Oligopeptides and free amino acids are absorbed along the intestinal tract by specialized membrane transporter proteins. Di- and tri-peptides are transported via the H+/coupled peptide transporter (PEPT1), which is located in the brush border membrane of intestinal epithelial cells. Free amino acids are absorbed by a variety of Na+- dependent and independent membrane transporters, frequently referred to as “systems”. Among the amino acid transporting proteins, the neutral amino acid transporter SLC6A19, also called system B(0) neutral amino acid transporter AT1 (B0AT1), is an integral plasma membrane protein responsible for the uptake of a broad range of neutral amino acids across the apical membrane of enterocytes and renal cells. In humans, the B0 system is associated with a severe neutral aminoaciduria known as Hartnup disorder. The objective of the present study was to examine the effect of diets with descending fish meal (FM) inclusion levels and the addition of salt to the diet containing the lowest FM level on growth performances, food conversion ratio, and intestinal SLC6A19 and PEPT1 transcript levels, in freshwater adapted European sea bass (Dicentrarchus labrax). We have firstly isolated by molecular cloning and sequencing a fulllength cDNA representing the neutral aminoacid transporter SLC6A19 in sea bass. The cDNA sequence was deposited in GenBank database (accession no. KC812315). The 12 transmembrane domains and the de novo prediction of the three-dimensional structure of SLC6A19 protein (634 amino acids) are presented. We then analysed diet-induced changes in the mRNA copies of SLC6A19 and PEPT1 genes in different portions of sea bass intestine using realtime RT-PCR. Sea bass were fed for 6 weeks on different diets, with ascending levels of fat or descending levels of FM, which was replaced with vegetable meal. The salt-enriched diet was prepared by adding 3% NaCl to the diet containing 10% FM. SLC6A19 mRNAs in anterior and posterior intestine of sea bass were not modulated by dietary protein sources and salt supplementation. Conversely, including salt in the diet containing low FM percentage, upregulated the mRNA copies of PEPT1 in hindgut. Fish growth correlated positively with the content of FM in the diets. Interestingly, the addition of salt to the diet containing 10% FM improved food intake, as well as specific growth rate and food conversion ratio.
Transcript quantification of intestinal neutral amino acids and oligopeptides transporters in European sea bass (Dicentrarchus labrax) reared in fresh water and fed fish and plant protein sources.
TEROVA, GENCIANA;RIMOLDI, SIMONA;BOSSI, ELENA;SAROGLIA, MARCO
2015-01-01
Abstract
Oligopeptides and free amino acids are absorbed along the intestinal tract by specialized membrane transporter proteins. Di- and tri-peptides are transported via the H+/coupled peptide transporter (PEPT1), which is located in the brush border membrane of intestinal epithelial cells. Free amino acids are absorbed by a variety of Na+- dependent and independent membrane transporters, frequently referred to as “systems”. Among the amino acid transporting proteins, the neutral amino acid transporter SLC6A19, also called system B(0) neutral amino acid transporter AT1 (B0AT1), is an integral plasma membrane protein responsible for the uptake of a broad range of neutral amino acids across the apical membrane of enterocytes and renal cells. In humans, the B0 system is associated with a severe neutral aminoaciduria known as Hartnup disorder. The objective of the present study was to examine the effect of diets with descending fish meal (FM) inclusion levels and the addition of salt to the diet containing the lowest FM level on growth performances, food conversion ratio, and intestinal SLC6A19 and PEPT1 transcript levels, in freshwater adapted European sea bass (Dicentrarchus labrax). We have firstly isolated by molecular cloning and sequencing a fulllength cDNA representing the neutral aminoacid transporter SLC6A19 in sea bass. The cDNA sequence was deposited in GenBank database (accession no. KC812315). The 12 transmembrane domains and the de novo prediction of the three-dimensional structure of SLC6A19 protein (634 amino acids) are presented. We then analysed diet-induced changes in the mRNA copies of SLC6A19 and PEPT1 genes in different portions of sea bass intestine using realtime RT-PCR. Sea bass were fed for 6 weeks on different diets, with ascending levels of fat or descending levels of FM, which was replaced with vegetable meal. The salt-enriched diet was prepared by adding 3% NaCl to the diet containing 10% FM. SLC6A19 mRNAs in anterior and posterior intestine of sea bass were not modulated by dietary protein sources and salt supplementation. Conversely, including salt in the diet containing low FM percentage, upregulated the mRNA copies of PEPT1 in hindgut. Fish growth correlated positively with the content of FM in the diets. Interestingly, the addition of salt to the diet containing 10% FM improved food intake, as well as specific growth rate and food conversion ratio.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.