Molecular cloning, characterization and expression analysis of glucose transporters fromRachycentron canadum

Abstract

It is very necessary to clarify the mechanism of carbohydrate metabolism in fish due to their poor ability to utilize carbohydrates. Glucose transporters (GLUTs) are important carriers involved in glucose transport across the plasma membrane, which is the first rate-limiting step of carbohydrate metabolism. In this study, five glucose transporters (RcGLUT1, RcGLUT2, RcGLUT3, RcGLUT5 and RcGLUT9) were obtained from Rachycentron canadum. The complete cDNAs open reading frames (ORF) of RcGLUT1, RcGLUT2, RcGLUT3, RcGLUT5 and RcGLUT9 were 1,476 bp, 1,530 bp, 1,551 bp, 1,539 bp and 1,578 bp (GenBank accession no. MK560257, MK560258, MH184460, MH184461 and MH184462), encoding 491, 509, 516, 512 and 525 amino acid (aa) residues respectively. All five RcGLUTs contained a Sugar_tr domain, which is an important feature of the GLUT gene family. The multiple sequence alignment and phylogenetic relationship analyses showed that RcGLUTs were highly conserved and homologous from fish to mammals. Examination of tissue distribution showed that RcGLUTs were expressed constitutively in R. canadum. RcGLUT1-5 and RcGLUT9 had the highest expression in the foregut, kidneys, haemocytes, muscles, liver and heart respectively. All six RcGLUTs first increased to peak levels of expression and then reduced both in the liver and muscle after fasting. The exception is that the expression levels of RcGLUT4 and RcGLUT5 in muscle were significantly lower than the control. In response to hypoxia, only RcGLUT2 in the liver and muscle and RcGLUT9 in muscle were expressed at a significantly lower level relative to the control. In sum, all RcGLUTs in the liver and muscles showed significant changes in response to fasting and hypoxia, suggesting that GLUT genes may play a role in the response to common physiological stresses.