Cloning of Yak (Bos grunniens) AQP1 Gene and Its Expression in Yak Testis of Different Ages
LIU Min-Qing, WANG Ya-Ying, HE Hong-Hong, PAN Yang-Yang, YANG Shan-Shan, GAO Ze-Chuan, ZHANG Tong-Xiang, ZHAO Ling, WANG Jing-Lei, CUI Yan, YU Si-Jiu*
Technology and Research Center of Gansu Province for Embryonic Engineering of Bovine and Sheep & Goat, College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China
Abstract:The aim of this study was to clone the gene sequence of Aquaporin-1 (AQP1) in yak (Bos grunniens) and study the expression of AQP1 in yak testes of different ages. Testicular samples of newborn (3 day), young (1 year old), adult (4 years old) and old (8 years old) yak were collected. The full-length cDNA sequence of AQP1 gene was amplified by RT-PCR technology and analyzed by bioinformatics. qRT-PCR, Western blott and immunohistochemistry methods were used to detect and locate at the gene and protein level. The qRT-PCR results showed that as the yak grew older, the expression level of the yak AQP1 gene showed a trend of rising first and then falling, reaching the highest in the young testis; the expression level in the young group was significantly different from other groups (P<0.05), while the difference between the middle-aged group and the elderly group was not significant (P>0.05). Western blot results showed that as the yak grew older, the expression level of AQP1 protein also showed a trend of first rising and then falling, reaching the highest in the young testis; the expression difference between the young group and other groups was significant (P<0.05), while the differences between the newborn group, middle-aged group and old group were not significant (P>0.05). Immunohistochemical results showed that AQP1 protein is positively expressed in yak testes of different ages, and the expression sites are basically same, mainly expressed in sperm, sperm cells, spermatogenic cells, testicular stromal cells, supporting cells, and peritubular muscles like cells and seminiferous tubules in the basement membrane. The results of showed that the expression of AQP1 in the young yak group was significantly higher than that in the other groups (P<0.05). It is speculated that AQP1 plays an important role in the young yak testis. This study provides theoretical basis for further research on the mechanism of AQPs in the yak testis.
刘敏清, 王亚营, 何翃闳, 潘阳阳, 杨珊珊, 高泽川, 张同享, 赵凌, 王靖雷, 崔燕, 余四九. 牦牛AQP1基因克隆及其在不同年龄牦牛睾丸中的表达[J]. 农业生物技术学报, 2021, 29(5): 933-942.
LIU Min-Qing, WANG Ya-Ying, HE Hong-Hong, PAN Yang-Yang, YANG Shan-Shan, GAO Ze-Chuan, ZHANG Tong-Xiang, ZHAO Ling, WANG Jing-Lei, CUI Yan, YU Si-Jiu. Cloning of Yak (Bos grunniens) AQP1 Gene and Its Expression in Yak Testis of Different Ages. 农业生物技术学报, 2021, 29(5): 933-942.
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