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Background  Sex determination processes vary widely among different vertebrate taxa, but no group offers as much diversity for the study of the evolution of sex determination as teleost fish. However, the knowledge about sex determination gene cascades is scarce in this species-rich group and further difficulties arise when considering hybrid fish taxa, in which mechanisms exhibited by parental species are often disrupted. Even though hybridisation is frequent among teleosts, gene based approaches on sex determination have seldom been conducted in hybrid fish. The hybrid polyploid complex of  Squalius alburnoides  was used as a model to address this question.    Methodology/Principal Findings  We have initiated the isolation and characterization of regulatory elements ( dmrt1 ,  wt1 ,  dax1  and  figla ) potentially involved in sex determination in  S. alburnoides  and in the parental species  S. pyrenaicus  and analysed their expression patterns by  in situ  hybridisation. In adults, an overall conservation in the cellular localization of the gene transcripts was observed between the hybrids and parental species. Some novel features emerged, such as  dmrt1  expression in adult ovaries, and the non-dimorphic expression of  figla , an ovarian marker in other species, in gonads of both sexes in  S. alburnoides  and  S. pyrenaicus . The potential contribution of each gene to the sex determination process was assessed based on the timing and location of expression.  Dmrt1  and  wt1  transcripts were found at early stages of male development in  S. alburnoides  and are most likely implicated in the process of gonad development.    Conclusions/Significance  For the first time in the study of this hybrid complex, it was possible to directly compare the gene expression patterns between the bisexual parental species and the various hybrid forms, for an extended set of genes. The contribution of these genes to gonad integrity maintenance and functionality is apparently unaltered in the hybrids, suggesting that no abrupt shifts in gene expression occurred as a result of hybridisation.

Sex Determination in the Squalius alburnoides Complex: An Initial Characterization of Sex Cascade Elements in the Context of a Hybrid Polyploid Genome