Molecular Human Reproduction, Vol. 5, No. 8, 742-747,
August 1999
© 1999 European Society of Human Reproduction and Embryology
Expression of oestrogen receptor-
and -ß in ovarian endometriomata
1 Department of Obstetrics and Gynecology, Gifu University School of Medicine, 40 Tsukasa-machi, Gifu City 500-8705, Japan
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Abstract |
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The contribution of oestrogen receptor (ER) isoforms, ER-
and ER-ß, in oestrogen-dependent development and growth of ovarian endometriomata, is unknown. Therefore, we examined the expression of ER- and ER-ß in ovarian endometriomata and normal uterine endometrium. ER- and ER-ß were shown to be dominantly expressed in the nuclei of the epithelial lining cells of ovarian endometrioma and of the glandular cells of normal uterine endometrium. ER-ß was expressed at a much lower level than ER- in the glandular cells of normal uterine endometrium, while ER-ß was expressed at a slightly lower level than ER- in the epithelial lining cells of ovarian endometrioma. In normal uterine endometrium, ER-ß mRNA was expressed at a much lower level than ER- mRNA, and the expression pattern of ER-ß mRNA during the menstrual cycle was similar to that of ER- mRNA. On the other hand, ER-ß mRNA expression was significantly higher and over a much greater range in ovarian endometriomata (P < 0.05) than in normal uterine endometrium during the menstrual cycle, while ER- mRNA expression was relatively lower and more random. Therefore, in ovarian endometriomata, oestrogen action via ER- cascades seems to be partially damaged, as the expression of ER- mRNA does not respond to endocrinological alterations during the menstrual cycle, while the relative over-expression of ER-ß might be related to a unique oestrogen-dependent growth and spreading of ovarian endometriomata.
ER-/ER-ß/ovarian endometrioma
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Introduction |
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It is well-known that endometriosis is an oestrogen-dependent disease, however the responsiveness of endometriosis to oestrogen is less stable than that of normal uterine endometrium. In endometrial tissue, diminished concentrations of oestrogen receptor (ER) were demonstrated by the oestrogen binding capacity and immunohistochemical antigen (Tamaya et al., 1979
; Janne et al., 1981; Gould et al., 1983; Bergqvist et al., 1985, 1993; Lessey et al., 1989; Nisolle et al., 1994). After ER- cDNA was cloned (Green et al., 1985, 1986; Ponglikitmongkol et al., 1988), the diminished expression of ER mRNA was also demonstrated in endometriosis (Misao et al., 1996). For example, ER exon 5 splicing variant contributes to spreading of endometriosis in dominant positive manner related to partial lack of oestrogen dependency (Fujimoto et al., 1997).
Although multiple forms had been identified for other members of the nuclear receptor superfamily, the existence of only one ER was believed for ~10 years following the cloning ER- cDNA (Parker, 1993; Mangelsdorf et al., 1995). Then novel rat ER-ß (rER-ß) (Kuiper et al., 1996) and human ER-ß (hER-ß) (Ogawa et al., 1998a; Messelman et al., 1996) were identified in cDNA libraries from rat prostate and human testis respectively. The rER-ß consists of 485 amino acids, and distinctly expresses in epithelial cells of prostate, granulosa cells of ovary (Kuiper et al., 1997), and osteoblastic cells of bone (Onoe et al., 1997). The hER-ß consists of 530 amino acids (Kuiper et al., 1997), and has a high affinity for oestradiol-17ß (Tremblay et al., 1997; Witkowska et al., 1997) and characteristics similar to ER- in specific binding to various oestrogenic substances and antagonists (Kuiper et al., 1997). Although the phosphorylation site for mitogen-activated protein kinase is conserved in the activation function (AF)-1 region of ER-ß as in ER- (Tremblay et al., 1997), both AF-1 and AF-2 regions of ER-ß are shorter than those of ER-. Furthermore, transcription at an AP1 element was inhibited by oestradiol and activated by anti-oestrogens via ER-ß cascades (Paech et al., 1997). This indicates some differences in the transcriptional efficiency and regulatory potential of the target genes. ER-ß is specifically expressed in testis, ovary, thymus, spleen (Messelman et al., 1996), osteoblasts (Arts et al., 1997) and fetus (Brandenberger et al., 1997).
