Follicle development in a period of negative energy balance (NEB), as experienced by the postpartum dairy cow, could be affected by undesirable metabolic changes, and may contain a developmentally incompetent oocyte with an impaired potential to establish a pregnancy. A differential feeding model in heifers was developed to evaluate the concurrent and carryover effects of reduced energy intake before insemination on pregnancy per artificial insemination (P/AI). Heifers were randomly assigned to either (i) control feed intake group (CF), n=68, 1.3 times estimated maintenance energy (M) requirements for 50 days and 2.0 M for 83 days or (ii) restricted feed intake (RF), n=88, 0.65 M for 50 days and 2.0 M for 83 days. Pregnancy per AI was determined by transrectal ultrasonography at day 30 following AI. Despite significant loss of live weight (LW; 5.8±2.1 vs 70.5±2.8kg, respectively) and body condition score (BCS; 0.05±0.03 vs 0.45±0.03) and a significant elevation in systemic concentrations of non-esterified fatty acids in RF heifers, there was no concurrent effect on P/AI (69 vs 72%) following AI at day 50. However, there was a carryover effect on P/AI as there was an 18 percentage point difference (64 vs 82%) between CF and RF heifers following AI on day 93. The results of the study indicate that a reduction in energy intake for a 50-day period pre-insemination had no concurrent effect but had a positive carryover effect on P/AI.