Targeted single cell expression profiling identifies integrators of sleep and metabolic state

The fruit fly, Drosophila melanogaster, suppresses sleep in response to acute food deprivation, presumably to forage for food. This process is dependent on a single pair of neurons, termed the Lateral Horn Leucokinin (LHLK) neurons, that secrete the neuropeptide Leucokinin. These neurons signal to insulin producing cells and suppress sleep under periods of starvation. Here, we use single-cell sequencing of LHLK neurons to examine the transcriptional responses to starvation. Overall design: LHLK neurons were genetically labeled by GFP expression (LK-GAL4>UAS-GFP). Later after behavioral purturbation, the targeted LHLK neurons were harvested on a custom rig, followed by CEL-Seq-based single cell RNA sequencing protocol. CEL-Seq data of LHLK neurons from female flies subjected to 24-hr starvation were compared to the control or fed group to identify differential expression in response to starvation.