Cell-type–resolved RNA-seq reveals molecular engrams of sexual experience in Drosophila neuromodulatory neurons

This study examined how different social/sexual experiences shape gene expression in defined neuromodulatory neurons of Drosophila melanogaster males. We profiled serotonergic (TRH), octopamine/tyramine (Tdc2), and neuropeptide F receptor (NPFR) neurons following three distinct conditions known to modulate motivational states: repeated successful mating, repeated sexual rejection, or social isolation (“naïve-single”). Cell-type-specific nuclei were isolated using the INTACT method, expressing an UNC-84-GFP nuclear tag under NPFR-Gal4, TRH-Gal4, or Tdc2-Gal4 drivers. RNA was extracted from 100 male brains per condition, with three biological replicates each, and subjected to bulk RNA-seq. Differential expression analysis revealed distinct transcriptional programs associated with mating outcomes and neuronal identity, identifying molecular pathways linked to chromatin regulation, metabolism, and synaptic remodeling. This dataset provides a resource for exploring how social experience is encoded in neuromodulatory circuits.