海洋科学2017，Vol.41Issue(2)：61-70,10.DOI:10.11759//hykx20160201003
凡纳滨对虾水孔蛋白-4的cDNA克隆及盐度胁迫对其肝胰腺mRNA表达水平的影响
Molecular cloning of aquaporin-4 (AQP4) gene in the Pacific white shrimp (Litopenaeus vannamei) and the effect of salinity stress on its expression in hepatopancreas
摘要
Abstract
In this study, to explore the function of aquaporin (AQP) in the osmoregulation of Litopenaeus vannamei, the full-length cDNA of L. vannamei aquaporin-4 (termed as LvAQP4) was cloned by RACE method, and the effect of salinity stress on the transcriptional expression of LvAQP4 in the hepatopancreas was examined. The results showed that the full length cDNA of LvAQP4 was 1048 bp in length, containing a 75 bp 5ˊ UTR, a 187 bp 3ˊ UTR, and a 786 bp ORF that encode for a deduced protein of 261 amino acids with the estimated molecular mass of 27.85 kDa and an isoelectric point of 8.11. The LvAQP4 protein shared identities from 48.8% to 97.3% with its counter-parts in other decapod species. Additionally, the phylogenetic analysis suggested that LvAQP4 belonged to AQP1-like subfamily and clustered with other AQP4s. Quantitative real-time PCR (qPCR) showed that the expres-sion of LvAQP4 was ubiquitous, with the highest expression in the gills. The expression levels of LvAQP4 were also high in the hepatopancreas, muscle, brain, and eyestalk but low in hemocytes, intestine, stomach, and thoracic gan-glion. Under hypersalinity stress (40), the expression levels of LvAQP4 mRNA in the hepatopancreas increased consistently from the beginning to 6 h, reaching the highest level at 6 h, and then decreased after that. Under hy-persalinity stress (4), on the contrary, the mRNA levels of LvAQP4 showed no significant change. In the hepato-pancreatic primary cells, the expression levels of LvAQP4 mRNA exhibited a dose-dependent upregulation when increased dosage of NaCl was added. Similar results were observed with addition of sucrose in the culture medium to increase the osmotic pressure, but the effect of sucrose on LvAQP4 transcripts was weaker than that with NaCl. These results suggested that the expression of LvAQP4 in the hepatopancreas was affected by salinity and LvAQP4 may play important roles in the osmoregulation of L.vannamei.关键词
凡纳滨对虾(Litopenaeusvannamei)/LvAQP4/盐度胁迫/渗透压调节Key words
Litopenaeus vannamei/LvAQP4/salinity stress/osmoregulation分类
农业科技引用本文复制引用
高沿,胡超群,任春华,钱璟,何香燕,江晓,黄文..凡纳滨对虾水孔蛋白-4的cDNA克隆及盐度胁迫对其肝胰腺mRNA表达水平的影响[J].海洋科学,2017,41(2):61-70,10.基金项目
广东省中国科学院全面战略合作专项(2013B091300020) （2013B091300020）
广东省省级科技计划项目(2014B030301064, 2015B020231007) （2014B030301064, 2015B020231007）
国家高技术研究发展计划("863"计划)(2012AA10A404-4) The Comprehensive Strategic Cooperation Project of The Chinese Academy of Sciences of Guangdong Province, No.2013B091300020 （"863"计划）
Science and Technology Planning Project of Guangdong Province, China, No.2014B030301064, 2015B020231007 （）
National High-tech Research &Development of China ("863" program), No.2012AA10A404-4 （"863" program）