Therefore, to study the contribution of ER isoforms ER- and ER-ß in oestrogen-dependent development and growth of ovarian endometriomata, we examined the expression of ER- and ER-ß and their mRNAs in ovarian endometriomata and the corresponding eutopic normal uterine endometrium.
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Materials and methods |
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Patients
Consent for the following studies was obtained from all patients and the Research Committee for Human Subjects, Gifu University School of Medicine. Patients (n = 20) aged 26–45 years underwent surgery for ovarian endometriosis and endometrial biopsy for the supply of normal uterine endometrium at the Department of Obstetrics and Gynecology, Gifu University School of Medicine, Japan, between March 1995 and October 1998. None of the patients had received any pre-operative therapy. The inner lining wall of the ovarian endometrioma was peeled and snap-frozen in liquid nitrogen. At least three parts of each wall of the ovarian endometrioma and eutopic normal uterine endometrium were collected and studied. Part of the uterine endometrium was submitted for endometrial dating (Noyes et al., 1950
), and oestradiol and progesterone in peripheral blood were measured to confirm the histological dating.
Immunohistochemistry
Sections (4 µm) were cut from formalin-fixed paraffin-embedded tissue with a microtome and dried overnight at 37°C on a silanized-slide (Dako, Carpinteria, USA). Samples were deparaffinized in xylene at room temperature for 80 min and washed with a graded ethanol/water mixture and then rinsed in distilled water. The samples for ER- and ER-ß were soaked in a citrate buffer and then microwaved at 100°C for 10 min. The protocol for Dako LSAB2 Kit, peroxidase (Dako) was followed for each sample. In the described procedure, rabbit anti-ER- (ER MC-20) [200 µg/ml, Santa Cruz, USA], and goat anti-ER-ß (ERß L-20) [200 µg/ml, Santa Cruz, USA] as the first antibodies were used at dilution of 1:100. The addition of the first antibody was omitted in the protocol for negative controls.
Preparation of internal standard recombinant RNA for competitive reverse transcription–polymerase chain reaction (RT–PCR) and Southern blot analysis
A scheme for the synthesis of internal standard recombinant RNA (rcRNA) is shown in Figure 1. DNA construction of the internal standard was originated and synthesized by PCR from a BamH/EcoRI fragment of V-erbB (Clontech Laboratories, Palo Alto, CA, USA) with two sets of oligonucleotide primer sequences. The sequences for the first set of primers for ER- mRNA (MIMIC ER- 5' and MIMIC ER- 3') and ER-ß mRNA (MIMIC ER-ß 5' and MIMIC ER-ß 3') in the first PCR were as follows:
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MIMIC ER-
5', 5'-ACAAGGGAAGTATGGCTATGCGCAAGTGAAATCTCCTCCG-3';
MIMIC ER- 3', 5'-CATCTCTCTGGCGCTTGTGTTCTGTCAATGCAGTTTGTAG-3';
MIMIC ER-ß 5', 5'-TGTTACTGGTCCAGGTTCAACGCAAGTGAAATCTCCTCCG-3';
MIMIC ER-ß 3', 5'-TCCTCTGTCTCCGCACAAGGTCTGTCAATGCAGTTTGTAG-3';
(Green et al., 1986; Vanden Heuvel et al., 1993; Mosselman et al., 1996).
