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Axonal compartmentalization may enable localized, presynaptic plasticity to alter neuronal output in a flexible, experience-dependent manner. Here, we show that olfactory learning generates compartmentalized, bidirectional plasticity of acetylcholine release that varies across the longitudinal compartments of\n <jats:italic>Drosophila<\/jats:italic>\n mushroom body (MB) axons. The directionality of the learning-induced plasticity depends on the valence of the learning event (aversive vs. appetitive), varies linearly across proximal to distal compartments following appetitive conditioning, and correlates with learning-induced changes in downstream mushroom body output neurons (MBONs) that modulate behavioral action selection. Potentiation of acetylcholine release was dependent on the Ca\n <jats:sub>V<\/jats:sub>\n 2.1 calcium channel subunit\n <jats:italic>cacophony<\/jats:italic>\n . In addition, contrast between the positive conditioned stimulus and other odors required the inositol triphosphate receptor, which maintained responsivity to odors upon repeated presentations, preventing adaptation. Downstream from the MB, a set of MBONs that receive their input from the \u03b33 MB compartment were required for normal appetitive learning, suggesting that they represent a key node through which reward learning influences decision-making. These data demonstrate that learning drives valence-correlated, compartmentalized, bidirectional potentiation, and depression of synaptic neurotransmitter release, which rely on distinct mechanisms and are distributed across axonal compartments in a learning circuit.\n <\/jats:p>","DOI":"10.7554\/elife.76712","type":"journal-article","created":{"date-parts":[[2022,3,14]],"date-time":"2022-03-14T09:15:20Z","timestamp":1647249320000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.76712","source":"Crossref","is-referenced-by-count":34,"title":["Associative learning drives longitudinally graded presynaptic plasticity of neurotransmitter release along axonal compartments"],"prefix":"10.7554","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3170-1101","authenticated-orcid":true,"given":"Aaron","family":"Stahl","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02dxx6824","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Neuroscience, The 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