Transgenerational epigenetic sex determination: Environment experienced by female fish affects offspring sex ratio

Sex determination is a complex process that can be influenced by environment in various taxa. Disturbed environments can affect population sex ratios and thus threaten their viability. Emerging evidences support a role of epigenetic mechanisms, notably DNA methylation, in environmental sex determination (ESD). In this work, using zebrafish as model and a transgenerational experiment comprising 4 successive generations, we report a strength link between the promotor methylation level of three genes in female gonads and population sex ratio. One generation of zebrafish was exposed throughout its lifetime to cadmium (Cd), a non-essential metal, at an environmentally relevant concentration. The subsequent generations were not exposed. At the first and the third generation a subset of individuals was exposed to an elevated temperature, a well-known masculinizing factor in zebrafish. While heat was associated to an increase in the methylation level of cyp19a1a gene and population masculinization, foxl2a/dmrt1 methylation levels appeared to be influenced by Cd and fish density leading to offspring feminization. Ancestral Cd exposure indeed led to a progressive feminization of the population over generations and affected the sex plastic response of zebrafish in response to heat. The effect of Cd on the methylation level of foxl2a was observed until the third generation, supporting potential transgenerational inheritance. Our results support (i) a key role of cyp19a1a methylation in SD in zebrafish in response to environmental cues and (ii) the fact that the environment experienced by parents, namely mothers in the present case, can affect their offspring sex ratio via environment-induced DNA methylation changes in gonads. [Display omitted] •Ancestral Cd exposure led to a progressive feminization of fish populations.•Methylation level of foxl2a/dmrt1 in females was affected by Cd and fish density.•Cd effect on foxl2a methylation was transmitted until the third generation.•foxl2a methylation level in mothers was highly correlated to offspring sex ratio.•Pollutant-induced changes in DNA methylation marks affect fish plasticity and sex. Pollution experienced by female fish transgenerationally affects the sex of their offspring via gonadal DNA methylation changes, leading to an increasingly biased sex ratio over generations.