A technique previously used to study placental transfer in pregnant rats, consisting of labeled tracer infusion through the left uterine artery, was employed to determine the utilization of lipogenic substrates by periuterine adipose tissue in the fed and 48-hr starved female virgin rat. After 20 min infusion with either D[U-14C]glucose, L-[U-14C]alanine, [U-14C]glycerol or L-[U-14C]lactate, the radioactivity appearing in periuterine adipose tissue 14C-labeled lipids from the left side was always higher than that appearing in tissue from the right side. Negligible radioactivity was detected in the tissue from either side when the infusion was done with non-metabolizable derivatives such as L-[1-14C]glucose or [1-14C]alpha-aminoisobutyric acid. Simultaneous infusion of L-[U-14C]alanine and an alanine transaminase inhibitor (aminooxyacetic acid) into the left uterine artery completely blocked the conversion of the alanine transaminase inhibitor (aminooxyacetic acid) into the left uterine artery completely blocked the conversion of the alanine into periuterine adipose tissue 14C-labeled lipids. The utilization of the infused substrate for fatty acids and glyceride-glycerol synthesis by the tissue was quantified by taking into account the infused radioactivity, the difference in the amount of 14C-labeled lipids appearing in periuterine adipose tissue on the left and the right sides, the arterial plasma concentration of the studied metabolite, and the uterine horn blood flow. In fed animals, the highest fatty acid synthesis was found with lactate, followed by glucose, alanine, and glycerol. This process was intensely decreased with all the substrates in 48-hr starved rats.(ABSTRACT TRUNCATED AT 250 WORDS)