Molecular characterization of fructose-1,6-bisphosphatase 1b in blunt snout bream Megalobrama amblycephala and the transcriptional response to glucose loading after the adaptation to high-carbohydrate diets.

Abstract

This study aimed to characterize the full-length complementary DNA (cDNA) of fructose-1,6-bisphosphatase 1b (FBP1b) from fish Megalobrama amblycephala, and investigate its transcriptional response to glucose administration after the adaptation to high-carbohydrate diets. The cDNA obtained covered 1435bp with an open reading frame of 1014bp. Sequence alignment and phylogenetic analysis revealed a high degree of conservation (76-96%) among most fish and other vertebrates, retaining one N-linked glycosylation site, one N-terminal acetylation site, 13 phosphorylation sites, one fructose-1,6-bisphosphatase (FBPase) active site, five metal-binding sites, four substrate-binding sites, and several AMP-binding sites. The highest messenger RNA (mRNA) level of FBP1b was observed in liver followed by intestine, whereas relatively low values were detected in heart, gill, and eye. Then, the mRNA levels of FBP1b and the FBPase activity were both determined in the liver of fish injected intraperitoneally with 1.67g glucose per kilogram body weight after being fed two dietary carbohydrate levels (30 and 42%) for 11weeks. After the glucose load, the mRNA levels of FBP1b in both treatments decreased significantly to the basal value at 8h and showed a slight increase afterward. However, the enzymatic activity showed no statistical difference during the first 4h, but increased remarkably with further increasing times. In addition, both the mRNA levels and activities decreased significantly with increasing dietary carbohydrate levels. The results indicated that the FBP1b of M. amblycephala shared a high similarity with that of the other vertebrates. Its mRNA expression in liver was downregulated remarkably by a glucose administration, as also held true after the long-term adaptation of a carbohydrate-rich diet.