Cloning, expression, and nutritional regulation of the glutamine synthetase gene in Ctenopharyngodon idellus

Comp Biochem Physiol B Biochem Mol Biol. 2017 Oct:212:70-76. doi: 10.1016/j.cbpb.2017.06.004. Epub 2017 Jun 19.

Abstract

Glutamine synthetase (GS) is considered a master enzyme that catalyzes ATP-dependent biosynthesis of glutamine from glutamate. In the present study, the GS gene was cloned from the intestine of grass carp (Ctenopharyngodon idellus). The full-length cDNA sequence of GS encodes a 371-amino-acid polypetide. Phylogenetic analysis of the C. idellus GS sequence reveals common carp (Cyprinus carpio) as its closest neighbor. GS mRNA was differentially expressed in different tissues, with high to low gradient expression the intestine, brain, muscle, heart, gill, liver, pituitary gland, and spleen. Additionally, GS exhibited a dynamic pattern of expression during embryonic development, reaching maximal and minimal levels in the organ and hatching stages, respectively, and constant low levels from 7 to 28days post-hatching. We also assessed dietary protein levels and feed sources in diet-regulated fish, and the results suggested that low crude protein (CP) and fish meal stimulate GS gene expression. Furthermore, intestinal GS mRNA expression was significantly increased by 0.2, 0.4, 0.6, 0.8mM concentrations of glutamine dipeptide in vitro. This study provides valuable knowledge about the regulation of GS expression in teleosts.

Keywords: Dietary protein; Glutamine dipeptide; Glutamine synthetase; Grass carp.

MeSH terms

  • Animal Nutritional Physiological Phenomena
  • Animals
  • Carps / embryology
  • Carps / genetics*
  • Carps / metabolism*
  • Cells, Cultured
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • Dietary Proteins / administration & dosage
  • Fish Proteins / genetics*
  • Fish Proteins / metabolism*
  • Gene Expression Regulation, Developmental
  • Gene Expression Regulation, Enzymologic
  • Glutamate-Ammonia Ligase / genetics*
  • Glutamate-Ammonia Ligase / metabolism*
  • Glutamine / biosynthesis
  • Intestines / enzymology
  • Phylogeny
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Tissue Distribution

Substances

  • DNA, Complementary
  • Dietary Proteins
  • Fish Proteins
  • RNA, Messenger
  • Glutamine
  • Glutamate-Ammonia Ligase