Expression Analysis of Hippo Signaling Pathway Related-genes in Yak (Bos grunniens) Ovaries During Different Reproductive Cycles
LI Yi-Juan1, FAN Jiang-Feng1,2,*, YU Si-Jiu1,2, ZHOU Ying-Cong1, QIAN Wen-Jie1, YAO Ying1
1 College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China; 2 College of Veterinary Medicine, Gansu Agricultural University/Research Center of Bovine and Sheep Embryo Engineering Technology, Lanzhou 730070, China
Abstract：Hippo signaling pathway is a key pathway regulating the growth and development of animal tissues and organs, and plays an important role in mammalian ovarian development. To investigate the Hippo signaling pathway related genes mammalian ste20-like protein kinase 1 (MST1), large tumor suppressor 1 (LATS1) and Yes-associated protein 1 (YAP1) in yak (Bos grunniens) ovaries during different reproductive cycles, in this study, the ovarian tissues of healthy female yaks (Bos grunniens) from different reproductive cycles (follicular, luteal, gestation) were selected as the test samples. qRT-PCR and Western blot were used to detect the expression differences of the MST1, LATS1 and YAP1 and their protein in yak ovaries of different reproductive cycles. Immunohistochemistry (IHC) was used to detect the distribution of genes related to Hippo signaling pathway in yak ovaries at 3 cycles. The results of qRT-PCR showed that there were significant differences in the expression of MST1 gene in ovarian tissues at different stages (P<0.05) with the highest expression in follicular stage, the second in gestation stage and the lowest in luteal stage. The expression levels of LATS1 and YAP1 genes in pregnancy were significantly higher than those in follicular and luteal phases (P<0.05). Western blot results showed that the protein expressions of MST1, LATS1 and YAP1 increased gradually, and their expression reached the peak in pregnancy phase, then decreased and maintained at a low level, which was significantly different from other stages of pregnancy phase (P<0.05). IHC results showed that MST1 was mainly distributed in granulosa cells, membrane cells, granular luteal cells, membranous luteal cells and ovarian germ epithelium of ovarian follicles. The distribution positions of LATS1 and YAP1 in different ovarian cycles were consistent with those of MST1. MST1, LATS1 and YAP1 were expressed in yak ovaries in different reproductive cycles, and the expression levels were significantly different, suggesting that MST1, LATS1 and YAP1 may be involved in the regulation of reproductive physiology related processes in yak. This study provides insights into the relationship between the expression of MST1, LATS1 and YAP1 in follicular granulosa cells of yaks at different developmental stages, which will provide a theoretical basis for further research on the reproductive performance of yaks by exploring Hippo signaling pathway.
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