The Expression of MST1 and LATS1 in the Formation, Maintenance and Degeneration of Corpus Luteum in Yak (Bos grunniens) and the Effect of HCG on Its Expression
ZHOU Ying-Cong1, ZHANG Tong-Xiang2, WANG Jing-Lei2, ZHAO Ling2, ZHANG Rui2, YAO Ying2, QIAN Wen-Jie2, CUI Yan2,3, YU Si-Jiu2,3, FAN Jiang-Feng2,3,*
1 Alxa Zuoqi Animal Disease Prevention and Control Center, Alxa Zuoqi 750300, China; 2 College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China; 3 Technology and Research Center of Gansu Province for Embryonic Engineering of Bovine and Sheep & Goat, Gansu Agricultural University, Lanzhou 730070, China
Abstract Mammalian STe20-like kinase 1 (MST1) and large tumor suppressor protein 1 (LATS1) are key members of the core kinase cascade of the Hippo pathway, which participate in the reproductive process of female animals and play an important role. This study aimed to explore the relationship between MST1, LATS1, and the formation, maintenance, and degeneration of yak (Bos grunniens) corpus luteum (CL). In this study, the corpus luteum of female yaks was collected and divided into 4 groups according to the different stages of corpus luteum formation (corpus rubrum stage, lacunose luteum stage), maintenance (pregnancy luteal phase) and degeneration (degenerative luteal phase). qPCR and Western blot were used to detect the differences of MST1, LATS1 mRNA and protein expression in the formation, maintenance, and degeneration of the corpus luteum. Immunohistochemistry (IHC) was used to analyze the localization of MST1 and LATS1 in the formation, maintenance, and degeneration of the corpus luteum. In addition, granulosa cells (GC) of the yak ovary were isolated and cultured, and luteinization of granulosa cells was induced by adding human chorionic gonadotropin (HCG). qPCR, Western blot and IF methods were used to analyze the quantity and localization of MST1, LATS1 mRNA and protein before and after luteinization. The results showed that MST1, LATS1 mRNA and protein were expressed in the yak corpus luteum, and there were differences in the expression of MST1 at different developmental stages of corpus luteum. The expression of MST1 was the highest in the pregnancy luteal phase, while the expression of LATS1 was the most abundant in the lacunose luteum stage, and the expression of MST1 and LATS1 was the lowest in the degenerative luteal phase. The results of IHC showed that MST1 and LATS1 were widely distributed in yak granular luteal cells, and MST1 was characterized by expression in vascular endothelial cells. After luteinization of granulosa cells induced by 5 IU/mL HCG, the expression of MST1, LATS1 mRNA and protein increased significantly. By IF staining, it was found that MST1 and LATS1 proteins were mainly located in the cytoplasm of granulosa cells, and the position did not change before and after luteinization. The results showed that MST1 and LATS1-mediated Hippo signaling pathway participates in the formation, maintenance, degeneration and functional operation of corpus luteum in female yaks by controlling granulosa cell differentiation, luteal cell proliferation and apoptosis, and which was closely related to various reproductive hormones in the physiological process of corpus luteum. This study provides basic data for further exploring the mechanism of the Hippo signal pathway regulating the size of the corpus luteum in yaks and regulating the reproductive physiological process of mammals.
ZHOU Ying-Cong,ZHANG Tong-Xiang,WANG Jing-Lei, et al. The Expression of MST1 and LATS1 in the Formation, Maintenance and Degeneration of Corpus Luteum in Yak (Bos grunniens) and the Effect of HCG on Its Expression[J]. 农业生物技术学报, 2024, 32(8): 1844-1856.
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