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Effect of Anti-Müllerian hormone (AMH) and bone morphogenetic protein 15 (BMP-15) on steroidogenesis in primary-cultured human luteinizing granulosa cells through Smad5 signalling

  • Reproductive Physiology and Disease
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

To determine if there is any effect of AMH and BMP-15 on estradiol and progesterone production from primary-cultured human luteinizing granulosa cells, to delineate what is the effect of FSH on their actions and which are the possible mechanisms involved.

Methods

Luteinizing granulosa cells (GCs), obtained from follicular fluid of 30 women undergoing in vitro fertilization, were cultured, after a short 24-h preincubation period, in serum-free medium for 24 or/and 48 h in the presence/absence of various concentrations of AMH, BMP-15 and FSH alone or in combinations. Estradiol and progesterone production, SMAD5 phosphorylation and StAR expression were studied in parallel. Steroids were measured in culture-supernatant using enzyme-immunoassays, while Smad5-signaling pathway activation and StAR protein expression were assessed immunocytochemically.

Result(s)

We found that the treatment of AMH in GCs for 24/48 h attenuated FSH-induced estradiol production (p < 0.001), had no effect on basal estradiol levels, decreased basal progesterone production (p < 0.001) and FSH-induced StAR expression (p < 0.001). On the other hand, BMP-15 decreased basal estradiol levels (p < 0.001) and attenuated FSH-induced estradiol production (p < 0.001). Furthermore, BMP-15 reduced progesterone basal secretion (p < 0.001), an effect that was partially reversed by FSH (p < 0.01), probably via increasing StAR expression (p < 0.001). FSH-induced StAR expression was also attenuated by BMP-15 (p < 0.001). FSH, AMH and BMP-15 activated Smad-signaling pathway, as confirmed by the increase of phospo-Smad5 protein levels (p < 0.001 compared to control).

Conclusion(s)

AMH and BMP-15 by interacting with FSH affect the production of estradiol and progesterone from cultured luteinizing-granulosa cells possibly via Smad5-protein phosphorylation.

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Acknowledgments

We wish to thank Dr Panagiotis Liakos, Assistant Professor in Department of Biochemistry in University of Thessaly and Anastasia Tzavella for their laboratory assistance.

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Authors and Affiliations

  1. Department of Obstetrics and Gynaecology, School of Health Sciences, Faculty of Medicine, University of Thessaly, 41110, Viopolis, Larissa, Greece

    Ermioni Prapa, Konstantinos Dafopoulos, Eleni Katsiani, Christina I Messini, George Anifandis & Ioannis E Messinis

  2. Department of Pharmacology, School of Health Sciences, Faculty of Medicine, University of Thessaly, Larissa, Greece

    Anna Vasilaki

  3. Department of Nuclear Medicine, School of Health Sciences, Faculty of Medicine, University of Thessaly, Larissa, Greece

    Panagiotis Georgoulias

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  1. Ermioni Prapa
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  2. Anna Vasilaki
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  3. Konstantinos Dafopoulos
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  4. Eleni Katsiani
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  5. Panagiotis Georgoulias
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  6. Christina I Messini
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  8. Ioannis E Messinis
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Corresponding author

Correspondence to Ioannis E Messinis.

Additional information

Capsule AMH and BMP-15 interact with FSH and affect the production of estradiol and progesterone from cultured luteinized granulosa cells possibly via Smad5-protein phosphorylation.

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Prapa, E., Vasilaki, A., Dafopoulos, K. et al. Effect of Anti-Müllerian hormone (AMH) and bone morphogenetic protein 15 (BMP-15) on steroidogenesis in primary-cultured human luteinizing granulosa cells through Smad5 signalling. J Assist Reprod Genet 32, 1079–1088 (2015). https://doi.org/10.1007/s10815-015-0494-2

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