The sequences for the second set of primers for ER- mRNA (MIMIC ER- P and MIMIC ER- 1049) and ER-ß mRNA (MIMIC ER-ß P and MIMIC ER-ß 559) in the second PCR were as follows:
MIMIC ER- P, 5'-TAATACGACTCACTATAGGACAAGGGAAGTATGGCTATG-3';
MIMIC ER- 1049, 5'-CATCTCTCTGGCGCTTGTGT-3';
MIMIC ER-ß P, 5'-TAATACGACTCACTATAGGTGTTACTGGTCCAGGTTCAA-3';
MIMIC ER-ß 559, 5'-TTCTCTGTCTCCGCACAAGG-3'.
The first PCR was constructed in a final volume of 50 µl containing PCR buffer (50 mmol/l, 10 mmol/l Tris–HCl, pH 8.3, 1.5 mmol/l MgCl2), 0.2 mmol/l deoxyribonucleoside triphosphates (dNTPs), 2 ng of BamH/EcoRI DNA fragment of V-erbB, 10 pmol each of the first set of PCR primers and 2.5 IU of Amplitaq DNA polymerase (Perkin-Elmer Cetus, Norwalk, CT, USA). The second PCR was conducted in a final volume of 100 µl containing PCR buffer, 0.2 mmol/l dNTPs, 50 pg of the first PCR products, 20 pmol each of the second PCR primers and 5 IU of Amplitaq DNA polymerase. The mixture was amplified for 28 cycles of PCR at 94°C for 45 s for denaturing, 55°C for 45 s for annealing and 72°c for 90 s or extension in a DNA Thermal Cycler (Perkin-Elmer Cetus).
The second PCR products were purified with a Gene Clean II kit (Bio 101 Inc, La Jolla, CA, USA) and transcribed using 100 IU of T7 RNA polymerase (Gibco BRL, Gaithersberg, MD, USA) in a final volume of 100 µl containing T3/T7 buffer [40 mmol/l Tris–HCl, pH 8.0, 8 mmol/l MgCl2, 2 mmol/l sepmidine-(HCl)3, 25 mmol/l NaCl], 0.1 mol/l dithiothreitol (DTT) 10 mmol/l ribonucleotide triphosphate, 40 IU RNAase inhibitor (Promega, Madison, WI, USA), 20 mmol/l template DNA and 10 µCi of [-32P]-UTP (New England Nucler Co, Boston, MA, USA), as a tracer. The reaction was incubated at 37°C for 1 h and then treated with 70 IU of RNAase-free DNAase (Takara Shuzo Co Ltd, Kyoto, Japan) at 37°C for 5 min to remove the DNA template. Subsequently, the products were extracted with water-saturated phenol/chloroform and passed through a Sephadex G50 column (Boehringer Mannheim, Mannheim, Germany). The amount of transcribed internal marker was calculated from total radioactivity of the transcribed RNA.
Competitive RT–PCR and Southern blot analysis
Total RNA was isolated from the tissues by the acid guanidium thiocyanate–phenol–chloroform method (Chomczynski and Sacchi, 1987). To obtain a standard curve each time, the total RNA (3 µg) and a series of diluted recombinant RNA for ER- mRNA (1 to 102 fmol) or ER-ß mRNA (10–2 to 1 fmol) were reverse-transcribed in 20 µl volume for 1 h at 37°C with a mixture of 200 IU Moloney murine leukaemia virus reverse transcriptase (MMLV–RTase, Gibco BRL) and the following reagents: 50 mmol/l Tris–HCl, pH 8.3, 75 mmol/l KCl, 3 mmol/l MgCl2, 40 IU RNAsin (Toyobo, Osaka, Japan), 10 mmol/l DTT, 0.5 mmol/l dNTPs and 30 pmol 3' end-specific primer as detailed below). The reaction was incubated for 5 min at 95°C to inactivate MMLV–RTase.
The sequences of primers to amplify the genes for ER- (ER- 740 and ER- 1049) and ER- (ER-ß 266 and ER-ß 559) were as follows:
ER- 740, 5'-ACAAGGGAAGTATGGTATG-3' (exon 2);
ER- 1049, 5'-CATCTCTCTGGCGCTTGTGT-3';
ER-ß 266, 5'-TGTTACTGGTCCAGGTTCAA-3';
ER-ß 559, 5'-TTCTCTGTCTCCGCACAAGG-3'.
