Figure 1.

Genetic disruption of developmental sleep (via npr-1 and aptf-1) increases DVB neurite outgrowth dependent on nrx-1

(A) Schematic of C. elegans developmental sleep stages between larval stages with timing of each sleep deprivation method indicated. aptf-1 (orange) and npr-1 (blue) animals experience impaired sleep during all larval sleep periods, while vibration (pink) and RIS silencing (green) are applied specifically during the L4 to adult transition/molt. Cartoon zoom in of the adult male tail with DVB shown.

(B) Schematic of C. elegans nrx-1 gene coding locus, red box indicates deletion in nrx-1(wy778) referred to as nrx-1(del), red star indicates premature stop in nrx-1(gk246237) referred to as nrx-1(stop). Quantification of DVB (C) total neurite length and (D) junctions in controls, nrx-1(del), aptf-1, and aptf-1; nrx-1(del) males at day 1.

(E) Representative images of DVB neuron in day 1 males using lim-6p::gfp. Red arrows indicate neurite branches. Black dashed lines represent body outline. White asterisk marks PVT neuron (not traced, see⁶⁴). Quantification of DVB (F) total neurite length and (G) junctions in controls, nrx-1(del), npr-1, and npr-1; nrx-1(del) day 1 males.

(H) Representative images of DVB at day 1 in npr-1 mutant males and all controls.

(I) Quantification of number of cla-1::gfp puncta in DVB in controls and sleep deprived aptf-1 and npr-1 males at day 1.

(J) Representative images of DVB (red) and cla-1::gfp puncta (green) in controls, aptf-1, and npr-1 day 1 males. Outlines of synaptic puncta generated from FIJI particle analysis are shown (black) superimposed on a trace of the DVB neuron (light red). Number of individual animals is indicated by “n”, p values from one-way ANOVA with Tukey’s post-hoc test shown, error bars represent SEM, scale bar size as labelled.