Metabolizable methionine (met) concentrations can be increased by feeding rumen-protected dl-Met (DL) or the isopropyl ester of 2-hydroxy-4-(methylthio)butanoic acid (HMB); however, hepatic response to these compounds have not been comparatively examined. The objective of this experiment was to examine the regulation of key genes in the Met and gluconeogenic pathways in response to doses of DL or HMB that mimic Met deficiency, adequacy, or supplementation in lactating dairy cows. Hepatocytes were isolated from 4 Holstein calves and were maintained as monolayer cultures in FBS for 24h before treatment. Treatments of DL or HMB (0, 10, 20, 40, 60 μM) were added to Met-free media in triplicate and after 24h, cells were collected for RNA isolation and quantification of gene expression by quantitative PCR. Data were analyzed in Proc Mixed of SAS 9.3. Analysis of covariance confirmed equivalent slopes of Met source and final model included source and dose, and random effect of calf within source. There was no main effect of Met source (P > 0.1) for any genes examined. Expression of BHMT and MTR, genes that catalyze generation of Met from betaine and homocysteine, decreased (P ≤ 0.05) with increasing Met concentration (BHMT: 0.7551, 0.7205, 0.6512, 0.582 ± 0.2563; MTR: 0.8897, 0.8481, 0.7648, 0.6815 ± 0.1763 arbitrary units). When concentrations of Met are high, Met can generate SAM, a methyl donor; however, expression of the gene that catalyzes this reaction, MAT1, was decreased (P ≤ 0.05) as Met concentration increased (0.7828, 0.7108, 0.5667, 0.4226 ± 0.1554 arbitrary units). Expression of CPT1a, G6Pase, and PEPCKm was not altered (P > 0.1) suggesting that increased Met was not used as a gluconeogenic precursor at these concentrations. The decrease in hepatocyte production of Met at higher treatment doses suggest that cellular Met requirements were being met. The lack of influence on gluconeogenic enzymes and the decrease in the enzyme responsible for SAM generation may reflect metabolic priority for Met use at these concentrations, an adequate glucose concentration, or the lack of a Met deficiency before treatment.