Incubation behavior in the turkey hen is associated with a large increase in prolactin secretion. Previous research using hourly sampling of incubating hens has shown that prolactin levels fluctuate widely throughout a 24-hr period, suggestive of pulsatile secretion. This study compared the prolactin secretory patterns of laying and incubating turkeys to determine if prolactin is secreted episodically and if the high prolactin levels characteristic of the incubating hen may result, at least in part, from a change in the amplitude or frequency of secretory pulses. Blood samples were collected from cannulated, unrestrained laying and incubating hens at 10-min intervals for up to 24 hr. Data were analyzed with the PULSAR program to determine baseline prolactin levels and to establish the magnitude, frequency, and duration of episodic secretory peaks. The results revealed tha prolactin is secreted in a pulsatile pattern in both laying and incubating turkey hens. Incubating hens had ninefold higher mean and baseline plasma prolactin levels than laying hens. The prolactin pulses were of approximately 12-fold greater amplitude in incubating hens than in laying hens, but the duration and frequency of pulses were the same in both groups. Therefore, the high prolactin levels required for incubation do not appear to result from an increase in the frequency of lactotroph stimulation, but rather from an increase in the prolactin secretion rate.