We compared the expression of bovine prolactin-related protein-1 (bPRP-1), placental lactogen (bPL), and pregnancy-associated glycoproteins-1 (bPAG-1) and -9 (bPAG-9) genes in artificially inseminated (AI) and nuclear transferred (NT) cows during the first trimester of gestation using real-time reverse transcription-polymerase chain reaction and in situ hybridization. Placentomal (cotyledonary, caruncular) and interplacentomal (intercotyledonary, intercaruncular) tissues of AI and NT cows carrying either motile (M) or immotile (IM) fetuses were examined. Transcripts for bPL and bPAG-9 were lower (P < 0.01) in the fetal membranes of NT (n = 4) cows at Day 30 of gestation, compared with AI (n = 4) cows. There was no difference in the mean (+/- SEM) levels of expressions of bPRP-1, bPL, and PAG-1 in the placentomal and interplacentomal tissues of AI (n = 5) and NT (M, n = 4) cows at Day 60 of gestation. The mRNAs for bPRP-1, bPL, bPAG-1, and bPAG-9 genes were higher (P < 0.01) in the caruncular tissue of AI cows, compared with NT (IM, n = 4) cows at Day 60 of gestation. Expression of bPRP-1, bPL, bPAG-1, and bPAG-9 in the placentomal and interplacentomal tissues of the NT (n = 3) group varied considerably more, compared with the AI (n = 4) group at Day 100 of gestation. These findings suggest defective binucleate cell-specific gene transcriptional commands in NT cows.