AMP-activated protein kinase α1 in Megalobrama amblycephala: Molecular characterization and the transcriptional modulation by nutrient restriction and glucose and insulin loadings

General and Comparative Endocrinology
Chao XuWen-Bin Liu

Abstract

This study aimed to characterize the full-length cDNA of AMP-activated protein kinase α1 (AMPKα1) from Megalobrama amblycephala and investigate the transcriptional response of this kinase to nutrient restriction and glucose and insulin loadings. The cDNA obtained was 3545-bp long with an open reading frame of 1710 bp encoding 570 amino acids. Multiple alignments and phylogenetic analyses revealed a high degree of conservation (80-100%) among most fish, retaining one kinase domain (KD), one auto-inhibitory domain (AID), one C-terminal domain (α-CTD), one regulatory-subunit-interacting motif (α-RIM), one serine/threonine-rich loop (ST loop), one α-hook, and several phosphorylation sites. AMPKα1 mRNA was predominantly expressed in white muscle, gill, and brain tissues, whereas little was expressed in the intestines. After a fasting-refeeding trial, phosphorylation and mRNA levels of AMPKα1 were significantly greater in fish fasted for 10 days, while in re-fed fish at 1 h after re-feeding, the levels of this kinase were intermediate between those of the fish in the fed and fasted groups. Further, AMPKα1 mRNA levels were quantified in the liver and muscle tissues of fish injected intraperitoneally with 1.67 g glucose per kg body wei...Continue Reading

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