisomers have been assumed to be the same. To evaluate this assumption, sorption of αE2 and βE2 was measured on seven surface soils representing a range in soil properties. Soils were autoclave-sterilized to minimize loss due to biotransformation, and both solution and soil phase concentrations were measured. Overall, E2 sorption is best correlated to soil organic carbon (OC) with an average log OC-normalized distribution coefficient (log , L ) of 2.97 ± 0.13 for αE2 and 3.14 ± 0.16 for βE2 with βE2 consistently exhibiting higher sorption than αE2 with the highest β/α sorption ratio of 1.9. Assuming that the two isomers sorb the same is not a conservative decision making approach. The lower sorption affinity of αE2 increases the likelihood that it will be leached from agricultural fields.
17β-Estradiol (E2) and 17α-ethinyl estradiol (EE2), which are environmental estrogens, have been determined with LC-MS in freshwater. Their sensitive analysis needs derivatization and therefore is very hard to achieve in multiresidue screening. We analyzed samples from all the large and some small rivers (River Danube, Drava, Mur, Sava, Tisza, and Zala) of the Carpathian Basin and from Lake Balaton. Freshwater was extracted on solid phase and derivatized using dansyl chloride. Separation was performed on a Kinetex XB-C18 column. Detection was achieved with a benchtop orbitrap mass spectrometer using targeted MS analysis for quantification. Limits of quantification were 0.05 ng/L (MS1) and 0.1 ng/L (MS/MS) for E2, and 0.001 ng/L (MS1) and 0.2 ng/L (MS/MS) for EE2. River samples contained n.d.–5.2 ng/L E2 and n.d.–0.68 ng/L EE2. Average levels of E2 and EE2 were 0.61 and 0.084 ng/L, respectively, in rivers, water courses, and Lake Balaton together, but not counting city canal water. EE2 was less abundant, but it was still present in almost all of the samples. In beach water samples from Lake Balaton, we measured 0.076–0.233 E2 and n.d.–0.133 EE2. A relative high amount of EE2 was found in river Zala (0.68 ng/L) and in Hévíz-Páhoki canal (0.52 ng/L), which are both in the catchment area of Lake Balaton (Hungary).
Purpose Whereas anti-oestrogen therapy is widely applied to treat oestrogen receptor (ER) positive breast cancer, paradoxically, oestrogens can also induce tumour regression. Upregulation of ER expression is a marker for oestrogen hypersensitivity. We, therefore, performed an exploratory study to evaluate positron emission tomography (PET) with the tracer 16 alpha-[F-18] fluoro-17 beta-oestradiol (F-18-FES) as potential marker to select breast cancer patients for oestradiol therapy. Methods Eligible patients had acquired endocrine-resistant metastatic breast cancer that progressed after >= 2 lines of endocrine therapy. All patients had prior ER-positive histology. Treatment consisted of oestradiol 2 mg, three times daily, orally. Patients underwent F-18-FES-PET/CT imaging at baseline. Tumour F-18-FES-uptake was quantified for a maximum of 20 lesions and expressed as maximum standardised uptake value (SUVmax). CT-scan was repeated every 3 months to evaluate treatment response. Clinical benefit was defined as time to radiologic or clinical progression >= 24 weeks. Results F-18-FES uptake, quantified for 255 lesions in 19 patients, varied greatly between lesions (median 2.8; range 0.6-24.3) and between patients (median 2.5; range 1.1-15.5). Seven (37 %) patients experienced clinical benefit of oestrogen therapy, eight progressed (PD), and four were non-evaluable due to side effects. The positive and negative predictive value PPV/NPV) of F-18-FES-PET for response to treatment were 60 % (95 % CI: 31-83 %) and 80 % (95 % CI: 38-96 %), respectively, using SUVmax >1.5. Conclusion F-18-FES-PET may aid identification of patients with acquired antihormone resistant breast cancer that are unlikely to benefit from oestradiol therapy.
It is now well accepted that sex hormones have immunoregulatory activity and may prevent exacerbations in multiple sclerosis during pregnancy. Our previous studies demonstrated that oestrogen (17β‐oestradiol; E 2 ) protection against experimental autoimmune encephalomyelitis ( EAE ) is mediated mainly through oestrogen receptor‐α ( ER α) and the membrane receptor G‐protein‐coupled receptor 30 ( GPR 30) and is abrogated in the absence of B cells and the co‐inhibitory receptor, Programmed Death‐1 ( PD ‐1). To critically evaluate the cell source of the E 2 and PD ‐1 co‐inhibitory pathways in EAE regulation, we assessed the requirement for ER s on transferred B cells and downstream effects on expression of PD ‐1/ PD ‐ligand on CD 4 + F oxp3 + regulatory T (Treg) cells in B ‐cell‐replenished, E 2 ‐treated B ‐cell‐deficient (μ MT −/− ) mice with EAE . The results clearly demonstrated involvement of ER α and GPR 30 on transferred B cells that mediated the protective E 2 treatment effect on EAE and further showed an E 2 ‐mediated B ‐cell‐dependent up‐regulation of PD ‐1 on CD 4 + F oxp3 + T reg cells. These findings identify regulatory B ‐cell populations as key players in potentiating T reg‐cell activity during E 2 ‐mediated protection against EAE .