A third member of the murine caudal-like gene family, Cdx-4, has been isolated. In situ hybridization and immunohistochemistry have been used to study the localization of mRNA and protein during murine embryogenesis. Cdx-4 is expressed transiently from 7.0 d.p.c. (days post coitum) until 10 d.p.c., starting at the beginning of gastrulation (7.0-7.5 d.p.c.) in the allantois and posterior tip of the primitive streak. At the mid-streak stage, Cdx-4 expression moves rostrally, and protein and mRNA are detected in all cells over the posterior half of the primitive streak. As development proceeds, Cdx-4 gene products continue to be restricted to the posterior of the embryo, including the remnants of the primitive streak. Cdx-4 is expressed in neurectoderm, presomitic and lateral plate mesoderm, and hindgut endoderm, but the anterior boundary in the paraxial mesoderm is staggered with respect to the other germ layers. At all stages analyzed, Cdx-4 exhibits a graded expression pattern with a posterior maximum, a distribution highly reminiscent of the Drosophila caudal gene. These data add to the recently described distributions of other vertebrate caudal-like genes, and further support the idea that members of this homeobox gene subfamily have regulatory roles in the specification of anteroposterior axial polarity in early embryos.