Total glucocorticoid binding sites were identified and quantitated in liver and brain of rainbow trout using an exchange method and [3H]dexamethasone as the ligand. Both tissues contained a predominantly cytosolic moiety that bound dexamethasone with high specificity. Binding was saturable, time dependent, and completely reversible. Scatchard analysis showed a linear relationship suggesting that receptors belong to a single class. Dexamethasone down-regulated both liver and brain receptors. Down-regulation was rapid (within hours) and dose dependent (ED50 = 1.5 mg/kg body weight). Dexamethasone-induced down-regulation was not a result of cytoplasm to nuclei translocation or due to increases in tissue concentrations of steroid. Dexamethasone administration resulted in a lowering of Bmax (82.3 +/- 2.5 to 20.6 +/- 10.5 fmol/mg protein) and an increase in Kd (15.6 +/- 0.2 to 44.3 +/- 5.0 nM) suggesting a conformational change in the receptor molecule as part of the mechanism. The brain and liver of the rainbow trout thus have glucocorticoid receptors similar to those described in the mammalian system. Further, these receptors are subjected to autologous regulation similar to their counterparts in other systems.