The study investigated the effects of maternal diets, varying in fat content, on lipid profiles and the expression of hepatic glucose transporter 2 (GLUT2) and glucokinase (GK) in neonatal Wistar rat offspring. Dams were maintained on diets of 10% (control), 20% (20F), 30% (30F) and 40% (40F) fat as energy throughout gestation; daily food intakes and weekly body weights were measured. Circulating fasting glucose, insulin and glucagon concentrations were determined in dams and their neonatal offspring. In neonates, total serum triglyceride, total and individual serum fatty acid concentrations, hepatic GLUT2 and GK mRNA and protein expression were determined. In dams, overall food intake of 20F (645.50 ± 25.26 g) and 40F (716.30 ± 14.15 g) dams was reduced compared to control (1007.00 ± 44.83 g) and 30F (924.50 ± 21.16 g) dams. The 20F neonates displayed elevated blood glucose concentrations (4.63 ± 0.153 mmol/l) compared to control neonates (4.14 ± 0.112 mmol/l). In 30F neonates, serum palmitoleic acid was reduced (1.63 ± 0.21% vs. 3.56 ± 0.38%) whereas stearic acid was elevated (10.05 ± 0.40% vs. 7.40 ± 0.72%) compared to control neonates. Further, the palmitoleic acid/palmitic acid ratio was reduced in 30F neonates (0.085 ± 0.009% vs. 0.165 ± 0.020% in control neonates). The 40F neonates displayed elevated GLUT2 immunoreactivity (22.86 ± 0.760%) compared to 20F (11.46 ± 2.701%) and 30F (6.45 ± 1.759%) neonates. Gestational programming with different dietary fat proportions minimally affects lipid profiles and hepatic GLUT2 immunoreactivity in neonatal offspring.