Previous studies have provided evidence indicating that the nuclear orphan receptor RTR plays an important role during embryonic development and in spermatogenesis. In this study, we examine the expression of RTR in murine placenta and several human placental choriocarcinoma cell lines. Northern blot analysis showed high expression of RTR mRNA in placental tissue. In contrast to murine testis, which contains 7.4 and 2.3 kb transcripts, placental tissue expressed only the larger transcript. Examination of RTR expression in murine placental tissue by immunohistochemistry demonstrated the presence of RTR protein in the nuclei of giant trophoblasts and spongiotrophoblasts. RTR mRNA was also expressed in rat choriocarcinoma Rcho-1 cells and in the human placental choriocarcinoma cell lines BeWo, JAR, and JEG-3. In trophoblasts, RTR was co-expressed with the estrogen-related receptors ERRα and ERRβ. Giant trophoblast differentiation in Rcho-1 cells, characterized by induction of placental lactogen I (PL-I), was accompanied by a steady decrease in the expression of RTR mRNA and down-regulation of ERRβ expression while levels of ERRα mRNA did not change significantly. RTR was able to inhibit ERRα-mediated transactivation through the consensus RTR-response element (RTRE) likely by competing with ERRα for binding to the RTRE. These results suggest the possibility of cross-talk between RTR and ERRα receptor signalling pathways in trophoblasts. © 2002 Elsevier Science Ltd.