The sizes of PCR products for ER- mRNA and its internal standard rcRNA were 309 and 440 bp respectively, and for ER-ß mRNA and its internal standard rcDNA, 293 and 440 bp respectively. PCR was carried out with reverse transcribed RNA as templates (1 µl) and 5 pmol of each specific primer using a DNA Thermal Cycler with 0.5 IU Amplitaq DNA polymerase in a buffer containing 50 mmol/l KCl, 10 mmol/l Tris–HCl, pH 8.3, 1.5 mmol/l MgCl2 and 0.2 mmol/l dNTP in 20 l volume. Amplification was performed for 38 cycles for ER- mRNA and 45 cycles for ER-ß mRNA at 94°C for 45 s for denaturing, 55°C for 45 s for annealing and 72°C for 90 s for extension.
Amplified PCR products (8 µl) were electrophoresed on a 1.2% agarose gel at 100 V. PCR products were capillary-transferred to an Immobilon transfer membrane (Millipore Corporation, Bedford, MA, USA) for 16 h. The membrane was dried at 80°C for 30 min and irradiated with UV to tightly fix the PCR products. The products on the membrane were prehybridized in a buffer of 1 mol/l NaCl, 50 mmol/l Tris–HCl, pH 7.6 and 1% sodium dodecyl sulphate at 42°C for 1 h and hybridized in the same solution with the biotinylated oligodeoxynucleotide probe (ER- DT, 5'-TGCTTCAGGCTACCATTATG-3' for ER- gene or ER-ß DT, 5'-TACGCATCGGGATATCACTA-3' for ER-ß gene), synthesized from the sequences of ER- or ER-ß cDNA between the specific primers, and the corresponding biotinylated internal standard gene-specific oligonucleotide probe (MIMIC-DT, 5'-GCAGATGAGTATCTTGTCCC-3') simultaneously to check gene specificity at 65°C overnight (Figure 2). They were also hybridized with the biotinylated ER- 5' or ER-ß 5' probe (10 pmol/ml) to determine the signal intensity under the same conditions (Figure 2). Specific bands hybridized with the biotinylated probes were detected with Plex Luminescent Kits (Millipore) and X-ray film was exposed on the membrane at room temperature for 10 min. Southern blotting quantification was carried out with BioImage (Millipore, Ann Arbor, MI, USA).
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In the competitive RT–PCR–Southern blot analysis for ER-
or ER-ß mRNA, only two predicted sizes of DNA fragments were detected with ER- DT or ER-ß DT and MIMIC-DT simultaneously to check their specificity and with ER- 5' or ER-ß 5' to determine their quantity respectively. As negative controls, neither ER- nor ER-ß bands were detected in samples without reverse transcription after 39 and 45 PCR cycles respectively. The levels of ER- and ER-ß mRNA were determined using a standard curve and a serial dilution of rcRNA in competitive RT–PCR–Southern blotting analysis (Figure 3).
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Statistical analysis
The amounts of ER-
and ER-ß mRNA were measured from three parts of the same tissue in triplicate. Statistical analysis was carried out using Student's t-test; P < 0.05 was considered to be statistically significant. |
Results |
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Immunohistochemical staining for oestrogen receptor (ER)-
and ER-ß revealed that ER- and ER-ß are expressed predominantly in the nuclei of the epithelial lining cells of all 20 ovarian endometriomata and of the glandular cells of all 20 normal uterine endometria, as shown in Figure 4. The intensity of immunohistochemical staining for ER-ß was much weaker than that for ER- in the glandular cells of all 20 normal uterine endometria, while the intensity for ER-ß was faintly weaker than for ER- in the epithelial lining cells of all 20 ovarian endometriomata (Figure 4).
