Rates of fatty acid synthesis from lactate and acetate and activities of selected lipogenic and NADPH-generating enzymes were determined in subcutaneous, intermuscular and intramuscular adipose tissues of cattle that were 11-19 months of age. Fatty acid synthesis from lactate and acetate increased from 11 to 13 months of age in subcutaneous and intermuscular adipose tissues; synthesis from lactate increased until 17 months of age. In intramuscular adipose tissue, synthesis from lactate also increased until 17 months of age while that from acetate continually increased. Activities of NADPH-generating enzymes increased in all three fat depots from 11 to 13 months of age, and little change occurred thereafter. Acetyl-CoA carboxylase activity was constant over entire growth period in all depots. Activity of ATP-citrate lyase increased from 11 to 13 months of age in subcutaneous and intermuscular adipose tissues, but did not increase until 19 months of age in intramuscular adipose tissue. In all cases, activities of ATP-citrate lyase were sufficient to support synthesis from lactate; therefore, lactate conversion to fatty acids in bovine adipose tissues seems to use the citrate cleavage pathway for generation of cytosolic acetyl-CoA.