De novo fatty acid synthesis was determined by the 3H2O method in numerous tissues and organs of TCDD-treated (125 micrograms/kg), pair-fed and free-fed male Sprague-Dawley rats to investigate if this important pathway of intermediary metabolism is altered by TCDD. Of the 12 tissues and organs examined, liver showed an increased, and interscapular brown adipose tissue (IBAT) a decreased de novo fatty acid synthesis when comparing TCDD-treated to pair-fed or free-fed control rats. De novo fatty acid synthesis was unaffected in other organs and tissues examined, with the exception that the concentration of 3H-fatty acids in plasma reflected the increased rate of synthesis seen in the liver of TCDD-treated animals. Increased de novo fatty acid synthesis in liver coincided with increased plasma triiodothyronine (T3) concentrations, whereas decreased de novo fatty acid synthesis in IBAT parallelled decreased plasma thyroxine (T4) levels. Thyroidectomy decreased de novo fatty acid synthesis, as expected, in both liver and IBAT. However, TCDD elicited no response in either of these organs in thyroidectomized rats. This finding suggests that changes observed in non-thyroidectomized rats are probably secondary effects. Indeed, known tissue-specific effects of T3 on liver and T4 on IBAT provide a likely explanation for the altered de novo fatty acid synthesis of these organs. It is suggested that increased de novo fatty acid synthesis in the liver of TCDD-treated rats might be responsible for the additional wasting away observable in these animals as compared to pair-fed controls.