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The levels of ER-ß mRNA were much lower than those of ER-
mRNA in ovarian endometriomata and normal uterine endometria (Figure 5). In normal uterine endometrium, the manner of ER-ß mRNA expression was similar to that of ER- mRNA during the menstrual cycle, as shown in Figure 5. On the other hand, ER-ß mRNA was expressed over a wider range and was significantly higher (P < 0.05) in ovarian endometriomata than in normal uterine endometrium, and ER- mRNA was relatively lower and randomly expressed (Figure 5). In addition, the ratio of ER-ß mRNA to ER- mRNA was stable in normal uterine endometrium, while it was over a wider range and was significantly higher (P < 0.05) in ovarian endometriomata than in normal uterine endometrium (Figure 5).
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Discussion |
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Whether ER-ß can compensate for ER-
functions is critical for understanding the regulatory system of oestrogen action in target organs. Although dysfunction of the genital tract was likely to occur only in female ER- knock-out mice, disruption of reproductive functional behaviour associated with sterility and reduction of bone density in both males and females has been observed (Lubahn et al., 1993; Korach, 1994). It was later demonstrated that ER- defects lead to abnormalities in spermatogenesis (Eddy et al., 1996) and sexual behaviour (Ogawa et al., 1997) in males and abnormalities in sexual behaviour in females (Rissman et al., 1997). The hER- defect disease has been discovered in only one man. His symptoms were unfused epiphyses in the knees, demineralized bone, low sperm viability, hyper-oestrogenaemia, slightly abnormal glucose tolerance, Acanthosis nigricans, etc. (Smith et al., 1994). As the ER- defect disease, including ER- knock-out state, has various symptoms, and ER-ß cannot compensate for ER- actions, ER-ß might not conserve the same physiological functions as ER-, and may play a role of co-function dependent on the presence of ER- (Couse et al., 1997). However, the different transcriptional status between ER- and ER-ß cascades with oestradiol and anti-oestrogens (Paech et al., 1997; Tremblay et al., 1997), and cross-inhibition of ER- and ER-ß signalling pathways have been demonstrated (Ogawa et al., 1998b).
In the present study, the expression of ER-ß was regulated in a similar fashion to that of ER- in normal uterine endometrium during the menstrual cycle, which implies that co-operation of ER- and ER-ß might lead to intact oestrogen action. Therefore, we assume that a characteristic stable ratio of ER-ß to ER- in each target organ may be necessary for intact oestrogen action via ER isoform cascades, and that ER-ß expression does not increase when ER- expression decreases in normal target organs. On the other hand, the expression of ER- mRNA was relatively lower in ovarian endometriomata than in normal uterine endometrium, and was not regulated in the same manner as in normal uterine endometrium during the menstrual cycle. Therefore, ER- cascades to support normal oestrogen-dependent growth and function might be damaged in ovarian endometriomata. The expression of ER-ß was relatively higher and wider in ovarian endometriomata than in normal uterine endometrium, and was not regulated in the same manner as in the normal uterine endometrium. Therefore, in ovarian endometriomata, ER-ß might fail to act with ER- for normal oestrogen dependency, which might thus be damaged. This endocrinological status might lead to unique oestrogen-dependent growth and spreading of ovarian endometriomata. Additionally, there is no evidence to indicate that endometriosis is a consequence or a cause of a change in ER isoform expression. We assume that typical endometriosis becomes established after transformation to an endometriotic lesion together with an alteration in the manner ER isoform expression.
In the future, to demonstrate the presence of functional ER isoforms and to understand the profound interaction of ER isoforms in target organs, introducing a reporter gene with oestrogen-responsive elements seems to be the most promising strategy (Hurst and Leslie, 1997).
