New alleles of nlp-2 , nlp-22 , and nlp-23 demonstrate that they are dispensable for stress-induced sleep in C. elegans

Sleep is ancient and genetically conserved across phylogeny. Neuropeptide signaling plays a fundamental role in the regulation of sleep for mammals, fish, and invertebrates like Caenorhabditis elegans . Developmentally timed-sleep and stress-induced sleep of C. elegans are controlled by distinct and overlapping neuropeptide pathways. The RPamide neuropeptides nlp-2 , nlp-22 , and nlp-23 , play antagonistic roles during the regulation of developmentally-timed sleep, however, their role in stress-induced sleep has not been explored. These genes are linked on the X chromosome, which has made genetic analyses challenging. Here we used CRISPR to generate new alleles of nlp-22 and nlp-23 , nlp-22 ; nlp-23 double mutants, and nlp-2 ; nlp-22 ; nlp-23 triple mutants. Confirming previous studies, we find that nlp-22 is required for developmentally-timed sleep, and show that nlp-23 is also required. However, all three genes are dispensable for stress-induced sleep.