We have analyzed the expression of four mouse homeobox genes related to two Drosophila genes expressed in the developing head of the fly. Two of these genes, Emx1 and Emx2, are related to empty spiracles, and two genes, termed Otx1 and Otx2, are related to orthodenticle. These genes are all expressed in the developing rostral brain of E10 mouse embryos and their expression domains can be compared. Otx2 is expressed in all dorsal and most ventral regions of telencephalon, diencephalon, and mesencephalon. The Otx1 expression domain is similar to that of Otx2, but smaller and contained within it. The Emx2 expression domain is comprised of dorsal telencephalon and small diencephalic regions, both dorsally and ventrally. Finally, Emx1 expression is exclusively confined to the dorsal telencephalon. At the time when regional specification of major brain regions takes place, the expression domains of the four genes appear to be continuous regions contained within each other in the sequence Emx1 < Emx2 < Otx1 < Otx2. The first appearance of transcripts of the four genes is also sequential: Otx2 is expressed first (E5.5), followed by Otx1 and Emx2 (E8-8.5), and finally by Emx1 (E9.5). It is tempting to speculate about a possible role of the four genes in establishing and/or signalling the limits of the various embryonic brain regions in a discrete progressive process with its center in the dorsal telencephalon.