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Notes |
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2 To whom correspondence should be addressed
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References |
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Arts, J., Kuiper, G.G., Janssen, J.M. et al. (1997) Differential expression of estrogen receptors
and ß mRNA during differentiation of human osteoblast SV-HFO cells. Endocrinology, 138, 5067–5070.Bergqvist, A., Jeppsson, S. and Ljungberg, O. (1985) Histochemical demonstration of estrogen and progesterone binding in endometriotic tissue and in uterine endometrium. J. Histochem. Cytochem., 33, 155–161.[Abstract]
Bergqvist, A., Ljungberg, O. and Skoog, L. (1993) Immunohistochemical analysis of estrogen and progesterone receptors in endometriotic tissue and endometrium. Hum. Reprod., 8, 1915–1922.
Brandenberger, A.W., Tee, M.K., Lee, J.Y. et al. (1997) Tissue distribution of estrogen receptors alpha (ER-) and beta (ER-ß) mRNA in the midgestational fetus. J. Clin. Endocrinol. Metab., 82, 3509–3512.
Chomczynski, P. and Sacci, N. (1987) Single-step method of RNA isolation by guanidium thiocyanate–phenol–chloroform extraction. Anal. Biochem., 162, 156–159.[ISI][Medline]
Couse, J.F., Lindzey, J., Crandien, K. et al. (1997) Tissue distribution and quantitative analysis of estrogen receptor- (ER) and estrogen receptor-ß (ER-ß) messenger ribonucleic acid in the wild-type and ER-knockout mouse. Endocrinology, 138, 4613–4621.
Eddy, E.M., Washburn, T.F., Bunch, D.O. et al. (1996) Targeted disruption of the estrogen receptor gene in male mice causes alteration of spermatogenesis and infertility. Endocrinology, 137, 4796–4805.[Abstract]
Fujimoto, J., Ichigo, S., Hirose, R. et al. (1997) Expression of estrogen receptor wild type and exon 5 splicing variant mRNAs in normal and endometriotic endometria during the menstrual cycle. Gynecol. Endocrinol., 11, 11–16.[ISI][Medline]
Gould, S.F., Shannon, J.M. and Cunha, G.R. (1983) Nuclear estrogen binding sites in human endometriosis. Fertil. Steril., 39, 520–524.[ISI][Medline]
Green, G.L., Gilna, P., Waterfield, M. et al. (1985) Sequence and expression of human estrogen receptor complementary DNA. Science, 231, 1150–1154.
Green, S., Walter, P., Kumar, V. et al. (1986) Human oestrogen receptor cDNA: sequence, expression and homology to v-erb-A. Nature, 320, 134–139.[Medline]
Hurst, B.S. and Leslie K.K. (1997) Uses of plasmid technologies. 1. The role of oestrogen in follicular development. Mol. Hum. Reprod., 3, 643–645.
Janne, O., Kauppila, A., Kokko, E. et al. (1981) Estrogen and progestin receptors in endometriosis lesions: comparison with endometrial tissue. Am. J. Obstet. Gynecol., 141, 562–566.[ISI][Medline]
Korach, K.S. (1994) Insight from the study of animals lacking functional estrogen receptor. Science, 266, 1524–1527.
Kuiper, G.G.J.M., Enmark, E., Pelto-Huikko, M. et al. (1996) Cloning of a novel estrogen receptor expressed in rat prostate and ovary. Proc. Natl Acad. Sci. USA, 93, 5925–5930.
Kuiper, G.G., Carlsson, B. and Grandien, J. (1997) Comparison of the ligand binding specificity and transcript tissue distribution of estrogen receptors and ß. Endocrinology, 138, 863–870.
Lessey, B.A., Metzger, D.A., Haney, A.F. et al. (1989) Immunohistochemical analysis of estrogen and progesterone receptors in endometriosis: comparison with normal endometrium during the menstrual cycle and the effect of medical therapy. Fertil. Steril., 51, 409–415.[ISI][Medline]
Lubahn, D.B., Moyer, J.S., Golding, T.S. (1993) Alternation of reproductive function but prenatal sexual development after insertional disruption of the mouse estrogen receptor gene. Proc. Natl Acad. Sci. USA, 90, 11162–11166.
