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Analysis of the Ovarian Marker Genes Expression Revealed the Antagonistic Effects of Serotonin and Androstenedione on the Functional State of Mouse Granulosa Cells in Primary Culture

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Abstract

Primary culture of granulosa cells is a prerequisite for a complete study of the normal functioning of the ovary and its pathologies. In this work, we selected the optimal protocol for obtaining a primary culture of mouse granulosa cells in the most functionally active state and revealed the effects of androstenedione and serotonin on the expression of ovarian markers that reflect the functional status of granulosa cells. The morpho-functional analysis of the ovary after PMSG stimulation revealed that 48 hours after PMSG stimulation is the optimal time for obtaining granulosa cells in the most active functional state. Using the set of 14 ovarian functional state marker genes we reveal that androstenedione inhibits cumulus and immature granulosa markers but stimulates genes characteristic of the mature state of granulosa. At the same time, granulosa cells express serotonergic receptors and transporter SERT. The ovarian marker genes expression analysis revealed that serotonin affects the expression of genes characterizing the differentiation of granulosa cells towards cumulus cells. Summarizing, we can conclude that serotonin and androstenedione have an antagonistic effect on the functional state of mouse granulosa cells in primary culture in vitro.

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ACKNOWLEDGMENTS

The study was carried out using equipment of the Core Centrum of Institute of Developmental Biology RAS.

Funding

In the part that regards the selection of optimal conditions for obtaining primary culture, the research was funded by the Government Program for Basic Research in the Koltzov Institute of Developmental Biology of the Russian Academy of Sciences in 2023 no. 0088-2021-0009 for DN, NA, LM and YN. In the part that regards the role of serotonin, the research was funded by Russian Science Foundation, grant no. 22-74-10009, for DN, NA, YN, VK and AE.

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Authors and Affiliations

  1. Koltzov Institute of Developmental Biology, Russian Academy of Sciences, Moscow, Russia

    N. M. Alyoshina, L. A. Malchenko, Yu. O. Nikishina & D. A. Nikishin

  2. Biological Faculty, Moscow State University, Moscow, Russia

    V. R. Rousanova, Yu. V. Khramova, V. V. Konduktorova, A. Y. Evstifeeva & D. A. Nikishin

Authors
  1. N. M. Alyoshina
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  2. V. R. Rousanova
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  3. L. A. Malchenko
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  4. Yu. V. Khramova
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  5. Yu. O. Nikishina
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  6. V. V. Konduktorova
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  7. A. Y. Evstifeeva
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  8. D. A. Nikishin
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Contributions

Conceptualization, DN; methodology, DN and YN; software, AE; investigation, NA, LM, VR, YK, YN, VK; writing—original draft preparation, NA; writing—review and editing, DN; visualization, DN; supervision, DN; project administration, DN; funding acquisition, DN. All authors have read and agreed to the published version of the manuscript.

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Correspondence to D. A. Nikishin.

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Conflict of interest. The authors declare no conflict of interest.

Statement on the welfare of animals. Experiments were performed in accordance with the Council of the European Communities Directive of November 24, 1986 (86/609/EEC). All protocols of animal experiments were approved by the Commission on Bioethics of the Koltzov Institute of Developmental Biology of the Russian Academy of Sciences (project identification code: no. 37, date: June 25, 2020).

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Alyoshina, N.M., Rousanova, V.R., Malchenko, L.A. et al. Analysis of the Ovarian Marker Genes Expression Revealed the Antagonistic Effects of Serotonin and Androstenedione on the Functional State of Mouse Granulosa Cells in Primary Culture. Russ J Dev Biol 54, 165–176 (2023). https://doi.org/10.1134/S1062360423030025

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  • DOI: https://doi.org/10.1134/S1062360423030025

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