It has been hypothesized that estrogen production may play a pivotal role in the sex determination of reptiles with temperature-dependent sex determination (TSD). This hypothesis has been furthered by studies that have shown higher aromatase activity in the developing ovaries in some reptiles. However, other studies have not consistently supported this hypothesis. In the current study we addressed this issue by cloning P450 aromatase cDNA in the turtle, Trachemys scripta, and developing a quantitative competitive RT-PCR for aromatase. This assay was then used to quantify aromatase mRNA levels in adrenal-kidney-gonad complexes (AKG) during TSD. Aromatase mRNA was detected in the AKGs at both male- and female-producing temperatures from the earliest stage of development sampled (stage 15), through hatching (stage 26). However, levels remained relatively constant during the thermosensitive period of TSD. Further, no significant difference was detected between male- and female-producing temperatures during the thermosensitive period. After the thermosensitive period, aromatase mRNA levels increased in females (this coincides with the period during which the ovaries are differentiating). These results are consistent with those of several previous studies of certain reptiles with TSD. The current results suggest that the expression of aromatase may not be a pivotal regulatory step in the sex determination cascade of this turtle. © 2002 Elsevier Science (USA). All rights reserved.