Figure 9.

Atg8a overexpression in the dorsal fan-shaped body neurons (dFB) rescues sleep defects. A, Sleep profiles of control and experimental genotypes for 3 AWs each comprising 3 d of data. Plots represent data from two independent replicate experiments. Expression of mHTT leads to reduction in predominantly nighttime sleep in the flies. Atg8a overexpression rescues the sleep defects in the flies. B, Plot represents quantification of mean total sleep quantified for a single replicate experiment for 3 AWs. Control genotypes show good sleep levels; however, the expression of mHTT leads to a significant decrease in total sleep in the initial two AWs (red bar). Expression of Atg8a with mutant protein rescues sleep defects, and the values are comparable to the control genotypes. n > 17 flies/genotype. *p < 0.05. C, Plot represents quantification of mean day sleep quantified for a single replicate experiment for 3 AWs. Control genotypes show good sleep levels; however, the expression of mHTT leads to a significant decrease in daytime sleep in the initial two AWs (red bar). Expression of Atg8a with mutant protein rescues daytime sleep, and the values are comparable to the control genotypes (green bar). n > 17 flies/genotype. *p < 0.05. D, Plot represents quantification of mean nighttime sleep quantified for a single replicate experiment for 3 AWs. Control genotypes show good sleep levels; however, the expression of mHTT leads to a significant decrease in nighttime sleep in the initial two AWs (red bar). Expression of Atg8a with mutant protein rescues nighttime sleep. n > 17 flies/genotype. *p < 0.05. E, Plot represents quantification of mean length of daytime sleep bouts quantified for a single replicate experiment for 3 AWs. Expression of mHTT leads to a significant decrease in daytime sleep bout length (red bar). Expression of Atg8a with mutant protein rescues length of daytime sleep episode. n > 17 flies/genotype. *p < 0.05. F, Plot represents quantification of mean length of nighttime sleep bouts quantified for a single replicate experiment for 3 AWs. Expression of mHTT leads to a significant decrease in nighttime sleep bout length (red bar). Expression of Atg8a with mutant protein rescues length of nighttime sleep episode. n > 17 flies/genotype. *p < 0.05. (+) and (–) in the bar graphs represent the presence or absence, respectively, of the gene in the fly. G, Plot represents quantification of nonaggregates and aggregated mHTT. Expression of mHTT in dFB leads to accumulation of nonaggregated mHTT in the axons (left, red bar). Atg8a overexpression in the presence of mHTT significantly reduces the nonaggregated mHTT from the projections (left, green bar). However, Atg8a overexpression does not lead to any significant change in the mutant aggregate number in the targeted neurons (right). n > 10 brains/genotype. *p < 0.05. H, Representative MIP images (day 3, adult brain) of dFB neurons depicting staining of HTT (red). Dotted area represents the projections from the neurons. Arow points to the neuropil region. Scale bar, 40 µm. Asterisk on individual genotypes indicates that the genotype is significantly different from all other plotted genotypes.