Different populations of midbrain neurons control the production of distinct acoustic categories of vocalization

Animals produce different acoustic types of vocalizations in different behavioral contexts. The midbrain periaqueductal gray (PAG) is obligatory for vocal production (Jurgens, 1994, 2002, 2009), but how midbrain circuits are organized to control the production of different vocalization types remains unknown. To test whether a single population of midbrain neurons regulates the production of different vocalization types, we applied intersectional methods to selectively ablate a population of midbrain neurons required for the production of ultrasonic vocalizations (USVs) in mice. We find that although ablation of these PAG-USV neurons blocks USV production in both males and females, these neurons are not required for the production of squeaks. Our findings provide strong evidence that distinct populations of midbrain neurons control the production of different acoustic categories of vocalizations.