The courtship song production circuit in the fruit fly Drosophila melanogaster provides an excellent animal model for studying decision-making and motor pattern generation that are important for mate selection in a number of species. With the small and genetically tractable nervous system of Drosophila, identification of the specific neurons involved in courtship song production allows interrogation of the principles underlying the transformation of complex patterns in the nervous system to a motor output. Though five neuronal classes capable of producing courtship song have been identified, previous screens for song production neurons only targeted neurons expressing the male-specific transcription factor fruitless. The functional properties and connectivity of these five neuronal classes also remains uncharacterized. To uncover more of the neural circuits underlying song production, we performed a neural activation screen for ventral nerve cord neurons that drive song production in headless males by expressing the temperature-sensitive ion channel TrpA1 in small subsets of neurons (labeled by the Janelia GAL4 collection). We identified seven GAL4 lines capable of song production when artificially activated. An intersectional approach where TrpA1 was restricted to neurons expressing either the fruitless or doublesex gene revealed that five of these GAL4 lines contain song production neurons that do not express either of these two genes. Chronic inactivation (using tetanus toxin light-chain (TNT), a genetically encoded protein that blocks synaptic transmission) of these candidate neurons during a courtship pair assay that measures both male and female movement while also recording song, allowed us to characterize these neurons’ effect on song patterning and female responses to song. Using in vivo calcium imaging of candidate song neurons from this screen combined with optogenetic activation of the song command neuron P1, we are currently examining whether or not the newly identified song neurons lie within an interconnected song circuit with those neurons previously identified.