Development of functional synaptic connections in the auditory system visualized with optical recording: afferent-evoked activity is present from early stages.

A comprehensive survey of auditory network formation was performed in the brain stem of the chicken embryo using voltage-sensitive dye recording. Intact medulla/brain stem preparations with the auditory branch of the eighth nerve attached were dissected from 5.5- to 8-day chicken embryos, and responses evoked by nerve stimulation were recorded optically. In the medulla of 7- and 8-day embryos, we identified four response areas, corresponding to ipsilateral Nucleus magnocellularis (NM) and Nucleus angularis (NA), which receive the auditory afferents, and ipsi- and contralateral Nucleus laminaris (NL), which receive projections from NM. The optical responses consisted of a fast spikelike signal followed by a long-lasting slow signal, which reflected the sodium-dependent action potential and glutamatergic excitatory postsynaptic potential (EPSP), respectively. In NM, NA, and NL, the EPSP-related slow optical signals were detected from some 6-day and all 7- and 8-day preparations, indicating that functional synaptic connectivity in these nuclei arises by the 7-day stage. In the pons of 7- and 8-day embryos, we identified two additional response areas, which evidently correspond to ipsi- and contralateral Nucleus lemnisci lateralis (NLL), the higher-order nuclei of the auditory pathway. Furthermore, we detected optical responses from the contralateral cerebellum, which possibly correspond to transient projections observed only during embryogenesis. The present study demonstrates that functional auditory circuits are established in the chicken embryo at stages earlier than previously reported. We discuss the possible role of afferent-evoked activity with reference to auditory neural network formation.