Locomotor activity and sleep behavior monitoring (1 minute resolution) of individual Drosophila melanogaster using DAM5H activity monitors

We conducted behavioral analyses using DAM5H Drosophila Activity Monitors (Trikinetics, Princeton, MA, USA). Using the DAM5H we tracked the locomotor activity of individual flies housed in a 5mm-diameter, 80mm-length tube. The system records activity when a fly crosses one of 15 infrared beams of each tube. The monitor quantifies movement by registering infrared beam disruptions as activity "counts." A fly that moves continuously through a beam zone or returns to the same beam multiple times generates several counts, whereas a fly that simply enters a beam area and stops moving is recorded as only a single count. The count data were registered per minute and were used to evaluate the locomotion activity and to identify sleep periods, which were defined as immobility bouts lasting at least 5 minutes. All behavioral experiments utilized 12-day-old mated female flies. Flies were individually housed in DAM tubes containing standard Bloomington cornmeal diet. All experiments were conducted at 25°C under a 12:12 light:dark cycle. To allow for acclimation, flies were placed in the monitoring tubes one hour before the start of a dark cycle. Data collection began at the onset of the following light cycle (designated as T0) and continued for three complete cycles (until T2). We performed three independent experiments, each including 8-12 flies per experimental condition.