The effects of 2-deoxy-D-glucose (2DG), oligomycin and theophylline on the in vitro production and metabolism of glycerol and its response to insulin and epinephrine were studied in epididymal fat pads from fed rats. 2-DG failed to affect basal or epinephrine stimulated glycerol production but it decreased the uptake of 1-14C-glycerol by the tissue and its conversion to glyceride-glycerol Oligomycin also failed to affect the basal production of glycerol but it in· hibited the effect of epinephrine on this parameter as well as the uptake and utilization of 1-14-C-glyceroL Theophylline enhanced the production of glycerol by the tissue and this effect was not further augmented by epinephrine. Theophylline also inhibited the uptake and utilization of 1-14C-glycerol; the most pronounced effect of theophylline was observed in the formation of 14C-fatty acids from 1.14c. glycerol in the presence of glucose. Insulin, but not epinephrine, decreased the inhibitory effect of theophylline on glycerol utilization. It is concluded that these compounds af. feet more intensely the ability of adipose tissue to metabolize glycerol than to release it through lipolysis. The pathway for glycerol utilization in adipose tissue appears to be more sensitive to changes in the availability of ATP than the mechanisms responsible for the release of glycerol fom the tissue.