Basal thyrotropin (TSH) levels in plasma and the TSH response to thyrotropin-releasing hormone (TRH) were inhibited after Leucine-enkephalin (L-EK) administration iv in rats. TRH and TSH responses to cold were inhibited after L-EK administration. In the L-DOPA, haloperidol or 5-hydoxytryptophan-treated rats, the inhibitory effect of L-EK on TSH release was restored. Findings suggested that L-EK acted both the hypothalamus and pituitary. Its inhibitory effects on TRH and TSH release at least partially mediated by interaction with amines in the central nervous system.