"GREEN FERTILITY": FEMALE FERTILITY AS A BIOSENSOR OF ECOSYSTEM POLLUTION

ABSTRACT PART I The first part of the thesis focuses on screening various compounds to evaluate their potential effects as endocrine disruptors and potential activators implementing human ovarian immortalized cell line hGL5, which shares characteristics with primary follicular granulosa cells. Utilizing hGL5 cells, we investigated the potential role of TAS2Rs in regulating steroidogenesis and delineated the molecular pathways influenced by different substances. The findings revealed that treatments with Biochanin A (BCA) and caffeine increased TAS2R14 abundance, altered mitochondrial function as indicated by mitochondrial footprint data. This was accompanied by a shift in steroid metabolism, with BCA promoting estrogen secretion while reducing progesterone production. Furthermore, BCA and caffeine influenced lipid droplet dynamics, suggesting their role in supporting granulosa cell differentiation and providing energy for follicle development. These attributes make hGL5 an ideal model for examining the effects of selected endocrine disrupting compounds (EDCs) on steroidogenesis. Building on the findings from hGL5 cells, the study further assessed the biological effects of the natural endocrine disruptor BCA on ovarian steroidogenesis in primary human granulosa cells. Extending these investigations to primary granulosa cells provided a clearer mechanism of BCA’s effects. By investigating TAS2Rs expression, we confirmed and validated the molecular changes induced by BCA, offering insights into the signal transduction mechanisms underlying this process. Our results provide a deeper understanding of the impact of endocrine disruptors on granulosa cell function, particularly the natural compound BCA. BCA activated TAS2Rs, leading to the upregulation of steroidogenic enzymes such as StAR and CYP17A1, increased estrogen production, and reduced progesterone levels. The use of TAS2R antagonists confirmed the essential role of these receptors in mediating the observed effects. Collectively, these experiments emphasize the significance of TAS2Rs in regulating steroidogenesis and mitochondrial function in granulosa cells, offering critical insights into how natural endocrine disruptors like BCA affect female fertility.