Synaptic Targets of Circadian Clock Neurons Influence Core Clock Parameters

Neuronal connectivity in the circadian clock network is essential for robust endogenous timekeeping. In the Drosophila circadian clock network, four pairs of small ventral lateral neurons (sLN_vs) serve as critical pacemakers. Peptidergic communication via sLN_v release of the key output neuropeptide Pigment Dispersing Factor (PDF) has been well characterized. In contrast, little is known about the role of the synaptic connections that sLN_vs form with downstream neurons. Connectomic analyses revealed that the sLN_vs form strong synaptic connections with a group of previously uncharacterized neurons, SLP316. Here, we show that silencing synaptic output in the SLP316 neurons via tetanus toxin (TNT) expression shortens the free-running period, whereas hyper-exciting them by expressing the Na[+] channel NaChBac results in period lengthening. Under light-dark cycles, silencing SLP316 neurons also causes lower daytime activity and higher daytime sleep. Our results revealed that the main postsynaptic partners of the Drosophila pacemaker neurons are a non-clock neuronal cell type that regulates the timing of sleep and activity.