Sex determination mechanisms, differing in their modality, are widely represented in all the various animal taxa, even at the intraspecific level. Within the highly diversified Class Insecta, Drosophila has been used to unravel the mechanistic molecular and genetic interactions that are involved in sex determination. Indeed, the molecularly characterized genes of the Drosophila sex determination hierarchy X:A > Sxl > tra > dsx have been fruitful starting points in the cloning of homologous genes from other insect species. This cascade seems to control sex determination in all Drosophila species. However, no sex-specific regulatory Sxl homologues have been isolated from the Mediterranean fruitfly (medfly), Ceratitis capitata, the housefly, Musca domestica, Chrysomya rufifacies nor from the distantly related phorid fly Megaselia scalaris. Moreover, all these other species use primary signals different from the intricate X:A counting system of Drosophila. However, dsx homologues isolated from these and other dipteran species as well as from the silkmoth, Bombyx mori, share a conserved sex-specific regulation based on alternative splicing. An understanding of the sex determination mechanisms in insects that are of agricultural or public health importance may help in the development of improved methods for their control using the sterile insect technique.