Identification and characterization of sexually dimorphic neurons that express the sex-determining gene doublesex in the brain of silkmoth Bombyx mori

Sexual differences in behavior are generated by sexually dimorphic neural circuits in animals. In insects, a highly conserved sex-determining gene doublesex (dsx) plays essential roles in the development of sexual dimorphisms. In the present study, to elucidate the neural basis of sexual differences in behaviors of silkmoth Bombyx mori, we investigated Bombyx mori dsx (Bmdsx) expression in the brains through development. In the brain, Bmdsx was differentially expressed in sex- and developmental stage-dependent manners. BmDSX protein-expressing cells were located in the dorsomedial region of the pupal and adult brains, and constituted two and one neural clusters in males and females, respectively. The number of BmDSX-positive cells was developmentally regulated and peaked at the early to middle pupal stages, suggesting that the sexually dimorphic neural circuits are established during this period. The detection of a neural activity marker protein BmHR38 suggested that the BmDSX-positive cells are not active during sexual behavior in both male and female moths, even though the cells in the vicinity of the BmDSX-positive cell clusters are active. These results imply that Bmdsx plays roles in the development of sexually dimorphic neural circuits, but the neural circuits are not related to sexual behavior in silkmoths.