Toxicological Sciences 54, 154-167 (2000)
Copyright © 2000 by the Society of Toxicology
Estrogenic Activity of Octylphenol, Nonylphenol, Bisphenol A and Methoxychlor in Rats
Endocrinology Branch, Reproductive Toxicology Division, National Health and Environmental Effects Research Laboratory, U.S. Environmental Protection Agency, Research Triangle Park, North Carolina 27711
Considerable attention has recently been focused on environmental chemicals that disrupt the reproductive system by altering steroid receptor function. Although numerous in vitro and in vivo methods have been shown to be useful approaches for identifying chemicals that can disrupt reproduction through a direct interaction with the estrogen receptor, it is imperative that the protocols selected be capable of detecting chemicals with a broad range of estrogenic activity. Here we evaluate the reliability of the 3-day uterotrophic assay for detecting chemicals with strong or weak estrogenic activity in both prepubertal and ovariectomized adult Long Evans rats. These data were compared to additional measures of estrogenic activity, which included the age of vaginal opening, the induction of cornified vaginal epithelial cells in ovariectomized adult rats, and estrous cyclicity in intact adult rats. Test chemicals selected for these studies included 17-ß-estradiol, ethynyl estradiol, methoxychlor, 4-tert-octylphenol, 4-nonylphenol and bisphenol A. Data from in vitro receptor binding assays compared the ability of the test chemicals to compete with [3H]-estradiol or [3H]-promegestone for binding to estrogen or progesterone receptors. As expected, the binding affinities for the estrogen receptor ranged from high to low, as reflected by Ki concentrations of 0.4 nM for 17-ß-estradiol and ethynyl estradiol, and 0.05–65 µM for 4-tert-octyphenol, 4-nonylphenol, and methoxychlor. Although none of the test chemicals demonstrated a high affinity for binding to the progesterone receptor, 4-tert-octylphenol and 4-nonylphenol exhibited a weak affinity, with Ki concentrations ranging from 1.2 to 3.8 µM. In vivo studies indicated that the 3-day uterotrophic assay in prepubertal rats was the best method for detecting estrogenic activity when compared with all other end points, based upon the dose-response data for ethynyl estradiol (0.01–0.1 mg/kg), 4-tert-octylphenol (50–200 mg/kg, oral), and 4-nonylphenol (25–100 mg/kg, oral). Although oral doses of ethynyl estradiol (0.01 mg/kg) and 4-nonylphenol (50 mg/kg) induced a significant increase in uterine weight in the prepubertal rats, these doses were ineffective for stimulating a similar response in ovariectomized adult rats. The age of vaginal opening was advanced following oral exposure from postnatal days 21–35 to ethynyl estradiol (0.01 mg/kg), methoxychlor (50 mg/kg), 4-tert-octylphenol (200 mg/kg), and 4-nonylphenol (50 mg/kg). Although bisphenol A (200 mg/kg, oral) induced a significant uterotrophic response within 3 days in prepubertal rats, doses up to 400 mg/kg failed to advance the age of vaginal opening. Monitoring changes in the vaginal epithelium of ovariectomized adult rats was the least effective method for detecting estrogenic activity for 4-tert-octylphenol and bisphenol A. The number of 4–5 day estrous cycles was reduced during a 25-day exposure to ethynyl estradiol (0.01 mg/kg), methoxychlor (50 mg/kg), 4-tert-octylphenol (200 mg/kg), 4-nonylphenol (100 mg/kg), and bisphenol A (100 mg/kg) by oral gavage. Although long periods of extended diestrus (7–14 days) were generally correlated with exposure to ethynyl estradiol and 4-tert-octylphenol, the cycling patterns following exposure to methoxychlor, 4-nonylphenol and bisphenol A were not as clearly defined, with shorter periods of extended diestrus (4–7 days) and/or estrus (3–5 days) intermittently observed throughout the exposure period. Together these data provide a comparison of the 3-day uterotrophic assay with alternative measures of estrogenic activity for a group of test chemicals with a broad range of affinities for the estrogen receptor. These data can be useful during the assessment and validation of methods for screening environmental chemicals for endocrine disrupting activity.
