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Detection of estrogen receptor (ER) in the rat brain using rat anti-ER monoclonal IgG with the unlabeled antibody method

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Summary

Application of Sternberger's unlabeled antibody enzyme method for detection of the estrogen receptor (ER) using a rat primary antibody with rat tissues has been discouraged, presumably because nonspecific staining of endogenous IgG was expected with the required anti-rat IgG bridging antibody. Because the blood-brain barrier greatly reduces immunoglobulin infiltration into the brain, we hypothesized that rat brain tissue could be specifically immunostained using rat IgG primary antibodies. A rat monoclonal anti-ER antibody (H222) specifically stained ERs in the brains of ovariectomized but not in ovariectomized estrogen-treated rats. In contrast, the uterus, a well-perfused target organ stained intensely in a nonspecific fashion. Dense populations of estrogen receptors were observed in the medial preoptic area, the bed nucleus of the stria terminalis, and the arcuate and ventromedial nuclei of the hypothalamus. A monoclonal rat IgG directed against alpha-tubulin labeled primarily cortical dendrites quite distinct from the neuronal nuclei that are the primary antigenic sites for the estrogen receptor antibody. These results confirm that the sensitive unlabeled antibody method can be applied to rat brain tissues, even when the primary antibody is rat IgG and that labeling of endogenous IgG may be used as a simple method to evaluate the integrity of the blood brain barrier.

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References

  • Anderson J, Orntoft TF, Poulsen HS (1988) Immunohistochemical demonstration of estrogen receptors (ER) in formalin-fixed, paraffin-embedded human breast cancer tissue by use of a monoclonal antibody to ER. J Histochem Cytochem 36:1553–1560

    Google Scholar 

  • Brown TJ, MacLusky NJ, Toran-Allerand CD, Zeilinski JE, Hochberg RB (1989) Characterization of 11B-methoxy-16a[125I]iodoestradiol binding: neuronal localization of estrogen-binding sites in the developing rat brain. Endocrinology 124:2074–2088

    Google Scholar 

  • Cintra A, Fuxe K, Harfstrand A, Agnati LF, Miller LS, Greene JL, Gustafsson JA (1986) On the cellular localization and distribution of estrogen receptors in the rat tel- and diencephalon using monoclonal antibodies to human estrogen receptor. Neurochem Int 8:587–595

    Google Scholar 

  • Fuxe K, Cintra A, Agnati LF, Harfstrand A, Wikstrom AC, Okret S, Zoli M, Miller LS, Greene JL, Gustafsson JA (1987) Studies on the cellular localization and distribution of glucocorticoid receptor and estrogen receptor immunoreactivity in the central nervous system of the rat and their relationship to the monoaminergic and peptidergic neurons of the brain. J Steroid Biochem 27:159–170

    Google Scholar 

  • Gorski J, Welshons WV, Sakai D, Hansen J, Walent J, Kassis J, Shull J, Stack G, Campen C (1986) Evolution of a model of estrogen action. Recent Prog Horm Res 42:297–329

    Google Scholar 

  • Greene GL, Jensen EV (1982) Monoclonal antibodies as probes for estrogen receptor detection and characterization. J Steroid Biochem 16:353–359

    Google Scholar 

  • Greene GL, Nolan C, Engler JP, Jensen EV (1980) Monoclonal antibodies to human estrogen receptor. Proc Natl Acad Sci USA 77:5115–5119

    Google Scholar 

  • Greene GL, Sobel NB, King WJ, Jensen EV (1984) Immunochemical studies of estrogen receptors. J Steroid Biochem 20:51–56

    Google Scholar 

  • Hsu S, Raine L, Fanger H (1981) Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques. J Histochem Cytochem 29:577–580

    Google Scholar 

  • King RJB (1987) Structure and function of steroid receptors. J Endocrinol 114:341–349

    Google Scholar 

  • King WJ, Green GL (1984) Monoclonal antibodies localize oestrogen receptor. Nature 307:745–747

    Google Scholar 

  • King WJ, Desombre ER, Jensen EV, Greene GL (1985) Comparison of immunocytochemical and steroid-binding assays for estrogen receptor in human breast tumors. Cancer Res 45:293–304

    Google Scholar 

  • Koch M, Ehret G (1989) Immunocytochemical localization and quantitation of estrogen-binding cells in the male and female (virgin, pregnant, lactating) mouse brain. Brain Res 489:101–112

    Google Scholar 

  • Medlock KL, Sheehan DM, Branham WS (1981) The postnatal ontogeny of the rat uterine estrogen receptor. J Steroid Biochem 15:285–288

    Google Scholar 

  • Medlock KL, Lyttle CR, Kelepouris N, Newman ED, Sheehan DM (1991) Estradiol down-regulation of the rat uterine estrogen receptor. Poc Soc Exp Biol Med 196:293–300

