Anim Reprod Sci. 2003 Sep 15;78(1-2):99-110.
Role of estradiol-17beta on nuclear and cytoplasmic maturation of pig oocytes.

Dode MA, Graves CN.

Animal Reproduction Laboratory, Embrapa Genetic Resources and Biotechnology, Parque Estacao Biologica, Avenue W5 Final Norte, Brasilia CEP 70770-900, DF, Brazil. margoenargen.embrapa.br

The role of estradiol-17beta on nuclear and cytoplasmic maturation of pig oocytes was investigated in the present study. To determine the estradiol effect, oocytes were cultured for 42 h in a steroid free medium composed of mTCM-199 supplemented with LH, FSH and 10% charcoal extracted follicular fluid. Estradiol receptor (ER), detected by a binding assay, were present in cumulus cells and oocytes during maturation with higher levels observed at 24 h of culture in the oocytes and at 36 h in the cumulus cells. To block estradiol action an antiestrogen (1-p-dimethylaminoethoxyphenyl-1,2-diphenyl-1-butene (tamoxifen)) was added to the maturation medium at various concentrations. The percentage of treated oocytes that underwent nuclear maturation was similar (P>0.05) to the control group. Cytoplasmic maturation, determined by the ability to form female pronucleus (FPN) and male pronucleus (MPN), was not different (P>0.05) among all groups. The presence of 4-hydroxy-4-androstene-3-17-dione (4-OHA) also did not influence nuclear (P>0.05) or cytoplasmic maturation (P>0.05). The results suggest that estradiol is not involved in maturation of pig oocytes. However, the present experiment used pronuclei formation as the endpoint, no studies were done in regard to estradiol's effects on the embryonic development.

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12753786&dopt=Abstract estradiol




Drug Metab Dispos. 2003 Jun;31(6):762-7.
The effect of incubation conditions on the enzyme kinetics of udp-glucuronosyltransferases.

Soars MG, Ring BJ, Wrighton SA.

Department of Drug Disposition, Lilly Research Laboratories, Drop Code 0710, Eli Lilly and Company, Indianapolis, IN 46285, USA.

Traditionally, the Michaelis-Menten equation has been used to determine kinetic parameters for in vitro glucuronidation assays. Recently, estradiol-3-glucuronide formation was shown to exhibit non-Michaelis-Menten kinetics consistent with autoactivation. A concern with the observation of nontraditional kinetics is that they may result as an artifact of the incubation conditions. To examine this concern, the formation of estradiol-3-glucuronide was investigated using human liver microsomes prepared by two different methods, a range of assay conditions, and activation by alamethecin, sonication, or Brij 58 (polyoxyethylene monocetyl ether). Interestingly, holding the other assay components constant, estradiol-3-glucuronide formation was up to 2.5-fold greater using microsomes prepared in phosphate buffer compared with those prepared in sucrose. Incubations activated by alamethecin consistently exhibited the highest rates of estradiol glucuronidation versus the other activators. Furthermore, estradiol-3-glucuronidation exhibited autoactivation kinetics in all of the conditions investigated (n = 1.2-1.7). Nontraditional kinetics were also observed when other UGT1A1 substrates such as ethinylestradiol, buprenorphine, and anthraflavic acid were studied with both human liver microsomes and recombinant UGT1A1. Naphthol, propofol, morphine, and androstanediol were used as probe UGT substrates selective for UGT1A6, UGT1A9, UGT2B7, and UGT2B15, respectively. Of these substrates, only androstanediol exhibited nontraditional kinetics using human liver microsomes. In conclusion, the Hill and/or Michaelis-Menten equations should be used to fit kinetic data to obtain an accurate assessment of in vitro glucuronidation.<




Neurosci Lett. 2003 May 22;342(3):201-5.
Estradiol reduces seizure-induced hippocampal injury in ovariectomized female but not in male rats.

Galanopoulou AS, Alm EM, Veliskova J.

