The patterns, and the cellular sites, of expression of the genes for retinol-binding protein (RBP) and transthyretin (TTR) were studied during early- to mid-stages of rat embryogenesis. In situ hybridization of single-stranded RNA probes to rat embryo sections revealed specific sites at which RBP messenger RNA (mRNA) and TTR mRNA were localized in rat conceptuses from 7 to 13 days of gestation. RBP and TTR mRNAs were both observed in the visceral endoderm as early as at 7 days of gestation. The transcripts were not expressed in the parietal endoderm. At 9 days of gestation, TTR mRNA was strongly detected in the visceral extraembryonic endoderm and in the foregut endoderm, whereas RBP mRNA was detected only in the visceral extraembryonic endoderm. From the 10th day to the 13th day of gestation, both transcripts were increasingly expressed in the visceral yolk sac endoderm and in the developing fetal liver, and gradually decreased in the foregut. At 11 days of gestation, TTR mRNA was first detected in the tela choroidea of the fourth ventricle in the brain; and at 13 days, the TTR mRNA was strongly evident in the developing choroid plexus of the fourth and lateral ventricles. These in situ hybridization studies with embryos at different developmental stages show that RBP and TTR mRNAs are transcribed quite early during embryogenesis. The protein products of these transcripts may play important roles in vitamin A and thyroid hormone metabolism, and in the functions that these regulatory compounds play, in the developing rat embryo.