Reproductive hormones can modulate communication-evoked behavior by acting on neural systems associated with motivation; however, recent evidence suggests that modulation occurs at the sensory processing level as well. The anuran auditory midbrain processes communication stimuli, and is sensitive to steroid hormones. Using multiunit electrophysiology, we tested whether sex and circulating testosterone influence auditory sensitivity to pure tones and to the natural vocalization in the green treefrog, Hyla cinerea. Sex did not influence audiogram best frequencies although sexes did differ in the sensitivities at those frequencies with males more sensitive in the lower frequency range. Females were more sensitive than males in response to the natural vocalization, despite showing no difference in response to pure tones at frequencies found within the advertisement call. Thresholds to frequencies outside the range of the male advertisement call were higher in females. Additionally, circulating testosterone increased neural thresholds in females in a frequency-specific manner. These results demonstrate that sex differences are limited to frequency ranges that relate to the processing of natural vocalizations, and depend on the type of stimulus. The frequency-dependent and stimulus-dependent nature of sex and testosterone influences suggests that reproductive hormones influence the filtering properties of the auditory system.