The stimulatory effect of TRH on prolactin (Prl) secretion by the anterior pituitary gland (APG) of the pseudopregnant (PSP) rat was studied in vivo and in vitro. TRH, 500 μg, did not increase Prl release during the Prl peaks which are generated daily between 01.00 and 12.00 for about 10 days (mean height of the peaks: 302±99 ng mL −1), nor did TRH induce Prl secretion during the phase of low (12±1 ng Prl mL −1) secretion (the “interphase”, between 20.00 and 01.00 h). 25 mg/kg b.w. of the dopamine- (DA) release blocking drug, 1-hydroxy-3-amino-pyrrolidone-2 (HA 966) did not itself innduce peaks of Prl during the interphase, but in 1 animal out of 6 which were treated with this dose of HA 966, TRH (500 μg) induced a peak of Prl with a height of 264 ng mL −. 100 mg HA 966/kg b.w. induced during the interphase peaks of Prl which were as high as spontaneous Prl peaks (343±96 ng Ml −1). TRH increased these Prl peaks to 844±48 ng mL −1. Apparently, HA 966 not only lowers DA concentrations but also somehow increased thesTRH-responsiveness of the lactotrophs. In vitro, the secretion of Prl by incubated pituitary glands was suppressed to the same extent by 5.10 −8 and 10 −6 M DA. TRH stimulated Prl secretion in the presence of 5.10 −8 M DA, but not in the presence of 10 −6 M DA or in the absence of DA. It is concluded that in vitro TRH acts as a physiological antagonist of DA. It is suggested that in vivo the effectivity of TRH as a Prl-releasing factor may be modulated by DA and that this effect of DA is independent from its Prl-release supprrssing effect.