FoxO1 in Micropterus salmoides: Molecular characterization and its roles in glucose metabolism by glucose or insulin-glucose loading

Forkhead box O1 (FoxO1), a nuclear transcription factor, plays an important role in insulin-mediated glucose metabolism. In this study, FoxO1 gene from largemouth bass (Micropterus salmoides) was cloned and characterized, and its effects on hepatic glucose metabolism regulated by insulin-AKT pathway were investigated in response to glucose or insulin-glucose injection. The full-length cDNA of FoxO1 consisted of 2541 bp and encoded 680 amino acids. Sequence alignments and phylogenetic analysis revealed that FoxO1 exhibited a high degree of conservation among teleost, retaining one forkhead domain, one transactivation domain, and three phosphorylation sites. FoxO1 mRNA was expressed in a wide range of tissues, and high in the brain and liver. Glucose loading resulted in persistent hyperglycemia, and plasma insulin levels remained unchanged except at 1 h. After the insulin-glucose injection, insulin levels were significantly elevated and glucose levels recovered to the basal value after 6 h, which indicated insufficient insulin secretion caused persistent hyperglycemia in this species. Compared with the glucose injection group, transcript levels and enzyme activities of hepatic glycolysis-related genes (GK and PK) were significantly activated, and gluconeogenesis-related genes (PEPCK and G6Pase) were significantly depressed at 3 h after the insulin-glucose injection. Besides, phosphorylation of AKT-FoxO1 pathway was significantly activated. Therefore, insulin improved glucose metabolism by activating the AKT-FoxO1 phosphorylation  to decrease hyperglycemia stress after the meal, which indicated insufficient insulin secretion was the reason for glucose intolerance in largemouth bass. Meanwhile, conserved S267 and S329 phosphorylation sites of FoxO1 were confirmed to be regulated by AKT and mediated the glucose metabolism. In conclusion, activation of insulin-AKT-FoxO1 pathway improved glucose tolerance through mediating glucose metabolism in largemouth bass.