Molecular mechanisms underlying steroid hormone action during sex determination in the red-eared slider turtle, Trachemys scripta elegans



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Many reptiles, including the red-eared slider turtle (Trachemys scripta elegans), exhibit temperature-dependent sex determination (TSD). Temperature determines sex during a temperature sensitive period (TSP), when gonadal sex is labile to both temperature and hormones -- particularly estrogen. Estrogen production is a key step in ovarian differentiation for many vertebrates, including TSD reptiles, and temperaturebased differences in aromatase expression during the TSP may be a critical step in ovarian determination. Steroidogenic factor-1 (Sf1) is a key gene in vertebrate sex determination and regulates steroidogenic enzymes, including aromatase. The biological actions of steroid hormones are mediated by their receptors, defined here as the classic transcriptional regulation of target genes. To elucidate the mechanism of estrogen action estrogen during sex determination, I examined aromatase, Sf1, ER[alpha], ER[beta], and AR expression in slider turtle gonads before, during and after the TSP, as well as following sex reversal via temperature or steroid hormone manipulation by administering exogenous estradiol (E2) or aromatase inhibitor (AI) to the eggshell. Sf1 is expressed at higher levels during testis development and following maleproducing temperature shift and AI treatment, while aromatase increases during ovary determination and feminizing temperature shift and E2 treatment. My results do not lend support to a role for Sf1 in the regulation of aromatase expression during slider turtle sex determination, but do support a critical role for estrogen in ovarian development. Estrogen receptor [alpha] and AR levels spike at the female-producing temperature just as aromatase levels are increasing during ovarian sex determination, while ER[beta] remains constant and only increases late in ovarian differentiation -- well after estrogen levels have increased, indicating that ER[alpha] and ER[beta] may have distinct roles in slider turtle ovarian development. Estrogen receptor [alpha] and ER[beta] are expressed along developing sex cords in the absence of estrogen (AI treatment). When shifted to female-producing temperatures, embryos maintain medullary ER[alpha] and AR expression while ER[beta] is reduced. By contrast, ER[alpha] and ER[beta] redirect to the cortex in E2-created ovaries. Warmer temperature and E2 result in the same endpoint (ovarian development), but may entail different steroid signaling patterns between temperature- and estrogen-induced feminization.