title: L-leucine, L-methionine, and L-lysine are involved in the regulation of intermediary metabolism-related gene expression in rainbow trout hepatocytes.
abstract: Using rainbow trout hepatocytes stimulated with l-leucine, l-methionine, or l-lysine in the presence or absence of bovine insulin, we investigated the ability of these amino acids to mimic the effects of a pool of amino acids on protein kinase B (Akt)/target of rapamycin (TOR) signaling pathways and expression of 6 genes known to be subjected to insulin and/or amino acid regulation [glucose-6-phosphatase (G6Pase), phosphoenolpyruvate carboxykinase (PEPCK), glucokinase (GK), pyruvate kinase (PK), fatty acid synthase (FAS), and serine dehydratase (SDH)]. Emphasis was placed on leucine, known to be a signaling molecule in mammals, and methionine and lysine that are essential amino acids limiting in plant-based diets for fish. In the presence of insulin, leucine (but not methionine or lysine) phosphorylated Akt and ribosomal protein S6 as previously observed with a pool of amino acids, suggesting that leucine might participate in the activation of the TOR pathway by amino acids in fish, as in mammals. G6Pase, PEPCK, GK, and SDH gene expression were higher in leucine-treated cells compared with control cells. Leucine combined with insulin reduced G6Pase gene expression by 90% and increased FAS gene expression > 4-fold compared with the control treatment. Methionine weakly decreased G6Pase, GK, and SDH gene expression and lysine weakly but significantly decreased the mRNA level of PEPCK. Thus, leucine regulated gluconeogenesis and lipogenesis, but not glycolysis, in the same way as a pool of amino acids. Methionine appeared to be involved in the regulation of specific genes, whereas lysine only had limited effects. These findings are particularly relevant regarding the involvement of amino acids in the regulation of metabolism-related gene expression.
authors.affiliation: INRA, UMR1067 Nutrition Aquaculture et Génomique, Pôle d'hydrobiologie, CD918, F-64310 St-Pée-sur-Nivelle, France.
authors.forename: Marine
authors.initials: M
authors.lastname: Lansard
authors.forename: Stéphane
authors.initials: S
authors.lastname: Panserat
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Plagnes-Juan
authors.forename: Karine
authors.initials: K
authors.lastname: Dias
authors.forename: Iban
authors.initials: I
authors.lastname: Seiliez
authors.forename: Sandrine
authors.initials: S
authors.lastname: Skiba-Cassy
mesh_terms: D000601:Amino Acids, Essential / Q000502:physiology*; D000818:Animals; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D005941:Glucokinase / Q000235:genetics; D005952:Glucose-6-Phosphatase / Q000235:genetics; D022781:Hepatocytes / Q000378:metabolism*; D007930:Leucine / Q000502:physiology; D008239:Lysine / Q000502:physiology; D008297:Male; D008715:Methionine / Q000502:physiology; D017686:Oncorhynchus mykiss / Q000378:metabolism*; D010729:Phosphoenolpyruvate Carboxykinase (GTP) / Q000235:genetics; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D019893:Ribosomal Protein S6 Kinases / Q000378:metabolism; D058570:TOR Serine-Threonine Kinases / Q000502:physiology
chemical_list: D000601:Amino Acids, Essential; D012333:RNA, Messenger; D008715:Methionine; D005941:Glucokinase; D051057:Proto-Oncogene Proteins c-akt; D019893:Ribosomal Protein S6 Kinases; D058570:TOR Serine-Threonine Kinases; D005952:Glucose-6-Phosphatase; D010729:Phosphoenolpyruvate Carboxykinase (GTP); D007930:Leucine; D008239:Lysine