GABAergic neurons participate in the brain's response to birdsong auditory stimulation

Birdsong is a learned vocal behaviour that requires intact hearing for its development in juveniles and for its maintenance during adulthood. However, the functional organization of the brain circuits involved in the perceptual processing of song has remained obscure. Here we provide evidence that GABAergic mechanisms are an important component of these circuits and participate in the auditory processing of birdsong. We first cloned a zebra finch homologue of the gene encoding the 65-kDa isoform of glutamic acid decarboxylase (zGAD-65), a specific GABAergic marker, and conducted an expression analysis by in situ hybridization to identify GABAergic cells and to map their distribution throughout auditory telencephalic areas. The results showed that field L2, the caudomedial nidopallium (NCM) and the caudomedial mesopallium (CMM) contain a high number of GABAergic cells. Using patch-clamp brain slice recordings, we found abundant GABAergic mIPSCs in NCM. Pharmacological antagonism of mIPSCs induced large EPSC bursts, suggesting that tonic inhibition helps to stabilize NCM against runaway excitation via activation of GABA-A receptors. Next, using double fluorescence in situ hybridization and double immunocytochemical labelling, we demonstrated that large numbers of GABAergic cells in NCM and CMM show inducible expression of the transcriptional regulator ZENK in response to song auditory stimulation. These data provide direct evidence that GABAergic neurons in auditory brain regions are activated by song stimulation. Altogether, our results suggest that GABAergic mechanisms participate in auditory processing and perception, and might contribute to the memorization of birdsong.