The objective was to investigate the effects of post-ruminal choline ion (CHO) supplementation to cows during a feed restriction-induced negative energy balance on metabolism, liver triacylglycerol (TG), and milk production. Ten primiparous rumen-cannulated Holstein cows (158 ± 24 DIM) were used in a replicated 5 × 5 Latin square design with 4 d treatment periods (d 1–4), and 10 d recovery periods (d 5–14). Treatments were (1) ad libitum intake with abomasal infusion of water (A0), (2) restricted intake (60% of net energy for lactation requirements) with abomasal infusion of water (R0), (3) restricted intake with abomasal infusion of 6.25 g/d CHO (R6.25), (4) restricted intake with abomasal infusion of 12.5 g/d CHO (R12.5), and (5) restricted intake with abomasal infusion of 25 g/d CHO (R25). Liver biopsies were performed in the morning on d 5 after the abomasal infusion ended, and tissue analyzed for TG content. Body weight and BCS were recorded on d 1 and 5. Blood was collected on d 1–5 for analysis of nonesterified fatty acids (NEFA) and β-hydroxybutyrate (BHB). Milk production was recorded daily and composite samples were analyzed for components. Two contrasts, CONT1 (A0 vs. R0) and CONT2 (R0 vs. the average of 6.25, 12.5 and 25 g/d CHO), and tests of linear and quadratic treatment effects of CHO dose were performed. BW, milk yield (MY) (P < 0.0001) and energy balance was lower in R0 and milk fat % greater compared with A0. Feed restriction also increased liver TG and plasma concentrations of NEFA and BHB. Supplementation of CHO vs. R0 decreased NEFA and milk fat % and increased MY. Supplemental CHO led to a linear increase in MY and a linear decrease in milk fat %. There was a linear decrease in liver TG with CHO supplementation (R0: 3.48 mg/g wet tissue, R6.25: 2.70 mg/g, R12.5: 1.96 mg/g, R25: 2.50 mg/g). These results suggest that supplementation of CHO at 25 g/d during a feed restriction-induced negative energy balance leads to the greatest improvement in MY, whereas 12.5 g/d CHO leads to greater improvement in liver TG storage. The mechanisms associated with these responses merit further research.