Lam system

Behavioral analysis was conducted according to the method reported in a previous study [4 (link)]. Behavioral analysis was performed using the LAM system (TriKinetics, Waltham, MA, USA) when Drosophila flies ingested a sucrose agar medium containing 1% GABA, 0.1% 5-HTP, or GABA/5-HTP (1% GABA and 0.1% 5-HTP). The group (minimum 20 flies) activity, which was measured by the LAM system, provided locomotor activity levels. All experiments were repeated five times.
Changes in locomotor activity by generation were measured using a Drosophila activity monitoring system (DAM, TriKinetics), and these measurements involved housing Drosophila flies individually in glass tubes. Drosophila flies were acclimated to the tube for 24 h, and all recordings were measured with an infrared detector every minute during the experiment in constant darkness at 25 ± 1 °C. Data were analyzed using the Actogram J software, and sleep parameters were calculated by summing the number of all activities recorded daily. Subjective nighttime activity was calculated as the sum of total activity, and subjective nighttime sleep was calculated as the sum of total sleep in the light and dark phases. Sleep was defined as a period of inactivity for at least 5 min (0 interruptions per minute). The number of sleep bouts was calculated by adding the sleep episodes.

To analyze total activity, a locomotor activity monitoring (LAM) system (TriKinetics, Waltham, MA, USA) was used to evaluate the sleep-promoting effects of GLE. Ten Drosophila were placed in a vial and acclimated for three days in constant darkness. After adaptation, vibration stress was applied for two days, and behavior patterns were analyzed for one day. Actogram J was used for data analysis, and sleep parameters were calculated based on the total number of activities recorded each day [15 (link)].