摘要： 为鉴定牦牛水通道蛋白8（Aquaporin，AQP8）基因，并分析其在不同组织及不同生长阶段肝脏中的表达差异，探讨AQP8蛋白的表达分布差异。采用RT-PCR技术克隆牦牛AQP8基因的CDS序列并进行生物信息学分析，利用qRT-PCR技术检测AQP8在成年牦牛8种组织和不同年龄段肝脏中的表达水平，采用免疫组织化学方法分析AQP8蛋白在不同年龄肝脏中的分布和表达。结果表明，AQP8基因的开放阅读框为792 bp，编码263个氨基酸，系统进化树分析显示，麦洼牦牛与野牦牛的亲缘关系最近，蛋白具有AQPs特有的MIP稳定结构域。AQP8在成年牦牛肝脏中的mRNA相对表达量最高，且显著高于心脏、脾脏和肾脏等组织（P<0.05），幼年时期（15月）肝脏中的AQP8表达量显著高于胎儿时期（1 d）和成年时期（5岁）（P<0.05）。免疫组化结果表明，AQP8蛋白在幼年肝脏组织中的表达量最高（P<0.05）。研究结果为深入探究牦牛AQP8在生长过程中作用机制提供基础依据，对高寒低氧环境中动物的适应性机制研究具有借鉴意义。

关键词: 牦牛, 水通道蛋白8, 基因克隆, 组织表达, 肝脏

Abstract: In order to identify the AQP8 gene of yak, analyze its expression differences in different tissues and livers at different growth stages, and explore the differences of expression and distribution of AQP8 protein, the CDS of yak AQP8 gene was cloned by RT-PCR and then the sequence was analyzed using bioinformatics. The RT-qPCR was used to detect the mRNA expression level of AQP8 in 8 tissues and liver at different growth stages. Immunohistochemistry was used to study the expression and distribution of livers in different ages. The results showed that the ORF of AQP8 gene was 792 bp, encoding 263 amino acids. The phylogenetic tree analysis showed that the AQP8 gene of Maiwa yak had the closest associated with that of Bos mutus. The AQP8 protein has a stable MIP domain specific to AQPs. The mRNA expression level of AQP8 in liver of adult yak was significantly higher than heart, spleen, kidney and other tissues(P<0.05), and the expression level of AQP8 in liver at juvenile(15 months) was significantly higher than fetus(1 day) and adult(5 years) (P<0.05). Immunohistochemical results showed that AQP8 protein expression was the highest in liver of juvenile(15 months) yak(P<0.05). This study provided relevant basis for further exploring the mechanism of yak AQP8 in the growth process, and provided implications for study of mammals in the alpine and hypoxic environment.

Key words: Yak, Aquaporin 8, Gene cloning, Tissue expression, Liver

中图分类号: 

• Q78

• S823