Short neuropeptide F regulates the starvation mediated enhanced locomotor activity in Drosophila
Geo, Anna et al. (2019), Short neuropeptide F regulates the starvation mediated enhanced locomotor activity in Drosophila, Dryad, Dataset, https://doi.org/10.5061/dryad.683b46g
Figure 1 Raw data
To assess the mRNA transcription profiles of snpf, three replicates were used for each time point. Each replicate contained 30 heads from w1118flies. Total RNA was extracted using QIAGEN RNAeasy Plus Mini kit; DNAase digestion was performed using QIAGEN RNase–free DNase. cDNA was synthesized using SuperScript™ III First-Strand Synthesis System (Invitrogen, Cat. No. 18080051) and real-time PCR was performed using Bio-Rad CFX96TM with the cDNA template, Power SYBR® Green PCR Master Mix (Cat. # 4368702) from ThermoFisher Scientific to check temporal transcription profile of snpf. mRNA levels were measured under LD cycle and constant darkness (DD). The molecular oscillation of mRNA levels was determined by normalizing the mRNA level of the gene of interest with the mRNA level of rp49 at each time point. Cosinor analysis was implemented in MATLAB-R2016a to test for rhythmicity, to estimate the peak phase and the amplitude (peak/trough ratio) of relative mRNA expression of snpf.
Figure 2 and 3 Raw data
For activity/rest assay, freshly emerged male flies were pre-entrained for two days in the LD cycle at 25 oC and a humidity of 75 ± 5% inside the incubator (MIR-154, Panasonic, Japan). Two-day-old virgin males were individually introduced into activity tubes. Flies were provided with standard cornmeal medium and their activity/rest behavior was recorded using Drosophila Activity Monitors (DAM, Trikinetics, USA). Following acclimatization in activity tubes for 24 hours, flies were transferred to 1% agar medium or to standard cornmeal medium in glass tubes at ZT12 (during the onset of dark phase) to provide starved or fed conditions, respectively, and locomotor activity was recorded for 36 hours at 25 oC. Each experiment was performed with 28-32 flies for each genotype and DAM readings were collected in every 1-minute interval and the activity counts/ hour is plotted in the locomotor activity waveform. Flies died within 24 hour of starvation were excluded and flies survived for 24-36 hours of starvation were used for the analysis. Change in activity under starvation was calculated as [(Activity under starvation-Activity under fed) /Activity under fed condition] x 100. Separate sets of flies were used for recording the locomotor activity under fed and starved flies condition. Hence the change in activity under starvation condition was calculated as [(Activity of individual fly under starvation-Average activity of flies under fed) / Average activity of flies under fed] x 100. The waveform of activity/rest rhythm, day time activity, nighttime activity, change in locomotor activity under starvation condition were analyzed by using ANOVA followed by Tukey’s post hoc multiple comparisons.
Figure S1 Raw data
Wellcome Trust/DBT India Alliance Fellowship, Award: IA/E/15/1/502329