In vivo and in vitro methodologies were utilized to study the regulation of placental lactogen (PL) secretion in the mouse. Chorioallantoic placental explants obtained from midpregnant (day 10 of gestation) C3H/HeN or Swiss Webster mouse conceptuses behaved differently in culture than explants of chorioallantoic placentas obtained later in pregnancy. Midpregnant placental explants maintained or showed an increasing amount of PL output per day for 5-6 days of culture, while gestationally older placental explants demonstrated a progressive decline in PL output per day over the 5-6-day culture period. On selected days of gestation, bilaterally ovariectomized C3H/HeN or Swiss Webster mice were shown to have significantly higher serum PL concentrations than sham operated mice. Reduction of serum progesterone concentrations by unilateral ovariectomy and/or partial lutectomy had no significant effect on serum PL concentrations. Progesterone significantly reduced PL production by midpregnant placental explants from both C3H/HeN and Swiss Webster mice, whereas 17 beta-estradiol did not significantly affect PL release in vitro. PL production by mouse chorioallantoic placentas has gestational specific characteristics in vitro and is responsive to regulatory influences of the maternal ovaries and progesterone.