Journal: bioRxiv

Article Title: Neurofibromin 1 mediates sleep depth in Drosophila

doi: 10.1101/2022.09.15.508161

Figure Lengend Snippet: (A-G). The Drosophila Arousal Tracking (DART) was used to probe arousal threshold and reactivity measurements. This system records fly movement while simultaneously controlling mechanical stimuli via a digital analog converter (DAC). All measurements included were taken from sleeping flies and determined hourly, starting at ZT0. A. Mechanical stimuli of increasing strength were used to assess arousal threshold and was determined by fly movement within 15 sec of stimulus delivery. B. Mechanical stimuli at maximum intensity was used to measure reactivity. A fly was considered reactive if it moved within 60 sec of stimulus delivery. C. There is a significant effect of genotype on arousal threshold (REML: F 2,94 = 37.62, P <0.0001; N = 29–35). Compared to control and heterozygote flies, arousal threshold significantly decreases in Nf1 P1 mutants and occurs during the day (+, P <0.0001; het, P <0.0001) and night (+, P <0.0001; het, P <0.0001). D. There is a significant effect of genotype on arousal threshold (REML: F 2,98 = 7.795, P <0.0007; N = 25–40). Compared to controls, panneuronal knockdown of Nf1 has mixed effects on arousal threshold during the day (nsyb>+, P <0.2138; Nf1 RNAi >+, P <0.0040) and significantly decreases arousal threshold during the night (nsyb>+, P <0.0271; Nf1 >+, P <0.00062). E. Linear regression of daytime reactivity as a function of time asleep in Nf1 P1 mutants, heterozygotes, and their respective control. The slopes of each regression line are not significantly different from each other (ANCOVA with time asleep as the covariate: F 2,2062 = 0.0254, P <0.9749). F. Linear regression of nighttime reactivity as a function of time asleep in Nf1 P1 mutants, heterozygotes, and their respective control. The slopes of each regression line are significantly different from each other (F 2,2100 = 57.05, P <0.0001). G. Linear regression of daytime reactivity as a function of time asleep in pan-neuronal Nf1 RNAi knockdown flies and their controls. The slopes of each regression line are not significantly different from each other (F 2,1285 = 0.5551, P <0.5741). H. Linear regression of nighttime reactivity as a function of time asleep in pan-neuronal Nf1 RNAi knockdown flies and their controls. The slopes of each regression line are significantly different from each other (F 2,1564 = 4.887, P <0.0077). For arousal threshold measurements, the median (solid line), as well as the 25th and 75th percentiles (dotted lines) are shown. For reactivity, error bars indicate ± SEM. The P -values in each panel indicates whether the slope of the regression line is significantly different from zero. White background indicates daytime, while gray background indicates nighttime. * p <0.05; ** p <0.01; *** p <0.001; **** p <0.0001.

Article Snippet: The vibrational stimulus, video tracking parameters, and data analysis were performed using the DART interface developed in MATLAB (MathWorks, Natick, Massachusetts).

Techniques: