title: Gluconeogenic enzymes are differentially regulated by fatty acid cocktails in Madin-Darby bovine kidney cells.
abstract: Expression of mRNA for pyruvate carboxylase (PC) is elevated at calving and during other physiological states when plasma NEFA concentrations are increased. The objective of this study was to determine the direct effects of fatty acids on expression of PC, cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C), mitochondrial PEPCK (PEPCK-M), and glucose-6-phosphatase (G6Pase) mRNA in Madin-Darby bovine kidney (MDBK) cells. Combinations of C14:0, C16:0, C18:0, C18:1n-6 cis, C18:2n-6 cis, and C18:3n-3 cis were created to mimic the profiles and concentrations in serum from far-off dry cows and late postcalving intervals (PRPT), the profile at calving (CALV), and the profile observed in cows induced to express fatty liver at calving (IFL). The MDBK cells were exposed to fatty acid mixtures for 24h at the following concentrations: 0.25 and 0.5mM for PRPT; 0.25, 0.5, and 1.0mM for CALV; and 0.5 and 1.0mM for IFL. Cells exposed to PRPT had greater PEPCK-C and tended to have greater G6Pase mRNA than control cells. Exposure of cells to 0.25mM PRPT increased expression of PEPCK-C compared with cells exposed to 0.5mM PRPT. Expression of PC and PEPCK-M did not differ with exposure to PRPT. Expression of PEPCK-C was decreased and that of PEPCK-M and G6Pase mRNA increased linearly in response to CALV. The ratio of PC:PEPCK-C mRNA was increased by the IFL mixture and in response to increasing amounts of the CALV fatty acid mixture. Treatment of cells with CALV or IFL increased the sum of PC 5' untranslated region (UTR) variants A, B, C, and F but did not alter PC 5' UTR D and E expression. The changes in PEPCK-C, G6Pase, and PC mRNA and the ratio of PC:PEPCK-C observed in MDBK cells in response to fatty acids suggests a role for fatty acid concentration and profile in mediating the expression of key gluconeogenic enzymes.
authors.affiliation: Department of Animal Sciences, Purdue University, West Lafayette, IN 47907, USA.
authors.forename: H M
authors.initials: HM
authors.lastname: White
authors.forename: S L
authors.initials: SL
authors.lastname: Koser
authors.forename: S S
authors.initials: SS
authors.lastname: Donkin
mesh_terms: D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D005227:Fatty Acids / Q000008:administration & dosage / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects; D005952:Glucose-6-Phosphatase / Q000187:drug effects* / Q000378:metabolism; D007668:Kidney / Q000187:drug effects / Q000201:enzymology*; D019764:Phosphoenolpyruvate Carboxykinase (ATP) / Q000187:drug effects* / Q000378:metabolism; D011766:Pyruvate Carboxylase / Q000187:drug effects* / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary
chemical_list: D005227:Fatty Acids; D005952:Glucose-6-Phosphatase; D019764:Phosphoenolpyruvate Carboxykinase (ATP); D011766:Pyruvate Carboxylase