Gluconeogenic enzymes are differentially regulated by fatty acid cocktails in Madin-Darby bovine kidney cells

• Published in: Journal of dairy science Vol. 95; no. 3; pp. 1249 - 1256
• Main Authors: White, H.M, Koser, S.L, Donkin, S.S
• Format: Journal Article
• Language: English
• Published: New York, NY Elsevier Inc 01.03.2012; Elsevier
• Subjects: Animal productions; Animals; Biological and medical sciences; blood serum; calving; Cattle; Cells, Cultured; cows; fatty acid composition; fatty acids; Fatty Acids - administration & dosage; Fatty Acids - pharmacology; fatty liver; Food industries; free fatty acids; Fundamental and applied biological sciences. Psychology; gene expression; Gene Expression Regulation - drug effects; glucose-6-phosphatase; Glucose-6-Phosphatase - drug effects; Glucose-6-Phosphatase - metabolism; Kidney - drug effects; Kidney - enzymology; kidney cells; messenger RNA; Milk and cheese industries. Ice creams; Phosphoenolpyruvate Carboxykinase (ATP) - drug effects; Phosphoenolpyruvate Carboxykinase (ATP) - metabolism; pyruvate carboxylase; Pyruvate Carboxylase - drug effects; Pyruvate Carboxylase - metabolism; Real-Time Polymerase Chain Reaction - veterinary; Terrestrial animal productions; Vertebrates; Carbon-carbon ligases; Bovine; Enzyme; fatty acid; Lipids; Fatty acids; Pyruvate carboxylase; Kidney; Vertebrata; Mammalia; Ligases; Dairy industry; Artiodactyla; Ungulata; Madin-Darby bovine kidney cell; Gluconeogenesis
• ISSN: 0022-0302; 1525-3198
• DOI: 10.3168/jds.2011-4644

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.