The purpose of these studies was to determine the effect of suckling on the plasma oxytocin (OT) concentration profile in conscious primiparous rats during midlactation. Comparisons were made with plasma prolactin (PRL) levels obtained in the same rats. OT levels in the majority of rats exhibited a single peak during the first 5-30 min, then fell rapidly during the course of a 45-min period of suckling. The plasma OT levels were sustained over a longer period in mothers suckling 8 rather than 6 pups; the amplitudes of the OT response were similar, however. By contrast, plasma PRL profiles indicated that a steady secretion of the hormone occurred throughout the suckling period, with suckling of 8 pups resulting in significantly higher plasma levels than suckling of 6 pups. A considerably greater increase in the peak plasma OT concentration resulted when hungry foster litters of 6 pups were suckled after the mothers' own 6 pups had been suckled. Plasma PRL levels during the two sucklings, though, were similar. The rapid onset of the OT response to suckling was seen more clearly in urethane-anesthetized rats following mammary nerve stimulation. Plasma OT levels rose to a peak within 5 s after the onset, then fell to prestimulus levels by the end of the 65-second stimulation period. These results suggest that different regulating mechanisms are involved in the secretory responses of OT and PRL to suckling and that different thresholds of activation are likely to exist for the two hormones.