Summary Deciphering how brains generate behavior depends critically on an accurate description of behavior. If distinct behaviors are lumped together, separate modes of brain activity can be wrongly attributed to the same behavior. Alternatively, if a single behavior is split into two, the same neural activity can appear to produce different behaviors [1]. Here, we address this issue in the context of acoustic communication in Drosophila . During courtship, males utilize wing vibration to generate time-varying songs, and females evaluate songs to inform mating decisions [2-4]. Drosophila melanogaster song was thought for 50 years to consist of only two modes, sine and pulse, but using new unsupervised classification methods on large datasets of song recordings, we now establish the existence of at least three song modes: two distinct, evolutionary conserved pulse types, along with a single sine mode. We show how this seemingly subtle distinction profoundly affects our interpretation of the mechanisms underlying song production, perception and evolution. Specifically, we show that sensory feedback from the female influences the probability of producing each song mode and that male song mode choice affects female responses and contributes to modulating his song amplitude with distance [5]. At the neural level, we demonstrate how the activity of three separate neuron types within the fly’s song pathway differentially affect the probability of producing each song mode. Our results highlight the importance of carefully segmenting behavior to accurately map the underlying sensory, neural, and genetic mechanisms.

Discovery of a new song mode in Drosophila reveals hidden structure in the sensory and neural drivers of behavior