Multiple alternative splicing of mouse Dmrt1 during gonadal differentiation

Evolutionarily conserved Dmrt1 encodes a transcriptional regulator that is expressed exclusively in the gonads and is required for testis differentiation. Here we report that four transcripts of the mouse Dmrt1 were generated in developing gonads and adult testis by alternative splicing. Dmrt1 a encodes the known protein with 374 amino acids. Dmrt1 b, Dmrt1 c, and Dmrt1 d encode predicted proteins with 212, 257, and 194 amino acids, respectively. Dmrt1 a2 and Dmrt1 a3 have anterior alternative polyadenylation signals in 3′UTR than the known Dmrt1 transcript (Dmrt1 a1). Dmrt1 a2 lacks 18 nucleic acids in 3′UTR. Interestingly, Dmrt1 b lacks exon5, Dmrt1 c lacks exon3, and Dmrt1 d lacks both exon1 and exon2 which encode DM domain. RT-PCR showed that all of Dmrt1 transcripts were only detectable in adult testis. However, during gonadogenesis, the multiple alternatively spliced transcripts all had gonad-specific and sexually dimorphic expression profiles. Northern blot and real time fluorescent quantitative RT-PCR further indicated that the expression of Dmrt1 a was dominantly higher than those of Dmrt1 b, c, and d, although they showed a similar up and down pattern of expression during embryo development. The expression of Dmrt1 transcripts climbed up to a climax at 13.5 dpc in both male and female gonad. Afterwards, expression in male gonad decreased to a low level and maintained, whereas female one reduced rapidly until undetectable in adult ovary. These results provide new insight into roles of regulation at level of splicing of the Dmrt1 in governing sex differentiation.