Expression profiles of sex differentiation-related genes during ontogenesis in the European sea bass acclimated to two different temperatures.

The European sea bass is a teleost fish that lacks sex chromosomes and for which temperature influences sex ratios. However, correlation between temperature, developmental stage at a given age and sex-specific gene expression is hampered by the lack of sex markers. To study this correlation, fish were exposed to feminizing (15 degrees C) or masculinizing temperature (21 degrees C) from 0-120 days post fertilization, throughout the thermosensitive period (TSP). Aromatase (cyp19a1a), 11beta-hydroxylase (cyp11b), androgen receptor (arb) and estrogen receptors (era, erb1 and erb2) were assessed by qPCR prior and during sex differentiation. Canonical discriminant analysis (CDA), with length--as proxy for developmental stage--and cyp19a1a expression as predictors, was validated and used to reliably assign gonadal sex to fish sampled within and outside the TSP. Differences in cyp19a1a and cyp11b expression could be detected 1-month before the first signs of histological sex differentiation. Cyp19a1a and cyp11b were significantly higher in future females and males, respectively, and revealed as robust molecular markers to predict future ovarian and testicular differentiation. In contrast, no association between phenotypic sex and arb, era, erb1 and erb2 expression was found, suggesting that these genes do not contribute to the differentiation of a particular sex. The CDA-based approach implemented here could be used to sex undifferentiated animals in species where genetic sex cannot be known owing to the lack of simple sex determining systems, as it is the case of many fish and reptiles with or without temperature-dependent sex determination, and provide a useful tool to relate gene expression and phenotypic sex.