Department of Neurology, Albert Einstein College of Medicine, Bronx, NY, USA. arisol.com

Estrogens protect ovariectomized rats from hippocampal injury induced by kainic acid-induced status epilepticus (SE). We compared the effects of 17beta-estradiol in adult male and ovariectomized female rats subjected to lithium-pilocarpine-induced SE. Rats received subcutaneous injections of 17beta-estradiol (2 microg/rat) or oil once daily for four consecutive days. SE was induced 20 h following the second injection and terminated 3 h later. The extent of silver-stained CA3 and CA1 hippocampal neurons was evaluated 2 days after SE. 17beta-Estradiol did not alter the onset of first clonus in ovariectomized rats but accelerated it in males. 17beta-Estradiol reduced the argyrophilic neurons in the CA1 and CA3-C sectors of ovariectomized rats. In males, estradiol increased the total damage score. These findings suggest that the effects of estradiol on seizure threshold and damage may be altered by sex-related differences in the hormonal environment.

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12757899&dopt=Abstract estradiol




Eur J Cancer. 2003 Jun;39(9):1302-9.
The activation of an extracellular signal-regulated kinase by oestradiol interferes with the effects of trastuzumab on HER2 signalling in endometrial adenocarcinoma cell lines.

Treeck O, Diedrich K, Ortmann O.

Clinic for Obstetrics and Gynecology, University of Lubeck, Ratzeburger Allee 160, 23538, Lubeck, Germany. treecmail.com

Cellular response to oestradiol stimuli is mediated both by oestrogen receptor (ER) binding to oestrogen response elements (EREs) and by non-nuclear actions like activation of mitogen-activated protein kinase (MAPK) signal transduction. Therefore, oestradiol stimuli might be able to interfere with the action of antitumoral substances directed against receptor tyrosine kinase signalling. We investigated the effect of oestradiol on the inhibition of HER2 signalling by trastuzumab (Herceptin) in two human endometrial adenocarcinoma cell lines. Activation of the extracellular signal-regulated kinase (ERK-1/2), a major mediator of HER2 signalling, was measured by means of western blotting experiments and ERE activation was determined in transient reporter-gene assays. In endometrial Ishikawa and HEC-1A adenocarcinoma cells, HER2 signalling was inhibited by trastuzumab only in the absence of oestradiol. We were able to demonstrate that oestradiol counteracted the inhibitory effects of trastuzumab by rapid phosphorylation of ERK-1/2, a kinase downstream of the HER2 receptor. The pure anti-oestrogen ICI 182,780 was able to restore both the trastuzumab-triggered inhibition of the ERK-1/2 pathway and the antiproliferative action of this substance in Ishikawa cells. Our data suggest that combinations of trastuzumab with anti-oestrogens may be effective in the treatment of endometrial cancers with a positive ER and HER2 receptor status.

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12763221&dopt=Abstract estradiol




Neuroendocrinology. 2003 Apr;77(4):223-31.
Repeated estradiol treatment prevents MPTP-induced dopamine depletion in male mice.

Ramirez AD, Liu X, Menniti FS.

CNS Discovery, Pfizer Global Research and Development, Groton, Conn., USA. Andres_d_Ramireroton.pfizer.com

Epidemiological data suggest that the steroid hormone 17beta-estradiol plays an important role in protecting the brain from neurodegenerative processes, including that causing the loss of dopamine (DA) neurons in Parkinson's disease. Determining the mechanisms of neuroprotection in experimental systems may facilitate the development of estrogenic therapies for these diseases. The present study sought to further investigate the mechanism of the neuroprotective effect of 17beta-estradiol in a murine model of Parkinson's disease, i.e. 1-methyl- 4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced striatal DA depletion. Consistent with previous findings, 17beta-estradiol was found to inhibit MPTP-induced DA depletion under a dosing regimen (repeated daily administration) that mimicked physiological levels of the steroid. However, high doses of the steroid administered repeatedly or acutely failed to inhibit toxicity, as did 17alpha-estradiol. These data suggest that the neuroprotective effect of 17beta-estradiol was mediated through an interaction with one of the nuclear estrogen receptors, and is not the result of an antioxidant action. In order to realize the therapeutic potential of the neuroprotective effect of 17beta-estradiol for Parkinson's disease, it will be necessary to identify synthetic estrogen receptor modulators that lack the activity of the steroid on peripheral tissue. In this study, raloxifene failed to mimic the neuroprotective effect of 17beta-estradiol against MPTP toxicity. Thus, exploration of new compounds with different pharmacological and/or physiochemical properties is warranted. Copyright 2003 S. Karger AG, Basel

