Determine estrogenic activity of environmental mixture and individual EDCs
Environmental samples belonging to different matrices (wastewater, soil) as well as selected food samples such as grains will be used as potential source of estrogenic EDCs. Samples will be subjected to extraction procedure to obtained total extracts, and estrogenic potency of the mixture and BPA will be tested in the ER-CALUX bioassay using E2 standard curve. Results from bioassay will be used to create experimental groups with different estrogenic activity and tested for potency to disrupt ovarian signaling and function.
Identify estrogenic EDC-sensitive signaling and follicular activation in developing ovary
Our working hypothesis is that exposure to estrogenic EDCs increases activity of the AKT signaling triggering premature activation of the primordial follicle pool. This hypothesis will be tested by exposure of neonatal ovaries to the estrogenic mixture and BPA followed by analysis of the AKT signaling and follicular activation in developing ovary.
Determine estrogenic EDC-sensitive signaling and luteinisation process in granulosa cells
Our working hypothesis is that direct exposure to estrogenic EDCs increase functional activity of the ERK signaling cascade triggering luteinisation of the GCs. Activation of ERK and transcriptional factor C/EBPβ will be assessed in the isolated immature GCs after in vitro exposure to estrogenic mixture and BPA.