Neuroscience. 2002;110(3):489-504.
17beta-estradiol enhances cortical cholinergic innervation and preserves synaptic density following excitotoxic lesions to the rat nucleus basalis magnocellularis.

Horvath KM, Hartig W, Van der Veen R, Keijser JN, Mulder J, Ziegert M, Van der Zee EA, Harkany T, Luiten PG.

Department of Molecular Neurobiology, Graduate School of Behavioural and Cognitive Neurosciences, University of Groningen, The Netherlands. k.m.horvatiol.rug.nl

Estradiol exerts beneficial effects on neurodegenerative disorders associated with the decline of cognitive performance. The present study was designed to further investigate the effect of 17beta-estradiol on learning and memory, and to evaluate its neuroprotective action on cholinergic cells of the nucleus basalis magnocellularis, a neural substrate of cognitive performance. Female rats were ovariectomized at an age of 6 months. Three weeks later they received injections of either a mid-physiological dose of 17beta-estradiol or vehicle (oil), every other day for 2 weeks. The effect of estradiol on cognitive performance was tested in two associative learning paradigms. In the two-way active shock avoidance task estradiol-replaced animals learned significantly faster, while in the passive shock avoidance test no differences were observed between the experimental groups. Subsequent unilateral infusion of N-methyl-D-aspartate in the nucleus basalis magnocellularis resulted in a significant loss of cholinergic neurons concomitant with the loss of their fibers invading the somatosensory cortex. Estradiol treatment did not affect the total number of choline-acetyltransferase-immunoreactive neurons and their coexpression of the p75 low-affinity neurotrophin receptor either contralateral or ipsilateral to the lesion. In contrast, cholinergic fiber densities in estradiol-treated animals were greater both in the contralateral and ipsilateral somatosensory cortices as was detected by quantitative choline-acetyltransferase and vesicul




Biol Reprod. 2002 Apr;66(4):1104-10.
Loss of luteinizing hormone surges induced by chronic estradiol is associated with decreased activation of gonadotropin-releasing hormone neurons.

Tsai HW, Legan SJ.

Department of Physiology, University of Kentucky, 800 Rose Street, Lexington, KY 40536, USA.

Chronic exposure of young ovariectomized rats to elevated circulating estradiol causes loss of steroid-induced LH surges. Such LH surges are associated with cFos-induced activation of GnRH neurons; therefore, we hypothesized that chronic estradiol treatment abolishes LH surges by decreasing activation of GnRH neurons. Regularly cycling rats were ovariectomized and immediately received an estradiol implant or remained untreated. Three days or 2 or 4 wk later, the estradiol-treated rats received vehicle or progesterone at 1200 h, and 7 hourly blood samples were collected for RIA of LH. Thereafter, all rats were perfused, and the brains were examined for immunocytochemical localization of cFos and GnRH. The GnRH neurons from untreated ovariectomized rats rarely expressed cFos. As reported, LH surges induced by 3 days of estradiol treatment were associated with a 30% increase in cFos-containing GnRH neurons, and progesterone enhanced both the amplitude of LH surges and the proportion of cFos-immunopositive GnRH neurons. As hypothesized, the abolition of LH surges caused by 2 or more weeks of estradiol was paralleled by a reduction in the percentage of cFos-containing GnRH neurons, and this effect was delayed by progesterone. These results suggest that chronic estradiol abolishes steroid-induced LH surges in part by inactivating GnRH neurons.

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11906931&dopt=Abstract estradiol




J Pharmacol Exp Ther. 2002 Apr;301(1):234-40.
Differential effects of sarcolemmal and mitochondrial K(ATP) channels activated by 17 beta-estradiol on reperfusion arrhythmias and infarct sizes in canine hearts.

Tsai CH, Su SF, Chou TF, Lee TM.

