Evaluation of the high-dose fsh effect on mice granulosa cells with hippo signal pathway proteins (YAP, TEAD and p73) and microRNAs (miR129-5p and miR145-5p)

Recombinant gonadotropins are widely used in assisted reproductive techniques for the development of many follicles. However, in cases requiring excessive FSH (follicle-stimulating hormone) use, follicle development is also impaired. Hormones, growth factors, or miRNAs in ovaries control cell proliferation and homeostasis by regulating the Hippo pathway. While TEAD/YAP genes in the pathway allow cells to survive; YAP/p73 controls progression to apoptosis. In this study, TEAD3/YAP1 expressions which also associated miR-129-5p and p73 expression which also associated miR-145-5p levels were investigated. The expressions of the casp3 and bax apoptosis markers were also evaluated for confirmation. It was aimed to reveal the effect of the Hippo pathway on granulosa cells after high-dose FSH administration and link with miRNA reflections. Eighteen, 6-8 w Balb/c female mice were divided into three groups including control, sham, and FSH groups. FSH group were injected by 10-10-5-5 IU/ml FSH respectively, every 12 hours, 4 doses total. The same amount of saline was applied to sham group. The animals were sacrificed 12 hr after the last injection. Ovary tissues and blood were taken for analysis by RT-PCR and immunohistochemistry technics. YAP1 and TEAD3 expressions were increased, while p73, caspase-3 expressions weren't changed in granulosa cells in ovary. There was no difference in the bax expression among the groups. MiR-129-5p and miR-145-5p were downregulated in FSH-treated group. The unchanged expression of p73, caspase-3 and bax in granulosa cells indicates no progression to atresia following administration of high-dose FSH. The increase in YAP1 and TEAD3 expressions and the decrease in miR-129-5p and miR-145-5p revealed that the proliferative process is dominant in the ovary and high-dose FSH exerts this effect through the Hippo pathway.