Pituitary glands of bullfrogs (Rana catesbeiana) were incubated in medium containing thyrotropin-releasing hormone (TRH) and/or dopamine in order to see their effects on the release and synthesis of prolactin, which was measured by a homologous radioimmunoassay. Prolactin synthesis was measured by monitoring the incorporation of [3H]leucine into prolactin. TRH (0.1-10 ng/ml) stimulated the release of immunoassayable prolactin and newly synthesized [3H]prolactin into the medium in a dose-dependent manner; however, it was ineffective in increasing total prolactin (medium plus pituitary) and the incorporation of [3H]leucine into the total prolactin during the experimental period (20 hr). The TRH-induced elevation of prolactin release was suppressed by the addition of dopamine (5 X 10(-7) M) to the medium.