Characterization and dietary regulation of oligopeptide transporter (PepT1) in different ploidy fishes

Abstract

The oligopeptide transporter (PepT1) is located on the brush-border membrane of the intestinal epithelium which has been regarded as a mediator of protein absorption. Here, we cloned and characterized PepT1 genes from diploid (red crucian carp), triploid and tetraploid fish. Then, the PepT1 expression pattern in different tissues and embryogenesis were assayed. Meanwhile, using real-time PCR and western blotting, we showed the expression profiles of diets with different protein levels, protein sources and additives (sodium butyrate) in triploids. The cDNAs of the three different ploidy fishes have a high sequence similarity of PepT1 among vertebrates. PepT1 mRNA expression was also developmentally regulated and showed the strongest expression around the 2-cell and 4-cell stage in all three kinds of fishes. The maternal transcripts were first detected in eggs and dropped from blastula stage to muscle contraction stage. Tissue expression studies showed higher expression of PepT1 genes in the intestines of fishes compared with other tissues. In adults, triploids showed significantly higher expression levels of PepT1 in the intestines of the three kinds of ploidy fishes during breeding season and non-breeding season. In addition, high or low protein level diets both promote PepT1 expression in the intestine. We also confirmed that fish meal showed a significant increase in PepT1 expression than soybean meal in triploid intestines. Furthermore, sodium butyrate additives induce PepT1 expression that may be mediated by CDX2 and CREB. This research provides a new insight into protein absorption and its regulation in triploid fish.