Immediate éarly genes such as the proto-oncogene c-fos can be expressed in neurons following synaptic excitation by sensory stimulation. C-fos immunocytochemistry has subsequently been shown to be a very sensitive marking technique for neuronal activity. Here, antibodies against the c-fos protein product Fos were used to map the tonotopic organization in the auditory system of adult and developing rats. After stimulating adult rats with pure-tone pulses, bands of Fos-immunoreactive neurons revealed the frequency representation in seven brainstem nuclei: all three subdivisions of the cochlear nucleus, the lateral superior olive, the medial nucleus of the trapezoid body, the ventral nucleus of the trapezoid body, the rostral periolivary nucleus, the dorsal nucleus of the lateral lemniscus and the inferior colliculus. With the exception of the dorsal cochlear nucleus and the inferior colliculus, tonotopicity has not been previously demonstrated in the brainstem nuclei of the rat. During development two striking results were obtained. First, beginning at postnatal day 14 (i.e. ∼2 days after physiological hearing begins in rats), not only low but also high frequencies were able to induce strong Fos immunoreactivity, indicating that gradual recruitment of formerly unresponsive high-frequency sites does not occur in the rat. Second, a gradual age-related shift of the position of isofrequency bands was not seen in any of the nuclei, suggesting that changes in frequency-place code do not occur after 2 weeks postnatally. These results indicate that the rat's auditory brainstem nuclei achieve their adult-like tonotopic organization early on, implying a somewhat different developmental time course than is found in other mammalian species.