Computational protocol: Modulatory Action by the Serotonergic System: Behavior and Neurophysiology in Drosophila melanogaster

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Protocol publication

[…] Flies were raised at 23°C, 12 : 12 LD cycle. The adult flies, females or males, were anesthetized by exposing them to CO2. The flies were left to recover for at least 16 hours. Then, the flies (less than 1 to 6 days old) were anesthetized by cold. Adult flies were transferred into empty vials, and they were placed inside an ice bucket for 20–30 sec. The flies (4-5) were transferred into a plastic Petri dish (9 cm diameter and 0.8 cm height), and a glass cover was put over it. The flies were left for 10–13 min to recover from cold anesthesia at room temperature. Afterwards, the flies were placed in a 37°C room. The fly locomotion activities were recorded with a webcam (WEBCAM HD4110, Hewlett-Packard Company, Palo Alto, CA), which was connected to a computer, and the activity was recorded at 5 frames per second (5 fps) for 10 min using VirtualDub-1.10.4 software (http://www.virtualdub.org/). The locomotion activity was analyzed for two time points, for 10 sec at the 5th min and for 10 sec at 10th min using manual tracking (http://rsbweb.nih.gov/ij/plugins/track/track.html) in ImageJ software (1.47 v) (http://rsbweb.nih.gov/ij/download.html) []. [...] All data are represented as mean ± SEM. One-way ANOVA test was carried out for multiple comparisons among treatments. Bonferroni t-test was performed to obtain significant levels (P values) of various groups of flies (SigmaPlot version 12.0). Paired t-test was employed to compare the number of EPSPs between the saline and agonist or saline-containing compound. Student's t-test was used to compare the treatment and control groups. The level of significance (P value) less than or equal to 0.05 is considered statistically significant. […]

Pipeline specifications

Software tools ImageJ, SigmaPlot
Applications Miscellaneous, Microscopic phenotype analysis
Organisms Drosophila melanogaster
Chemicals Fluoxetine, Serotonin, Tryptophan