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      Immunocytochemical Detection of Estrogen Receptor-α in the Female Rabbit Forebrain: Topography and Regulation by Estradiol

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          Abstract

          Two antibodies (H222 and Zymed) directed towards different sites of the estrogen receptor-α (ERα) were used for the following objectives: (1) to map the ERα in the forebrain of ovariectomized (ovx) rabbits by immunocytochemistry and (2) to determine the effect of endogenous (intact non-pregnant animals) and exogenous (ovx, estrogen-treated animals) estradiol (E2) on the population of ERα in the forebrain. Similar results were obtained with both antibodies used: dense aggregations of ERα-immunoreactive (IR) neurons were found in the infundibular nucleus (IN), the medial preoptic area (POA), the bed nucleus of the stria terminalis (BNST), and some nuclei of the amygdala. By contrast, no ERα-IR neurons were present in the ventromedial hypothalamic nucleus (VMN), but a dense aggregation of ERα-IR neurons occurred lateral to it in nucleus X. Numerous ERα-IR neurons were observed in the paraventricular hypothalamic nucleus, but not in the supraoptic or suprachiasmatic nuclei. The hippocampus proper lacked ERα-IR neurons, but the ventral subiculum in the hippocampal formation had a dense group of such cells. Intact non-pregnant rabbits showed less ERα-IR neurons in all regions tested than ovx animals. This difference was particularly clear in the medial POA, amygdala and BNST, while the IN showed only a marginal decrease. The dorsal, but much less the ventral, part of nucleus X also showed a decrease in the number of ERα-IR neurons compared with ovx animals. E2 benzoate (5 µg/day for 5 days) reduced even further the number of ERα-IR neurons in all regions except in a circumscribed area of the IN and the ventral part of nucleus X. These results show the existence of both sensitive and insensitive neurons to the down-regulatory effect of E2 on the presence of ERα. Sensitive neurons are located in the telencephalon, POA and several hypothalamic nuclei (PVN), while insensitive neurons are mainly restricted to the IN and the ventral part of nucleus X in the basal hypothalamus.

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          Most cited references 19

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          Cell Membrane and Nuclear Estrogen Receptors (ERs) Originate from a Single Transcript: Studies of ER  and ER  Expressed in Chinese Hamster Ovary Cells

           M Razandi (1999)
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            Distribution and hormone regulation of estrogen receptor immunoreactive cells in the hippocampus of male and female rats.

            Estrogen regulates the synaptic plasticity and physiology of the hippocampus as well as learning behaviors that are mediated by the hippocampus. The density of dendritic spines and synapses, the number of N-methyl-D-aspartate (NMDA) binding sites, the levels of NMDA receptor subunit NR1 protein, muscimol binding to the gamma-amino butyric acid (GABA)A receptor, and levels of glutamic acid decarboxylase message in the CA1 region of the hippocampus are altered with estrogen treatment. In addition, some of these parameters exhibit sex differences in their response to estrogen treatment. To establish that estrogen can have a direct effect on the hippocampus and to determine whether or not sex differences in estrogen responsiveness are due to sex differences in estrogen receptor (ER) levels, we used immunocytochemistry with the AS409 antibody to map the location of ER-immunoreactive (ER-ir) cells in the hippocampus of male and female rats. We found that (1) the ERs appear to be in interneurons rather than pyramidal or granule cell neurons, (2) ER-ir cells are located in greatest concentration in the hilus of the dentate gyrus and the stratum radiatum of the CA1 region, (3) the density of ER-ir cells exhibits a rostral to caudal gradient in the hilus and the CA1 regions, (4) there are no sex differences in either the number or immunostaining intensity of ER-ir cells in the hippocampus, (5) the ER levels are down-regulated by estrogen in both male and female rats, and (6) the mean intensity of staining for the ER-ir cells in the hippocampus is about 25% of that in the ER-ir cells of the hypothalamus. From this, we can conclude that estrogen can have a direct effect on hippocampal neurons and that any sex differences in estrogen responsiveness is due to something other than sex differences in ER levels or function in the hippocampus.
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              A consideration of neural counting methods

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                Author and article information

                Journal
                NEN
                Neuroendocrinology
                10.1159/issn.0028-3835
                Neuroendocrinology
                S. Karger AG
                0028-3835
                1423-0194
                2003
                March 2003
                03 April 2003
                : 77
                : 3
                : 208-222
                Affiliations
                aLab. Biologia Reproducción Instituto de Investigaciones Biológicas, Universidad Veracruzana, Xalapa, Ver., bCentro de Investigación en Reproducción Animal, CINVESTAV-Universidad Autónoma de Tlaxcala, Ixtacuixtla, Tlax., México; cCenter for Molecular and Behavioral Neuroscience, Rutgers, The State University of New Jersey, Newark, N.J., USA
                Article
                69508 Neuroendocrinology 2003;77:208–222
                10.1159/000069508
                12673054
                © 2003 S. Karger AG, Basel

                Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher. Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug. Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

                Page count
                Figures: 7, Tables: 1, References: 82, Pages: 15
                Categories
                Cellular Neuroendocrinology

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