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Revealing the organization and function of neural circuits is greatly facilitated by viral tools that spread transsynaptically. Adeno-associated virus (AAV) exhibits anterograde transneuronal transport, however the synaptic specificity of this spread and its broad application within a diverse set of circuits remains to be explored. Here, using anatomical, functional, and molecular approaches, we provide evidence for the preferential transport of AAV1 to post-synaptically connected neurons and reveal its spread is strongly dependent on s