Diet has the ability to influence phenotypes, not just in an individual, but also in the progeny. This can be due to developmental programming in the growing offspring in utero, especially during critical stages of plasticity during foetal growth and development. Our prior studies have demonstrated that different diets show varying phenotypes, which can be observed in successive generations (Adedeji et al., 2019). Thus, we hypothesized that these diets could be causing the early-life programming changes by altering the activity of particular key rate-limiting enzymes of different aspects of metabolism. Eighty (80) Wistar rats of both sexes (F0 generation) were divided into control, high carbohydrate (HCD), high fat (HFD) and high protein diets (HPD) of twenty (20) rats each.  They were fed for a period of nine (9) weeks on the diets after which they were mated. F1 sibling pairs from each dietary group were selected randomly (10 males and females respectively from each group) and fed for a period of nine (9) weeks on the experimental diets after a three-week weaning period to give an F2 generation. Liver samples were collected from ten (10) randomly selected weaned pups from the F2 generation of each dietary group. Real-time qPCR was used to determine enzyme gene expression, in F2 generation not fed the experimental diet. Data obtained were evaluated using analysis of variance (ANOVA) and expressed as mean±SEM. The means (diet vs control) were compared using Tukey-Kramer multiple comparison test. P<0.05 was regarded as statistically significant. In F2 generation, HFD (Glucokinase 0.98±0.04 vs 1.49±0.07; Pyruvate kinase 0.88±0.04 vs 1.04±0.02; Glycogen synthase 1.02±0.06 vs 1.33±0.07) and the HPD (Glucokinase 0.54±0.08 vs 1.49±0.07; Pyruvate kinase 0.78±0.08 vs 1.04±0.02; Glycogen synthase 1.03±0.07 vs 1.33±0.07) groups showed a significant decrease (P<0.05) in glycolysis and glycogen synthesis gene expression, while the HCD was upregulated (Glucokinase 3.96±0.04 vs 1.49±0.07; Pyruvate kinase 3.64±0.09 vs 1.04±0.02; Glycogen synthase 4.01±0.14 vs 1.33±0.07). HFD (Phosphoenolpyruvate carboxykinase 3.11±0.05 vs 1.02±0.04) and HPD (Phosphoenolpyruvate carboxykinase 3.54±0.09 vs 1.02±0.04) also caused an upward shift in gluconeogenic enzymes gene expression, while HCD (Phosphoenolpyruvate carboxykinase 0.38±0.06 vs 1.02±0.04) reflected downregulation. The HCD (AcetylcoA carboxykinase 3.88±0.06 vs 1.08±0.12; Fatty acid synthase 4.27±0.07 vs 2.00±0.03) and HFD AcetylcoA carboxykinase 3.79±0.06 vs 1.08±0.12; Fatty acid synthase 4.78±0.03 vs 2.00±0.03) reflected significant upregulation (P<0.05) in expression of fatty acid biosynthesis gene expression. The HPD (AcetylcoA carboxykinase 0.34±0.05 vs 1.08±0.12; Fatty acid synthase 0.28±0.05 vs 2.00±0.03), on the other hand, showed downregulation. HPD (Carnitine palmitoyltransferase 1.60±0.04 vs 1.28±0.07; acyl CoA oxidase I 0.82±0.06 vs 0.83±0.04) and HFD (Carnitine palmitoyltransferase 1.48±0.04 vs 1.28±0.07; acyl CoA oxidase I 0.45±0.08 vs 0.83±0.04) groups expressed upregulation of β-oxidation genes, while HCD (Carnitine palmitoyltransferase 0.87±0.05 vs 1.28±0.07; acyl CoA oxidase I 0.70±0.07 vs 0.83±0.04) showed downregulation. The results of this study suggest that prenatal parental diet affects the expression of genes rate-limiting enzymes involved in metabolism, with an effect observed in F2 generation.