Graduate Thesis Or Dissertation
 

The role of phosphoenolpyruvate carboxykinase in the periparturient and ketotic dairy cow

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  • Although the occurrence of ketosis is a postpartum phenomenon, recent studies have focused on the prepartum period as key in the development of the disorder. Indicators of prepartum energy status, such as depressed dry matter intake (DMI) and elevated plasma non-esterified fatty acid (NEFA) concentrations have been associated with the occurrence of ketosis. The objective of this study was to investigate the role of phosphoenolpyruvate carboxykinase (PEPCK) in the periparturient and ketotic cow. The enzyme PEPCK catalyzes the rate limiting step in gluconeogenesis in hepatocytes. Whereas, in adipocytes, it has been suggested that PEPCK functions in the synthesis of glycerol for the formation of triacylglycerol (TAG) when plasma glucose concentrations are low. Thirty-four pregnant multiparous Holstein dairy cows were fed a single prepartum ration that consisted of 50% oat hay, 18% corn silage and 32% grain mix (DM basis). The ration was formulated to meet or exceed NRC requirements of 14% CP and 1.6 Mcal/kg NE[subscript L]. At calving, cows were transitioned onto one of two postpartum diets: control (n=14) or 3.5% supplemental fat (n=20). The postpartum diets, fed from wk 1 to 3, were formulated to isonitrogenous and to meet NRC requirements. Both diets consisted of 25% alfalfa, 25% corn silage and 50% grain mix. The control and fat diets contained 17.2 and 17.6% CP and 1.67 and 1.74 Mcal/kg NE[subscript L] respectively. Liver biopsies from 28 cows and adipose tissue biopsies from 6 cows were collected at -14, 2 or 3 and 14 d relative to calving. Tissue samples were analyzed for PEPCK mRNA and activity. All results were analyzed by period: prepartum (-21 to -2 d), freshening (2 to 7 d) and postpartum (8 to 21 d). In a previous study in our lab, 25 and 75% cows on the control and fat diets, respectively, experienced ketosis. In the current study there a 40% occurrence of ketosis for both control and fat diet groups. The high occurrence in both diets may be attributed to the rapid transition from the dry cow ration (70:30 forage to concentrate ratio, DM basis) to the lactating cow ration (50:50 forage to concentrate ratio, DM basis). The cows on the fat diet had lower serum glucose at freshening. Cows with ketosis had higher prepartum body weights (788 kg) than non-ketotic cows (743 kg; P<.1). No prepartum differences were seen in body condition score, DMI, NE[subscript L] balance, NEFA, glucose or β-hydroxybutyrate concentrations were detected between ketotic and non-ketotic cows. Expression of adipose PEPCK mRNA was not different between ketotic and non-ketotic cows. However, hepatic PEPCK mRNA expression was higher in non-ketotic cows at freshening when compared to ketotic cows. Cows that experienced ketosis had lower hepatic PEPCK activity prepartum (6.6 vs. 9.3 units /min/g protein) and postpartum (7.6 vs. 10.2 units/min/g protein; P<0.5) when compared to non-ketotic cows. Our data indicated that hepatic PEPCK is a useful prepartum predictor of a cows susceptibility to ketosis.
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