GSE202407
Dopamine and GPCR-mediated modulation of DN1 clock neurons gates the circadian timing of sleep
The metronome-like circadian regulation of sleep timing must still adapt to an uncertain environment. Recent studies in Drosophila indicate that neuromodulation not only plays a key role in clock neuron synchronization but also affects interactions between the clock network and brain sleep centers. We show here that the targets of neuromodulators, G-Protein Coupled Receptors (GPCRs), are highly enriched in the fly brain circadian clock network. Single cell sequencing indicates that they are not only enriched, but they are also differentially expressed and contribute to clock neuron identity. We generated a comprehensive guide library to mutagenize individual GPCRs in specific neurons and verified the strategy by introducing a targeted sequencing approach. Combined with a behavioral screen, the mutagenesis strategy revealed a novel role of dopamine in sleep regulation by identifying two dopamine receptors and a clock neuron subpopulation that gate the timing of sleep. Overall design: GFP-labeled neurons (nSyb>GFP) and GFP-labeled clock neurons (clk856>GFP) were FACS sorted and SmartSeq2 libraries were prepared at ZT2 and ZT14. 2 Repeats each.