Negative energy balance-related metabolic disorders suggest that the balance between available lipogenic and glucogenic nutrients is important. The objectives of this study were to compare the effects of a glucogenic or a lipogenic diet on liver triacylglycerides (TAG), metabolites, and metabolic hormones in dairy cows in early lactation and to relate metabolite concentrations to the determined energy retention in body mass (ER). Sixteen dairy cows were fed either a lipogenic or glucogenic diet from wk 3 prepartum to wk 9 postpartum (pp) and were housed in climate respiration chambers from wk 2 to 9 pp. Diets were isocaloric (net energy basis). Postpartum, cows fed a lipogenic diet tended to have higher nonesterified fatty acid concentration (NEFA; 0.46 +/- 0.04 vs. 0.37 +/- 0.04 mmol/L) and lower insulin concentration (4.0 +/- 0.5 vs. 5.5 +/- 0.6 microIU/mL). No difference was found in plasma glucose, beta-hydroxybutyrate, insulin-like growth factor-I, and thyroid hormones. Liver TAG was equal between both diets in wk -2 and 2 pp. In wk 4 pp cows fed the glucogenic diet had numerically lower TAG levels, although there was no significant dietary effect. Negative relationships were detected between ER and milk fat and between ER and NEFA. A positive relationship was detected between ER and insulin concentration. Overall, results suggest that insulin plays a regulating role in altering energy partitioning between milk and body tissue. Feeding lactating dairy cows a glucogenic diet decreased mobilization of body fat compared with a lipogenic diet. The relative abundance of lipogenic nutrients, when feeding a more lipogenic diet, is related to more secretion of lipogenic nutrients in milk, lower plasma insulin, and higher plasma NEFA concentration.