A 9-week feeding trial was conducted to estimate the dietary isoleucine requirement of juvenile blunt snout bream. Six isonitrogenous and isoenergetic experimental diets were formulated to contain graded isoleucine levels ranging from 5.3 to 20.1 g kg À1 dry diet. At the end of the experiment, weight gain (WG), specific growth rate (SGR), feed efficiency ratio (FER) and protein efficiency ratio (PER) increased with increasing dietary isoleucine level up to 11.1 g kg À1 dry diet, and dietary isoleucine level above 14.2 g kg À1 dry diet declined these performances. Dietary isoleucine levels (14.2 and 17.3 g kg À1 dry diet) significantly improved whole-body protein content, but decreased whole-body lipid, plasma triglyceride and cholesterol contents. Significantly lower visceral fat index (VFI) in fish fed with 14.2 g kg À1 dietary isoleucine was observed compared to those fed with deficient or excessive isoleucine. Dietary isoleucine supplementation significantly increased plasma isoleucine concentration, while plasma valine and leucine concentrations showed a reversed trend. Dietary isoleucine levels regulated the target of rapamycin (TOR) gene expression and improved plasma superoxide dismutase (SOD) activity in juvenile blunt snout bream. Based on second-order polynomial regression model analysis of SGR and FER, the optimum dietary isoleucine requirement was estimated to be 13.8 g kg À1 dry diet (40.6 g kg À1 dietary protein) and 14.0 g kg À1 dry diet (41.2 g kg À1 dietary protein), respectively.
This study evaluated the mechanisms governing insulin resistance, glucose metabolism and lipogenesis in juvenile fish fed with graded levels of dietary arginine. The results showed that, compared with the control group (0.87%), 2.31% dietary arginine level resulted in the upregulation of the relative gene expression of IRS-1, PI3K and Akt in the insulin signaling pathway, while 2.70% dietary arginine level led to inhibition of these genes. 1.62% dietary arginine level upregulated glycolysis by increasing GK mRNA level; 2.70% dietary arginine level upregulated gluconeogenesis and resulted in high plasma glucose content by increasing PEPCK and G6P mRNA level. Furthermore, 2.70% dietary arginine level significantly lowered GLUT2 and increased PK mRNA levels. 1.62% dietary arginine level significantly upregulated ACC, FAS and G6PDH mRNA levels in the fat synthesis pathway and resulted in high plasma TG content. These results indicate that 1.62% dietary arginine level improves glycolysis and fatty acid synthesis in juvenile blunt snout bream. However, 2.70% dietary arginine level results in high plasma glucose, which could lead to negative feedback of insulin resistance, including inhibition of IRS-1 mRNA levels and activation of gluconeogenesis-related gene expression. This mechanism seems to be different from mammals at the molecular level.
Blunt snout bream (Megalobrama amblycephala) is a widely favored herbivorous fish species and is a frequentlyused fish model for studying the metabolism physiology. This study aimed to provide a comprehensive illustration of the mechanisms of a high-starch diet (HSD) induced lipid metabolic disorder by identifying microRNAs (miRNAs) controlled pathways in glucose and lipid metabolism in fish using high-throughput sequencing technologies. Small RNA libraries derived from intestines, livers, and brains of HSD and normal-starch diet (NSD) treated M. amblycephala were sequenced and 79, 124 and 77 differentially expressed miRNAs (DEMs) in intestines, livers, and brains of HSD treated fish were identified, respectively. Bioinformatics analyses showed that these DEMs targeted hundreds of predicted genes were enriched into metabolic pathways and biosynthetic processes, including peroxisome proliferator-activated receptor (PPAR), glycolysis/gluconeogenesis, and insulin signaling pathway. These analyses confirmed that miRNAs play crucial roles in glucose and lipid metabolism related to high wheat starch treatment. These results provide information on further investigation of a DEM-related mechanism dysregulated by a high carbohydrate diet.
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