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12766322&dopt=Abstract estradiol




Neuroendocrinology. 2003 Apr;77(4):239-45.
The effect of estradiol in the striatum is blocked by ICI 182,780 but not tamoxifen: pharmacological and behavioral evidence.

Xiao L, Jackson LR, Becker JB.

Department of Psychology, The University of Michigan, Ann Arbor, Mich., USA.

Estradiol in the striatum enhances amphetamine (AMPH)- or KCl-stimulated dopamine (DA) release and the pacing of sexual behavior in the female rat. These effects of estradiol in the striatum are rapid, steroid specific and thought to be mediated by a G protein-coupled membrane receptor for estradiol. In the current experiments, we examined whether two antiestrogens, ICI 182,780 (ICI) and tamoxifen (TAM), affect the enhancement by estradiol of (1) AMPH-induced DA release from striatal tissue in vitro, and (2) paced mating behavior in the female rat. The steroidal antiestrogen ICI significantly blocked the effect of estradiol on AMPH-induced striatal DA release. In contrast, the nonsteroidal, triphenylethylene antiestrogen TAM did not block the effect of estradiol, even when a concentration 10 times the concentration of estradiol was delivered to the tissue. Neither of the compounds showed estrogenic action when tested in the absence of estradiol. When implanted into the dorsolateral striatum, ICI inhibited the effect of estradiol on pacing of sexual behavior, while TAM did not. The specific effects of ICI, but not TAM, on striatal DA release and pacing behavior provide further information about the binding site for estradiol in the striatum. Copyright 2003 S. Karger AG, Basel

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12766324&dopt=Abstract estradiol




Comp Biochem Physiol C Toxicol Pharmacol. 2003 Jan;134(1):45-55.
Effects of octylphenol and 17beta-estradiol on the gonads of guppies (Poecilia reticulata) exposed as adults via the water or as embryos via the mother.

Kinnberg K, Korsgaard B, Bjerregaard P.

Danish Centre for Environmental Oestrogen Research, Institute of Biology, University of Southern Denmark--Odense, Campusvej 55, DK-5230 Odense M, Denmark. kinnberiology.sdu.dk

Endocrine disrupting alkylphenolic compounds have been found in the aquatic environment, and concern has arisen over the ability of these compounds to affect the reproductive system of fish. In this study, the effects of exposure to an environmentally relevant concentration of octylphenol or 17beta-estradiol on the gonad structure of fish were examined. Viviparous guppies (Poecilia reticulata) were exposed as adults via the water or as embryos via the mother to 26 microg/l octylphenol or 0.85 microg/l 17beta-estradiol (mean measured water concentrations). Histological examinations revealed effects of the exposures on the gonads of the fish exposed as adults. Indications of blocked spermatogonial mitosis were seen in the testis structure of adult males after exposure to octylphenol or 17beta-estradiol. The post-parturition ovaries of adult females exposed to 17beta-estradiol showed effects suggesting an inhibited yolk deposition. At the tested concentrations, exposure to octylphenol or 17beta-estradiol via the mother fish did not significantly affect the weight, length, gonopodium index or sex distribution of the offspring. However, histology revealed effects on the liver structure, suggesting some effect of maternal exposure to octylphenol or 17beta-estradiol. These findings indicate that although octylphenol and 17beta-estradiol affect the gonad structure of adult male and female guppies, these substances have no significant effects on the sexual differentiation of the embryos at the tested concentrations.

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12524017&dopt=Abstract estradiol




Hypertension. 2003 Jul;42(1):82-7. Epub 2003 May 27.
Methylation of 2-hydroxyestradiol in isolated organs.