    Google Scholar 

  • Pelletier G, Liao N, Follea N, Govindan MV (1988) Mapping of estrogen receptor-producing cells in the rat brain by in situ hybridization. Neurosci Lett 94:23–28

    Google Scholar 

  • Press MF, Greene GL (1984) An immunocytochemical method for demonstrating estrogen receptor in human uterus using monoclonal antibodies to human estrophilin. Lab Invest 50:480–486

    Google Scholar 

  • Press MF, Greene GL (1988) Immunocytochemical localization of estrogen and progesterone receptors. In: DeLellis RA (ed) Advances in immunohistochemistry. Raven Press, New York, pp 341–361

    Google Scholar 

  • Rapoport SI (1976) The blood-brain barrier in physiology and medicine. Raven Press, New York, p 98

    Google Scholar 

  • Scallet AC, Lipe GW, Ali SF, Holson RR, Frith CH, Slikker W Jr (1988) Neuropathological evaluation by combined immunohistochemistry and degeneration-specific methods: application to methylenedioxymethamphetamine. Neurotoxicology 9:529–538

    Google Scholar 

  • Sheehan DM, Branham WS, Gutierrez-Cernosek R, Cernosek SR Jr (1984) Effects of continuous estradiol administration by polydimethylsiloxane and paraffin implants on serum hormone levels and uterine responses. J Am Coll Toxicol 3:303–316

    Google Scholar 

  • Sternberger LA, Sternberger NH (1986) The unlabeled antibody method: comparison of peroxidase-antiperoxidase with avidinbiotin complex by a new method of quantification. J Histochem Cytochem 34:599–608

    Google Scholar 

  • Sternberger LA, Hardy PH Jr, Cuculis JJ, Meyer HG (1970) The unlabeled antibody enzyme method of immunohistochemistry. J Histochem Cytochem 13:315–333

    Google Scholar 

  • Stumpf WE, Sar M (1982) Histochemical approaches for the localization of steroid hormone receptors. Acta Histochem Cytochem 15:560–571

    Google Scholar 

  • Stumpf WE, Sar M, Keefer DA (1975) Atlas of estrogen target cells in rat brain. In: Stumpf WE, Grant LD (eds) Anatomical neuroendocrinology. Karger, Basel, pp 104–119

    Google Scholar 

  • Stumpf WE, Morin JK, Ennis BW, Zielinski JE, Hochberg RB (1987) Utility of [16a-125I] iodoestradiol for autoradiography for the study of cellular and regional distribution of receptors. J Histochem Cytochem 35:87–92

    Google Scholar 

  • Tse J, Goldfarb S (1988) Immunohistochemical demonstration of estrophilin in mouse tissues using a biotinylated monoclonal antibody. J Histochem Cytochem 36:1527–1531

    Google Scholar 

  • Umans RS, Weichselbau RR, Johnson CM, Little JB (1984) Effects of estradiol concentration on levels of nuclear estrogen receptors in MCF-7 breast tumor cells. J Steroid Biochem 1984; 20:605–609

    Google Scholar 

  • West NB, McClellan MC, Sternfeld MD, Brenner RM (1987) Immunocytochemistry versus binding assays of the estrogen receptor in the reproductive tract of spayed and hormone-treated macaques. Endocrinology 121:1789–1800

    Google Scholar 

  • Yamashita S, Korach KS (1989) A modified immunohistochemical procedure for the detection of estrogen receptor in mouse tissues. Histochemistry 90:325–330

    Google Scholar 

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Authors and Affiliations

  1. Division of Reproductive and Developmental Toxicology, Department of Health and Human Services, Food and Drug Administration, National Center for Toxicological Research, 72079, Jefferson, AR, USA

    W. W. Henry Jr., K. L. Medlock, D. M. Sheehan & A. C. Scallet

  2. Department of Biochemistry, University of Arkansas for Medical Sciences, 72205, Little Rock, AR, USA

    D. M. Sheehan

  3. Department of Pharmacology and Interdisciplinary Toxicology, University of Arkansas for Medical Sciences, 72205, Little Rock, AR, USA

    D. M. Sheehan & A. C. Scallet

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  1. W. W. Henry Jr.
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  2. K. L. Medlock
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  3. D. M. Sheehan
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  4. A. C. Scallet
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Henry, W.W., Medlock, K.L., Sheehan, D.M. et al. Detection of estrogen receptor (ER) in the rat brain using rat anti-ER monoclonal IgG with the unlabeled antibody method. Histochemistry 96, 157–162 (1991). https://doi.org/10.1007/BF00315987

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  • DOI: https://doi.org/10.1007/BF00315987

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