Department of Surgery, Cardiology Section, National Taiwan University College of Medicine, National Taiwan University Hospital, Taipei, Taiwan. tmlea.mc.ntu.edu.tw

We have demonstrated the effects of estrogen on modulation of ATP-sensitive K(+) channels; however, the subcellular location of these channels is unknown. The purpose of the present study was to investigate the role of the sarcolemmal and mitochondrial ATP-sensitive K(+) channels in a canine model of myocardial infarction after stimulation with 17 beta-estradiol. Anesthetized dogs were subjected to 60 min of the left anterior descending coronary artery occlusion followed by 3 h of reperfusion. Infarct size was markedly reduced in estradiol-treated dogs compared with controls (14 +/- 6 versus 42 +/- 6%, P < 0.0001), indicating the effective dose of estradiol administrated. Pretreatment with the mitochondrial ATP-sensitive K(+) channel antagonist 5-hydroxydecanoate completely abolished estradiol-induced cardioprotection. The sarcolemmal ATP-sensitive K(+) channel antagonist 1-15-12-(5-chloro-o-anisamido)ethyl-methoxyphenyl)sulfonyl-3-methylthiourea (HMR 1098) did not significantly attenuate estradiol-induced infarct size limitation. In addition, estradiol administration significantly reduced the incidence and duration of reperfusion-induced ventricular tachycardia and ventricular fibrillation. Although 5-hydroxydecanoate alone caused no significant effect on the incidence of reperfusion arrhythmias in the presence or absence of estradiol, the administration of HMR 1098 abolished estrogen-induced improvement of reperfusion arrhythmias. Pretreatment with the estrogen-receptor antagonist faslodex (ICI 182,780) did not alter estrogen-induced infarct-limiting and antiarrhythmic effects. These resul




Climacteric. 2000 Dec;3(4):271-7.
Comparison of the effects of continuous combined and sequential combined medroxyprogesterone acetate-estradiol treatment on the proliferation of MCF-7 cells.

Lippert C, Seeger H, Wallwiener D, Mueck AO.

Section of Endocrinology and Menopause, Department of Obstetrics and Gynecology, University of Tuebingen, Schleichstr. 4, 72076 Tuebingen, Germany.

OBJECTIVE: The aim of the study was to examine the effects of two different, clinically relevant, hormone replacement therapy (HRT) regimen types, continuous combined and sequential combined, on breast cancer cells by means of an in vitro model. The study was carried out using the C21-progestin medroxyprogesterone acetate (MPA) in combination with estradiol, and with the proliferation of MCF-7, a human breast cancer cell-line, as end-point. METHODS: Proliferation of MCF-7 cells was measured by means of a crystal violet staining technique. Growth was triggered using a constant estradiol concentration of 10(-10) mol/l, while varying the MPA concentration from 10(-11) to 10(-6) mol/l. RESULTS: The continuous combined model of treatment led to the inhibition of estradiol-induced growth of MCF-7 with MPA concentrations of 10(-10) mol/l and upwards, compared with estradiol-alone-induced growth. The sequential combined model showed a greater inhibition at the higher MPA concentrations of 10(-8)-10(-6) mol/l, with reduced sensitivity to inhibition at the lower MPA concentrations tested, of 10(-11)-10(-9) mol/l. The different treatment types resulted in significantly different sensitivities of the MCF-7 cells to inhibition of estradiol-induced proliferation at the higher MPA concentrations of 10(-8)-10(-6) mol/l. CONCLUSIONS: The results demonstrate the importance of considering in vivo factors in an in vitro model with regard to improving the cell culture techniques used, to obtain a clearer picture of the possible mechanisms involved in the potential breast cancer risk with different HRT regimens.

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=119&dopt=Abstract




Climacteric. 2000 Mar;3(1):17-24.
Early follicular phase serum FSH as a function of age: the roles of inhibin B, inhibin A and estradiol.

Burger HG, Dudley E, Mamers P, Groome N, Robertson DM.

Prince Henry's Institute of Medical Research, PO Box 5152, Clayton, Victoria 3168, Australia.

