For more than 100 years, the fruit fly Drosophila melanogaster has been one of the most studied model organisms. Here, we present a single-cell atlas of the adult fly, Tabula Drosophilae , that includes 580,000 nuclei from 15 individually dissected sexed tissues as well as the entire head and body, annotated to >250 distinct cell types. We provide an in-depth analysis of cell type–related gene signatures and transcription factor markers, as well as sexual dimorphism, across the whole animal. Analysis of common cell types between tissues, such as blood and muscle cells, reveals rare cell types and tissue-specific subtypes. This atlas provides a valuable resource for the Drosophila community and serves as a reference to study genetic perturbations and disease models at single-cell resolution.

Motion detection is a fundamental neural computation performed by many sensory systems. In the fly, local motion computation is thought to occur within the first two layers of the visual system, the lamina and medulla. We constructed specific genetic driver lines for each of the 12 neuron classes in the lamina. We then depolarized and hyperpolarized each neuron type and quantified fly behavioral responses to a diverse set of motion stimuli. We found that only a small number of lamina output neurons are essential for motion detection, while most neurons serve to sculpt and enhance these feedforward pathways. Two classes of feedback neurons (C2 and C3), and lamina output neurons (L2 and L4), are required for normal detection of directional motion stimuli. Our results reveal a prominent role for feedback and lateral interactions in motion processing and demonstrate that motion-dependent behaviors rely on contributions from nearly all lamina neuron classes.

Ventroposterior medialis parvocellularis (VPMpc) of thalamus, the thalamic relay nucleus for gustatory sensation, receives primary input from parabrachial nucleus, and projects to insular cortex. To reveal unique properties of gustatory thalamus in comparison to archetypical sensory relay nuclei, this study examines the morphology of synaptic circuitry in VPMpc, focusing on parabrachiothalamic driver input and corticothalamic feedback. Anterogradely visualized parabrachiothalamic fibers in VPMpc bear large swellings. At electron microscope resolution, parabrachiothalamic axons are myelinated and make large boutons, forming multiple asymmetric, adherent and perforated synapses onto large caliber dendrites and dendrite initial segments. Labeled boutons contain dense-core vesicles, and they resemble a population of calcitonin gene-related peptide containing terminals within VPMpc. As typical of primary inputs to other thalamic nuclei, parabrachiothalamic terminals are over 5 times larger than other inputs, while constituting only 2% of all synapses. Glomeruli and triadic arrangements, characteristic features of other sensory thalamic nuclei, are not encountered. As revealed by anterograde tracer injections into insular cortex, corticothalamic projections in VPMpc form a dense network of fine fibers bearing small boutons. Corticothalamic terminals within VPMpc were also observed to synapse on cells that were retrogradely filled from the same injections. The results constitute an initial survey in describing unique anatomical properties of rodent gustatory thalamus.

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