The effect of dietary restriction (half of the control ration) on cholesterol biosynthesis was investigated in rabbits fed either standard or cholesterol-rich diets. Accompanying the amplification of hypercholesterolemia, additional disturbances of cholesterol metabolism were observed when cholesterol feeding was associated with dietary restriction. In the intestine, underfed rabbits showed a more marked inhibition of duodenal cholesterol biosynthesis from [14C]acetate following cholesterol feeding than rabbits on normal caloric ration. In contrast liver cholesterogenesis was equally suppressed in both groups receiving cholesterol-rich diets. Cholesterol biosynthesis from [14C]mevalonate was also inhibited by cholesterol feeding particularly in the duodenum of underfed rabbits. In addition cholesterol feeding induced a marked increase of the labeled esterified: free cholesterol ratio in the liver, demonstrating intensive esterification, this was enhanced by dietary restriction. The additional cholesterol which accumulates in the plasma and in various tissues in underfed rabbits is of dietary origin since the feedback control of cholesterogenesis by exogenous cholesterol was shown to be very effective in these animals.