Postnatal auditory stimulation influences early perceptual learning. Previously we reported morphological effects of prenatal auditory stimulation by species-specific and sitar musical sounds on the chick brainstem auditory nuclei-nucleus magnocellularis and nucleus laminaris. At hatching, these two nuclei of auditory enriched embryos showed higher neuronal numbers, amongst other morphological changes. There were also increases in synaptophysin and syntaxin1 expressions in the sound enriched groups and modulation of the developmental expression of transcription factors c-Fos and c-Jun. We hypothesized that prenatal auditory enrichment may have reduced embryonic apoptosis in these nuclei with possible alteration of molecular mechanisms enhancing the postsynaptic neuron's ability to survive. In the present study, therefore, we examined apoptotic cell death by TUNEL technique and Bcl-2 expression using immunohistochemistry and immunoblotting. In the controls, a peak percentage in the TUNEL-positive cells was noted in the auditory nuclei at embryonic day 12, which was reduced at embryonic day 16. Bcl-2 immunoreactivity decreased from embryonic day 8 to embryonic day 12 overlapping the period of embryonic cell death in these nuclei. The stimulated groups, however, showed fewer apoptotic neurons and higher Bcl-2 level than that in the controls. On the other hand, Bax immunohistochemistry showed correlated reverse changes compared to Bcl-2 expression. Thus prenatal extra-acoustic stimulation appears to alter Bcl-2 and Bax expression to support cell survival and differentiation, thereby augmenting the development of auditory nuclei.

Recognition of mother's voice by human neonates and behavioral responses of birds and animals to sounds experienced prenatally emphasize the role of sensory inputs in auditory system development. Spontaneous and experience driven neural activity influence the neural circuits' refinement in developing brain. However, cellular mechanisms endowing plasticity for such structural refinement during critical developmental periods are less understood. Sensory stimulation induces fluctuating expression of transcription factors (TFs) of Fos, Jun, and Krox families in the related brain nuclei to activate genes to synthesize proteins such as those needed for cytoskeletal structures, ion channels, and regeneration. To understand the cellular mechanism of response to prenatal auditory stimulation, we studied the expression of c-Fos and c-Jun in brainstem auditory nuclei, nucleus magnocellularis, and nucleus laminaris of the domestic chick. The chick brainstems, five each of E8 (embryonic day 8), E12, E16, E20, and posthatch day 1 were processed for immunohistochemistry as well as Western blotting and quantified using image analysis systems. In controls, c-Fos and c-Jun expression in both the nuclei was developmentally up-regulated. Reduced c-Fos expression and increase in c-Jun was temporarily observed between E12-16. In the stimulated groups, c-Fos expression was elevated while c-Jun showed a reduction matched to controls. This diametrically opposing pattern of c-Fos and c-Jun expression in response to stimulation is indicative of cell survival. Thus the expression of TFs in the auditory nuclei shows a relationship beyond a simple stimulation-activity-expression. While developmental signals control the expression of TFs, extra sensory stimulation modulates their expression to possibly support neuronal survival and enhance synthesis of other proteins.