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

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.