The brain regions that control the learning and production of song and other learned vocalizations in songbirds exhibit some of the largest sex differences in the brain known in vertebrates and are associated with sex differences in singing behavior.Song learning takes place through multiple stages: an early sensory phase when song models are memorized, followed by a sensorimotor phase in which auditory feedback is used to modify song output through subsong, plastic song, to adult crystalized song. However, how patterns of neurogenesis in these brain regions change through these learning stages, and differ between the sexes, is little explored. We collected brains from 63 young male and female zebra finches (Taeniopygia guttata) over four stages of song learning. Using neurogenesis markers for cell division (proliferating cell nuclear antigen), neuron migration (doublecortin), and mature neurons (neuronspecific nuclear protein), we demonstrate that there are sex-specific changes in neurogenesis over song development that differ between the caudal motor pathway and anterior forebrain pathway of the vocal control circuit. In many of these regions, sex differences emerged very early in development, by 25 days post hatch, at the beginning of song learning. The emergence of sex differences in other components of the system was more gradual and had specific trajectories depending on the brain region and its function. In conclusion, we found that sex differences occurred early and continued during song learning. Moreover, transitions from the different phases of song development do not seem to depend on large changes in neurogenesis in the vocal control areas measured.
The role of myelination in the development of motor control is widely known, but its role in the development of cognitive abilities is less understood. Here, we examined sex differences in the development of myelination of structures and tracts that support song learning and production in songbirds. We collected brains from 63 young male and female zebra finches (Taeniopygia guttata) over four stages of development that correspond to different stages of song learning. Using a myelination marker (myelin basic protein), we measured the development of myelination in three different nuclei of the vocal control system (HVC, RA, and lateral magnocellular nucleus of the anterior nidopallium [LMAN]) and two tracts (HVC‐RA and lamina mesopallium ventralis [LMV]). We found that the myelination of the vocal control nuclei and tracts is sex related and male biased. In males, the patterns of myelination were age‐dependent, asynchronous in rate and progression and associated with the development of song learning and production. In females, myelination of vocal control nuclei was low or absent and did not significantly change with age. Sex differences in myelination of the HVC‐RA tract were large and emerged late in development well after sex differences in the size of vocal control brain regions are established. Myelination of this tract in males coincides with the age of song crystallization. Overall, the changes in myelination in the vocal control areas and tracts measured are region‐, age‐, and sex‐specific and are consistent with sex differences in song development.
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