兽类学报 ›› 2021, Vol. 41 ›› Issue (1): 51-58.DOI: 10.16829/j.slxb.150433

• 研究论文 • 上一篇    下一篇

牦牛水通道蛋白AQP1和AQP3基因克隆及在不同组织中表达和定位

李娟1, 王利1, 罗晓林2, 官久强2, 张翔飞2   

  1. 1 西南民族大学, 青藏高原动物遗传资源保护与利用教育部和四川省重点实验室, 成都 610041;
    2 四川省草原科学研究院, 成都 611731
  • 收稿日期:2020-04-24 出版日期:2021-01-30 发布日期:2021-01-21
  • 通讯作者: 王利,E-mail:qinxin916@aliyun.com;罗晓林,Luoxl2004@sina.com
  • 作者简介:李娟(1995-)女,硕士研究生,主要从事动物遗传学研究.E-mail:1134323458@qq.com
  • 基金资助:
    国家肉牛牦牛产业技术体系(CARS-37);第二次青藏高原综合科学考察研究项目(2019QZKK0302);国家十三五重点研发计划项目(2018YFD0502304);四川省科技计划(16ZC2530,20YSZH0018);四川省留学人员科技活动择优资助项目(2019)

Cloning of AQP1 and AQP3 genes and their locations and expressions in different tissues of yak(Bos grunniens)

LI Juan1, WANG Li1, LUO Xiaolin2, GUAN Jiuqiang2, ZHANG Xiangfei2   

  1. 1 Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Vtilization, Ministry of Education and Sichuar Province, Southwest Minzu University, Chengdu 610041, China;
    2 Sichuan Academy of Grassland Sciences, Chengdu 611731, China
  • Received:2020-04-24 Online:2021-01-30 Published:2021-01-21

摘要: 水通道蛋白(Aquaporin,AQP)广泛存在于生物体的各组织部位,影响着生物体水代谢的过程。为进一步研究水通道蛋白1(AQP1)和水通道蛋白3(AQP3)生物学功能,本文对牦牛(Bos grunniens)不同组织中AQP1和AQP3基因的表达与定位进行了研究。采用PCR方法扩增牦牛AQP1和AQP3基因,对其序列进行生物信息学分析,采用qPCR方法检测2个基因在牦牛不同种组织中的表达量,采用免疫组织化学的方法分析AQP1和AQP3蛋白在组织中的定位和表达。结果表明,牦牛AQP1和AQP3基因CDS区分别为816 bp和840 bp,与野牦牛的同源性最高为99%。AQP1和AQP3基因的表达量在肾脏中最高,显著高于心脏、肝脏、脾脏、肺脏、肌肉、小肠和瘤胃,AQP1基因在各组织中的表达均高于AQP3基因的表达。AQP1和AQP3蛋白定位发现其主要分布于肾脏近曲小管、小肠固有层细胞和瘤胃颗粒层细胞中,均在肾脏中的表达量最高。AQP1蛋白在脾脏、小肠和肌肉组织中的表达极显著高于AQP3蛋白表达。该研究结果对高寒低氧环境中动物的研究具有借鉴意义,有助于牦牛AQP1和AQP3功能研究。

 

关键词: 牦牛, 水通道蛋白1, 水通道蛋白3, 免疫组织化学

Abstract: Aquaporin(AQP) is widely expressed in tissues of organisms and affects the process of water metabolism.In order to investigate the biological function of AQP1 and AQP3, the expression patterns and location of AQP1 and AQP3 in different yak tissues were clarified in this study.The PCR was used to amplify the AQP1 and AQP3 genes of yak, and their sequences were analyzed by bioinformatic techniques.The qPCR was used to detect the expression of AQP1 and AQP3 in different tissues of yak.Immunohistochemistry was used to study the expression and distribution of AQP1 and AQP3 proteins in tissues.The results showed that the CDS of AQP1 and AQP3 genes of yak were 816 bp and 840 bp.And the homology with the wild yak was at most 99% of AQP1 and AQP3.The expression levels of AQP1 and AQP3 genes were the highest in the kidney, which were significantly higher than those of the intestine, muscle, rumen and other tissues(P<0.01).The expression of AQP1 gene is higher than AQP3 in all tissues.AQP1 and AQP3 proteins were mainly distributed in the renal proximal tubules, intestinal lamina propria cells and rumen granulosa cells, and both showed the highest expression in the kidney.The results of this study have implications for the study of mammals in the alpine and hypoxic environment, and contribute to the study of the functions of AQP1 and AQP3 in yak.

Key words: Yak, Aquaporin1, Aquaporin3, Immunohistochemistry

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