草鱼SGLT1/2基因克隆及葡萄糖处理对其mRNA表达的影响

作为重要的转运载体,SGLTs(钠依赖性葡萄糖协同转运蛋白,sodium-glucose co-transporters)在物质的吸收过程中发挥着关键的作用.SGLT1和SGLT2作为SGLTs家族的重要成员,参与调节体内的葡萄糖吸收,从而维持血糖的稳态.为研究SGLT1和SGLT2在草鱼葡萄糖吸收过程中的作用,本实验...

Cloning of grass carp (Ctenopharyngodon idella) SGLT1/2 genes and effect of glucose on its mRNA expression

As the co-transporters, SGLTs play a pivotal role in the absorption of the substance. As the important member of SGLTs, SGLT1 and SGLT2 are involved in keeping blood glucose homeostasis by regulation of glucose uptake. To investigate the role of SGLT1 and SGLT2 in the glucose uptake of grass carp (Ctenopharyngodon idellus), a series of experiment was performed. In this study, the grass carp SGLT1 and SGLT2 were cloned in foregut and kidney by RT-PCR, and the sequences were analyzed by bioinformatics and the mRNA expression were detected by Real-time PCR. The results showed that the ORF of grass carp SGLT1 and SGLT2 were 1977 bp and 1989 bp, which encode 658 and 662 amino acids. Based on amino acids sequence, the structure of SGLT1 and SGLT2 protein were 14 transmembrane, and the results of sequence alignment and phylogenetic tree showed that the grass carp SGLT1 and SGLT2 gene were closest to that of goldfish and sinocyclocheilus. The Real-time PCR results indicated that the higher expression of SGLT1 was in the intestine and kidney, and the low expression of SGLT1 was in other tissues of grass carp. The higher expression of SGLT2 was in the kidney, and the low expression of SGLT2 was in other tissues of grass carp. In OGTT experiment of grass carp, the SGLT1 mRNA expression was significantly increased in grass carp foregut, and the SGLT2 mRNA expression was markedly increased in grass crap kidney by glucose treatment 1h. In in vitro experiment, the SGLT1 and SGLT2 mRNA expression were markedly increased in the CIK cells by glucose treatment. The results will provide the data base for consummating SGLTs functions and theoretical foundation for investigating the regulatory of glucose homeostasis in fish.