Determination of the relationship between C-ABL tyrosine kinase and yap in response to DNA damage in mouse granulosa cells

Granulosa cells are the crucial type of cell in the ovarian tissue. These cells provide the physical support and microenvironment necessary for the development and maturation of oocyte. Granulosa cell proliferation and function are very curicial for a healthy follicle development, maturation, ovulation and atresia. Yes-associated protein (YAP) translocates to the nucleus and becomes activated when Hippo pathway is inactivated. c-Abl is a tyrosine kinase which can be activated by DNA damage and functioning in apoptosis, stress response and adhesion. It has been known that under the DNA damage, c-Abl alters the oncogenic function of YAP to tumor supressor. In this thesis study, by performing double immunofluorescence staining and immunoblotting assay, we aimed to exert the changes in the expression of c-Abl and YAP in response to 2.5μM, 5μM, 10μM, 20μM, 50μM hydrogen peroxide (H2O2) concentrations and 100μM, 200μM and 400μM hydrogen peroxide (H2O2) concentrations in mouse GCs. According to the results, it was obvious that the nuclear expression of c-Abl decreased in 5μM, 10μM, 20μM, 50μM H2O2 concentrations. Similarly, nuclear localization of c-Abl decreased in 100μM, 200μM and 400μM H2O2 treated GCs in compare with positive control which meant that cytoplasmic c-Abl could be activated in response to increasing H2O2 concentrations. According to the results, this cytoplasmic c-Abl activity caused to preserved nuclear YAP and increased p73 protein expression in normal GCs. Our findings indicated that to the fill the gap between oxidative stress and c-Abl/YAP activation-dependent GCs proliferation, migration and apoptosis, the determination of signaling relation between c-Abl and YAP is very important. Key words: Granulosa cell (GC), Yes-associated protein (YAP), Abelson tyrosine kinase (c-Abl), Hydrogen peroxide (H2O2)