Anteroposterior patterning by mutual repression of orthodenticle and caudal-type transcription factors

Members of the Otx (orthodenticle) and Cdx (caudal) families of homeodomain transcription factors are expressed in similar embryonic regions in all animal groups and have been shown to be directly involved in anteroposterior patterning in a number of species. In the amphibian Xenopus laevis, the Otx family gene Xotx2 and the Cdx family gene Xcad3 are both expressed within the early dorsal organizer. We show that they have mutually repressive activities, suggesting that they play a crucial role in the early regionalization of the organizer into anterior and posterior territories. Xotx2 can act both as an activator and repressor of gene expression depending on context. A form of Xotx2 that acts exclusively as a repressor (OtxEn-R) was made by fusing the Xotx2 homeodomain to the Drosophila melanogaster engrailed transcriptional repressor domain. Overexpression of this protein in vivo indicates that OtxEn-R antagonizes the activating function of endogenous Xotx2 for anterior marker genes such as XCG and goosecoid but retains the ability to repress the expression of posterior markers such as Xcad3 and Xbra. OtxEn-R overexpression causes a severe derangement of anterior development, resulting in the loss of cement gland, eyes, stomodeal opening, and pharynx. The specification and development of anterior neural structures is dramatically abnormal up to and including the isthmic signaling center at the midbrain/hindbrain junction. This study provides good evidence that Xenopus Otx2 is required for normal head patterning and the process of anterior neural specification. We propose that a mutually antagonistic relationship between Otx and Cdx factors is a basic aspect of anteroposterior patterning in all vertebrates.