OBJECTIVE: Reproductive aging in regularly cycling normal women is characterized by a gradual decline in ovarian follicle number and a progressive increase in serum follicle stimulating hormone (FSH), particularly over the age of 40 years. The lack of any consistent decrease in circulating estradiol and progesterone has led to the hypothesis that the FSH increase results from decreasing ovarian inhibin production. The aim of this study was to investigate the relationship between serum inhibins A and B, FSH and estradiol in normal women between the ages of 20 and 50 years. DESIGN AND PATIENTS: Serum from 66 regularly cycling subjects, aged 20-50 years, was collected on days 3-5 of the menstrual cycle for this cross-sectional study. MEASUREMENTS: Serum inhibin A and inhibin B levels were measured by specific enzyme-linked immunosorbent assays (ELISAs). Alpha subunit forms were determined by an immunofluorometric assay which detects all known monomeric and dimeric forms of inhibin A and inhibin B and free alpha subunit. FSH and estradiol levels were measured by immunoassay. Data were log transformed before analysis. RESULTS: Serum FSH, inhibin A and estradiol, but not inhibin B, were positively correlated (p < 0.05-p < 0.001) with age between years 20 and 50. Between 40 and 50 years, serum FSH was negatively correlated with inhibin B (r = -0.61, p < 0.001) and alpha subunit forms (r = -0.47, p < 0.05) and with estradiol (r = -0.39, p < 0.05), but not with inhibin A (r = -0.21, not significant). When log(FSH) was modelled as a function of log(inhibin B) and log(estradiol) with age fitted as a covariate, inhibin B only was a significant independent predictor of FSH (beta = -0.30, p < 0.01).




Biochem Pharmacol. 2002 Mar 1;63(5):985-92.
Effect of clofibrate administration on the esterification and deesterification of steroid hormones by liver and extrahepatic tissues in rats.

Xu S, Zhu BT, Conney AH.

Laboratory for Cancer Research, Department of Chemical Biology, College of Pharmacy, Rutgers, The State University of New Jersey, Piscataway, NJ 08854-8020, USA.

Treatment of rats with clofibrate markedly stimulated the liver microsomal esterification of estradiol, testosterone, pregnenolone, dehydroepiandrosterone, and corticosterone by acyl-CoA:steroid acyltransferase. This enzyme catalyzes the esterification of estradiol with long-chain fatty acids in both liver and extrahepatic tissues. In untreated control rats, brain had the highest acyltransferase activity per milligram of microsomal protein for estradiol esterification (3- to 4-fold higher than in the liver). Although, treatment of rats with clofibrate stimulated the esterification of estradiol by 9- to 14-fold in the liver, estradiol esterification in kidney, lung, brain, uterus, fat, and mammary glands was not increased, indicating that liver may be uniquely sensitive to induction of acyl-CoA:estradiol acyltransferase by clofibrate. In additional studies, esterase activity for hydrolysis of the oleoyl ester of estradiol was determined in control and clofibrate-treated rats. Clofibrate administration increased esterase activity by an average of 107% in fat and 70% in liver. The results indicate that treatment of rats with clofibrate stimulates the hepatic formation of highly lipophilic fatty acid esters that can be hydrolyzed in the liver and in extrahepatic tissues to the parent steroid hormone by a clofibrate-inducible esterase.

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11911851&dopt=Abstract estradiol




Mol Hum Reprod. 2002 Apr;8(4):318-25.
Transforming growth factor-beta1 inhibits steroidogenesis in human trophoblast cells.

Luo S, Yu H, Wu D, Peng C.

Department of Biology, York University, 4700 Keele St, Toronto, Ontario, M3J 1P3, Canada.

Transforming growth factor-beta (TGF-beta) is an important regulator of placental development and function. In this study, we have investigated the effect of TGF-beta1 on steroidogenesis, as well as its sites of action in the steroidogenic pathway by using a choriocarcinoma cell line, JEG-3, and a normal trophoblast cell line (NPC). The effect of TGF-beta1 on progesterone and estradiol production was evaluated in the absence or presence of a membrane-permeable analogue of cholesterol and some intermediate substrates of steroidogenic enzymes. The effect of TGF-beta1 on P450 aromatase (P450arom) mRNA levels was determined by Northern blot analysis. TGF-beta1 significantly decreased progesterone production in both NPC and JEG-3 cells. The inhibitory effect of TGF-beta1 on progesterone production was reversed by addition of 22R-hydroxycholesterol, a membrane-permeable analogue of cholesterol, or pregnenolone. In JEG-3 cells, TGF-beta1 also inhibited estradiol production when androstenedione, but not estrone, was added to the culture. Estradiol production was too low to be detected in NPC cells. Treatment with TGF-beta1 also suppressed aromatase mRNA levels. This study has demonstrated that TGF-beta1 inhibits progesterone and estradiol production by trophoblast cells, and that the sites of TGF-beta1 action on progesterone and estradiol production are likely to be cholesterol transport and P450arom respectively.

