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Vocal learners, including humans and songbirds, acquire their complex vocalizations by accurately memorizing and imitating the vocal patterns of other individuals. In songbirds, the caudomedial nidopallium (NCM), considered the secondary auditory region, has been suggested to play a critical role in memorizing and recognizing the songs of tutors. However, the mechanisms by which NCM neurons encode the acoustic information of tutor song are not yet fully understood. Here, we investigate the neural representation of tutor song information in NCM neurons by examining their sensitivity to spectral changes in song structure, using electrophysiological recordings in anesthetized male zebra finches. We manipulated the acoustic structures of both tutor songs and unfamiliar conspecific songs by shifting the fundamental frequency (FF) of harmonic syllables by various frequency steps and recorded neural responses to those FF-shifted and original songs. Our results demonstrate that NCM neurons are highly sensitive to FF shifts in tutor song but much less in unfamiliar conspecific song, providing novel evidence for neural encoding of tutor song information in NCM neurons. Moreover, we find that the effects of FF shifts on neural responses depend on the direction of FF shifts. These findings suggest that NCM neurons encode detailed information of tutor song, which can serve as a tutor song template required for song learning.