Determining the presence of the hippo signal pathway in the spermatogenesis process

In the seminiferous tubule of mammalian testis consists of germ cells at various stages of development and Sertoli cells. Mitosis and meiosis, proliferation and differentiation occur in this epithelium. Sertoli cells are attached to both each other and to germ cells by unique junctional units. Especially, these connecting junctions, called ectoplasmic specializations, are specific to the testis. It is known that cytoskeleton, actin-containing proteins and ectoplasmic are effective during sperm secretion. However, the mechanism of this secretion has not been fully elucidated. The Hippo signaling pathway, which was discovered for the first time in Drosophila and also preserved in mammals, has been demonstrated that it is very important in many stages where the fate of cells is determined, such as cell division, death or proliferation. Studies have shown that it is a signaling pathway that is highly influenced by actin proteins and cytoskeleton. However, the effect of this signaling pathway on the testis is not fully known. For this reason, we tried to determine the effectiveness of the Hippo signaling pathway in adult Wistar albino male rats in our study. Therefore, we created an experimental model in which the actin cytoskeleton and actin-associated proteins are affected by adjudine (n=10). In order to compare this experimental model, the control group that without any substance administration (n=10) and the methylcellulose group, which is dissolved in the adjudine, were formed (n=10). In order to be compatible with the literature, two animals from each group were sacrificed at 0 th, 6th, 12th, 24 th and 96 th hours, and the testicles were collected. Our study was evaluated in terms of Hippo signaling pathway by immunohistochemistry, immunofluorescence, Western blotting and QRT-PCR methods. Control and methylcellulose groups showed similar characteristics at each hour, thus proving the effect of adjudin and achieving success in creating an experimental group. The experimental group showed similar properties to the control and methylcellulose groups at the 0th hour. When the experimental groups are compared with the control and methylcellulose groups, it is evident that the distribution of actin-related proteins Arp3 and EPS 8 and F-actin begin to deteriorate as of the 6th hour with immunofluorescent techniques. At the 96th hour, it is quite clear that this distribution has declined. In the experimental groups, especially at the 96th hour, the expression of YAP and p-YAP was decreased in immunohistochemical, immunofluorescent and Western blot analyzes. The QRT-PCR experiments also show a parallel features to these results. In conclusion these data show us that the Hippo signaling pathway may be effective in the testis, especially in ectoplasmic specialization and sperm secretion.