GnRH Antagonists Produce Differential Modulation of the Signaling Pathways Mediated by GnRH Receptors
Abstract
:1. Introduction
2. Results
2.1. Gonadotropin Releasing Hormone (GnRH) Antagonist-Induced Inhibition of Intracellular Ca2+ Increase
2.2. Assessment of cAMP Accumulation
2.3. Evaluation of pERK1/2 and pCREB Activation
2.4. Analysis of β-Catenin Activation and Localization
2.5. Analysis of GnRH-Induced Lhb Gene Expression
3. Discussion
4. Materials and Methods
4.1. Materials
4.2. Cell Culture
4.3. Transfection Methods
4.4. BRET Measurement of Intracellular Ca2+ Increase and cAMP Production
4.5. Western Blotting Analysis
4.6. Immunofluorescence Analysis
4.7. Lhb Gene Expression
4.8. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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No Antagonist | 109,340 ± 13,866 | ||
---|---|---|---|
Cetrorelix | Ganirelix | Teverelix | |
1 pM | 78,113 ± 29,979 | 76,189 ± 18,564 | 85,301 ± 21,321 |
10 pM | 124,981 ± 33,684 | 91,896 ± 21,766 | 98,939 ± 27,373 |
100 pM | 126,243 ± 34,982 | 101,943 ± 27,388 | 58,864 ± 27,101 |
1 nM | 71,127 ± 25,268 | 66,665 ± 16,035 | 81,702 ± 18,920 |
10 nM | 21,482 ± 6718 * | 73,164 ± 16,237 | 74,321 ± 17,569 |
75 nM | 18,632 ± 4423 * | 43,870 ± 4193 * | 42,401 ± 8974 * |
100 nM | 8946 ± 3296 * | 34,734 ± 13,731 * | 29,931 ± 8731 * |
1 µM | 3264 ± 3024 * | 3346 ± 2977 * | 4107 ± 3804 * |
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Sperduti, S.; Limoncella, S.; Lazzaretti, C.; Paradiso, E.; Riccetti, L.; Turchi, S.; Ferrigno, I.; Bertacchini, J.; Palumbo, C.; Potì, F.; et al. GnRH Antagonists Produce Differential Modulation of the Signaling Pathways Mediated by GnRH Receptors. Int. J. Mol. Sci. 2019, 20, 5548. https://doi.org/10.3390/ijms20225548
Sperduti S, Limoncella S, Lazzaretti C, Paradiso E, Riccetti L, Turchi S, Ferrigno I, Bertacchini J, Palumbo C, Potì F, et al. GnRH Antagonists Produce Differential Modulation of the Signaling Pathways Mediated by GnRH Receptors. International Journal of Molecular Sciences. 2019; 20(22):5548. https://doi.org/10.3390/ijms20225548
Chicago/Turabian StyleSperduti, Samantha, Silvia Limoncella, Clara Lazzaretti, Elia Paradiso, Laura Riccetti, Sara Turchi, Ilaria Ferrigno, Jessika Bertacchini, Carla Palumbo, Francesco Potì, and et al. 2019. "GnRH Antagonists Produce Differential Modulation of the Signaling Pathways Mediated by GnRH Receptors" International Journal of Molecular Sciences 20, no. 22: 5548. https://doi.org/10.3390/ijms20225548