Mangelsdorf, D.J., Thummel, C. and Beato, M. (1995) The nuclear receptor superfamily: the second decade. Cell, 83, 835–839.[ISI][Medline]
Messelman, S., Polman, J. and Dijkema, R. (1996) ERß: identification and characterization of a novel human estrogen receptor. FEBS Letts, 392, 49–53.[ISI][Medline]
Misao, R., Fujimoto, J., Nakanishi, Y. et al. (1996) Expression of estrogen and progesterone receptors and their mRNAs in ovarian endometriosis. Gynecol. Endocrinol., 10, 303–310.[ISI][Medline]
Misao, R., Nakanishi, Y., Sun, W.S. et al. (1999) Expression of oestrogen receptor and ß mRNA in corpus luteum of human subjects. Mol. Hum. Reprod., 5, in press.
Nisolle, M, de Menten, Y., Casanas-Roux, F. et al. (1994) Immunohistochemical analysis of estrogen and progesterone receptors in endometrium and peritoneal endometriosis: new quantitative method. Fertil. Steril., 62, 751–759.[ISI][Medline]
Noyes, R.W., Hertig, A.T. and Rock, J. (1950) Dating the endometrial biopsy. Fertil. Steril., 1, 3–5.
Ogawa, S., Lubahn, D.B., Korach, K.S. (1997) Behavioral effects of estrogen receptor gene disruption in male mice. Proc. Natl. Acad. Sci. USA, 94, 1476–1481.
Ogawa, S., Inoue, S., Watanabe, T. et al. (1998) The complete primary structure of human estrogen receptor ß (hERß) and its heterodimerization with ER in vivo and in vitro. Biochem. Biophys Res. Commun., 243, 122–126.[ISI][Medline]
Ogawa, S., Inoue, S., Orimo, A. et al. (1998) Cross-inhibition of both estrogen receptor alpha and beta pathways by each dominant negative mutant. FEBS Letts, 423, 129–132.[ISI][Medline]
Onoe, Y., Miyaura, C., Ohta, H. et al. (1997) Expression of estrogen receptor ß in rat bone. Endocrinology, 138, 4509–4512.
Paech, K., Webb, P., Kuiper, G.G. et al. (1997) Differential ligand activation of estrogen receptors ER alpha and ER beta at AP1 sites. Science, 277, 1508–1510.
Parker, M.G. (1993) Steroid and related receptors. Curr. Opin. Cell Biol., 5, 499–504.[Medline]
Ponglikitmongkol, M., Green, S. and Chambon, P. (1988) Genomic organization of the human oestrogen receptor gene. EMBO J., 11, 3385–3388.[ISI][Medline]
Rissman, E.F., Early, A.H., Taylor, J.A. et al. (1997) Estrogen receptors are essential for female sexual receptivity. Endocrinology, 138, 507–510.
Smith, E.P., Boyd, J., Frank, G. et al. (1994) Estrogen resistance caused by a mutation in the estrogen-receptor gene in a man. N. Engl. J. Med., 20, 1056–1061.
Tamaya, T., Motoyama, T., Ohno, Y. et al. (1979) Steroid receptor levels and histology of endometriosis and adenomyosis. Fertil. Steril., 31, 396–400.[ISI][Medline]
Tremblay, G.B., Tremblay, A., Copeland, N.G. et al. (1997) Cloning, chromosomal localization, and functional analysis of the murine estrogen receptor ß. Mol. Endocrinol., 11, 353–365.
Vanden Heuvel, J.P., Tyson, F.L. and Bell, D.A. (1993) Construction of recombinant RNA templates for use as internal standards in quantitative RT–PCR. Biotechniques, 14, 395–398.[ISI][Medline]
Witkowska, H.E., Carlquist, M., Engstrom, O. et al. (1997) Characterization of bacterially expressed rat estrogen receptor ß ligand binding domain by mass spectrometry: Structural comparison with estrogen receptor . Steroids, 62, 621–631.[ISI][Medline]
Submitted on February 8, 1999; accepted on May 11, 1999.
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