Drosophila melanogaster activity and climbing distance assays
Wild-type Drosophila melanogaster was maintained on standard medium (50 g/L yeast, 50 g/L glucose, 0.8% agar, and 1% soy flour) at 25-26°C in 50% relative humidity under a 12:12 h light-dark cycle. The 7-day-old flies were loaded into plastic tubes containing standard food or with GS (1 mg/mL) for 7 days. Then, the sleep-waking activity of 32 flies was consecutively monitored for 24 h using the Drosophila Activity Monitoring System (Trikinetics, Waltham, MA, USA) and recorded as beam-breaks in 1-min bins for generating the number of activities. For the climbing ability assay, 30 flies in the vertical vials (110×10 mm) were acclimatized for 15 min and simultaneously tapped to the bottom of the vial for 5 sec. The climbing distance of each fly from control or GS pretreatment was recorded to obtain the average of the distance (Algarve, Assmann, Aigaki & da Cruz, 2020).