The objective of this study was to describe fluctuations in leptin concentrations during late pregnancy and lactation and to investigate how those fluctuations are related to energy balance, milk yield, milk components, dry matter intake, live weight, first postpartum luteal activity, and first observed estrus during lactation. Live weight, dry matter intake, energy balance, and milk yield were measured weekly on 304 primiparous Holstein cows for the first 80 d of lactation. The first postpartum luteal activity was determined by measuring milk progesterone, and independently, first observed estrus. For measuring leptin concentrations from 30 d before until 80 d after calving, blood samples were taken at 2-wk intervals at a fixed time of the day after milking but before feeding. Leptin concentrations were high during pregnancy and declined to a nadir at parturition. It seems that leptin concentrations reflect the state of energy balance during lactation; plasma leptin concentrations were lower in cows with a mean negative energy balance during lactation. Those cows usually produced more milk, consumed less feed, and had a lower live weight compared with cows having a mean positive energy balance. The recovery of leptin concentrations from the leptin nadir at parturition seemed to depend on the extent and duration of the negative energy balance, thus probably on the amount of fat that was re-accumulated. Although there was lack of a relationship between leptin and first postpartum luteal activity, higher leptin concentrations associated with shorter intervals to first observed estrus might indicate a relationship between leptin and expression of estrus.