These files contain data supporting all results reported in Ziobro et al., Different vocalization types in mouse are controlled by distinct populations of midbrain neurons. In Ziobro et al., we tested whether a single population of midbrain neurons regulates the production of different vocalization types. To this end, we applied intersectional methods to selectively ablate a population of midbrain periaqueductal gray (PAG) neurons important for the production of ultrasonic vocalizations (USVs) in mice. We found 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 distress calls. Our findings suggest that distinct populations of midbrain neurons control the production of different vocalization types.