Two spliced isoforms of the sex-determination gene fruitless in the Chinese mitten crab Eriocheir sinensis.

The fruitless (fru) gene plays an important role in the sex determination pathway and courtship behavior of Drosophila melanogaster. In the present study, two fru isoforms (Esfru1 and Esfru2) were identified from the Chinese mitten crab Eriocheir sinensis. Sequence analysis showed that Esfru1 and Esfru2 were encoded by the same genomic locus and generated by alternative splicing of pre-mRNA. Esfru1 had all introns completely spliced out, while Esfru2 had a longer exon1 with an additional 78bp sequence. They both contained a conserved BTB domain which was also found in D. melanogaster Fru isoforms. Analysis on temporal expression profiles of Esfru1-2 (a common region of Esfru1 and Esfru2) and Esfru2 (a specific region of Esfru2) showed that they expressed in similar patterns during embryonic development but in different patterns during larval development. The expression of Esfru1-2 decreased gradually from zoea III stage, however, Esfru2 increased from zoea IV stage and reached the peak at megalopa stage. Expression distribution in tissues and in situ hybridization analysis revealed that they showed sexually dimorphic expression in gonads, hepatopancreas and brains. Esfru1-2 showed significantly higher expression in female gonads and hepatopancreas than in males, but highly expressed in male brains than that in females. Interestingly, Esfru2 was displayed in a male-specific manner in each tissue, especially in gonads and hepatopancreas. Our results indicate that Esfru1 might be involved in both sexual brain neuronal structure development and related to female-specific character development. Esfru2 may participate in male-specific character development. This is the first report that characterizes two spliced variants of fru in crustaceans and provides basic information for further functional studies of the crab sex-determination mechanism.