Voltage imaging reveals circuit computations in the raphe underlying serotonin-mediated motor vigor learning.

As animals adapt to new situations, neuromodulation is a potent way to alter behavior, yet mechanisms by which neuromodulatory nuclei compute during behavior are underexplored. The serotonergic raphe supports motor learning in larval zebrafish by visually detecting distance traveled during swims, encoding action effectiveness, and modulating motor vigor. We tracked the raphe's input-output computations at millisecond timescales using voltage and neurotransmitter imaging and found that swimming opens a gate for visual input to cause spiking in serotonergic neurons, enabling the encoding of action outcomes and filtering out learning-irrelevant visual signals. Specifically, swim commands initially inhibited serotonergic neurons via γ-aminobutyric acid (GABA). Immediately after, membrane voltage increased via post-inhibitory rebound, allowing swim-induced visual motion to evoke firing through glutamate, triggering serotonin release to modulate future motor vigor. Ablating local GABAergic neurons impaired raphe coding and motor learning. Thus, serotonergic neuromodulation arises from action-outcome coincidence detection within the raphe.