Opposite Roles of Estrogen Receptor (ER)- and ERβ in the Modulation of Luteinizing Hormone Responses to Kisspeptin in the Female Rat: Implications for the Generation of the Preovulatory Surge

Supplemental Data

Supplemental Data

Files in this Data Supplement:

• Appendix A - Pharmacological and pharmacokinetic characterization of the Organon proprietary compounds, with activity as ERα agonist (ERA-45), ERα antagonist (ERA-90), ERβ agonist (ERB-79), ERβ antagonist (ERB-88), or PR antagonist (Org 31710), used in this study.
• Appendix B - Uterus weight and basal (<12:00 proestrus) serum LH levels in cyclic female rats treated s.c. with vehicle, ERα antagonist (aERα), ERβ antagonist (aERβ) or PR antagonist (aPR), as described in Materials and Methods for Experiments 1 and 2 (acute blockade). For each group, the animals were divided in three groups that received vehicle, kisspeptin-10 (K) or GnRH (G) at 12:00 proestrus. Groups with different superscript letters are statistically different (P<0.01; ANOVA followed by Student-Newman-Keuls multiple range test).
• Appendix C - Ovulatory rates (determined at termination of experiments at estrus) in female rats pre-treated s.c. with vehicle, ERα antagonist (aERα) or ERβ antagonist (aERβ), and injected at 12:00 proestrus with vehicle, kisspeptin-10 (Kp10) or GnRH, as described for Experiments 1 and 2. In addition to quantitative data (mean ± SEM; upper panel), representative photomicrographs from ovarian and oviductal sections of relevant experimental groups are shown. While discernible formation of corpora lutea and released oocytes at the oviduct were detectable in vehicle- and aERβ-treated groups, in female rats pre-treated with aERα only preovulatory follicles (but not ovulation) were detected in those animals injected with vehicle or Kp10. In clear contrast, GnRH injection to aERα-treated rats rescued ovulation, with a mean number of released oocytes per ovary similar to that of control animals.
• Appendix D - Uterus weight and basal (<12:00 proestrus) serum LH levels in female rats treated s.c. for one complete cycle with vehicle, ERα antagonist (aERα) or ERβ antagonist (aERβ), as described in Materials and Methods for Experiment 3 (chronic blockade). For each group, the animals were divided in two groups that received either vehicle (V) or kisspeptin-10 (Kp) at 12:00 proestrus (last day of treatment). Groups with different superscript letters are statistically different (P<0.01; ANOVA followed by Student-Newman-Keuls multiple range test).
• Appendix E - Ovulatory rates (determined at termination of experiments at estrus) in female rats pre-treated s.c. for one complete cycle with vehicle, ERα antagonist (aERα) or ERβ antagonist (aERβ), and injected at 12:00 proestrus with vehicle or kisspeptin-10 (Kp10), as described for Experiment 3. In addition to quantitative data (mean ± SEM; upper panel), representative photomicrographs from ovarian and oviductal sections of relevant experimental groups are shown. While discernible formation of corpora lutea and released oocytes at the oviduct were detectable in vehicle- and aERβ-treated groups, in female rats pre-treated with aERα only preovulatory follicles (but not ovulation) were detected in those animals injected with vehicle or Kp10 (data not shown).
• Appendix F - Uterus weight and basal serum LH levels in OVX female rats treated s.c. for 4-days with vehicle (V), ERα agonist (α), ERβ agonist (β), progesterone (P), or their combination. In addition, data from cyclic female rats at diestrus-1 (D1) are presented as reference controls. All the animals were injected with kisspeptin-10 4-h before termination of experiments and tissue (uterus) sampling. Hormonal data are presented for basal (pre-injection) levels. Groups with different superscript letters are statistically different (P<0.01; ANOVA followed by Student-Newman-Keuls multiple range test).