Zacharia LC, Dubey RK, Mi Z, Jackson EK.

Center for Clinical Pharmacology, University of Pittsburgh Medical Center, 623 Scaife Hall, 3550 Terrace Street, Pittsburgh, Penn 15261, USA.

Vascular smooth muscle and glomerular mesangial cells in culture express a biochemical pathway that methylates 2-hydroxyestradiol (17beta-estradiol metabolite) to produce 2-methoxyestradiol, a cell growth inhibitor that may mediate the cardiorenal protective effects of 17beta-estradiol. Whether this pathway exists in intact organ systems is currently unclear. Accordingly, the purpose of the present investigation was to characterize the methylation of 2-hydroxestradiol in intact organs from both male and female rats. No significant differences were detected in the ability of male and female tissues to methylate 2-hydroxyestradiol. In isolated hearts, kidneys, and mesenteries perfused with Tyrode's solution, Km values for 2-hydroxyestradiol methylation were 0.175+/-0.021, 0.387+/-0.054, and 0.495+/-0.089 micromol/L, respectively, and Vmax values were 21.0+/-1.58, 24.9+/-1.49, and 1.01+/-0.148 pmol 2-methoxyestradiol x min(-1) x ml(-1) per gram, respectively. The catalytic efficiency (Vmax/Km) was greatest in the heart compared with the kidney and mesentery (132+/-14.3, 78.4+/-15.1, and 2.30+/-0.263 pmol 2-methoxyestradiol x min(-1) x mL(-1) x micromol/L(-1) per gram, respectively). In the kidney, the catechol-O-methyltransferase inhibitor quercetin and norepinephrine (10 micromol/L) reduced methylation of 2-hydroxyestradiol by approximately 90% and 41%, respectively. Importantly, methylation in the kidney was inhibited by an average of 16.6+/-1.80% by endogenous norepinephrine released by renal artery nerve stimulation. Our results indicate that a robust 2-hydroxyestradiol methylation pathway exists in the kidney and heart, but not in the mesentery, and that this pathway is mediated by catechol-O-methyltransferase. Our f




Reproduction. 2003 Jun;125(6):847-54.
Role of oestradiol in growth of follicles and follicle deviation in heifers.

Beg MA, Meira C, Bergfelt DR, Ginther OJ.

Department of Animal Health and Biomedical Sciences, University of Wisconsin, Madison, WI 53706, USA; and The Eutheria Foundation, Cross Plains, WI 53528, USA.

Follicle deviation is characterized by continued growth of the largest (developing dominant) follicle and reduced growth of the smaller (subordinate) follicles. The aim of the present study was to test the following hypotheses: (1). oestradiol contributes to the depression of circulating FSH encompassing follicle deviation and (2). oestradiol plays a role in the initiation of deviation. Heifers were treated with progesterone (n = 5) or antiserum against oestradiol (n = 7) or given no treatment (control; n = 6). On the basis of previous studies, progesterone treatment would decrease LH and thereby the circulatory and intrafollicular concentrations of oestradiol and the antiserum would reduce the availability of oestradiol. Progesterone was given in six 75 mg injections at 12 h intervals beginning when the largest follicle of wave 1 first reached >or=5.7 mm (t = 0 h). Oestradiol antiserum (100 ml) was given in a single injection at t = 12 h. Follicles of the wave were defined as F1 (largest) and F2, according to the diameter at each examination. Blood samples were collected at 12 h intervals during t = 0-72 h. Treatment with progesterone lowered the circulatory concentrations of LH by 12 h after the start of treatment (P < 0.05), and concentrations remained low compared with those of controls during the treatment period. Treatment with oestradiol antiserum had no effect on LH. Both progesterone and the antiserum treatments increased the FSH concentrations compared with controls (P < 0.05), which supports the first hypothesis. The interval from t = 0 h to the beginning of deviation was longer in the progesterone- (51.0 +/- 7.6 h; P < 0.06) and antiserum (51.4 +/- 6.3 h