Yak (Bos Grunniens) AQP1 And AQP11 Gene Cloning And Their Expression In The Testes At Different Developmental Stages

The yak,as a cattle species unique to the Qinghai-Tibet high pressure that can withstand high cold and hypoxia,which is an important species for the development of agriculture and animal husbandry in high-altitude areas.AQP1 and AQP11 are members of the AQPs family and play an important role in the reproductive process of sperm production,concentration and movement in male animals.However,there are no reports about the expression of AQPs in male yak reproductive organs at home and abroad.Therefore,we selected yak testicles of different developmental stages as the test materials,Real-time fluorescent quantitative PCR（quantitative Real-time PCR,q RT-PCR）,Western-blot（Western-blot,WB）,and immunohistochemistry（Immunohistochemistry,IHC）were used to analyze the different development of yak AQP1 and AQP11 genes and proteins.The expression location in the testis tissue of the yak at the stage was analyzed.The results are as follows:1.The yak AQP1 and AQP11 genes successfully cloned and obtained the coding sequence（CDS）of,Its Gen Bank accession numbers are:MN892521 and MN892522.The AQP1 gene is 703 bp in length,encoding 161 amino acids,molecular weight:32.16KD,molecular formula:C₇₆₉H₁₂₁₂N₂₀₄O₂₂₆S_4.The AQP11 gene is 462 bp in length,encoding 144 amino acids,molecular weight:29.78KD,molecular formula:C₇₁₀H₁₁₃₅N₁₈₇O₁₉₁S₁₂,Genetic analysis shows that AQP1 and AQP11 genes are highly conservative.2.AQP1 gene and protein are expressed in yak testes at different developmental stages.Among them,the expression level was the highest in youth,followed by middle-aged age,second in old age,and lowest in the newborn period,showing an overall trend of increasing first and then decreasing.The expression and location of AQP1 protein in yak testes at different developmental stages are basically the same,mainly expressed in sperm,sperm cells,spermatogenic cells at all levels,testicular stromal cells,Leydig cell and peritubular muscle-like cells.3.AQP11 gene and protein are expressed in yak testes at different developmental stages.Among them,the expression of AQP11 gene was the highest in youth,followed by middle-aged and old age,the lowest in the first birth period,and the overall trend is to increase first and then decrease.The expression of AQP11 protein was the lowest at birth,and remained basically the same in youth,middle age and old age,and showed a trend of increasing first and then stabilizing.The expression and location of AQP11 protein in yak testis at different developmental stages are basically the same,mainly expressed in sperm cells,sperm,Sertoli cells,Leydig celland peritubular muscle-like cells.The results showed that AQP1 and AQP11 proteins are commonly expressed in yak testes at different developmental stages,and the expression sites are the same in yak testes at different developmental stages.However,with the gradual increase of yak age,there are significant differences in the expression levels of AQP1 and AQP11proteins.It is revealed that it has important biological significance for the regulation of male yak’s reproductive physiology.This study helps to further clarify the mechanism of AQPs involved in the reproductive physiology of male mammals.