Dmrt1 has been implicated as an important factor in sex determination in all classes of vertebrates, including reptiles with temperature-dependent sex determination (TSD). Specifically, early embryonic expression of Dmrt1 appears to be an integral part of normal testicular development in vertebrates. Recently, a number of TSD studies have placed Dmrt1 expression at the top of a short list of putative temperature-sensitive events for TSD. Dmrt1 expression has been shown to be up-regulated at male-producing temperatures during the thermosensitive period (TSP) of several TSD reptile species. An interesting finding in Dmrt1 studies of fish and amphibians has been that in species where exogenous steroids can stimulate sex reversal, Dmrt1 expression can also be manipulated by these steroid treatments. In the current study, we examine the effects of exogenous 17β-estradiol treatment on Dmrt1 expression at male-producing temperature (26°C) in a reptile with TSD, the red-eared slider turtle,Trachemys scripta. Previous studies have demonstrated that exogenous estrogens can stimulate sex reversal (at male-producing temperatures) in this species of turtle. In the current study, T. scripta embryos that received estradiol treatment displayed a significant decrease in Dmrt1 expression during the TSP, while embryos in the control group (no estradiol treatment) showed significant increases in Dmrt1 expression during the TSP. Furthermore, the pattern of Dmrt1 expression for the estradiol-treated group was similar to the pattern we previously reported for Dmrt1 at female-producing temperature (31°C). These findings indicate that exogenous estrogen is acting at or before the Dmrt1 event in the sex determination/sex differentiation cascade of T. scripta. Further, the results are consistent with the hypothesis that exogenous estrogen could be exerting its feminizing effect by down-regulating the expression of Dmrt1. © 2005 Wiley-Liss, Inc.