High-throughput assessment of vocal modulation following chemogenetic inhibition in songbirds

Chemogenetic tools that enable reversible manipulation of neuronal activity are essential for studying the neural bases of behavior, yet their application in songbirds remains limited. Here, we evaluated inhibitory DREADD (designer receptors exclusively activated by designer drugs)-mediated suppression in two widely studied songbird species: zebra finches (Taeniopygia guttata) and Bengalese finches (Lonchura striata domestica). Using the ligand deschloroclozapine (DCZ), we demonstrated effective suppression of neuronal activity both in vitro and in vivo. A high-throughput system for recording and analyzing birdsongs revealed that HVC inhibition in zebra finches reduced song production for approximately 90 min, while Area X inhibition altered phonological features over longer time courses. Bengalese finches exhibited similar but distinct song effects, suggesting species-specific differences in song production mechanisms. Notably, higher DCZ doses were required than in mice. These findings refine chemogenetic approaches in songbirds and aid in revealing further insights into the neural mechanisms underlying vocal communication.