Key Words: female reproductive toxicology; endocrine disruptors; ethynyl estradiol; 4-tert-octylphenol; 4- nonylphenol; bisphenol A; methoxychlor; rat; uterotrophic assay; estrous cyclicity; vaginal opening assay.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  W Karmaus, J R Osuch, I Eneli, L M Mudd, J Zhang, D Mikucki, P Haan, and S Davis Maternal levels of dichlorodiphenyl-dichloroethylene (DDE) may increase weight and body mass index in adult female offspring Occup. Environ. Med., March 1, 2009; 66(3): 143 - 149. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M.-J. Lee, H. Lin, C.-W. Liu, M.-H. Wu, W.-J. Liao, H.-H. Chang, H.-C. Ku, Y.-S. Chien, W.-H. Ding, and Y.-H. Kao Octylphenol stimulates resistin gene expression in 3T3-L1 adipocytes via the estrogen receptor and extracellular signal-regulated kinase pathways Am J Physiol Cell Physiol, June 1, 2008; 294(6): C1542 - C1551. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Miyawaki, S. Kamei, K. Sakayama, H. Yamamoto, and H. Masuno 4-Tert-Octylphenol Regulates the Differentiation of C3H10T1/2 Cells into Osteoblast and Adipocyte Lineages Toxicol. Sci., March 1, 2008; 102(1): 82 - 88. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Caserta, L. Maranghi, A. Mantovani, R. Marci, F. Maranghi, and M. Moscarini Impact of endocrine disruptor chemicals in gynaecology Hum. Reprod. Update, January 1, 2008; 14(1): 59 - 72. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Sabo-Attwood, J. L Blum, K. J Kroll, V. Patel, D. Birkholz, N. J Szabo, S. Z Fisher, R. McKenna, M. Campbell-Thompson, and N. D Denslow Distinct expression and activity profiles of largemouth bass (Micropterus salmoides) estrogen receptors in response to estradiol and nonylphenol J. Mol. Endocrinol., October 1, 2007; 39(4): 223 - 237. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. K. Kuester and I. G. Sipes Prediction of Metabolic Clearance of Bisphenol A (4,4 '-Dihydroxy-2,2-diphenylpropane) using Cryopreserved Human Hepatocytes Drug Metab. Dispos., October 1, 2007; 35(10): 1910 - 1915. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Rasier, A.-S. Parent, A. Gerard, M.-C. Lebrethon, and J.-P. Bourguignon Early Maturation of Gonadotropin-Releasing Hormone Secretion and Sexual Precocity after Exposure of Infant Female Rats to Estradiol or Dichlorodiphenyltrichloroethane Biol Reprod, October 1, 2007; 77(4): 734 - 742. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. K. Gupta, G. Aberdeen, J. K. Babus, E. D. Albrecht, and Jodi. A. Flaws Methoxychlor and Its Metabolites Inhibit Growth and Induce Atresia of Baboon Antral Follicles Toxicol Pathol, August 1, 2007; 35(5): 649 - 656. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. P. Hernandez, W. Huang, L. M. Chapman, S. Chua, D. D. Moore, and W. S. Baldwin The Environmental Estrogen, Nonylphenol, Activates the Constitutive Androstane Receptor Toxicol. Sci., August 1, 2007; 98(2): 416 - 426. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. H. Dang, T. H. Nguyen, K.-C. Choi, and E.-B. Jeung A Calcium-Binding Protein, Calbindin-D9k, Is Regulated through an Estrogen-Receptor Mediated Mechanism following Xenoestrogen Exposure in the GH3 Cell Line Toxicol. Sci., August 1, 2007; 98(2): 408 - 415. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Savabieasfahani, K. Kannan, O. Astapova, N. P. Evans, and V. Padmanabhan Developmental Programming: Differential Effects of Prenatal Exposure to Bisphenol-A or Methoxychlor on Reproductive Function Endocrinology, December 1, 2006; 147(12): 5956 - 5966. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. C. Laws, S Yavanhxay, R. L. Cooper, and J. C. Eldridge Nature of the Binding Interaction for 50 Structurally Diverse Chemicals with Rat Estrogen Receptors Toxicol. Sci., November 1, 2006; 94(1): 46 - 56. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. D. Anway, M. A. Memon, M. Uzumcu, and M. K. Skinner Transgenerational Effect of the Endocrine Disruptor Vinclozolin on Male Spermatogenesis J Androl, November 1, 2006; 27(6): 868 - 879. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. P. Miller, R. K. Gupta, and J. A. Flaws Methoxychlor Metabolites May Cause Ovarian Toxicity Through Estrogen-Regulated Pathways Toxicol. Sci., September 1, 2006; 93(1): 180 - 188. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Daidoji, M. Ozawa, H. Sakamoto, T. Sako, H. Inoue, R. Kurihara, S. Hashimoto, and H. Yokota SLOW ELIMINATION OF NONYLPHENOL FROM RAT INTESTINE Drug Metab. Dispos., January 1, 2006; 34(1): 184 - 190. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. C. Kwekel, L. D. Burgoon, J. W. Burt, J. R. Harkema, and T. R. Zacharewski A cross-species analysis of the rodent uterotrophic program: elucidation of conserved responses and targets of estrogen signaling Physiol Genomics, November 17, 2005; 23(3): 327 - 342. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Can, O. Semiz, and O. Cinar Bisphenol-A induces cell cycle delay and alters centrosome and spindle microtubular organization in oocytes during meiosis Mol. Hum. Reprod., June 1, 2005; 11(6): 389 - 396. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Aravindakshan and D. G. Cyr Nonylphenol Alters Connexin 43 Levels and Connexin 43 Phosphorylation Via an Inhibition of the p38-Mitogen-Activated Protein Kinase Pathway Biol Reprod, May 1, 2005; 72(5): 1232 - 1240. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y.-W. Jung, E.-J. Hong, K.-C. Choi, and E.-B. Jeung Novel Progestogenic Activity of Environmental Endocrine Disruptors in the Upregulation of Calbindin-D9k in an Immature Mouse Model Toxicol. Sci., January 1, 2005; 83(1): 78 - 88. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E.-J. Hong, K.-C. Choi, and E.-B. Jeung Induction of Calbindin-D9k Messenger RNA and Protein by Maternal Exposure to Alkylphenols During Late Pregnancy in Maternal and Neonatal Uteri of Rats Biol Reprod, August 1, 2004; 71(2): 669 - 675. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. S. Cupp, M. Uzumcu, H. Suzuki, K. Dirks, B. Phillips, and M. K. Skinner Effect of Transient Embryonic In Vivo Exposure to the Endocrine Disruptor Methoxychlor on Embryonic and Postnatal Testis Development J Androl, September 1, 2003; 24(5): 736 - 745. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Daidoji, H. Inoue, S. Kato, and H. Yokota GLUCURONIDATION AND EXCRETION OF NONYLPHENOL IN PERFUSED RAT LIVER Drug Metab. Dispos., August 1, 2003; 31(8): 993 - 998. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Pocar, R. Augustin, F. Gandolfi, and B. Fischer Toxic Effects of In Vitro Exposure to p-tert-Octylphenol on Bovine Oocyte Maturation and Developmental Competence Biol Reprod, August 1, 2003; 69(2): 462 - 468. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Wright, A.C.O. Evans, N.P. Evans, P. Duffy, J. Fox, M.P. Boland, J.F. Roche, and T. Sweeney Effect of Maternal Exposure to the Environmental Estrogen, Octylphenol, During Fetal and/or Postnatal Life on Onset of Puberty, Endocrine Status, and Ovarian Follicular Dynamics in Ewe Lambs Biol Reprod, December 1, 2002; 67(6): 1734 - 1740. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. W. Tyl, C. B. Myers, M. C. Marr, B. F. Thomas, A. R. Keimowitz, D. R. Brine, M. M. Veselica, P. A. Fail, T. Y. Chang, J. C. Seely, et al. Three-Generation Reproductive Toxicity Study of Dietary Bisphenol A in CD Sprague-Dawley Rats Toxicol. Sci., July 1, 2002; 68(1): 121 - 146. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. B. Wetherill, C. E. Petre, K. R. Monk, A. Puga, and K. E. Knudsen The Xenoestrogen Bisphenol A Induces Inappropriate Androgen Receptor Activation and Mitogenesis in Prostatic Adenocarcinoma Cells Mol. Cancer Ther., May 1, 2002; 1(7): 515 - 524. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Masuyama, H. Inoshita, Y. Hiramatsu, and T. Kudo Ligands Have Various Potential Effects on the Degradation of Pregnane X Receptor by Proteasome Endocrinology, January 1, 2002; 143(1): 55 - 61. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. D. Papaconstantinou, B. R. Fisher, T. H. Umbreit, P. L. Goering, N. T. Lappas, and K. M. Brown Effects of {beta}-Estradiol and Bisphenol A on Heat Shock Protein Levels and Localization in the Mouse Uterus Are Antagonized by the Antiestrogen ICI 182,780 Toxicol. Sci., October 1, 2001; 63(2): 173 - 180. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Elsby, J. L. Maggs, J. Ashby, and B. K. Park Comparison of the Modulatory Effects of Human and Rat Liver Microsomal Metabolism on the Estrogenicity of Bisphenol A: Implications for Extrapolation to Humans J. Pharmacol. Exp. Ther., April 1, 2001; 297(1): 103 - 113. [Abstract] [Full Text]
|
|