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11912279&dopt=Abstract estradiol




Reproduction. 2002 Apr;123(4):601-11.
Effect of adrenocorticotrophic hormone on the development of oestrogen-induced changes and hyperplasia formation in the mouse uterus.

Gunin AG, Emelianov V, Tolmachev AS.

Department of Histology Medical School Chuvash State University, PO Box 86, 428034, Cheboksary, Russia. drguniahoo.com

The aim of this study was to examine the role of adrenocorticotrophic hormone (ACTH) in proliferative and morphogenetic reactions in the uterus under continuous oestrogen treatment. Ovariectomized mice received injections of oestradiol dipropionate or vehicle once a week (2 microg per 100 g), and injections of ACTH (10 microg per 100 g) once a day or once a week for 30 days. Additional control mice received oestradiol and saline, vehicle of oestradiol, and ACTH once a day or once a week, or vehicle of ACTH, for 30 days. This study shows for the first time that ACTH affects oestrogen-dependent reactions in the uterus. Treatment with ACTH once a day resulted in a decrease in uterine mass, in cell proliferation (assessed by the number of mitotic and bromodeoxyuridine (BrdU)-labelled cells) and in the incidence of endometrial hyperplasia, in particular complex and atypical hyperplasia. Treatment with ACTH once a week led to a marked reduction in the incidence of endometrial hyperplasia, a slight increase in uterine mass and had almost no effect on cell proliferation. Daily treatment with ACTH reduced the concentration of oestrogen receptor alpha in all uterine compartments, but weekly ACTH administration had the opposite effect. Expression of glucocorticoid receptors, beta-catenin and glycogen synthase kinase-3beta in uterine tissues was lower in animals treated with oestradiol and ACTH once a day or once a week. When olive oil was used instead of oestradiol, treatments with ACTH did not produce detectable changes in all parameters examined. Thus, glucocorticoid receptor, oestrogen receptor alpha, beta-catenin and glycogen synthase kinase-3beta are involved in the eff




Reproduction. 2003 Apr;125(4):579-83.
Effects of food supplementation on litter size and oestradiol concentration during gestation and oestrous cycle of capybaras (Hydrochaeris hydrochaeris) in captivity.

Becker GK, Pederassi GC, Santos EA, Colares EP.

Programa de Pos-graduacao em Fisiologia Animal Comparada, Departamento de Ciencias Fisiologicas, Fundacao Universidade Federal do Rio Grande, RS, Brazil.

The present study analysed the response of adult female capybaras (Hydrochaeris hydrochaeris) to different dietary supplementation in relation to litter size per parturition and oestradiol secretion profile during pregnancy and the oestrous cycle. All four experimental groups received 'camerum' grass (Pennisetum purpureum) and water ad libitum and three of the groups were also fed a protein, lipid or protein and lipid supplement. Litter size per parturition did not show any significant difference among treatment groups, but was significantly higher (P < 0.05) than in the control group. There was no significant difference in oestradiol concentrations among treatments and the control group, during each phase of the oestrous cycle or during gestation. The average oestradiol concentrations in dioestrous, oestrous and metoestrous phases were not significantly different from each other, but were significantly lower (P < 0.05) than the average oestradiol concentration in the pro-oestrous phase. In addition, average oestradiol concentrations increased after the second gestation month, but were significantly higher (P < 0.05) only after the fourth gestation month, achieving maximum value by the end of gestation. Dietary supplementation had no significant effect on hormonal concentrations during the oestrous cycle and gestational period. However, an increase in litter size per parturition was observed among female capybaras that received dietary supplementation.

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12683928&dopt=Abstract estradiol